Ministry

The Seventhof Agriculture,
Forestry and Fisheries(MAFF),
International
Workshop on Genetic Resources

Part iWild Legumes

National

Institute of Agrobiological
Tsukuba, Ibaraki, Japan

13th-15th

Research

October

Resources

1999

Sponsored byCouncil Secretariatof MAFFandNational

Institutein cooperationof Agrobiological

Resources

with other MAFF institutes

in Tsukuba

Japan

CONTENTS
Welcome address
N. Katsura
Keynote addresses
Research on wild legumes with an emphasis on soybean germplasm
Y. Shimamoto
Wild legume diversity

Questions

Sessionand domestication- insights from molecular methods
P. Gepts, R. Papa, S. Coulibaly,
A. Gonzalez Mejia and R. Pasquet

and Answers

1 Biosystematics

and ecogeographic

Biosystematics

and genetic

resources

Ecogeographic

techniquesand conservation:

Questions

SessionBiodiversity,

techniquesof the genus Vigna subgenus Ceratotropis
N. Tomooka, Y. Egawa and A. Kaga
Case study for the legume genus Vigna in
Africa.
N. Maxted, P. Mabuza and S. Kelland Answers

2 Genetic

diversity,

ecologyevolutionand genetic

and conservation

resources of Phaseolus

beans - Seven answered andunanswered questions

D. DebouckViciafaba

L. and related

species:

Genetic

The genetic structure of natural populations
RFLP's of mitochondrial
DNS's: possible
demography.

95

diversity and evolution
E. Potokina, D. A. Vaughan, N. Tomooka,
and S. Bulyntzev
of wild soybeans revealed
influence of seed dispersal,
J. Abe1 25

by isozymes andcross pollination
and143

The Vigna angularis

Questions

SessionEvaluation

Allelopathy

complex in Japan

Closing159

in breeding
M.Ishimoto

1 83

legume species.
Y. Fujii

191

and Answers

3 Research,evaluation

and useand use of wild Phaseolus

speciesin Vicia, Mucuna and related

The use of wild relatives

QuestionsD. A. Vaughan, N. Tomooka, R.-Q. Xu,
A. Konarev, K. Doi, K. Kashiwaba,
and A. Kagaand gamma radiation in mungbean and blackgram breeding
P. Srinives, N. Hual-alai, S. Saengchot, andS. Ngampongsai

205

and Answers

Remarks
M.Kumamoto

225

Participants

Photograph

226

Participants

List

229

Introduction
Welcome address
Keynote addresses

Welcome address

Naoki

KatsuraDirector

General,National

Instituteof Agrobiological

It is a great pleasure
Ministry of Agriculture,

Resources

for me to extend a warm welcome to all the participants

Forestry and Fisheries(MAFF)

like to extend a special welcome to participantsof thisworkshop on genetic resources. I would

who have travelled

a great distanceto be with us

here in Tsukuba. Welcome to Japan and welcome to Tsukuba. We hope that you will feel when
you leave that your time in Japan has been well worthwhile.
workshop. I believe I am right in saying we have participantsThis reallyis an internationalfrom more than twelve countries inthis room. Weare also very pleased to welcome students from Hokkaido University.
I would like to acknowledge
workshop possible.

the contributionof several organisations

The workshop has been sponsored by the Agriculture,

in making thisForestry

Research Council. Here in Tsukuba we have been supported by our sister institutes,
National InstituteJapan Internationalof Agro-environmental

Sciences,

National Agricultural

Research Center for Agricultural

Sciences.

and Fisheries
including,theResearch Center and

We are gratefulfor their kind

support.
This year the workshop has two themes, but two themes which I think will have a measureof complementarity.
research"*.

The first theme is "Wild Legumes"* the second "In-situ

conservation

Some of the workshop speakers are giving talks in both themes since their researchoverlaps with both topics.
I would like to say a few words about my instituteResources (NIAR). The core activities
areas of biology
within- biodiversity

the National

Instituteof Agrobiologicalof NIAR revolve around two related and rapidly advancing

and biotechnology.

NIAR, as well as with other institutes

Both research areas require strong linkages
in Japan and around the world. We hope thisworkshop in a small way helps develop these linkages.
Our biodiversity

research involves

plants,microorganisms

and animals. We are also

involved in conservationof useful biologicalmolecules in the DNA bank and genetic resourcesand genome informatics.

Adjacent

to this room is the almost complete Genome Information

Center.
Our biotechnology

research extends from basic sequencing to genome function and wholeplant physiology.
I cannot describe

our activities

in the detailJapanese of our last annual report are available

outsidetwo days you will have a chance to learn a little
Finally

this workshop is an excellent

I would like. However, copies in English

for any one who wishes. I hope in the nextof the research

opportunity

under way here.

to exchange ideas and develop linkages.

I hope that the exchanges both in this room and outside will be helpful

to all. To all participants

I would encourage you all to ask many questions.

of this workshop

publishedandThe proceedings

will beso that an audience beyond this room will benefit from the ideas and research presented.

Again thank you for your participation.
.(* Wild

legumes is publishedpublishedsubsequently

here separately

as Part 1. In-situ

as Part 2 of the proceedings.

Editors

note)

conservation

research will beResearchon wild legumes

with an emphasis

on soybean

germplasm
Yoshiya Shimamoto
Laboratoryof Plant

SapporoGenetics

and Evolution,

Facultyof Agriculture,

Hokkaido

University,

060, Japan

Abstract
Soybean is the major legume crop and after maize, wheat and rice the most important
crop. The genus Glycine, consistsof two subgenera, subgenus Soja, of which the cultigen

form of soybean are members, and subgenus Glycine, which includes
greatly differentiatedfood

and the wild

many wild species

which arefrom subgenus Soja. Subgenus Soja is a gene complex, and the wild soybean has

greater diversity than the cultigen, and it is an important primary genetic resource for the cultigen.
Wild soybeans are distributed in semi-disturbed
habitats of East Asia, China, Korea, Japan and Far
East Russia. The center of diversity for wild soybeans is in the southern region of China and thenorthern region of Honshu, Japan, for both isozymes and the organelle genome. The cytoplasmic
genometype is different

and allele frequency varies, amongvarious regions of East Asia. Most of thevariation in the cytoplasmic

genome detected in the cultigen

has been observed in the wild soybean.

1. Introduction
The soybean,

Glycineproduction

which is aboutwild relatives

has recently

into two subgenera,composed
Glycinespecies1).

speciesThe

compatible

reviewed

main grain

tons annually.

The taxonomy of the soybean

and subgenus Soja. The subgenus

mainlyindigenous

speciesIslands

Glycineto Australia.

are perennial

Glycineand its

is divided
is currently

However, some subgenus

and as far north as Miyako

with each other. The members of subgenus

and inbreeding

Island,Japan (Table

and are not crossSoja are annual inbreeding

and can beoffspring.

The subgenus Soja, which includes

the cultigen,andformosana

four taxa can be crossed

and together

The genus Glycine

Glycineof subgenus
fertilelegume crop in terms of total

by Hymowitz et al. (1998).

are found on Pacificcrossed to produce
gracilis

140 million

subgenusof 16 wild

max, is the worlds(Table1). Theseshare a common primaryis composedof three wild forms, soja,
to produce

gene pool and form a single

biologicalfertileoffspring

species.

The

Table 1. Species in the genus Glycine,
Hymowitz et al., 1998).

Fig. 1 Geographic

distribution

chromosome number and distribution(modifiedof known sites of wild soybean (Glycinesoja).

fromwild forms composed of G soja, gracilis

and formosana are definedas the wild soybean

the G. soja complex, and are the likely

progenitor

of the soybean(Hymowitz,

complex isresource

for soybean

ecologicalan important

plant

distribution,

genetic

soybeans is intensively

required.

2. Ecologicaldistribution

genetic

diversity,of Russia

in lowland

and is found in semi-disturbed
crop fields.
cultivated1). The wild soybean

sitessuch as roadsides,al.(1997)

surveyedwithsome collection

riverbanks,

is a typicalhas prolificmostlythe allelic
sites

plant

waste land and the fringe

vine around relatively

tall herbs

wild soybeans

ruderalofsuch as Miscanthus
and occasionallyshow a prostrateor bush formand seed production

and shows1994).

soybeanhabitof the wildareas (Fig.

communis, Artemisia montanas and Solidago altissima,(Ohara and Shimamoto,inbreeding

and domesticationon thein East Asia, China, Korea, Japan and the Farsoybean. When there are no tall herbs,
The wild

Research

distributed

It grows mainly as a twining

sinensis, Phragmites

differentiationThisin the wild soybean.

The wild soybean is widely
East region

genetic

breeding.

1970).

orvegetative

growthcleistogamous

flowers

types

loci of the wildof isozyme

there is higher

within-population

divergenceamong populations

furthermore,

the mean multi locus outcrossing

in field
genetic

than would be expected

trials.

and found that at

variationand lower genetic

for an inbreeding

the four populations

along the Omono River in Akita Prefecture,

rate was supported

by observation

visitsetsoybeanrate was 13%, ranging

of frequent

However, Fujita

plantspecies,from 9.3-19%

and
among

Japan. The high outcrossing

by honeybees

and carpenter

bees toflowers of the wild soybean.
3. Diversityof the wild soybeans

We have made morphological,

cytological,

wildsoybeansand elucidated

wildsoybean,among our observations

earlierthan specimensfrom low latitude,

a distalor proximal

region on the chromosome, variants

mutant of soyasapogenol

theirgenetic

isozyme and moleculardiversityfrom various

have been that specimens
the fluorescentsignalregions

marker studies

of East Asia. In

from high latitudeof 5S rDNA loci is locatedof seed proteins

of
ripen
at

and a deficientA.

3a. Isozyme lociAllelic

types at 15 isozyme

loci for the wild soybean collectedfrom 770 growing

sitesLo cus S pe cie s

A co l
A co2
A co3
A c o4A c o5Ap
D ia lE np

w ild form
cu ltig e n
w ild form
cu ltig en
w ild fo rm
cu ltig en
w ild fo rm
cu ltig en
w ild fo rm
cultige nw ild fo rm
c ultige nw ild fo rm
c ultige nw ild fo rm
c ultige n

A c o l‑aA c o l‑aA c o2 ‑aA c o2 ‑a
A co 3 ‑a
A co 3 ‑a
A co 4 ‑a
A co 4 ‑a
A co 5 ‑a
A co 5 ‑a
A p ‑aA p ‑aD ia l‑a
D ia l‑a
E np ‑a
E n p ‑aA co l‑b
A co l‑b
A co 2‑b
A co2 ‑b
A co 3‑b
A co3 ‑b
A co4 ‑b
A co4 ‑b
A co5 ‑b
A co5 ‑b
A p ‑b
A p ‑bD ia l‑b
D ia l‑b
E np ‑b
E np ‑b

A llele

o20
wbs
ith
%erve
mfreq
odreuall
tha
ency
e len

A co l‑cA c o l‑aA c o l‑a
A co 2‑b
A co 2‑b
A co 3‑aA co 3‑aA co 4‑b
A co 4‑b
A co5 ‑aA co5 ‑aA p ‑a ,A p ‑c
A p ‑bD ia l‑a ,D ia l‑b
D ia l‑a ,D ia l‑b
E np ‑a , E np ‑b
E np ‑a , E np ‑b

A co2 ‑cA c o3 ‑c A co3 ‑d A co 3 ‑n
A co 4 ‑c A co4 ‑d A co 4‑n
A co 4 ‑c
A co4 ‑n
A co 5 ‑c A co5 ‑n
A p ‑cA p ‑cD ia l‑c

A p ‑dD ia l‑d
D ia l‑d

E np ‑c
E np ‑dE stl

w ild fo rm E s tl‑a E s tl‑b E stl‑c E stl‑nc ultig e n E s tl‑a E s tl‑b
Id h l w ild form Idh l‑a Idh l‑b
Id h l‑c
cu ltig e n Idh l‑a Idh l‑b
Id h2
w ild form Idh 2 ‑a Idh2 ‑b Idh 2 ‑c
Idh 2‑ncu ltig e n Idh 2 ‑a Idh2 ‑b Idh 2 ‑c Id h2 ‑d
Lap l w ild fo rm L ap l‑a Lap l‑b L ap l‑c Lap l‑d
cu ltig e n L ap l‑a Lap l‑b L ap l‑c
M piw ild fo rm M p i‑a
M p i‑b
M p i‑c M p i‑d
cu ltig e n M p i‑a
M p i‑b
M p i‑c
P g m l w ild fo rm P g m l‑a P gm l‑b P g m l‑c
cu ltig e n P g m l‑a P gm l‑b
P g m 2 w ild fo rm P g m 2 ‑a P gm 2 ‑b P g m 2 ‑c P g m 2 ‑dcu ltig en P g m 2 ‑a Pg m 2 ‑b

Table 2. Observed

alleles

N a tio n

R e g io n

R u s s ia

C h in a

at 15 isozyme

E s tl‑a, E stl‑b
E s tl‑a, E stl‑b
Idh l‑b
Idh t‑a , Idh l‑b¥dh2 ‑hIdh2 ‑b
Lap l‑b
Lap l‑b
M pi‑b, M p i‑c
M p i‑b, M pi‑c
Pg m l‑a
P gm l‑a ,P g m l‑b
P gm 2 ‑b
Pg m 2 ‑bloci in wild and cultivatedsN aumm pblee rs oitef s

N a

HT

A m rs k a y a
K h a b a ro v s k
U s s u ri

16
17
18

1 .6 0
1 .8 7
1 .9 30 .14 1
0 .2 0 4
0 .2 3 5

to ta l51

2 .0 70 .2 3 1

N o rth E a s t
H ua ng R .V .u ppe r
lo w e r
Y a n g tz e R . V . u p p e r
lo w e r
S o u th

86
26
30
17
47
22

2 .8 02 .3 32 .2 0
2 .0 7
2 .7 3
2 .4 00 .2 7 6
0 .2 2 2
0 .2 3 8
0 .2 2 5
0 .2 6 3
0 .3 0 4

to ta l2 28

3 .2 00 .2 7 3

S o u thK o re a

W e stC e n te r
E a stS o u thC h e ju

9
20
13
13

2 .4 02 .6 02 .4 02 .5 3
2 .3 30 .2 6 2
0 .2 6 9
0 .2 5 7
0 .2 6 3
0 .2 7 3

to ta l64

3 .0 00 .2 7 9

J ap a n

H o k k a id o
N o rth e rn H o n s h u
C e n tra l H o n s h u
S o u th e rn H o n s h u
S h ik o k u
K yush u

35
12 1
15 1
49
31
40

1 .8 7
2 .8 02 .7 3
2 .4 72 .3 3
2 .3 30 .2 1 2
0 .2 7 7
0 .3 0 3
0 .2 9 6
0 .2 5 9
0 .2 7 5

to ta l427

3 .2 00 .2 9 2

soybean.

Table 3. Mean number of alleles per locus (Na) and gene diversity(HT) for 15 isozyme
in the wild soybean collected from China, Japan, Russia and South Korea.

lociacross East Asia were determinedshown (Table2) including

and the 57 alleles

the cultigen

soybean could be observed

for reference.

and differences

loci compared with the cultigen.
soybean in contrast

Idhl

and a allele at Pgml are predominant

to the b allele

were detectedwhichand populationsin the wild

have conserved

covering

several regions

variedamong various regions

regions

and countries.

distances

higherplant

is shown (Table
withinfrequenciesof threeare predominant

in thein the cultigen.

The b allele

atto the other alleles

and gene diversity(Tablein various regions

than the cultigen

The allelic4). Thoughallelic

frequencies

at a few isozyme locithe genetic

distanceswere small among theamong the regionalfrequencies

for four countries

populations

using

frequencies

of isozyme loci is shown (Fig.

with Cheju Island,South Korea and Shikoku

cluster.

West and Central

coastalregionsof South Korea and the northeast

Russia.

The Yangtze River Valley

cluster.

Hokkaido,

regions

China constituted

the genetic2). Honshu,

and east and south

a cluster,which includes

and the lower region of the Huang River Valley
respectively,of the Huang River Valley
and are independent

Japan

and Kyushu formed aof South Korea made a cluster

Japan, and the upper region
clusters,3), that

species.

separate

China formed different

to wild

soybean in contrast

diversity

countries,

relationships

based on the allelicformed a separate cluster

specificin the allelicis predominantas

in the cultigen.

of other inbreeding

The phylogenetic

were detected

More than 20 alleles

It was found as shown by mean number of allele
wild soybean populations

a null allele

At the Ap locus the a and c alleles

wild

which are more frequent

includingformed aand the southernfrom each other and fromother clusters.
3b. RFLPs of chloroplast
RFLP analysesof chloroplast

the maternal phylogeny
collected(Hatfield,

et al. 1985)

DNA were carriedof the wild soybeans(cp) typesout to determine

for about 2000 specimens

were distinguishedby the probe of the H2genes between 5y-rpsl2 and rpsl9 with EcoBI or Clal digestion

mitochondrial

DNA (mt)combinationof seven different

13 different

profilesassociatedwith a specificthe

21 different

profileof the atp6 with BamHl digestionof mitochondrial

types

have been

size fragments of the cox2 with Hindlll

chloroplast(Fig.

and chloroplast

So far 24 cytoplasmic
types

(Fig.

3). Indetected

in a

(Fig.

4) anddigestion5). Mitochondrial

type, but there are three exceptions,

mtVb, which have two types of chloroplast.
combinationDNA

and mitochondrialin East Asia. Three chloroplast

clone containing
theand mitochondrial

type is usually
mtlVb, mtlVc andgenome types based on thehave been detected

in wild

soybeans.

Table 4. Allele frequencies
Russia and South Korea.

of isozyme loci in wild

Fig. 2. Dendrogram based on allele frequencies
from Russia, China South Korea and Japan.

soybeanscollected

from China, Japan,

in 1 5 isozyme loci in the wild soybean collected

Fig. 3. Southern blots of EccRl- and Claldigested
DNAs hybridized
to the H2 clone
probe of chloroplastobtained from the wild
soybean. Numbers indicate
the different
patterns identified:
Lane 1, 2 and 3 corresponds
to cpl, cpll and CPIII, respectively.

Fig. 4. RFLP profiles in digest of Hindlll withthe probe containing mitochondrial
gene coxl.
Roman numeral on each lane indicatesthemitochondrial
type.

Fig. 5. RFLP profiles in digest of BamRl with the probe containing
Alphabetic
letter on each lane indicates the mitochondrial
type.

mitochondrial

gene atp6.

12

Their frequencies

in various regionsof the cultigen
frequent

as a referencecytoplasmic

cytoplasmic

types

soybeans

5). The geographicalof wild

is shown (Fig.

in the cultigen,predominantsoybeanwith CPIII

are widely(Tablehas occasionallychloroplast

Yangtze river valley,

distribution

distributedin East Asia,

been found in restricted

the Korean Peninsula

but rare or absent

Russiaand is the predominant

type ofin southern

except for southernwith cpl, the predominant
areas of Japan. CpIII,

and is frequently

in China north of theand Russia. In Japan mtlVa is frequentJapan.

in northern

mtlVb is common except for Hokkaido

type south of the Yangtze river valley,

Island,is thefound in the cultigen.

genome mtlVa is common and predominantregionsof the more

6). The predominant

5). A few wild soybeans

type in wild soybeans

In the mitochondrial

type frequencies

among the regions.

China where cpll is most frequenttype

the cytoplasmic

is shown (Table

genome is different

Wildof East Asia, including

northern

on Cheju

predominant.

mtVa type was found in East Asia. MtVa, mtlc and mtld were exclusively

found in wild

soybeans.

However, the most commonmitochondrial

MtlVa and mtlVb, are frequently
cytoplasmic

types

of the cultigen

are composed mainly of those in the cultigen
and in contrast,

CPIII+

mtVIIIc,

observed

relationships

mitochondrialin wild soybean,

are shown (Table(82%),South Korea (76%)

analyzed

types

restriction

endonucleases

phylogenetic

cladogram

using

the

phylogenetic

among thecox2, atpl, atp6 and atp9)

BamHl and EcoBI)

the NJ method (Fig.

7). The result

and constructed

distinguishedare the mitochondrial

types

associatedwith cpl or cpll.

The latter

threeor CPIII, the fourth

genome.

relationships(Hindlll,

7). The first and second clades

type of the cultigen,of cytoplasmic

types based on RFLPs of five mtDNA probes (coxl,

and three

and Russia

accessions.

among the differental.(1998a)5), of which

genome types of the wild soybean

only 26% in Japan. However, the cytoplasmic

Kanazawa et(Fig.

in China

could be found in only Japanese

4. Phylogenetic

types

where mtld isfound in the cultigen.

seven types are found in the wild soybean. The cytoplasmic(75%),

regions

Mtlc type ispredominant

The eight

Korea and Japan except

China.

andfour clades
only with CPIII,

the thirdwith cpll

cultigen,while the first clade consistsof rare types of wild soybean (Table

5 for references).

The wild soybean

populationsin the second

clade

in, and limited

Japan.

The CPIII+mtVIIIc

is uniqueto the cultigen

and has been observed(Table5). The wild

populations

northernChinasoybeanclassified

and South Korea. The CPII+mtVb

are dominant

into the third

clade

a

cladesincludestheto,

only in Japanare predominant

may be assumed to be derivethroughinTable 5. Cytoplasmic
genome types based on the combinationof chloroplast(cp) andmitochondrial(mt) types in the wild soybean and their geographic
distribution
and genetic
diversity (H value) (after Shimamoto et al., 1997 and 1998; Tozuka et al., 1998; Shimamoto,1999 and unpublisheddata).

Fig. 6. Distributionof frequent types of cytoplasmic
Russia, China, South Korea and Japan.

genome in the wild soybean collected

from

14

mutationfrom CPIII to cpll

on the chloroplast

genome of the CPIII+mtVb.

thought

to have occurred in South Korea after the third

soybean of the mtVb with cpll and CPIII is restricted
In the fourthwith CPII+mtlVc

clade

differentiated,clade the wild soybean with CPII+mtlVb

observed

and that

uncommonin South Korea and south of the Yangtze River Valleyof China

as genetic

The presumed
cytoplasmic

resources

is frequently

pathwayscpll are containedin the fourthcultigens,

CPI+mtlVb

derivedfrom the wild

and places

chloroplastis shown (Fig.

and CPII+mtlVc.

including

The cultigens

or theof the cox2 or atp6 region

with CPII+mtlVb

events are thought
China,

on the8). Five typesof cultigen

the currently

first and second leadingare presumed to be

CPII+mtlVb

through(singlesubstitution)in theKanazawa et al., 1998b).

The wildwith

et al., 1989;

with cpl or

cpl or cpll

and mutation

may be a progenitor

based

base

of the cultigenchangesin the fourthclade.

inThese

to have occurred after the fourth clade was established.

The cultigen
northeast

domestication

withcultigengenome from cpll to cpl (Closesoybeanof soybean

clade in Fig.7,

soybeanin Japan.

for the cultigen

genome of wild soybeans

recombination

because the wild

to South Korea.

and the other members with cpl have been found only occasionally
5. Wild soybean

The change iswith CPIII+mtlVa

is in the third

the Huang river valley,

5). The source of the cultigen

China

clade

and is observed

frequently

and South Korea and but not in Japan

with CPIII+mtlVain the thirdclade

genome. This domestication

may have occurred in northernor the Korean Peninsula,

becauseand cultigen

soybeans(Tablemay be the wild soybean

with the same cytoplasmic

both wild

in

China

with CpIII+mtlVa

arefound commonly in both regions.
In the second clade there are two cultigens,can be found in wild soybeans

mtle and mtVIIIc

and both are restricted

northern

Honshu, Japan. Therefore,

the cultigen

in these

two regions

form. The cultigen

Japanese

cultivarsoybeans.

However, the wild

CPIII+mtVIIIc

grow widelyfrom its wildgroup

"AKIDAIZU"in Japan.

CpIII+mtle

to the Yangtze river valley,

Chinawith CPIII+mtle(Nagata,soybeans

with CPIII.

classifiedThus the genetic

may have been domesticatedwith CPIII+mtVIIIc

1960)
into

and

is specificto theand has not been observedin wild

theincludingthewith the CPIII+mtVIIIc

may

and it is an annual inbreeding

plant.

resources

secondclade

be found in Japan.
Conclusions
1. Wild

soybeans

Populations

grow in East Asia in lowlandwith a high outcrossing

habitats

rates have been observed at some sites.

Fig. 7. (above) Cladogram constructed using the NJ method
amongmitochondrial genome types in the wild soybean basedon mtDNARFLP data analysis of five mtDNA probes (coxl ,cox2, atpl, atp6 and atp9) and three restriction
endonucleases (HindJR, BamHl and EcoBI) after Kanazawa
et al.(1 998) with some modifications.

Fig. 8. (right) Assumed pathways and places of soybean
domestication
based on the cytoplasmic genome of wild
soybean after Shimamoto (1 999) with some modifications.
Broken lines indicate cytoplasmic genome types involved indomestication. Two dotted lines on the wild form side

indicate a change in the mitochondrial
genome to attain the genome
type (arrowed) through a single event at the cox2 region of themitochondrial DNA.A thin solid line joining the same genometype on
the cultigen side indicates dissemination to the arrow point between
China and Japan. Thick solid lines indicate a mutation in thechloroplast genome fromcpll to cpl or from CPIII to cpll.

16

2. Geographic

differentiation

including

isozyme3. Higher

genetic

and differences

loci and the cytoplasmic

diversity

is observed

Honshu, Japan for isozyme
4. Duringsoybeanprovided

from the cultigen

cytoplasmic

for various

traitsgenome.

in southern

China,

loci and the cytoplasmic

domestication

are observed

severalwild

South Korea, northernand central

genome.

soybeanpopulations

are thought

to havegenomes to the cultigen.

References
Close,

P.

S., R. C. Shoemakerpolymorphism

withinand P. Keim.

the chloroplast

1989.

Distributionof restriction

genome of the genus Glycine,

site

subgenus

Soja.

Theor. Appl. Genet. 77:768-776.
Fujita,Hatfield,R., M. Ohara, K. Okazaki

and Y. Shimamoto.

1997.

in wildsoja).

88: 124-128.

soybean(Glycine

Jour, of Heredity

P. M, R. C. Shoemaker and R. G. Palmer.
DNA within

the genus Glycine

subgenus

Hymowitz, T. 1970. On the domestication
Hymowitz, T., R. J. Singhin Plant

Breeding

Kanazawa, A., A. Tozuka,
relationshipsof the soybean.

Vol. l6.(J.

S. Akimoto,of the mitochondrial

Maternal

inheritanceof chloroplast

Soja. J. Hered.76:373-374.

and K. P. Kollipara.
Reviews,

1985.

The extent of natural-pollination

1998.

Econ. Bot. 23:408-421.

The genomes of the Glycine.

Janick,ed.) John Wiley

p289-317&Sons, Inc.

J. Abe and Y. Shimamoto.

1998a.

Phylogenetic

genomes in the genus Glycine

subgenus

Soja. Genes

Genet. Syst. 73:255-261.
Kanazawa, A., A. Tozuka and Y. Shimamoto.
that involves
Syst.
Nagata,EcoRl and CM restriction

Sequence

variationsite polymorphismsof chloroplastin soybean.

DNA

Genes Genet.

73:111-119.

T. 1960.

Studieson the differentiation

Hyogo Univ. Agric,Ohara,

1998b.

in Japanand the world.

Mem.

Agron. Ser. 4:63-102.

M and Y. Shimamoto.
characteristics

of soybeans

1994.

A studyof two growth forms of wildof somesoybeanecological(Glycineandsoja Sieb.

demographic
et Zucc.)Can.

Polymorphisms

andJ. Bot. 72:486-492
Shimamoto,Y.,

A. Tozuka, M. Yamamoto, M. Ohara and J. Abe. 1997.

differentiationsof the8th

Breeding
Shimamoto,of cytoplasmic
SABRAO
Society:41

Y., H. Fukushi,

chloroplast

Generalgenome in wild
Congresssoybeanand thegrowingAnnualin Korea. Proceeding

Meetingof theKorean1-412.
J. Abe, A. Kanazawa, J. Gai, Z. Gao and D. Xu. 1998. RFLPs

and mitochondrial

DNA in wild

soybean,Glycinesoja, growing

in China.

of17
Genet. Resour. Crop Evol. 45:255-261.
Shimamoto,

Y.

1999.

mitochondrial

Polymorphism

andoriginofsoybeanfromchloroplastandDNA. JARQ submitted

Tozuka, A., H. Fukushi,

T. Hirata,

Shimamoto

Composite

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by RFLP analysis

M. Ohara, A. Kanazawa, T. Mikaru,
and clinalof mitochondrial

distributionof Glycine

DNA. Theor. Appl.

J. Abe and Y.
soja in Japan

Genet. 96: 170-176.

revealed

19

Wild Legume Diversity
Molecular Methods

and Domestication

P. Gepts1,

R. Papa2,

A. Gonzalez

departmentofAgronomy and Range Science, University

CA 95616-8515,

S. Coulibaly3,

USA. 2 Permanent

Ambientali,

Universita

degli

Vegetable

Crops, University

Darwin, NT, Australia.

Studi

address:

Mejia4,- Insights

fromand R. Pasquet5of California,

1 Shields

Avenue, Davis,

Dipartimento

di Biotecnologie

Agrarieeddi Ancona, Ancona, Italy.

3Current address:

Department

ofof California,

Davis.

5Current address:

IRD-ICIPE,

4Current address:
Nairobi,

CSIRO

Plant

Industry,

Kenya.

Abstract

In the last decade, increased attention

has been devoted to genetic resources of wild relativein general, and wild legumes, in particular.
studies:1 ) they provide a geographic

domestication;
traits.

Wild relatives

have two major functions

framework of reference to elucidate

and 2) they constitutean increasingly

importantof crop plants,

in genetic

resources

patterns of genetic diversity

source of diversity

We will provide examples fromour research on 10 the phylogeography

andfor a wide range ofof Phaseolus vulgaris L.,

the organizationof genetic diversity

in Vigna subterranea (L.) Verde, and V. unguiculata (L.) Walp.; 3)

the inheritanceof the domestication

syndrome in P. vulgaris;

wild and cultivated

P. vulgaris. hi all these studies,molecular markers, including

provided a wealth of data, which, together with phenotypic
our understanding

and 4) an assessment of gene flow between

and ecological

DNA sequences, havedata have significantly

increasedof the genetic resources of these legumes.

Introduction
Over the last decade there has been increased
generaland of legumes in particular.

conservationof genetic

framework of reference
testedto identify

and domesticatedincreasingly

againstpresumed
gene pools.

importantbeen used primarily

resourcesinterest

These wild relatives
and their utilization.

which the diversity

centers of domesticationin the wild relative

play a two-foldrole in thethe constitutea geographic

First,of the domesticatedsource of genetic

gene pool can be

and analyze differences

Second, the gene pools of wild relatives

as additional

diversity.

as a source of major genes for diseaseof crops in

between the wild

are becoming

Whereas wild relatives

or pest resistance(Stalker

have
1980;

20
(Frey

1975;

Frey et al 1984;

opportunities

to identify

Stalker1980).

Recent progress

and transfer genes for quantitative

traitsin molecularmapping provides(Tanksley

and McCouch 1997;

Xiao et al 1996).
In this contribution,of wild relatives,

we will highlightsome of our research findings

namely the study of patternsof genetic

more recent results from studies in Phaseolus
sequences, are providingthesea remarkableallowing

us to make significant

during domestication
Phylogeographyinferences

Wild P. vulgaris

Debouck 1991;

from northern

diversity

analyses,in

they arephenomena such as genetic bottlenecks
beans after domestication.

in the Americas. It has been reported

Mexico to northwestern

information

distribution

is not a continuous

conditions.

Two gaps are particularly

and northernDNAvulgarishas a very broad distribution

with furtherof genetic

statisticaland gene flow between wild and domesticatedof Phaseolus

an arc extending

data and multivariateonmarkers, including

to our understanding

about biologicalthe first role

We will focus particularly

and Vigna spp. Molecular

contribution

genera. Combined with phenotypic

diversity.

regarding

Argentina(reviewed

along

in Gepts

andin Debouck et al 1993 and Freyre et al 1996).

This

one but one with several gaps due to unsuitable
significant

environmental

in the Andean. The first is in southern Colombia

Ecuador and the second one in northern

Peru. Wild P. vulgaris

populations

in

Ecuador and northern Peru grow on the western slope of the Andes, whereas north of Ecuador
and in the southern

Andes they grow on the eastern slope.

Analyses with molecular markers have revealed the existencegroups, Andean, Intermediate,
Debouck et al 1993;
comprises

and Mesoamerican (Koenig.and

Freyre et al 1996;

populations

distributedof three major geographic

Gepts 1 989;

Koenig et al 1 990;

R. Papa and P. Gepts, unpubl. data). The Andean groupin southern

Peru, Bolivia,

and northwestern

Argentina.

The

Intermediategroup is found in Ecuador and northern Peru. The Mesoamerican group includes

populations

from Mexico, Central America, Colombia,

Until
first

clue

the relationships

was provided

Intermediate(Freyrerecentlygroup to have a combination

et al. 1996)

Intermediategroup to be distinct

to two possible

hybridizations

between these three groups was not well established.
analyses(Deboucket al. 1993),

whichof Andean and Mesoamerican alleles.

and AFLP data (R. Papa and P. Gepts, unpubl.

The intermediatenessprioriby allozyme

and Venezuela.

data)

A

showed the
Further

RAPD

also showed thefrom the Andean and Mesoamerican groups.
of the Ecuadoran-northerncauses.

First,

this

group

Peruvian group could be attributedof populations

is actuallyathe resultofbetween the Andean and Mesoamerican groups. Second, this group is ancestral

to

21

the two other groups. Information
been provided
Intermediate

us to distinguishby two sets of data leading

between these two hypotheses

to consistent

inferences

has

about the nature of thegroup.

An analysis
providedallowingof the DNA sequences of the phaseolin

the first evidence

that the Intermediate

Mesoamerican groups (Kami et al. 1 995).

seed proteingroup is actually

Phaseolin

multigeneancestral

to the Andean andgenes from accessionsof the Intermediategroup do not have direct tandem repeats observed in at least one of three locations
genes in accessions
P. vulgaris

from the Andean or Mesoamerican genes. The most closely- P. polyanthus

family hasof phaseolinalliedand P. coccineus - do not show these repeats either

speciessuggesting

to
that

the absence of the three tandem direct repeats is an ancestral

state and their presence is a derived

state.

P. vulgaris

These

observations

circumscribedsuggestedthat

the species

originatedin a fairly

based on DNA sequence analysis

of a second

area in the central Andes.

Furtherdata has been recently

obtained

multigene family coding for seed proteins,

namely the a-amylase inhibitor

P. Gepts, unpubl. results).

is actuallya sub family

et al. 1994;

Moreno and Chrispeels

This family

includesthe lectins

1 986).

To analyze genes of the a-amylase inhibitor

identified

and arcelins

that provided(Mirkov

family (S. Coulibalyof a larger family,
1 989;

andwhich also
Osborn et al.

family alone, a sub-sequence of the gene wasa PCR fragment of diagnostic

length(200bp) and could thereforebecloned and sequenced separatelyfrom lectin or arcelinsequences.

in a sample of two domesticatedP. vulgaris(one Mesoamerican and one Andean),five wild P. vulgaris(3 Mesoamerican,

1 Andean, and one Intermediate),

P. costaricensis(a novel Phaseolus

accessions

accession each ofP. polyanthus,
1 996),

accessions

and P. coccineus. From each accession 10 amplified

This fragment was amplified
and one wild

taxon; Freytag and Deboucksequences were cloned and sequences

at random.
The results of the sequence analysis
group is indeed distinct

confirmingthat the Intermediate

and P. coccineus formed distinct

clusters

and molecular evidence (Llaca et al. 1 994; Marechal

and Debouck 1991 ; Schmit
group had higher

further confirmationfrom the Andean and Mesoamerican groups and presumably

them. Sequences from P. polyanthus
previous morphologicalprovidedet al. 1993).

levels of genetic diversity

previous observations

These analyses

furtherprevious observations(summarizedin Gepts 1 993)

Schmitshowed that the Mesoamerican(Freyre et al 1 996; Tohme et al. 1 996).

was comparable in wild and domesticatedconfirming

et al. 1 978;(as measured by n, the average nucleotide

the level of diversity

ancestral toaccessions.

and may reflect

Surprisingly,

diversity)
however,This is in contrast

the rather

small germplasm

to22
sample used in this study.
Molecular

clock estimates

based on a mutation rate of 2 x 109 per site per

year (Nei

1987),

polyanthusof 2 x 106 years and between the Andean and Mesoamerican gene pools of some

500,000

suggestof divergenceyears. Neutrality

a time of divergencetests (Fu and Li 1 993)

group in which a departure
P. vulgaris.

between P. vulgaris

from neutrality

provideand P. coccineus or P.

the interestingobservationthat the only

was observed was the wild Mesoamerican group of

There are several, not mutually exclusive

explanations

to account for this observation.

It may be that an insect predatorof wild beans is distributedin the Mesoamerican region (but notin the Andes) and this predator

effects some type of selection

on seeds of the bean plant. A recent

genetic bottleneck

could have led to strong modification

absence of neutrality

is due to linkage

disequilibrium

genes subject

to selection.

tightly

to a-amylase genes (Osborn

linkedaccessions(Osborn

It is significant

et al. 1988).

on the magnitudeof linkage

gene, controlling

primarily

short,

to note that arcelin

et al 1986),

are found only in wild Mesoamerican

no information

drag. Recent results

selfing

species,

To summarize this section,
ancestorhas been identified

Peru. From this nuclear ancestral
the south to generate
independent

hitchhikingof P. vulgaris

for P. vulgaris

the hitchhiking
It is likely

distance

is verythat in common

will be longer.

available

so far strongly

suggeststhat anin the Andean mountains of Ecuador and northernarea, wild beans were disseminatedboth towards the north andIn a subsequent

stage,

took place in these two gene pools to give rise to the Andean and(reviewed

in Gepts

1 998).

are that the two gene pools are likely

genes. This contrasts with earlier

thinking

according

The implications
to containfor genetic

different

sets ofto which the Andean gene pool was derived

fromthe Mesoamerican gene pool and would therefore
lattertrait,

distances

all the information

Mesoamerican groups of domesticatesconservation

is available

the wild Andean and Mesoamerican gene pools.

domestications

genes, which arefrom maize, however, show that for the tbl

a domestication

for P. vulgaris

thein this respectof the order of a few hundred base pairs (Wang et al. 1999).

bean, a predominantly

Alternatively,

between a-amylase genes and neighboring

To our knowledge,

branch length,of gene frequencies.

only contain a subset of the genes of thegene pool.

Organizationof genetic

diversity

in Vigna spp.

Tworecent studies

illustrate

the challenge

relationships

between wild and domesticateddeterminationof the area of domestication.

posed by gene flow to our understanding
types of crop plants,

in particularof thewith regard to23
Bambara groundnut
subterranea)(Vigna subterranea

is the Africancountries it has actuallyespecially(I.) Verde,peanut, in that, like Arachis

41 loci was conducted

it remains locally

An isozyme analysis

in 21 wild or weedy accessions

data were then combined with morphological

known as Voandzeia

hypogaea, it buries

been replaced by peanut althoughfrom Senegal to Ethiopia.

previously

its fruit.

In many

importantin Africa,

using 23 enzyme systems representing
and 79 domesticatedobservations

accessions.

on the same accessions

These(Pasquet,R.

etal1999).
As observed in most other crops, domestication
Ht in wild = 0.087; Ht in domesticatedclusters.
with

A principal
eigenvalues

domesticated=0.052).

component analysisof 72 and 11%,has induced a genetic

bottleneck(e.g.,A UPGMA analysis of isozyme data lead to fiveof the morphologicalrespectively.

data resulted

The first

in two major axes

component separated

wild

and

forms, but also recognizedsome intermediate,weedy forms. The latter

forms

showed large seeds and tough, wrinkled

pods, but smaller

leaves and longer internodes

thandomesticatedtypes. A principal

gradient

in wild to domesticatedcloserto domesticateddifference

suggesttypes with one important difference.

for considering

traits are obviously

subjectthat the wild types included

groundnut,
specific

showed a similar

The weedy types were muchthe existencethat isozymes

to selection.

are generally

Whereas Nei distancesin this study are the progenitor

of weedy types suggests

data. This

neutral,

whereasbelow 0. 1 stronglyof domesticatedcaution when attempting

Bambarato identify

moreareas of domestication.
The same concern was highlighted

in a study of cowpea, Vigna unguiculata

Coulibalyand P. Gepts, unpubl. results).

distribution

and current taxonomy of V.unguiculata

were evaluatedfor AFLP diversity(Fig.

A total of 1 1 7 accessions,

1 14 (61%)

reflecting

subsp. unguiculata

1). The accessions

wild and weedy annuals, as well as 18 perennial

included(L.) Walp (S.

the geographicaland closely

relatedtaxa

47 cultivatedannuals, 52

forms. A total of 1 88 loci were scored, of whichwere polymorphic.

UPGMA clustering
Southern Africa),
perennialof isozyme Nei distances

types based on isozyme data than based on morphologicalcan be accounted

morphologicalcomponent analysis

analysisand three perennialof domesticatedtypes, wild materials

taxa (pubescens,tenuis, and alba),

from the annual types (wild or domesticated).

(Western,

showed separationin that region. However, a more detailed

story. All wild cowpea from Western Africa are distributedof theThe wild types closest to the domesticatedtypes were those from Western Africa. One might conclude (Vaillancourt
cowpea was domesticatedEastern, andand Weeden 1992) thatanalysisin disturbed

suggests

a different

environments,

includingfields

or field margins. None were observed in natural vegetation.

In addition,

when subdividing

the Eastern

Africanwild cowpea

group

accountfortopopulations

wildof Northeastern Africa,

the UPGMA clusteringshows thelatter group to be also closely
to thedomesticatedalliedgroup.

Th

Eastern African group also showed
higherlevelsof diversity

higherfrequency

and aof unique alleles

compared to Western and Southern
Africa.

Further

explorations

needed,however,

additional

geraiplasm

NortheasterntoAfricanobtainfrom theregionandperform a more detailed

analysisand test the hypothesisthat

region may actually

Figure 1. Autoradiographshowing AFLP banding patterns
generated from genomic DNA of 33 Vigna accessions using primer
combination (EcoRI+ACA, Msel+ACA). Arrows indicate putative
wild and perennial-specificproducts.

aredomestication

thisbe the center ofof cowpea

Kenya, Sudan, Ethiopia).

(e.g.,It should

be noted here that a domestication
for sorghum has also been proposed

specific

Africa (Morden et ah 1990).

The discussionof these two Vigna species

domestication

centers with molecular markers should proceed with caution. Additionaldata, such as ecological

data, and different

approaches,

are needed to distinguishdomesticatedforms of a species,

domesticated-wildInheritancedomestication

clearlyapproaches,indicatesthat the identificationsuch as phylogeographlcal

between two causes of close identity

namely progenitor-descendant

relationshipofor cladistic

between wild and(domestication)orgene flow.

of the domestication

Molecular

analysisin Northeastern

syndrome

mapping tools have allowed us to pursue a much more detailed

syndrome than was possibleon the basis of phenotypicof this kind was performed in maize by Doebley

and Stec (1991

analysisof theanalysesalone. The first, 1993)and Doebleyet25
al (1 990).

Their studies

maize were actually

revealed

that the marked phenotypic

differences

between teosinte

andcontrolled

by a limited

number of genes, many of which have a major

phenotypic

effect, and are linked

in a limited

number of regions of the maize genome. Earlier on,

Pernes (1983)

had suggested

outcrossing

speciesrecombinationthat linkage

of domesticationsuch as maize if the entirecaused by outcrossing

genes would have to be expected

domesticationinsyndrome were to survive

between the first domesticates

and their sympatricwild

relatives.
Subsequentstudiesvulgaris(Koinange

et al 1996),

1999),

have shown that domestication

mere coincidenceof this nature in predominantly
tomato (Grandillo

selfing

species,

however, including

and Tanksley

1996),

and rice (Xiong et al

genes in these species

or is there a biologicalmay be that outcrossing(at whateverdevelopmentof a domesticate.

it is unlikely

domestication

syndrome would appear in the same lineage.

on the developmenttightdomestication

Indeed,and jointnecessary condition

accommodating

segregationof domestication.

genes may therefore

be a desirable

trait such as rachis brittlenessindependently

in Mesoamerica

domesticatedsuggestsside effect,

linkageAfrica{e.g., dominance)

Doebley

et al 1 995).

genes (see above),or enhance gene expression

Furthercharacterization

not aof crops, if not aelement

of Paterson et al (1 995)

(sorghum)to generate mutations with maj or phenotypic

actionalthoughin the early stages ofand Asia (rice).

This

during domestication

thanof such genes would be. Suggested

to other domestication

effects

are homologous in cereals that werethat some genes maybe more prone to selection

others. It is not clear what the attributes
the ability(maize),would provide

An additional

is the observation

thecontradictory

attribute

potential."

for theconstituting

syndrome. Linkage,in the later stages

that genes for a domestication

mutability,

it has therefore

the need of recombination

fashion - to such a potentialcondition

Because outcrossingthat is a key element of a "domestication- in a preliminary

observation

is a necessaryathe species justthat all the mutations

and maintenance of the domestication

Linkage of domestication
pointing

level)

mutations into a single lineage,

one, may be a solution

Is this linkage

reason for it? The commonpoint linking

mentioned

away to recombine individual

are also linked.

P.

of domestication

includeeffect and without deleterious

and the possibility

in different

attributes

genetic

to modify genebackgrounds{e.g. ,

genes is needed to answer thisquestion.
Gene flow between domesticatedFrom the preceding

sections,

and wild P. vulgaris
it is clear that gene flow between wild and domesticatedtypes is an importantfactor in shaping

more popular given the potential
1994;

Mikkelsen

et al. 1 996).

crop genetic diversity.

release of transgenes

This topic has recently

become also

in a number of crops (Linderand Schmitt

Commonbean is consideredwith usual outcrossing

rates below 5%. Occasionally,

have been reported

that can reach 50-60% (reviewed

hybriddomesticatedx wild swarms have been reported

1993;

Freyre et al 1996;

Beebe et al. 1997).

to be a predominantly

however, much higher levelsof allogamy

in Ibarra-Perez

In addition,

et al. 1997).

on several occasions(Debouck

What is not clear from these observations,is the frequencyof outcrossing

the organizationof genetic diversity

in both the wild and domesticatedRecently,

we have initiateda series of studies

Papa, A. Gonzalez-Mejia,

selfing species,

et al.

however,in centers of origin and to what extent this gene flow has shaped

J. Acosta, A. Delgado-Salinas,

gene pools of P. vulgaris.

to help us elucidate

thesequestionsand P. Gepts, unpubl. results).

(R.

An AFLP

Figure 2. Principal coordinate analysis of ISSR data from a sample of P. vulgaris wild anddomesticatedindividuals
from Puebla, Mexico.

27

survey of genetic

diversityof 25 domesticated,

55 wild and 6 wild-weedy

Mesoamerican gene pool was analyzed. Data, subjected
domesticatedand wild materials

Mejia, A. Delgado-Salinas,
was a small cluster

were generally

to UPGMA clustering,grouped into two distinct

and P. Gepts, unpubl. results).

containing

accessions

both domesticatedthat more detailed

revealedthatclusters(A. Gonzalez-

One of the exceptions

to this pattern

and wild forms from the state of Puebla,

Mexico, hi that region, the two forms grow in close-range sympatry (withinraised the possibility

from theanalyses of sympatricsituations

the same field),

which

would reveal higher levelsof outcrossing.
ISSR (Inter Simple
wild and domesticatedaxisof a principal

Sequence Repeats)

analysis

was successful

types and among domesticatedtypes (Gonzalez

betweenet al. 1998).

The first

analysisseparatedwild from domesticatedthe four domesticatedtypes (Fig.

2). Calculations

on the presence of dominant alleles(presenceof a band) at loci that were normally homozygous

recessive,

outcrossing

rates

axes separated

coordinate

in differentiating

revealed

cumulative

domesticatedand wild P. vulgaris.

observation

made earlier,

was tentatively

coccineus and P. polyanthus
Furtherdomesticatedoutcrossing,high level of outcrossing,which confirmed theto the presence of allogamous

P. vulgaris

are distributedin varying

In contrast

speciesdegreesof sympatry.

Overallbetween wild and domesticateddomestication

syndrome genes identified

reductionin domesticatedin the vicinity
a limitedof the fields

in otherwise

gene flow, it was observed thattypes was strongest

by Koinange et al. (1996).

for loci of QTLs linked

types compared to wild types. In contrast,

in diversity

Further analyses are being conducted to provideof the gene flow between wild and domesticatedtoThese loci also showed a

did not show this population

pictureofstudied.

genes of QTLs for domestication
upon domestication.

levels

1 0%. The lower level compared to Pueblawasthat suggest

differentiationin diversity- P.

Mexico, where wild andusing the frequency of dominant alleles

were of approximately

with these findings

species

sympatry with P. vulgaris.
in the state of Chiapas,

to the absence of allogamous

based

20 and 50% in bothwere conducted

as measured by AFLP diversity

frequencies

betweenattributed- in close-rangeof outcrossing

analyseshomozygous recessive,
attributed

This fairly

ranging

types, whereas the other

differentiationloci unlinked

tonor the reduction
a more detailed

P. vulgaris.

Final comments
Analyses of genetic diversity

are only as good as the plant and genome samples on which

they are based. In recent years a substantial

amount of effort has led to increased

availability

of

28

wild

crop relatives,in particular

germplasm is providing

wild legume relatives.

us a with a wealth of information

crops as illustrated

by our data on Phaseolus

directfield

additional

geographic

regions.

Literature

cited

Combined

explorations

with molecular

about the origin

efforts

Beebe, S., O. Toro, A. Gonzalez, M. Chacon, and D. Deouck, 1997.

and their implications
Debouck,

thisof legumehelps

to potentiallyinteresting

Wild-weed-cropcomplexes

vulgaris

L., Fabaceae)

in the Andes of Peru and Colombia,

for conservation

and breeding.

Genet. Res. Crop Evol. 44: 73-91.

D.G., O. Toro, O.M. Paredes,

and ecologicaland evolution

spp. and Vigna spp. In turn this informationand other conservationof commonbean (Phaseolus

analyses,

distribution

W.C. Johnson, and P. Gepts,of Phaseolus

vulgaris

1993.

in northwestern

Genetic

diversity

South America. Econ.

Bot. 47: 408-423.
Doebley,J., and A. Stec, 1 99 1. Genetic
and teosinte.

Doebley,

comparisonof resultsfor epistasisof the morphologicalfor two F2 populations.
1995.

Teosinte

and the volutionJ., A. Stec, J. Wendel,a maize-teosinteof the morphologicaldifferences

between maize285-295.

J., A. Stec, and C. Gustus,
evidence

Doebley,129:

J., and A. Stec, 1 993. Inheritanceteosinte:

Doebley,

Genetics

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S. Beebe, and M.C. Duque, 1996.

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AFLP analysis

genetic

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31
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R.E., and N.F. Weeden, 1 992. Chloroplast

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Leguminosae.

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1999.

Identificationof geneticof rice using an F2 population

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1.

factorsof a cross between

33

Questionsand answers:

Questions

Keynotesessionto Professor Y.Shimamoto

Q. Howmanywild legume populations
A. Based on cytoplasmic
soybeanwere involved in the domestication

process?

genome data we believe that several wild populations

domestication.

At least

three

wild

soybean(Debouck)were involved in

populations

became soybean

landraces.
Q. Is there any information

on how wild soybean was domesticated

A. I do not know what the process of domesticationin China or Japan?(Vaughan)

was from small seeded indeterminate

to large

seeded determinate.
Questionsto Professor P. Gepts

Q. I was surprisedthat your data showed that the wild type is less polymorphic

Do you think it is due to selection
A. We were surprisedby man during domestication.

by this observation

reduction in genetic

diversity

domestication.

sequences from the Mesoamerican cultivars
subcluster

within the Phaseolus

under cultivation
increasedvulgaris(not necessarily

reproductive

isolationcluster.

resultsalso indicatedA neighbor-joining

includedconsciousatree shows thein our study to form a distinct

This suggests
selection

due to enhanced selfing,

divergence for a amylase inhibitor(Morishima)also. Most of our earlier

duringthan the cultigen.

that increasedby humans),

selection

combined

may have accelerated

sequences. However, this is a hypothesis

withsequence

which remains

to be tested.
Q. I had thoughtthat the structureand functionof a amylase inhibitor

resembled those in commonbean. Your results indicate
in P. coccineas is far from that in P. vulgaris.
A. For the structureand function

a amylase inhibitor

What is your explanation?

of a amylase inhibitor

those ammino acids (and hence correspondingin Phaseolusof the two species,
nucleotides)

involved

coccineas
gene family(Ishimoto)it is sufficientforin the structure

andfunction of these enzymes to be conserved. All other amino acids can vary as long as theconformation of the enzyme is not affected. This is particularly
used in our study, which represents
Presumably,

therefore,

this region

polymorphism

among accessions.

Q. Do you have any information

the case with the sequencepart of the gene not part of the activeis less subject

on the a amylase inhibitor

to conservation

and will show moregene family in Phaseolus(Ishimoto)
A. Wehave no information

about a amylase inhibitors

in Phaseolus

site.

acutifolius.

acutifoliusl

34

Q. What is the possibility

and how soon will we have appropriate

the geneticof genebank collections

priorities

composition

for ex-situ collecting

A. Genomics (i.e. high throughput)
These techniques

technologies

developedthat will adequately

there

is a positive

political

challenges.

general, willing
eliminatingof in-situ reserves?

that affect their application

to gene banks.

in that the particularsequences have to be identified

describe

diversity

the genetic

this technology

is still

return on investmentof the species

is a financial

expensive.

Regardingought to be able to develop

Finally,

there areprograms infrom genetic analyses, for example by

the first challenge,

the appropriate

costs

It is not clear at what pointin genomics technologies.

obtainedandand will beone. Although

To what point are gene banks and genetic conservationto act upon the findings

duplicates?

in terms ofso far mostly to "model species" such as humans andamenable to genomic approaches. A second challenge
are expected to decrease,eitheranalysealready exist to assess DNA sequence diversity.

yeast. There are several types of challenges
challenge

to routinely

to aid conservation,routes and the location

have been applied

There is a biological

techniquesa reasonably

methodologies

well funded projectin 3-5 years.

Session

Biosystematics

1and ecogeographic

techniques1Crop37

Biosystematics

and genetic

resourcesof thegenus

Vigna subgenus Ceratotropis
N. Tomooka1, Y. Egawa2 and A. Kaga1

Evolutionary
Tsukuba 305-8602,
2JIRCAS

Dynamics
Ibaraki Japan

Laboratory,

Genetic

Resources

Okinawa branch, Maesato Kawarabaru 109 1- 1 , Ishigaki,

Department,

NIAR,

Okinawa 907-0002

Abstract
A brief review of the taxonomic history
given and principle

morphologicalcharacteristics

Recent research which sheds light
discussed

which distinguishon the relationships

and this leads to several suggestions

our recommendations is to divideof the genus Vigna subgenus Ceratotropis

this subgenus are described.

among taxa within this subgenus

for revisingand Aconitifoliae.

boundaries

between these taxa is problematic.

arenomenclature of the group. Amongthe subgenus into three sections

Radiataeisnamely sections Angulares,

Due to a lack of germplasm for several
This is particularly

taxa delimitationoftrue for close relativesof V

hirtella.
Recent germplasm collecting
in collaboration

with scientists

are discussed.

Genetic variationand island

to island

Ceratotropis

variation

for subgenus Ceratotropis

by MAFF(Japan)

scientists,

in other Asian countries,

and research on collected

germplasmin species

and temperate regions are comparedelaborated.

from tropical
Evaluation

species level core collection

for bruchid

resistancehas revealed a high proportion

using a subgenus
of potentially

usefulaccessions for plant breeding.
1. Introduction
1970)

The genus Vigna subgenus Ceratotropis
currently
to 17 recognised
species (Marechal
et aL, 1978;

naturally

distributed

sevensubgenera

characteristics

inacross

Asia

the

genus

and thus often
Vigna,

which make it of particularthecalledconsists
Tateishi,of 16 (Verdcourt,
1985)
which arethe AsianVigna. Among thesubgenus

Ceratotropis

hasinterest:a) Biosystematics
The Asian Vigna are considered

to be recently

evolvedand morphologicalseveral38differentiation
between taxa is limited (Baudoin & Marechal, 1988). As a consequence
distinguishing
characters between taxa are limited and the subspecificclassificationofsome of the Asian Vigna is complex. The group provides germplasm with which tounderstand speciation processes,b) Domestication
Half of the Asian Vigna are domesticated,semi-domesticatedor cultivatedforfood, forage and/or cover crops (Table 1). The various stages of domesticationanddifferentiation
between ancestral wild species and domesticatesprovides germplasm
with which to clarify domestication
processes.

Table

1. Eight

cultivatedspeciesin the 17 Ceratotropis(adapted

sp e c ie s
V . ra d ia ta

speciesfrom Tomooka et al, 2000 b, c)/c olom c ma l on namn ae m eu sew ild fo rm

d istrib ut io n o f w ild fo rm

m u n gb e a n ,fo o dv a r. su b lo b a ta

E ‑ A fric a , M ad a g a sc a r,

g re e n g ram
V. m ung o

V . a n g u la ris

blac k g ram ,b lac k m atp e ,u rdaz uk i b e a n ,A sia, N e w G u in e a , A u stra lia
fo o dv a r. silve str is

In dia , M y an m arfo o dv ar . n ip p o n e n sis

H im a lay a s, N . M y an m ar,re d b e an

C h in a , K o re a , Ja p a n

V . u m b e lla ta

ric e b e an

fo o dv ar. g ra cilis

E ‑In
In
d o ‑Cdiah ,inT ah, ailan
C h in da,

V. a c o n itif o lia

m o th b ea n ,m at b e a n

fo o dc o v er c ro p

in distin g uish a b le

P ak ista n , In d ia

V . tr ilo b a ta

p illip se sa ra b e an , fo ra g e
jun g li be a n
c o v e r c ro p
(In d ia n )fo o din d istin gu ish a ble

In dia , Sri L a n k a

V . stip u la c e a

V . refl e x o ‑p ilo sa L en tille de c re o le fo ra ge
v a r .g la b ra
(M a uritiu s)(= V . g la b re sc e n s )V . trin er via
tua p e e (T h a i)v ar # ref lex o ‑p ilo sa S .E . A sia , S . Ja p an ,f‑‑ dc o v e r c ro po c ea n ia
in distin g u ish ab leTM hda aod ila
Innageansia
scd, ar,,MP aInlap udyaiasia
N, Srie, w GL uanink eaa

c) Biotechnology
Among legumes the Asian Vigna have particularcharacteristics
which makethem useful with respect to new biotechnology
methods. A highly
efficienttransformation
system has been developed for azuki bean (V angularis)(Sato,
1995) aprocess which is generally
reported to be difficultin legumes. The genome size of
species in this subgenus that have been measured are among the smallest for legumes
ranging from 470 to 560 Mb for mungbean (Arumuganathan & Earle, 1991). Thus thepotential
for map-based gene cloning exists. Asian Vigna can be used as a model forgene(s)function analysis cloned from the diverse wild relatives
found in various
habitats in Asia.
In this paper we present a brief historicalreview of the taxonomic treatments ofthe subgenus Ceratotropis and some of our own research and its implications
related tobiosystematicsof the subgenus. We then discuss the efforts by the MinistryofAgriculture,
Forestry and Fisheries(MAFF), Japan, in collaboration
with partners inAsia, to collect, geneticallycharacterise
and evaluate Vigna subgenus Ceratotropis
genetic resources.
2. Biosystematics
2.1. Taxonomy
a. History
The history of the taxonomic treatments of the Asian Vigna at the genus andsubgenus level is summarized (Table 2). The Asian Vigna were initiallyclassified
into
Table

2. Historyof taxonomic treatment

Names in parenthesisof the Asian Vigna (the subgenus(after Tateishi& Ohashi,

Ceratotropis)
1990)

refers to the subgenus or section of the genus in the same box

by

40

the genus Phaseolus by
Candolle (1825).
A Japanese taxonomist, Ohwi (1953)
proposed a new genus Azukia for this group. Maekawa (1955) further divided this group
into two genera, Azukia and Rudua, mainly on the basis of seedling characteristics.
However, these treatments that consider the Asian Vigna constitute a distinct genus or
genera has not gained general acceptance.
Verdcourt (1970)
proposed a restrictedconcept for Phaseolus,
limiting
itexclusively
to those American species with a tightly
coiled style and pollen grains
lacking coarse reticulation.
As a consequence the concept of Vigna was enlarged tocontain the Asian Vigna group as a subgenus, Ceratotropis.
Verdcourt listed 16 species
in the subgenus Ceratotropis.
Marechal et al. (1978)
wrote a monograph on the Phaseolus-Vigna
complex,which has become the standard taxonomic system for this group. In their monograph,they recognized seven subgenera in the genus Vigna, namely, Ceratotropis, Haydonia,
Lasiospron, Macrorhyncha, Plectotropis,
Sigmoidotropis
and Vigna. The relationships
and geographicaldistributionof these subgenera are summarized (Fig. 1). Marechal etal. (1978)

recognised

17 species

in the subgenus

Ceratotropis.

Fig. 1. Phylogenetic

relationshipand geographicaldistribution

genus Vigna and the genus Phaseolus
Lines between each group indicate
Size of the box corresponds(modifiedof seven subgenera

from Marechal et al 1978).

a close relationship.

to the number of species.

in the41

Tateishi(1985)
proposeda revisionof the Asian Vigna based both on
examining specimens in European and Asian herbaria and intensive field studies inmany Asian countries.
The taxonomic systems of Marechal et al (1978)
and that of Tateishi (1985)
are compared (Table 3). Both recognize
17 species
in the subgenus Ceratotropis.
However, there are some differences.
Tateishi's
revision contains two new species and 9
newcombinations. The new treatments by Tateishi can be summarized as follows.
1) V. grandiflora
and V pineryia were separated from V radiata var. sublobata as
distinct
species.
2) V. subramaniana was distinguishedfrom V. radiata and the key morphologicalcharacters of V.subramaniana were described.
3) Among specimens formerly recognized
as V. trilobata,a new species wasrecognized V stipulacea.
4) V nakashimae and V. riukiuensis
were included within the species limits of V
minima as V minima subsp. nakashimae and V. minima subsp. minima var. minor,
respectivel y.
5) V umbellata var. gracilis was transferred to V minima as V minima subsp. gracilis.
6) The tetraploid
cultigen V glabrescens was considered a subspecies of the tetraploidwild species V reflexo-pilosa and called subsp. glabra.
7) V. nepalensis was described as a new species based on the materials collectedbyTateishi in Nepal.
8) V. exilis was described
as a new species based on specimens from Thailand
formerly identified
as V.dalzelliana.
9) V. bourneae was transferred to V trinervia as var. bourneae.
Synonymy of V. malayana and V.papuana listed in Marechal et al. (1978)
arenot mentioned by Tateishi (1985).
However, the type specimen of V. papuana wasidentified
by one of us (NT) as V umbellata.
b. Key morphologicalcharacteristicsof the Asian Vigna
According to the monograph presented by Marechal et al. (1978),usefuldiscriminating
characters
between the genus Vigna and the genus Phaseolus are as
follows (Fig. 2):
1) Stipule: In the genus Phaseolus, the stipule is attached to the stem by its basal part(Fig. 2-(a) Sb-0) and does not spread underneath the attachment point. By contrast,in the genus Vigna the stipule is attached in several ways but in the subgenus
Ceratotropis, the stipule is attached to the stem by its central part (Fig. 2-(a) Sb-3).
2) Knob : Knobs are reduced from the inflorescence
branch in the genus Vigna (Fig. 2

Table 3. Species and infraspecifictaxa of the Asian Vigna , the subgenus Ceratotropis
reconsnized by Marechal et al (1978) and Tateishi (1985)* species marked with * were not examined but listed in Marechal et al. (1978)
followingthetreatment of Verdcourt ( 1 970).
** V. mungo (L.) Hepper var. silvestris
Lukoki, Marechal & Otoul was described in Lukoki et al. (1980).
Synonyms between two systems are listed in the same row.

dFig.

2. Key traits
which distinguish
Phseolus(modifiedfrom Marechal et al. 1978 )Traitof subgenus

Fig. 3. Key traits

Ceratotropis

in theand Vigna

Vigna box is surroundedof Ceratotropis
species(modifiedfrom Tateishi& Ohashi,1990)

by box

44

-(b)

kb+),

but knobs are not seen in the genus Phaseolus

3) Keel and Style
(d)

6),

while

subgenus: In the genus Phaseolus,

the keel and stylein the genus

and stylemorphology(Fig.

3, (d) 3).

Ceratotropis

4) Style-beakVigna keel

they are incurved: In the genus Phaseolus,

2-(c)stigma is positioned(e) Bk 0). In the genus Vigna, the tip of the style
style

beak,

(Fig.2-(e)(Fig.2-(b)so that the stigma

is situatedkb-).

are coiled(Fig.

varied2-(c)but in theat the end of stylesometimes

elongateson a somewhat lateral

4,

(Fig.

2-

to form apart of the styleBk-2).

All the above mentioned Vigna characters are represented
in the subgenus
Ceratotropis with a high degree of expression. Subgenus Ceratotropis
species always
have flowers colored various shades of yellow, but are never purple, violet, blue or
white as is often found in other Vigna subgenera (Baudoin and Marechal, 1988).
The Asian Vigna are considered to be a morphologicallyhomogeneous groupwhich have a very specialized
and complex floral organs. The key morphologicalcharacters recognized for the subgenus Ceratotropis were summarized by Tateishi andOhashi (1990)
as follows:1) peltate stipule (Fig.3-a).
2) standard with a protuberance near the center of the inner surface of the lamina (Fig.
3-b).

3) keel petals
4) pocket

curved to the left in the upper part (Fig.3-c).

on the left keel petal

5) style

extending6) pollen

grains

(Fig.

3-d).

beyond the stigma as a beak (Fig.3-e).
with a coarse reticulate

Mungbean group

sculpture.

V. aconitifolia

Azuki bean group

group

Fig. 4. Three subgroups in the Asian Vigna based on the seedling
Mungbean group : epigeous
V. aconitifolia

cotyledonsand sessile

firstcharactersand second leaves

group : epigeous

cotyledonsand petiolate

firstand second leaves

Azuki bean group : hypogeous

cotyledonsand petiolate

firstand second leaves

Although the Asian Vigna have a rather homogeneous morphology, Maekawa(1955)
pointed out that the first and second leaves are sessile in V radiata and V mungo,
while those of V angularis and V umbellata are petiolate.
He used this character todivide these species into two genera Rudua and Azukia. These two groups of species
differ in the position of the cotyledon on germination. Rudua species have epigeous
cotyledonsand Azukia hypogeous. Baudet (1974)
found V. aconitifolia
has epigeal
cotyledonsand petiolate
first and second leaves. This seedlingtype is intermediate
between Rudua type and Azukia type. Based on these results, Tateishi(1996)
recognised
three subgroups in the subgenus Ceratotropis,
bean group s. str.9 and V aconitifolia

the Mungbean group s. stn, the Azuki
group showing intermediate
seedling

characteristics(Fig. 4).
The revision of the subgenus Ceratotropis
by Tateishi(1985)
is the mostcomprehensive and thorough to date. Recent germplasm collecting,herbarium studies
and laboratory based research have added to our knowledge of the Asian Vigna and are
discussed below.
2.2 Species relationships
in the Asian Vigna
Genetic relationships
among Asian Vigna have been analysed using isozymes(Jaaska & Jaaska, 1990), protease inhibitors(Konarev et al, 2000), RAPD (Kaga et al.9
1996; Tomooka et al, 1996) and RFLP (Kaga, 1996).
These analyses have allconfirmed that the Azuki bean group and Mung bean group are distinct. To illustratetherelationships
among species revealed based on DNA level and gene product levelvariation RFLP and protease inhibitor
results will be discussed.

Fig. 5. A dendrogram showing
the subgenus Ceratotropis

genetic relationships
among 25 accessions
inbased on RFLP analysis,(adaptedfrom Kaga, 1996)

Kaga (1996) determined genetic dissimilarities
among 25 accessions of 12 taxa
in the subgenus Ceratotropis based on RFLP data. A dendrogram based on these data ispresented (Fig. 5). The accessions were clearly divided into 3 groups (I, II and III)
which correspond to the Azuki bean group s. str., Mung bean group s. str. and V
aconitifolia
group, respectively.
This dendrogram reveals the clear genetic distinction
between taxa particularly
at the species level.

Fig. 6. A species relationships
estimated from polymorphismof proteinase
inhibitors(trypsin,chymotrypsin,
subtilisin,cysteine
protinaseinhibitors).
A phenogram
constructed by the UPGMA method, (modified
from Konarev et ah, 2000).

47

Using a more comprehensive set of accessions Konarev et al. (2000) showedthat combined data on diversity
in four types of proteinase
inhibitors(trypsinandchymotrypsin
inhibitors,
subtilisin
and cysteine proteinase
inhibitors)
also clearly
separated species into three groups which corresponds to the Azuki bean group, Mung
bean group and V aconitifolia
group (Fig. 6). While generally
diversity
analysisgrouped accessions according to taxa, clear differences
can be seen between analysis ofproteinase
inhibitors(Fig. 6) compared to RFLP analysis(Fig. 5). For example
proteinase inhibitors
did not show much variation among the taxa of the V minima
complex (V minima, V. riukiuensis
and V. nakashimae)
whereas many genetic
differenceswere found by RFLP analysis.
Proteinase
inhibitors
were particularly
revealing with respect to accessions identified
as V hirtella and revealed several groups
within this complex species (discussed
below).
2.3 Genetic differentiationin V. hirtella
V. hirtella
is distributedin the region of maximum species diversityofsubgenus Ceratotropis, northeast South Asia and mainland Southeast Asia. Collections
which have been assigned to this species have a very diverse morphology for several
characters (Table 4).
Recently, 21 accessionsof this species considered
to be V hirtella
wereanalysed by RAPDs (unpublisheddata). A principal coordinate (PC) analysis plot of thedata is shown (Fig. 7). This shows that groups based on morphology correspond well togroups based on the polymorphism revealed at the DNA level by RAPDs. Broadly PCI
separated group C and D while PC2 separated group C and the "nipponensis like"group. The intermediate group was separated from other groups by both PCI and PC2.
The accessions of V. hirtellaused in this study while from a very wide
geographic
range were not evenly distributed
and there are many areas from whichsamples were not available
for analysis,
particularly
Myanmar. Accessions fromThailand in group C and D were from a restricted area of northern Thailand (Mae HongSon, Chiang Mai and Chiang Rai) (Tomooka et at., 1997). At some sites group C and D
accessions
were growing sympatrically.
Differences
found in morphology,
genetic,
protease inhibitor(Konarev et al., 2000) and cross compatibility(discussed
below)
studies strongly suggest group C and D are separate taxa within the Azuki bean group.
Since group C accessions correspond morphologicallyto the type specimen of V
hirtella we consider this V hirtella. Group D accessions are currently being comparedwith V dalzelliana
to determine their relationshipwith this species. Accessions whichare "nipponensis
like" are being compared to V nepalensis.
Intermediate
group
accessions require further investigation
to determine whether they fall within the range

Table4. Characteristics
complex.

and accessionsof each morphologicalgroups in the V. hirtella

Fig. 7. Grouping of the Vigna hirtella complex by means of principal
coordinate analysis from RAPD data

49

of variationof V hirtella(C group accessions)or D group accessions(cf. V.
dalze lliana).
Studiesof germplasm assigned to V. hirtella
highlight
the difficultyindistinguishingseveral species in the Azuki bean group which have overlapping
areas ofdistribution
in mainland Asia, such as, V.angularis, V dalzelliana,
V hirtella,
V.minima
and V. nepalensis. Further clarificationof these species from throughout their range is apriority.
2.4 Cross compatibility
studies
Wehave studied cross compatibilityof azuki beans with other species of the
Azuki bean group. So far this research has revealed three groups of species. The firstgroup shows no major barriers to hybridizationand fertile Fj progeny can be formed
using these species as either the male (pollen)or female (seed) parent. The species inthis group are the V. minima complex (V. minima, V. riukiuensis,
V. nakashimae),Vnepalensis and an accession of V. hirtella in group D (cf. V. dalzelliana).
The second group consists only of an accession of V. hirtella(group C). This
accession produced fertile Fj progeny when it was the female (seed) parent but not the
male (pollen)
parent with azuki bean. The third group consists of V umbellata which
can only produce fertile F, progeny with azuki bean when it is the seed parent andembryo rescue procedures are employed. (Ahn & Hartman, 1978). The rescued ¥l plants
show regular chromosome pairing at meiosis but linkage disequilibriumon segregation(Ahn & Hartman, 1978; Kaga et al, 2000).
These results suggest that all these species share the same homolgous genome
and thus gene exchange is possible.
2.5 Biosystematics
studies of Vigna subgenus Ceratotropis- new proposals andfuture research directions.
a. Genepools.
Lawn (1995)
proposed that the Asian Vigna consists of three more or lessisolated
genepools based on cross compatibility
studies. These genepools correspondwith groups based on seedling characters proposed by Tateishi (1996)
as shown below:
Genepool: angularis-umbellata;
radiata-mungo; aconitifolia-trilobata(Lawn, 1 995)
Group:
Azuki bean s. str. ; Mungbean s. str:, V.aconitifolia(Tateishi,
1996)
Our own genepool studies have focussed on V angularis (azuki bean) the mostimportant of the Asian Vigna crops in Japan (see Vaughan et al. these proceedings).
From our studies of cross compatibility
between V angularis and other taxa in the
angularis-umbellata
genepool (discussed
above) we propose a genepool classification
for azuki bean (Fig. 8).

The primary genepool (GP-1) of azuki bean consists of its wild, weedy andcultivatedforms, V. angularis var. angularisand var. nipponensis. The secondary
genepool (GP-2) consists of two parts. Secondary genepool (a) consistsof species
which cross relatively
easily with V. angularis - Vminima, V nakashimae, V nepalensis,
V riukiuensis
and cf. V. dalzelliana(V hirtellagroup D). Secondary genepool (b)consists of V umbellata and V hirtella (group C) and possibly
other species in the
angularis-umbellata
genepool. The tertiary genepool (GP-3) would be species in taxa inthe radiata-mungo genepool and aconitifolia-trilobata
genepool.

Fig. 8. Gene pool classificationof Azuki beanb. Revised list of Ceratotropis taxa
Wepropose a revised list of taxa in the subgenus Ceratotropis based on pasttaxonomic treatments and the results of recent research (Table 5).
The main differences from previous lists are as follows:(a) Many studies (for example Kaga, 1996; Konarev et aL, 2000) have confirmed thevalidityof the 3 groups of species suggested by Tateishi (1996) within the subgenus
Ceratotropis based on seedling characteristics.
We have revised the species withinTable 5. Preliminary

revised list of taxa in the subgenus Ceratotropis

1. The taxonomic rank section is suggested for the Azuki bean group sensu stricto, Mung bean group sensu stricto
and V. aconitifolia
group and series for V. minima , V. nakashimae and V. riukiuensis.
These names are in theprocess of being formalised (Tomooka et al. in preparation).
2. Currently not in the MAFF germplasm collections.

52

these groups in the lightof new germplasm (Table5) and suggest

these groups are

given the taxonomic rank section and propose they be called Section Angulares(Azuki bean group), Section Radiatae (Mungbean group) and Section Aconitifoliae(b)(V aconitifolia
group).
V. minima, V.nakashimae

and V riukiuensis

are closely

morphological(Tateishi,

genetic

differences

1985),

related.

However, based ondivergence(Yoon et al., 2000)

and distinct
areas of distribution
we consider these three taxa warrant specificranking. Given the close relationshipbetween these taxa we suggest they arerecognised

as a series

withinSectionAngulares

and suggest

the name Series

Minimae (Tomooka et al., in preparation).
(c)Many workers have applied the term V. umbellata var. gracilis to the wild form of
rice bean (Lumpkin & McClary, 1994; Smartt, 1990; Lawn, 1995; Kaga et al, 1996)
in recentyears. We therefore

form of V. umbellata.
(d)feel it is logicalto conserve

this name for the wild

However, we should note that the specimens

listedas Vumbellata var. gracilis in Marechal et al. (1977)
are V minima.
Based on various studies the two tetraploidtaxa in the subgenus

Ceratotropis

belongEgawa et al., 1996;

to the same species,

Konarev et al., 2000).

V reflexo-pilosa(Tateishi,

The wild and cultivatedappear to have greater divergence

1985;

shouldforms of V. reflexo-pilosa

does notfrom each other than wild and cultivatedforms ofazuki bean, rice bean, mungbean and black gram (Konarev et al., 2000). Since wild
and cultivatedazuki bean, rice bean, mungbean and black gram are all distinguishedby the taxonomic rank of variety
reflexo-pilosa
pilosawe consider that the wild and cultivated

are best given varietal

and V.reflexo-pilosa

rank namely - V.reflexo-pilosa
topositionof V. exilis;delimitationof V. hirtella(c) species
hirtella,relationships
among V angularis var. nipponensis,
V. minima (especiallysubsp. gracilis in Tateishi's

nepalensis;(d) treatment of V. trinervia

Furthercollections

related(b) species(e) taxonomic(f) taxonomic

position
positionprogressvar. reflexo-

var. glabra.

There remain many unansweredquestionsbiosystematics.
Further studies are require in relation to:(a) taxonomic

form of V.

Ceratotropis

complex;
V.dalzelliana,Vsystem) and V.

var. bourneae (living

material

not available);of V. subramaniana;of V. khandalensis(living

material

not available).

in biosystematics

and their study which are discussed

will
below.

come from improved

germplasm

53

3. Genetic

resources
3.1 Collection
Since 1989, systematic explorationand collection of wild Asian Vigna species
have been conducted by MAFF, Japan, researchers in collaboration
with the Departmentof Agriculture,
the National Seed Bank, Katsesart University of Thailand,
Agricultural
Universityof Malaysia (UPM), Agricultural
Universityof Sabah (UMS), Plant Genetic
Resources Center, Department of Agricultureand Lands, Sri Lanka and Vietnam
Agricultural
Science
Institute(VASI)(Table6). During these explorations,Vriukiuensis,
V reflexo-pilosa, V angularis var. nipponensis and V nakashimae have beencollected in Japan. In Thailand, V grandiflora, V umbellata (wild form), the V hirtella
complex, V minima, V trinervia,
V mungo var. silvestris,
and V exilis have beencollected.
In Malaysia, V. trinervia, V. reflexo-pilosa, and the V hirtella complex werecollected.
V.stipulacea,
V trilobata
and V.trinervia were collected in Sri Lanka and V
reflexo-pilosa (var. reflexo-pilosa and var. glabra) was collected in Vietnam. Accessionsof all of these species are maintained in the MAFF genebank, Japan.
Table 6. Collectionof wild Ceratotropis

c ollab orator

Y ear

A rea

M A F F (Jap an )

198
19 99
.I9 ‑ I OｻJA
T ,kA .iP̲,An AawNIVTa,K ago shim
, . a,

by MAFF and Collaborators
C ollected sp ecies

R eferen ces

V1/t/. refV.
n ・,ul kiu
exo
・en
‑p si
il
.,s・
o sa

M A F F collection repVv o rt, 199 10 *V: mtriv.
hgra
irtell
un ng
nd
ervia
oaifl ,var.siloV.
ra m, V.
inim
v um
estri
a bellatas, V .exilis

MT o Am Fo Fokco
a,llection
199 1 rep o rt ,20, 19 00
97

re‑ r^: ;.‑.? ‑.n

KD asetsart(TOh ailan
A , T ISTd)U nR iv.

19
!9 96
9891
94

T h ailan d

19 99
u p M ,(MU Malay
S sia) lf
m t r92
19
93
v2
vｻ
19 97

Ma * alay
i (p
, sia
enin
・ . su lar, ..
M alay, sia V .h
. trinerv
irtella" ia , V. reflexoJ‑pF ilosaBM ujan
AJ F gBF e?
co a/.
llecti19o94
n rep ort, 199 3& Sab ah)V A SI(V ietnam ^19 97

V ietn am^V '. refl exo ‑pP ^oilo sa var‑ref
var.g lla bra
exo ‑p ilo sa M A F F co llectio n rep ort , 199 4

P G RC

20 00

S ri L an k av. trilo ba ta , V. stip ulacea(Sri L an ka)* MAFF collection

M A F F co llectio n rep ort,2 00 0

y trin ervia, V ign a sp .

report: Annual Report of Exploration(ed.) NIAR, MAFF

and Introductionof Plant

Genetic

Resources

54

3.2 Genetic characterizationa. Comparative intra-specificvariation in temperate and tropical Vigna species
Genetic diversityof 33 accessions of V. angularis var. nipponensis from Japan,
27 accessions of wild V umbellata from Thailand and 23 accessions of V trinervia from
Malay Peninsula were analyzed and variation in 4 isozymes (6PGDH, SDH, Lap, GOT)
were compared (unpublisheddata). No intra specific variation was detected for 6PGDH(Fig. 9a). For SDH, 2 phenotypes, 4 phenotypes and 3 phenotypes were detected for V
angularis var. nipponensis, V umbellata and V. trinervia, respectively.
For Lap, 1, 4 and
4 phenotypes were found for V angularis var. nipponensis, V umbellata and V.trinervia,
respectively.
For GOT, 2 phenotypes
were detected
only for V. angularisvar.
nipponensis.
Combining the phenotypes of four enzymes, a total of 3, 8 and 8 phenotypes
were detected
for V angularisvar. nipponensis,
V. umbellata,
and V. trinervia,
respectively(Fig. 10). Judging
from the number of phenotypes
and frequency of each
type, the genetic diversity
was considered to be the highest in the tropical
species V
umbellata and V. trinervia and the lowest in the temperate species V. angularis var.
nipponensis. The results suggest that collecting
strategies
for temperate and tropicalVigna species may need to be different with a smaller interval between collecting
sites
for tropical Vigna species. To what extent this is a general phenomenon requires further
study.
b. Intra specific variation in island populations
To clarify the genetic diversity
within the diploid
species V. riukiuensis
andthe wild tetraploid
V. reflexo-pilosa
var. reflexo-pilosa
and its related cultigen
var.
glabra, 52, 19 and 1 accessionof these three taxa, respectively,
were analyzed(unpublisheddata). Germplasm of both wild species were collected from the Nansei
islands,
Japan and Taiwan (China)(Fig. ll). An accession of V. reflexo-pilosa
var.
glabra was collectedfrom the Philippines.
The intra specificgenetic variation was
estimated by means of polymorphism
in SDS-PAGE banding of total seed protein andtwo kind of enzymes (SDH and Lap).
For V. riukiuensis,
no variation was detected for SDH and total seed protein
banding pattern, but 5 phenotypes were found for Lap (Fig. ll; Table 7). In contrast, all
the methods could detect polymorphism
in V. reflexo-pilosa (Fig. 1 1 ; Table 8).
Lap phenotype A in V. riukiuensis
was the most commonand found in all the
Nansei islands
but not Taiwan. On the other hand, Lap type C was found only inaccessions from Iriomote and types D and E were found only in accession from Taiwan.
For SDH in V. reflexo-pilosa, only one accession from Okinoerabu showed type B,

Fig.

9. Zymograms

showingvariationand (d) GOT for V angularisand V.trinervia

observed

var. nipponensis

in (a) 6PGDH,(nip)

V. umbellata

(b)

SDH,
(umb)

(tri)

Fig. 10. Isozyme phenotypes
among accessions(histogramaxes : left = no. of accessions,of 3 Vigna species,
bottom = phenotypes(c) Lap

Fig.

ll.

Collection

and var. glabra)

locationand theirof V riukiuensis
isozymeand V reflexo-pilosa

phenotypes(var. reflexo-pilosa(a. SDH, b. Lap) and seed protein

electrophoregrams(c).
Table 7. Lap phenotypes in Vigna riukiuensis

Table

8. Isozyme and seed protein types in Vigna reflexo-pilosa
and V. reflexo-pilosa
var. glabra

var. reflexo-pilosa

57

while the other 19 accessions
includingvar. glabra showed type A. For Lap in V
reflexo-pilosa, 4 phenotypes were detected. Lap type B was found only in accessions
from Momote and Yonaguni, type C was found only in an accession from Yonaguni,and type D was found only in two accessions, one of the wild var. reflexo-pilosa
inTaiwan and the other in the cultigen var. glabra from the Philippines.
For total seed
protein banding, only one wild accession of V reflexo-pilosa
from Okinoerabu showed
type B, while all the other wild accessions showed type A. The cultigen var. glabra
showed a different pattern (type C), but the pattern was similar to that of var. reflexopilosa.
These results suggested
that V riukiuensis
and V. reflexo-pilosa in the Nansei
Archipelago,
Japan, and Taiwan (China) contains considerable
island to island genetic
variation.
The results also revealed that in the wild variety of V. reflexo-pilosa thedetected variation contains some of the phenotypes of the cultigen var. glabra. These
data confirms the results of others that V reflexo-pilosa var. reflexo-pilosa is the wildprogenitor of the cultigen V. reflexo-pilosa var. glabra (Egawa et al, 1996; Jaaska &Jaaska, 1990).
3.3 Evaluation
Wild species core collection and bruchid resistance
To date only one source of complete resistance to bruchid has been identified
in the genus Vigna subgenus Ceratotropis
and used in breeding
programs. This
resistance source to Callosobruchus
chinensis (azuki bean weevil) and C. maculatus(cowpea weevil) is a major gene (Br) and was found after screening only 3 accessions ina wild mungbean collection(V. radiata var. sublobata - V.sublobata in the original
paper) (Fujii & Miyazaki, 1987; Fujii et al, 1989; Kitamura et al, 1988; Tomooka et al,1992). Since this resistance source is a single major gene new sources of resistance are
an important focus of Vigna genetic resources evaluation. A subgenus Ceratotropis wild
species core collection
was evaluated for new sources of resistance to bruchid beetles
and compared with evaluation of cultivatedaccessions.
The species in this core collection represent 76% (19 taxa) of the known taxa inthe subgenus Ceratotropis
and this was evaluated for resistanceto two species ofbruchid beetles, Callosobruchus
chinensis and C. maculatus. Seven taxa (37% of testedtaxa) consistingof 29 accessions(46% of screened accessions)were found to beresistant to C. chinensis and 4 taxa (26%) consistingof 24 accessions (38%) were foundto be resistant to C maculatus (Table 9) (Tomooka et al, 2000a). On the other hand, noeffective
resistancesources were found when 426 and 330 mungbean landraces
accessions were evaluated for C chinensis and C. maculatus resistance,
respectively.

Results of others who have screened landrace accessionsof mungbean and cowpea(Vigna unguiculata)
have also found very few resistant accessions (Epino & MoralloRejesus, 1990 ;Singh et al, 1985).
Table 9. Effectivenessof finding new resistance sources to bruchid beetles using
species level collectionin the genus Vigna. (adapted from Tomooka et al.> 2000)

The wild species core collection
for the Asian Vigna has been found to beeffective in revealing new bruchid resistance sources. Consideringthat bruchid strainsor biotypes may overcome a single resistance gene (Credland,1990), incorporationofmultiple resistance into the crops is likely to be more durable. Species level collections
may also be effective for finding other types of novel genes.

Acknowledgements
Wewould like to thank collaborators
we have worked with in relation
the work presented
Plant

Breeding

Professor

in Japan and around the world with whom

various aspectsof the work presented

here. Some ofwas conducted while one of us (AK) was a graduate student

laboratory,

Kobe University,and acknowledge

the kindin thesupportof

Kamijima.

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Maekawa,

F. 1955.

Topo-morphologicalcomplexed'especes(English

Piper,

C.V. 1926.

Sato, T. 1995.
Ohashi).
Singh,

Studies
study

traitees

published

Smithsonian

Five oriental

Basictaxonomical
1978.

des genres Phaselolus

editionWalker (Eds.)
C.V. 1914.

and& F. Stainier.

de donnees morphologiques,

Piper,

production

and uses. CAB

studiesinPhaseoliae,

Jap. J. Bot. 15: 103-116.

R., J. M. Mascharpa

Ohwi, J. 1953.

Azuki bean: Botany,

268 pages.

Leguminosae.
Marechal,

1994.

speciesin Americantaxonomiqued'un groupe

et Vigna (Papilionaceae)

sur la basepar Fanalysis

in 1965)

Flora

informatique.
of Japan.

DC, USA.

of beans, Bull.

U.S.D.A. 119,

Phaseoleae,

28.

1-32.

Contr. U. S. Nat. Herb. 22, 663-701

in Adzuki

Prefectural

Boissiera

Meyer, F. G. & E. H.

Inst., Washingtonof Biotechnology

Report of Hokkaido

Etudebean (Vigna

Agricultural

87. (in Japanese with English summary).
B. B., S. R. Singh & O. Adjadi. 1985. Bruchid

resistanceangularis

Experimentin cowpea.

Ohwi &

Stations.

No.

Crop Sci.

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25:736-739.
Smartt, J. 1990.

Grain legumes. Evolution

and genetic

resources.

Cambridge

University

Press. 379 pages.
Tateishi,Y. 1985.

A revisionof the Azuki bean group, the subgenus

genus Vigna (Leguminosae).

Ph. D. Dissertation,

Ceratotropisof theTohoku University,

Japan.

292pages.
Tateishi,Y. 1996.

Systematics

24 in P. Srinives,
Collection,
Tateishi,of the species

C. Kitbamroong

evaluationof Vigna subgenus& S. Miyazaki

and utilization

Ceratotropis.

(eds.)

for breeding

Pages 9-Mungbean germplasm:

program. JIRCAS.

Y. & H. Ohashi. 1990. Systematicsof the Azuki bean group in the genus Vigna.
pp. 189-199.
In Fujii, K. et at. (eds.) Bruchids
and Legumes: Economics,
Ecologyand Coevolution.

Tomooka, N. 1991.
radiata

Genetic(L.)

Kluwer Academic Publishers,

Diversity

Wilczek,

Ceratotropis)

andas breeding

Agriculture

Research

Development

evaluation

Tomooka, N., C. Lairungreang

wild

relatives(thesubgenus

No.28,

pp.1-64.

TropicalY. Egawa & C. Thavarasook.

resistantmungbean

Plant Breeding.
& Y. Egawa. 1996.

in Thailand

based

line

on RAPD analysis.

C. Kitbamroong
evaluation

and utilization& S. Miyazaki
for breeding

Tomooka, N., S. Chotechuen,

N. Boonkerd,

B. Taengsan,(eds.)

Egawa, T. Yokoyama & Y Tateishi.

1997.

nodulesamplesCeratotropis

Centraland Northernand Introduction

with English

effectiveness(Ed.)

tolerant

AnnualResources,

Vigna

31-40

in P.

Pages

Mungbean

germplasm:

S. Nuplean,Collection

D. A. Vaughan, Y.

of seedspeciessamplesandVigna)

in(genus

In: NIAR, MAFF, Japan (Ed.)

AnnualReportof Plant

13:189-206.

(in

Genetic

wild species;

searchingin the genus Vigna subgenus

Tomooka, N., D. Boonmalison,temperatureof wild

program. JIRCAS.

Resources

D. A. Vaughan, M. Ishimotoof evaluation

beetles

mungbeansummary).

Tomooka, N., K. Kashiwaba,
bruchidsubgenus

Thailand.

wild

Taxonomic position

Srinives,

from wild

using

1992.

109:60-66.

Collection,

Japanese

Vigna

In: Tech. Bull.

P. Nakeeraks,of bruchidof Explorationof mungbean,

Center, Tsukuba, Japan.

germplasm in Thailand.
collected

differentiationof itsmaterials.

Tomooka N., C. Lairungreang,speciesand landrace

Netherlands.

P. Srinives

on Exploration

1 6 (in preparation).

Ceratotropis.

& Y. Egawa. 2000b.

Asian Vigna species

Report& Y. Egawa. 2000a.
for sourcesin Thailand.

Euphytica
Field

The

of resistanceto(in press)

survey of the high

In: NIAR, MAFF, Japanand Introductionof Plant

Genetic

62

Tomooka, N., D. A. Vaughan, Ru-Qiang Xu, K. Kashiwaba
nativeVigna genetic

resources.

Japan& A. Kaga. 2000c.

Agricultural

ResearchJapanese

Quarterly,

(in

the

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Verdcourt,

B. 1970.
TropicalStudies

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Yoon, M. S., H. Jinno,

in theLeguminosae-Papilionoideae

Flora

IV. Kew Bull. 24:507-569.

A. Kaga, N. Tomooka & D. A. Vaughan. 2000. Genetic

in the Vigna minima complex and related(in process).

forspeciesin East Asia. Plant

diversity
Research.

63

Ecogeographic

Techniquesand Conservation:

A Case Study for the Legume Genus Vigna in Africa.
Nigel Maxted \ Phumzile
Schoolof BiologicalB15 2TT, U.K.
236 Broad Street,

Mabuza1 and Shelagh

Sciences,

The University

Kell 2of Birmingham,

Edgbaston,

Birmingham

Lyme Regis, Dorset DT7 3QF, U.K.

Abstract
An ecogeographic
been established;

thismethodology

for identificationof centres of plant diversity

has recently

will be discussed

and illustratedusing a case study for the legume genus

Vigna Savi in Africa.
Ecogeographic
studies involve the collation
and analysis of large andcomplex data sets obtained from the literature and from the passport data associated with herbarium
specimens and germplasm accessions.
geographic

and taxonomic.

The data compiled

are of three basic

These data can be synthesisedkinds:

ecological,

to produce three basic products:

thedatabase - which contains the raw data for each taxon; the conspectus - which summarisesthe data
for each taxon; and the report - which discusses the contents of the database and conspectus, as wellas proposing

future collection

and conservation strategies.

Ecogeographic

studiesare an essential

component of any in situ conservation programme, not only because they aid the location oftaxonomic and genetic diversity, but also because they provide critical assistance in the formulationof appropriate
1

and effective

conservation

programmes for target plant species.

Introduction

1.1 Why undertake

ecogeographic

The

inhabitedlocationsenvironmental

and geographicalpassport

data associatedto predict

where plantparticularspecies

limestone

scree slopes

perennial

gardenthese constraints

surveys?
byplant

constraints.

with herbarium
populations

or phenotype
above 2000

pea relative
are clearlyspeciesare

The analysis
specimens

definedbydifferingof ecologicaland germplasm

sets

of

and geographicalaccessions

can be used

may currently

be located.

For instance,

if such data for a

indicates

previously

it hadbeen

thatmetres in South

Vavilovia

onlyWest Anatolia,

formosa (Stev.)

A.Fed.,

foundon

as is the case for thethen locations

where one should survey first. The financial

matching

and practical

64

resources available
the application
is essential

for conservationof ecogeographicof genetic

diversity

will always be limited;

techniques

to identify

to make the best use of available

resources.

needefficiency

of conservation(CBD).

ParticularlyThis

for increased

Convention on Biological

Diversity

optimaltherefore

locationsof genetic

reserves

effortis underlinedin thein the field of in situ conservation,

where the CBD calls on nations to:"Develop,where necessary,

management of protected

guidelines

for the selection,areas or areas where special

taken to conserve biologicalestablishment

measures need to be

diversity."
ArticleEcogeographic

techniques

providethis CBD article(Maxtedet al, 1997a).

a partial"An ecogeographic

surveyinformationand synthesis

gatheringbe used to assist

and8 - CBD (UNCED, 1992)

but essential

means of meeting the objective

An ecogeographic

survey can be defined thus:

is an ecological,
process.

in the formulation

geographical
The resultsof collectionofand taxonomic

are predictive

and conservationand canpriorities."

Maxted e* al (1995)
1.2

Survey or Study?
The difference

less detailed
in their

between a survey and a study is one of degree;

data analysis

ecogeographicof the climaticand soil characteristics

throughout

relatedto annual rainfall

list

Syria.

recently

and that populations

priorities

been completed

by the author.

activities,

ecogeographic

collatingexisting

1.3

for forage species

ecogeographic

exploration

the basisof the ecogeographic

because of the very detailed

For instance,

were

A similarly

and conclusions
data gathered

as a routine

a comprehensive
detailedprerequisitehas

tree genus Leuceana.
derivedto plant

were only

the jute genus Corchorus L. in Africa,
data.

collection

as they focus onrather than collecting
passport

study

over fifteen years of field

to prove more applicable,

by other plant collectors,

Methodology

and seed production

data analysis

specimen and germplasm accession

An Ecogeographic

theyon the leguminous

reason,surveyed

analysesof the species

workingsurveys are likely

Edmonds (1990)

multivariate

data, they proposed

in Syria.

ecogeographic

For thisdata recordedof herbarium

Ehrman & Cocks (1990),

the distribution
diversityain the drier areas face greater threat of genetic

by Hughes (1998)

However, the detailed
possiblethat influenced

They found that species

Based on their analysis

of conservation

phase than a study.

study of the annual legumes of Syria, presented

samplederosion.

and interpretation

a survey involves

fresh data.
purelyon

65

In practice,precedessome form of ecogeographic

all conservation

activities.

survey is outlinedin the simple

This methodology

will be discussed

Vigna Savi.
collection

One procedure

methodology

The model

and analysis,

and production.

Project

2.1

Project Commissioning
The projectcommission

ecogeographic

necessarily

an ecogeographic

by Maxted et al. (1995)(Figureusing a case study for the legume genus

is divided

into

three

phases:may vary from an individual

data prior to a collectingin Pretoria,

relatives,to international

Institute(IPGRI),

trip, to a national

South Africa, planning

project

design,data

conservation

agencies

study of a particulartargetarea may also vary from one species
Vigna L. species

in Africa,

throughout

agency e.g. National

Biodiversity

conservation

e.g. International
Conservation

in a restricted

Plant

Union

taxon world-wide.

of a group of crop
Genetic

Resources

commissioning

The target

a fulltaxon and target

area to a whole genus worldwide,
Africastatement

area to be investigated,

outlining

any specificwill outline

or Vigna unguiculata

the objectives,

breadthof the target

whether a survey or study is to be undertaken,

requirements,

such as whether

thefor in situ and ex situ conservation

An example of a project
Saharan Africa

commission

by the International

not alreadyproducts

strategies

shouldor plantof utilisation

Vigna species
agriculture.

Genetic Resources

Institute.

areas

containnovel genetic

in selection

or breedingof African agriculture.

strategy
local

of immediate"

for the genus Vigna L. in Subthat

contacts.

It should

The

The report should containfor the genus, includingcollecting

also attempt

and medium-term potential

routes,to identify

value to African

taxon

focus

for the legume genus Vigna :conserved that could be utilised

conservationand suitable

Plantof identifying

programmes for the benefit
a detailed

is provided

survey is commissioned

survey has the objective
diversityL.

as well as

potential.
"An ecogeographice.g.

range.

and target

recommendations

gathering

Vigna sect. Catiang in Southern

its natural

The commission

conservationist

the systematicor IUCN - The Worldecogeographic

timing1).

designInstitute(Walp.)

and analysis

for undertaking

proposed

and illustrated

in Africa.

2.0

data collection

those

on

PHASE 1

PROJECT

1. 1 Projectcommissioning

1.2 Identification

1.3 Selection

1.6 Designing
PHASE 2

DESIGNof taxon expertiseof target taxon taxonomy

1.4 Delimitationof the target area

1. 5 Identificationof taxon collections

and building

the ecogeographic

DATA COLLECTION

2. 1 Listing

database structure

AND ANALYSIS

of germplasm conserved

2.2 Media survey of geographical,
2.3 Collection
2.4 Selection

ecologicaland taxonomic dataof ecogeographicof representativedata
specimens

2.5 Data verification

2.6 Analysis

of geographic,

ecologicaland taxonomic data

PRODUCTION

PHASE 3

3. 1 Data synthesis

3.271
Ecogeographic
database

3.2.2

3.2.3

Ecogeographic
conspectus

Ecogeographic
report3.3 Identification

Figure1.

AnEcogeographic

Paradigm

of conservation

priorities(from Maxted et aL, 1995)

67
2.2

Identificationof Taxon Expertise

The inherent

difficulties

many identification

aids,

in the identificationof wild

species,with

identifying

olderherbariumof material.

Emphasisshouldthereforebe placed

undertakeor supervise

ecogeographic

surveys.

and problems

associatedthe poor qualityofspecimens,

can lead to mis-identification

on finding

appropriate

specialists

to either

The specialist

should,if possible,

have a good understandingof the geographyof the area tobe studied,especiallyin thecase of regionalor localstudies.

Local

geographicalknowledge

may also prove invaluable

when trying to decipher

location

detailsfrom aspecimen label
continent
citieswrittenby hand several decades ago. Such knowledge

such as Africa,

in awhere there have been many name changes of villages,

towns andliterature,in recentyears.

The specialist

will

recommend local

florasand monographs,

suggestvisited,may be essential

put the researcher

in contact

be ableto suggest

which

appropriate

herbaria

and/orwith other specialists

and provide

and phytogeographic

specialists

gene banks be

generalassistancebased on their own experiences.
The appropriate
identified

from the authorship

phytogeographic
lists

taxonomic

studiesof taxonomicof the target

the major herbariaof Africa

region.

and gives names of researchersgroup that they study.

maintainedof Helsinki

available

over the Internet.

lists

http ://www. hel sinki. fi/kmus/botmenu.

taxa,

and

more than 1,500

floras(Holmgren

et al., 1990)

working in those herbaria.

On the Internet,

The web site is updated

can beof target

Index Herbariorum

It may also list the specialist
by the University

treatments

to approacha World Wide Web site

links

to botanicalresources

weekly and can be found at:

html

Contact name: Dr. R. Lampinen, email: Raino.Lampinen@helsinki.fi
2.3

Selectionof Target

The importanceto undertaking(1995).

Taxon Taxonomyof having a good taxonomic understanding

an ecogeographic

Such knowledge

recent classifications

study is underlined

can be obtained

and revisions,florasand monographs,

classificationby Marechal

These classificationsof thetargetidentification
details.

taxon

is that proposed

If there is no generally

can be traced by consulting

acceptedclassification,

an expert in the particulartaxon specialists,

and taxonomic studies.

by Verdcourt

synonyms,

as well as providing

group priorand Maxted et al.

targetlist the taxa currently

and commonly associatedaids for the taxa included,by IBPGR (1985)

from various sources:

genus Vigna, the accepted
et al (1978).

of the target

delimit(1971),

and amended

consideredmembers

theprovidestudy,

some generalecogeographic

the various classifications
plant

For thegroup or by a literaturethat exist
search.

68More obscure groups may lack a recent revisioncollatethe available

publishedtaxonomic

or monograph,

data as this will

but the researcher

must

form the taxonomic backbone

tothe study.
The majorityof morphologicalof wild species
characteristics

have been described(as is the case for Vigna).

less often considered

the degree

tend to be restricted

to well-known

relationshipsof genetic

crop plants

well understood,or Ng and Padulosi(1991)

2.4

Delimitationof the Target

The target

area of the taxon beingcommission,

throughout

its range.

The commissioning

geographicaland is likely

survey.

Once the region

is delimited,

distribution

or ecologicalVigna, the project

preferences
Florascommission

to limit

the predictive

detailed
by Frodin

Index Herbariorum records the location

both major (international)
passport

data.

herbarium

for the researcher

herbaria

The relative

partialthestudies

on localgeographicalfrom local Floras.

and Davis et al. (1986).

For

and germplasmofthe survey to Africa.

will need to visit the major herbarium

will be able to suggest

restricting

value of the ecogeographic

information(1984)

regionand/or time andHowever, unnecessarily

The researcher
and specialists

definedof the target taxon can be obtained

are provided
limitsclearlyofbe studied

the survey to a specific

if it is fairly
study.

by the terms

the taxon should

agent may restrictof Taxon Collections

taxon.

for wheat (Triticum

may be restricted

Identification

the target

the genetic

prove to be a false economy if multiple

undertaken,

to appropriate

where

and the make-up of the genebut if it is unspecified

for a complete

survey may ultimately

are subsequently

2.5

studiedregion of Africa or all Africa)

are not available

studiedallies,Such studies

Areaof the project

Guidesand their

havefor cowpea (Vigna unguiculata).

referenceresources

taxonomists

between species.

for example, Feldman et al. (1995)

spp.)(e.g. Zambesiaca

using a combination

Traditional

differentiation

among the taxa have been extensively

pool is relatively

and classified

which herbaria
and local

advantages
are as follows:(national)

collectionsof major dried plantcollections,and libraries

should be visited.

Ideally,

herbaria

would be visited

to recordand disadvantagesof the two categories

of

69
A

Major international
Herbaria

dvantages

1.

Broad
taxonomic
coverage, 1. Predominance of old collections,makingpossibly
material
used in theextraction of passport data more difficult
productionofrevisionsandand likely predictive value lower.
monograph s.
2. Geographical names associated with older
Broad international
geographicalcollection
sites may have changed morecoverage, possibly material used inrecently.
the production of local Floras.
Skilledresearchersavailable
toprovide general advice.
Appropriate
taxonomic
andgeographicalspecialists.
Type material of target taxa.
Good botanical library.

2.
3.
4.
5.
6.
Regional
herb aria

1.

Good local regional
coverage ofl.
target area.
Better documented material, as the2.
herbarium is likely to have been3.
more recently established.
Regional specialists
present, who
can assist in deciphering
localgeographical names.

2.
3.

Table 1.

Disadvantages

Limitedresourcesforherbarium
maintenance.
Lack of target taxon specialists.
L
imited botanical library.

The relative advantages and disadvantages
and local (national)
herbaria.

For the ecogeographicof major (international)

survey of Vigna in Africa,

ideally

the researcher

Museum, U.K., Museum

the Royal Botanical

Gardens,

Kew, U.K., the Natural

History

Nationald'Histoire

Naturelle,

Paris,

France and Jardin

Botanique

Brussels,

Belgium.

These herbaria

have large collectionera.

were collectedduringthe coloniallargelyin these

institutions

writtenspecimens used. Within
Herbarium locatedfrom Central

in the National
specimens

Institute,

Ibadan,Nigeriafrom Southernwhichecogeographer

shouldalso visit

Vigna, notably

the International
the sampling

germplasm, the more likely
While

few herbaria

commonpracticeherbaria

Institute,from Africa

sets of thesuch as the East African

Pretoria,

500,000

specimens

South Africa which

Africa, and the National

Forestry

Research

contains

100,000

specimensfrom west Africa.

the institutes

thathouse large

germplasmof Tropical

Agriculture(IITA),

Instituteof ecogeographic

The

collections
Ibadan,specimens

the data will prove ecologicallyand geographically

predictive.

have databases

data for theirof the passportand may therefore

be possible

collections,of

Nigeria.

with herbarium

among germplasm collections

data associatedthathave been

complete

Museums of Kenya, which contains
Botanical

de Belgique,parts of Africa

have retained

might visit

would visit

Nationalof specimensof various

and the herbaria

and East Africa, the National

900,000

The broader

The Floras

Africa, the researcher

contains

herbariaor
it is

to obtain the70required

passport

data without

recordingdetailed

passport

with

germplasm

specimens.

willoften be of a higher

relatively

recently.

data is unavailable
Maxtedsignificant

the gene bank.

data by germplasm

However, systematicestablisheddata.

visitingand so herbariumet al(1997a)collections

qualitythan that

acquisition

how to select(Bettencourt,& Damania, 1989),

be noted any publishedcatalogues,

catalogues

have only beenwhichgene banks have

which gene banks need to be visited.

such as the volume on Food Legumes
which

includeswill quickly

Vigna collections

become out of date),sites,

oftheholdings(HTTP ://www. cgiar.org/singer/index.
(GRIN)of the United

World Information
Organisation

the

CGIAR

htm),

or the Genetic

StatesDepartment

and Early

Warningof the United

Nations

centresof Agriculture
Systemin the(though

databases,

Programme on Genetic Resources (ECP/GR),asdatabase

Resources Information

Network(HTTP://www.ars-grin.gov)

or the(WIEWS)of the Food and Agriculture(http://apps.fao.org:8080/wiews.new).

Each of thesewhere Vigna germplasm from Africa is conserved

2.6

Designing

the Ecogeographic

Databasea databaseshouldbe designed

discussion

on databasedata collection,

retrievalof data.

For a detailedreferred

to Maxted et al (1995).

3.0

Data collection

3.1

ListingsafelyIf sufficient
eitheron full-scale
genetic

data collation,material

structurefor efficientstorageand design,

the reader is

Details

obtainedfrom botanic

gardens,

national

and international

experts

may be able to guide the researcher
Instituteof the target

current conservation

activities

andof what material

for Tropicalis currently

being

gene banks and in situ conservation

catalogues,

databases

Agriculture

and web sites

to the appropriate(IITA)

shouldtaxon from the target area is alreadyin situ and / or ex situ, there may be no justification

activities.

International

Structureof Germplasm Conservedconservedconservation

ex situ.

and analysis

Before embarking
be reviewed.

such

SINGER

prove useful for locating
Beforeitand internet

shouldand Building

passport

the only source of passport

as by the European Cooperativesuchherbarium

germplasm accession

in detailpublished

given towithprogrammes

may provideof a target taxon and therefore

Konopka

associatedspecimens

discuss

generallymeans that the data associatedfor many species

They recommend using
shouldcollectors

germplasm

As a result,

The importancefor further

conserved

can beareas, as well as from(see

catalogues

section

2.5).

Taxon

or databases.

The

has the mandate for Vigna whilst

the

71

collection

held by Meise, Belgium

has been designatedthe world Vigna base collection

by

IPGRI.
However, care must be taken when interpreting
reserve, gene bank or botanic
1. The material

garden holdings,

because:held may be incorrectly

identified,the identificationby consulting

information
thoughon current genetic

it may be possible

voucher material or identifying

2. It may not contain a representative

sample of the genetic

livingdiversity

to checkmaterial.
of the sampled

population.
3. The actual quantities

of germplasm

gene banks and botanic
gene banks
impression

or botanic
shouldcould be misleading.

are encouraged

gardens,of the actual genetic

4. The researcher
the material

gardens

availableconsiderto duplicate

thus

duplicateddiversity

conserved.

that although

accessions

accessions

may for various reasons be unavailable

false impressionof a taxon's conservation

5. Some collections

databases

their

Genetic reserves,collection

in othercan givea falsemay be held in a gene bank,

to potential

users and so create astatus.

may not be efficiently

managed and therefore

records

may not be current or contain errors.
3.2

Media Survey of Geographical,
Increasingly,

as new technologies

media other than traditional
Maxted et al. (1995)
and the expanding

printedstressedpotential

sourcesof data willbooks, such as microfiche,

diskettes

and CD-ROMdiscs.

informationfrom these sources

the importance
The kinds

Note that the asterisks

efficiently

be storedof obtaining
databases,

as well as from herbariumof data that might

and the world wide web are listed

in Africa.

and Taxonomic Datadevelop,network of wild species

and gene bank accessions.
databases

Ecologicalbe obtained

specimensfrom the literature,in Table 2, with an example for a Vigna species

in Tables

2 and 3 indicatesas codes rather than full text,

data thatmight

as TAN, the standard

country code, thus saving space in the computer.

The collation

may be undertaken

which often have good botanicalattached.

major herbaria,

morefor example the name of the country"Tanzania" would commonly be stored in the database
while visiting

includethreeletterof much of this information
libraries

72

E cogeog ra ph ie d ata obtain ed from a m edia
surveyaccepted tax on nam e*
locally u sed taxon nam e*
D istrib ution*
tim ing of localflow eri n g and fruitin g*

E xam p le d ata for a Vig n a sp ecies (F rom M oss,
1985)
V ig na sch im p eriB ak.
C ong o,E thiop ia,K enya,Su dan,T anzania & U gan da,
O ctob er‑January but also A pril‑A u gust in E ast
A fricaC oarse grasslan d0‑2250 m
U nk now n
N on e ob served
U nk now n
U nk now n
N on e ob served
N on e ob served
N on e available
A lso know n as E m aret,M angw ia,or N jaziin K enya,
O rubore or O m ah arakuk u in U G A , A djuru or
O kw orokw oro in Z A R
N o currently threatened

habitatiat eren
prerceA ltitude
soilp referen ce*geologicalp referen ce*
clim atic an d m icro ‑clim atic preference*
breed in g system *ge notypic an d phenotyp ic variation
biotic interaction s (pests,p athogen s,herbiv ores)
A rchaeological inform ation
E thnobotanicalin form ation

C on servation status*(e.g .R ed D ata B ook status)

Table 2.

3.3

Ecogeographic
data types that might be recorded from a media survey, withan example for a Vigna species in Africa.
Those items marked with anasterisk could be coded in the database.

Collectionof ecogeographic

data

The kinds

of informationthatassociatedwith herbarium

specimens

an example for a Vigna speciesof potential
recordedcategories

Other

data that may be recordedof grazing,for the study to yieldbioticand abiotic

appliedshouldthroughout

always be checked

obvious effectwill inevitably

specimens.

data from herbariumin Tableand it is unlikely

results,data
3, withThis listingthat all willbedata items thatand these are shown in bold.

site includes

slope,

aspect,land use,etc.

scientificnames written

to ensure the taxonomic

on herbarium

sheets;theconcepts are consistently

the study.

The final databasefor particularpredictive

factors

are listed

however, there are certain

at the collection

Care must be taken in accepting
identification

this list,

from passport

data that could be coded).

data is extensive

Withinbe recorded

evidence(* indicatesof ecogeographic

shouldcan obtainand germplasm accessionsin Africa

for a single specimen.

the researcher

In general,

specimens,on the analysis

containmany gaps, where passport

it is much easier to record

than it is to recordand must be considered

ecologicalcuratorial
data.

when interpreting

data is missing
or geographic

This will
the results.

have an73

Table 3.
Ecogeographic
data types that might be recorded from passport dataassociated with a herbarium specimen or a germplasm accession, with an example for aherbarium specimen of a Vigna species in Africa.
Those items marked with an asterisk
could be coded in the database.
E c o g e o g r a p h ic d a ta o b ta in e d fr o m h e rb a ri u m sp e c im e n s
o r g e rm p la sm ac c e ssio n s
S a m p le id e n tific a tio n *H e r b a ri u m , g e n e b a n k o r b o ta n ic g a r d e n w h er e
sp ec im e n / a c ce s sio n is d e p o site d *C o llec to r 's n a m e a n d n u m b e r
C o llec tio n d a te * (to d e ri v e fl o w e r a n d fr u itin g tim e )P h e n o lo g ic a l d a ta * (d o e s sp e cim en h a v e fl o w e r o r fru it? )E x a m p le d a ta fo r a
V isn a sp e cim en
V ig n a v ex illa ta v a ri e ty a n g u stif o lia
K
W ilso n , 1 7 7 6
A u g u st 1 9 7 2
F lo w e r ‑ y e s
F ru it ‑ y e s
C ou n try ‑ U g an d a
P rov in c e ‑ U l ‑K a ram oj a
L o c ation ‑ B o k o ta co u n ty , N a k isu m et
L a titu d e 2 9 S ‑4 4 1
L o n g itu d e 3 0 E ‑2 3 1
7 50 m
D ry , ro c k y v eld in M l sun
B la ck
C la yA ca c ia" E c h o rek ilen g "T ap ro o t e d ib le

P a r tic u la r a re a o f p r o v en a n c e *A ltitu d e*
H a b itat*
S o il co lo u r
S o il ty p e *V e g e ta tio n *L o c al n am es
P la n t u se s*

3.4

Selectionof Representative

Specimens

The scope of all ecogeographic

investigations

and materials.

of the world containfacilities

The herbaria

will be limited

number of specimensof any one target taxon can be vast.

forces the researcher

to select

the survey or study.

Duringspecimensof the target

identification

representative

specimensof thosetime,of specimens

and theA combinationof the target

the course of an ecogeographic

taxon may be seen.

and a proportion

millionsby the available

seen will

be selected

placesemphasistaxon for inclusion

project

Each of theseof these factors
severalspecimens

to have theirinthousandwill

requirepassport

data

recorded in the database.
It is importantthat the researcher

provenance data for those
selected

if they

particulartaxonomic,

specimens

have detailed
ecologicalcome from unusual environments
Maxted (1995)
Vicia were finallyto be included.

on obtaining
Specimens

specimenare more likely

to beecogeographic

passport

data or if theyor geographic

interest,

e.g. they are odd forms or rare taxa,

show features

or are found on the edges of their natural distribution.

found that data from about a third of the specimens
includedreliablein the ecogeographic

database.

seen during a study of

of

74

Ideally,

only specimensthat either

have latitude

and longitude

data available

or forshouldbe selected

for inclusionin the database,as thisdata is so important

for the ecogeographic

analysis.

In practice,advisable

to include

specimens

longitudedetails

which these data can be establishedsubunitdetailgeographicalwith two levels

can be obtained(possiblyof detail,thoseusing a gazetteer)(i.e. province

or state)

data should

not be included

however, it may be

for which full latitude

andand those with major country

alone is known. Specimens that lack even this levelunlessthey are particularlyofnoteworthy,e.g.

of geographicandtaxonomic off-types.
Specimens
ecologicalshould be positively

conditions

detailed

passport

data alreadyselectedunder which the target

to represent

the breadthtaxon is found.

data from a broad range of representative

includedin the databasethan one specimen sampledfrom previous

It is desirable

specimens,

specimens.

from the same population

to collectrather than duplicate

Entering data from moreis likely

to add little

to the predictive

value of the data set as a whole.
Recentlycollected

specimens

often have higher

frequently

type written and easier to read.

current.

Extensiveuse of specimens

collected

details

about historical

distributions,but is likely

qualitypassport

data, which

These data are also more likely
severalhundredto yieldisto have remained

years ago may provide

less useful

information

about

contemporary populations.
The researcher
soil,

altitude,etc.)

may be able to infer various features (latitude

of collection

whether this is possible,
topography

willof the collection

sitesfrom location

dependon the precisionarea, and the precision

example, if the collection

site is situatedand longitude,

data by reference

geology,

to appropriate

maps;of the location

data available,

theof the environmental

data required.

For

on a gently undulating

plain,

then a crude estimateof altitude

may be gained

from the locationdata,

significantlyin the vicinityof the collecting

However, if the collection

site is situatedto vary markedly

relatively

in a mountainous

area, then the altitude

distancesand so estimates

extremely

precisely

the databaseto distinguishpossible,

specimens

availablein the originalmeans giving
coordinates

based on location

This kind of secondarily
it from data derived

should also be flagged
record or derived

a code for the scale(Rhoadesis likelyof site altitude

specified.

site.

as the altitude

is unlikely
withinwill be misleading
deriveddirectly

data shouldfrom herbarium

to denote the accuracy of locationfrom other locality

information;of map used to pin-point

and Thompson, 1992).

a locality

to vary
shortunless this is
be flaggedlabels.

inWhendata, whether
in practice,

thisand read off

75

Researchers

are faced with the question,

into the database
not increase

before the amount of extra information

the predictive

question.
ecogeographic
full

value of the data set?

However, the compiler

latitudinal/

combinations
longitudinal

have been recorded

shouldgainedThere is no specific
beingof the species

nichesbe enteredanswer to thisfor the point

no longer occur in the specimens
and ecologicalshouldfrom each specimen does

be on the look-out

extent of the distribution

range of geographical3.5

how many specimens

when novel

examinedand thehas ceased to expand:that the taxon inhabits

thewill then probably

in the database.

Data Verification
Before the databasecan be deemed complete

errors.

the databasecorrect

Indexing(i.e.

the researcher

re-arranging

the records

numerical

order)

on each fieldin turn may highlight

Mapping

latitudeand longitude

data may reveal errors if particular

as obvious

outliers

in impossible

often send duplicate
researcher

shouldplaces.

sets of their

Herbariummaterial

search the databasetypingerrorsin alphabeticalor invalid

localities

international

duplicatesorentries.

are shown up

specimen and germplasm

to different

for thesemust search for andcollectors

collections.

Theand be aware of their

possibleeffect on data analysis.
3.6

Analysisof Geographic,

The quality

through

the analysis

Geographicalduring the survey or study.

The more detailed

the data, theSystem (GIS)

databaseis a relatively

throughout

databasevariate,mapping,

and more complex multi-variate

analysis.

using Vigna data contained
approximately

6,000

be carriedout with thosethe data analysis

to extractfor import into any data analysis

time, but also avoid the possible

in a recently

accessions.

Althoughecological

data andlarge one, very few specimens had completeof data should

be possible

varies between simple

will be illustrated

for African taxa containing

as such, interpretation

re-typingby the quality

Information

developed

ecogeographic

will be dictated

The analysisof the data analysis

shouldanalysis

possible.

The results
the databaseand Taxonomic Dataand scope of the ecogeographic

and scope of the data collated
more detailed

Ecologicalconstraints

the data

in mind.

directly

from theprograms, this will obviously

introductionof errors.

the relative

frequencyItsave

Pie charts
Pie charts
places.

can be used to display

They are commonly used to show allelicgeographicallocations.

specimens with physicalPiechartscharacteristics,

frequencies

of a characterin populations

in different

at different

can also be used to compare the distribution
e.g. altitude,

temperatureor soil type.

ofFigure2 shows the proportion

on the African continent.
data (aboutof all Vigna specimens

This is basedon the 2500

collected

at various altitudes

Vigna specimens that carried

altitude

40 % of the number studied).

Figure 2.

Proportionof all Vigna specimens

collected

at various altitudes

in Africa.

TablesEcogeographic

data interpretation

indicating

the number (or percentage)

ecologicalunits (e.g. climate

arrangedin this fashion

If the ecologicalcan be aided by the use of tablesof specimens

type, soil type,will help

identify

factor is continuous,

aspect,seen from different

geographicalshading

habitat).

the particularsuch as altitude,

characters

ecotypic

adaptation,

both in wild and cultivated

To identify

the centre of taxonomic (species)

contained

in a databasedegree grid squares.
gridsquares,case the highest

was analysed

latitudeenvironmental

east of the Democratic

Congo.

Data

taxon prefers.

can be calculated.
will

help

of

Correlation

indicatepossiblematerial.
diversity

for Vigna in Africa, the dataon the basis of presence in anyone of 72 map 5 x 5are includedin the table.

For ease of presentation,
By comparing

each square the areas with high taxon diversity
concentrationorand soil pH, correlation

gradients

Table 4 shows part of this table.

and 26 species

taxa found within

withcharacteristics,

niche that the target

the frequency of occurrence of specimens with the gradientof morphologicaland bar charts

only, 18

the total number of

can be identified.

is found in squares 3 1 and 32, which are locatedIn thisin the North-77

Table 4.

Species

S p e cie sof Vigna found in 1 8 grid squares scored for presence of species

G R ID S
29

+

+

N o.
31

32

+

+

+

4‑

+ 蝣

V .fi s c h e r i

+

+

+

V . fr ie s io r u m

+

+

+

+

+

+

V . a m a b ac e n sis

2 8

V . c o m o sa

3 0

+

+

V .f r u te s c e n s
V . g r a c i lis++

V . k ir k ii++

V . lu t e o l a

+

+

33

34

36

+

+

+

+

37

+

38

39

+
+
蝣

V . m a c ro rh y n ch a

+

+

+

+

+

+

+

+

41

42

43

44

4 5

+

+

+

+

+

+

13

+

+

+

+

+

14

+

+

+

+

+

+

+

+

+

+

+

12

+

+

+

+

+

+

+

+

+

15

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

V . n ig r iti a

+

+

+

V .nud a
V . o b lo n g if o li a+

V .p a rk e ri
V . p ra e c ox

+
+

+

4‑

+

+

15
5

+

5

+

+

7
14

4‑
+

+

+

+

+

+

+

+

+

+

+

+

+

+

15

+

+

+

9
2

+

V.pyg m aea

+

+
+

4

+

+

+

+

+

+

+

+

9

V . m em b ra n a c e a

V . m u lt in e r v is5+

V . m a r in a

V . m o n o p h y l lag ri d s

+
+

+

40

3
蝣

+

V . h e te r o p h y l la

35

8
8

+

+

2

+

V . ra c em o sa++

V . r e tic u l a ta

+

+

+

+

V . s c h im p e r i
V . tr ip h y l la!蝣V . w it te i+

T o t a l n o . S p e c ie s

8

1
+

+

+

+

+

+

+

!

+

+

S

4

19

+

+

+

4‑

+

+

+

+

+

+

+

+

+

14

+

+

+

+

+

15

+

+

+

16
6
2

+
蝣

19

+

!

+

10

+

3

a
10

ll

ll

13

16

蝣
12

3

+

9

8

9

ll

ll

jr e s e n t in g ri dApart from the ease with which high diversity
facilitateareas can be defined,easy recognitionof endemic

taxa or thosewith restricted

need to be given priority

in identificationof importantsites for conservation.

pygmaea, which only occurs in one square,conservation

which in itself

such tables

distribution

that

also
may

In this case, V.

means that the species

warrants

priority.

GraphsEcogeographic
collection
Figuresite could

data is also commonly plottedgraphically;

one factor concerning

be plottedagainstanother,

for example longitude

3 the number of species

is plottedagainstlatitude.

of Vigna species

between 10° N and 10° S.

againstaltitude.

theIn

The graph shows a concentration

Figure 3. Number of Vigna species
Vigna data set).

plottedagainstlatitude.

(Basedon the African

Maps
Another approachcollection

sites

topographical,

climate,mean of visuallyenclosingof different

to the study
taxa.

These distribution

geologicaldisplaying

data involves

distribution.

line (see Figure

preparingmaps ofmaps can be used in conjunction

or soil maps. Stace (1989)

plant

an area with a singleof ecogeographic

stresseswiththe importanceof theThis can take two forms: (1)shading4), or (2) using

maps (seedot distribution

or

Figure5).
The use of an enclosing

line can be ambiguous,frequencyof the taxon withinsuggestthat the taxon is continuouslyof a species
line.

the region.
present

is often sparse at the peripherypresencein thisoutlying

throughout

no indication

specimenmight

an entire region.

of its range and there is rarely

manner alsomeans thatand geo-morphologicalfactors

the individual

cannot be shown. The problems

associatedecological

Indicating

A singleas it provides

withinwith enclosedof theerroneously

The occurrencea distinct

cut-off

any variation

due to local

provincesor countries

line maps can be illustrated

with

79

Figure4.

Enclosedline distribution

map for V.juncea in Africa.

the Vigna juncea distribution

in Africa drawn above. The crude enclosed

Figurearea including

4 encloses

Republicbring

the entireof the Congo, extending

majorityof records

are from Zambia.

perhaps

its identification

shouldV.gracilis

south-eastern

into north-west Malawi and Tanzania.

out the fact that only one specimen

commonly used to indicate

most of Zambia,

line map shown inhas been collected

The single

also be checked.

individual

species

specimen

This map does notfrom Tanzania,

while

the

may be a genuine outlierbut

As a result dot distribution

distribution,

Democratic

maps are more

as is shown for two subspeciesofshown in Figure 5.

Enhanced Dot distribution
Maps can also be used to study geographic
with the taxon under study.
is plotted
Figurecharacters

associatedFor example, if the flower colour of various specimens

on a map with corresponding
6, it can be clearly

trends in various

co-ordinatesas has been done for V. ambacensis inseen that west African forms are yellow

African forms are predominantly

blue flowered.

studiedfloweredwhilsteast

FigurePractically,5.

Dot distribution

programs or geographicalAtlas GIS, Strategic

Mapping

ArcView 3.0 for windows
available,

even digitised
longitudein Africa.

maps can be draw by hand but are more commonly now drawn using

one of the mapping

readily

map of V. gracilis

Inc. (1995);(ESRI,

co-ordinates

Mappingsystems currently

Atlas Mapmaker, Strategic

1996).

but if more detailed
from scratch.

information
Digitisedmaps are requiredlarge-scale

database,

Mapping(e.g.

Inc. (1991);

maps of the worldarethey may need to be customisedor

programs allow the directfrom the ecogeographic

available

importation

which are plottedof latitude

andonto customisedmaps.
Morphological,

ecologicalonto a dot distribution

map. Figure

degreegrid squares

pointof the grid square,present.

or taxonomic diversity

in Africa.
while

can be superimposed

7 shows an enhanced dot distribution

The position
the typeThus, areas with high species

informationof the symbolof symbol

diversity

map for 5 x 5on the map indicatesthe mid-shows the number of Vigna species

can be visuallydemonstrated.

FigureFigure7.

6.

Flower colour of V. ambacensis specimens

collected

Enhanced dot distribution
map of Vigna specimensx 5 degree grid squares in Africa.

collected

from Africa.

from various 5

Isoflor maps
Enclosing
distribution
line

line

maps can alsoof species.

is a contour

illustrated

Such isoflordelimiting

passport

to indicate

maps do not show actuala greaterusing accession

be usedor lesser

concentration

data for herbarium

concentration
speciesratherdistributions:of species.

than
each

This

may beof three speciesof Vignafrom the three speciesof Vigna

specimens

Africa.
Figuresection8 shows the dot distribution

Haydonia in Africa

distribution

data.

distributions

are superimposed

and Figure

To produceof specimens

9 shows the isoflor

the isoflorline

on the same

maps for the

species

onto a single map, then contour lines are drawn around areasof the map with the same number of species.
enclosedmap, enclosed

map based

The centre of diversity

is locatedin the areaby the most contour lines and in the example for Vigna section Haydonia this is ineastern Zambia,

northern Zimbabwe,south-western Tanzania.

Isoflorsouth-eastern

Democratic

maps can be produced(e.g. subspecies

or crop land races) as well as for species

FigureDot distribution

8.

map of species

Republicof the Congo, andfor infra-specifictaxa within

withinor genera.

sectionsof Vigna Section Haydonia.

species

83

Figure 9.
Isoflor map for Vigna Section Haydonia of subgenus Haydonia in Africa.
The central isoflor contains all three species in the section and each subsequent isoflor
contains two more taxa than the previous line.

Geograph ical Information Systems
More sophisticatedof such geographic
analysis

mapping

informationof ecogeographic

programmes have built-in
systems (GIS)

data. Haslett(1990)

will

databases.

increasingly

providesanalyseand displaythat can be related
GIS packages

spatiallyreferenced

prove invaluable

the following"GIS are computer hardware and software packages

The development
summary of a GIS:

designed

to store,

data; they deal with information

to some form of map".

generally

includea graphics

programme for handling

cartographic

data, interfacedwith a Database Management System (DBMS)

manipulating

data associatedwith particularmapping system, which can easily
biological

data sets.
The researcher

environmental

map features.

Once cartographic

adaptable

ecologicalandmaps, aerial

photographs,

field

form, they can be manipulated

and analysedusingdata from existing

can also use GIS to infer features

data is available

digitised

for storing and

The whole is a highly

cope with a wide range of geographical,

surveys, and remote sensing are in digital
GIS.

in theof collecting

and locate areas with particular

sites

for which no

combinationsof ecological(Nix,

for example,

characteristics.
The BIOCLIM programme of Nix, Busby and Hutchinson1986),

84

uses interpolation

surfaces to estimate

mean climaticconditions

at Australiansites.

interpolation

surfacesare available

for Africa.

temperature

variables

represented

by a given set of sites have been calculated,such as AFRMAP (Boothconditions

are found.

be introduced,
particular

can be used to display

This was developed

to identify

taxon might be expectedWORLDMAP(Williams,
the selection1992),

area

areas for conservation.

extensive

review of the paper

currently

available.

all the regionsof climatic

previously.

combines taxonomic and geographicalof priority

Multivariate

from that

andprogrammes

context to identify

to be found on the basis

have been collected

rainfallwhere suchareas where new crop species

but it can also be used in the conservationno specimens
assistet al, 1989)

Once the ranges of six important

Similarareas where a

conditions,even if

Another

program,distribution

data toGuarino et al,

and computer based ecogeographic

might(1997)

providedata analysisanmethods

analysis

One of the most thoroughly

statisticallyreportedby Cocks and Ehrman (1987),

Ehrman and Cocks (1990)

and Ehrman and Maxted(1990)

for the annual legumes in Syria.

These authors undertook

comprehensive

over several years, during which they gathered
to use this data to predictclimatic

regions,

potential

extensive

ecogeographic

the percentage

dataecogeographic

areas for active conservation.

and then recorded

annual legume species

testedof sites

sets is thatfield work

data and were able

They divided

Syria

into

for each region where each

was found. The authors also studied

the percentage

of sites of eachsoil type in which various taxa were found. The influenceof both climatic

factors

alkalinity

demonstrated.

on the distributionof various speciesSuchmethods

deal

with

morphologicalvariable.

Ecogeographic

was clearly

one environmental

factordata, however, is multivariate,

items of data are available

for each record {e.g. each collecting

herbarium

Ehrman and Cocks (1990)

specimen).

environmental

data to classify

members of which had climates
one single

variable),the collecting

sites

or a single

in that two or moresite, germplasm accession

performed

cluster

into groups

or classes

which were more similar

overall(ratheranalysis

Products

4.1

Data Synthesis
The final phase of the project
The researcher

commences with the synthesisoron their(clusters),

thethan as regards

anythan they were to members of any other class.

4.0

during the study.

at a time,and soilof all data collectedshould be aware of the degree of completenessof the85

database,or the collections

been effectively
be because
becauseon which it was based, in terms of how fully the target area hascovered.

If a particular

the taxon is absent

the habitat

habitator has not been recognised

has not been sampled.

amount of data duplicationin the database

4.2

The Ecogeographic

Database,

The ecogeographic

database,of an ecogeographic
project.
information

for the target

discusses

from that habitat,

The researcher

shouldor possiblyof the results.

and Reportand report

The ecogeographic

database

containssummarises the available

ecological,

geographicalthe collated

it mayalso be aware of theas this may bias interpretation
Conspectus

taxon throughand interpretsin the database,conspectus

survey.

The conspectus

is under-represented

are the three essential

part or the whole of its range.

data and provides

products

the raw data of theand taxonomic
Finally,

the reporta clear summary of conservation

priorities.
The conspectus
The conspectus
alphabeticallyis commonly arranged

or systematically.

included.

The conspectus

shouldliterature

survey. When possibleby plant

names, which can be listed

In both cases it is helpful

to provide

summarise informationfrom both the database

the informationin Tabledetailed

eitheran index to the taxa

5 should

and thebe included

foreach taxon:
E c o g e o g r a p h ic c o n sp e c tu s
a c ce p te d ta x o n n a m e , au th o r(s), d ate o f p u b lic atio n ,p la ce o f p u b lic atio n
R efe re n ce to p u b lish e d d e sc ri p tio n s an d ico n o g rap h y

E x a m p le d a ta fo r a V ig n a sp e cie s
V ig n a h e te rop h y lla A . R ich a rd T e n t. F l. A b y ss. 1 :2 18 ( 18 4 7 )F lo ra o f T ro p ic a l A frica , 2 :1 9 7 (1 8 7 1); L .T .A .: 4 1 0(1 9 2 9 ); F .C .B . 6 :3 7 7 (19 5 4 )P h e n o lo g y , fl o w e rin g se a so n
M ar ch ‑ A u g u st
L o ca l n am e s
U m h o ro ro (R W A )C A R C M R , E T H , K E N , M W I, N G A , R W A , T Z A ,g eo g rap h ic al d istri b u tio n
U G A ,Z A R ,7M B .
d istrib u tio n m a p s
S e e F ig u re s 4 ‑9
E co lo g ic a l n o tes : in c lu d in g a ltitu d e (m in im u m an d A lt. 1 6 8 0 ‑2 0 10 m ;m a x im u m ), h ab ita t, to p o g rap h ic , so il, g eo lo g ic al, H ab ita t: g ra ssla n d s a n d cu ltiv ate d (w h e at) fie ld s
clim ate an d m ic ro ‑c lim a tic p refe ren ce , b io tic w h e re it g ro w s a s a w e edin tera ctio n s an d o th er h a b itat d etails
G eo g ra p h ic al n o tes : in clu d in g a n in te rp reta ti o n o f T h e d istrib u tio n o f th e sp ec ie s e x ten d s f ro m th e flo ra
th e ta x o n 's g eo g ra p h ica l d istrib u tio n
Z a m b e sia c a R e g io n to W e st T ro p ic a l A fr ic a
T a x o n o m ic n o tes : in clu d in g n o te s o n an y d istin ct T h e sp ec ie s is clo sely re la ted to V . a m b a ce n sis fro m
g en o ty p ic a n d p h e n o ty p ic v a ri ati o n w ith in th e ta x o n w h ic h it ca n b e d istin g u ish ed b y its lo n g e r an dn ar ro w e r c aly x lo b es an d b ra cte o le s
C o n se rv atio n n o te s : c o n tain in g an as sessm e n t o f th e T h e sp ec ie s is o n ly rep re se n ted b y tw o sp e cim e n s,
v a ri ab ility cu rre n tly co n serv e d , th e p o ten tial g e n eti c d u p lic ates o f a sin g le a c ce ssio n o f K en y an o rig in ,ero sio n fac e d an d th e c o n se rv atio n sta tu s o f th e w h ic h are h e ld a t IIT A an d M e ise. T h is rar e sp e cie s
is h ig h ly en d an g er ed .
ta x o n in th e fie ld

Table 5.

The ecogeographic

conspectus,with an example for a Vigna species.

86

If the scope of the investigation
may be necessary

to provide

is broada summary of the ecogeographic

ecologicalsubunit.

geographicalregiondifferences.

In this case it would increase

dates were listed

in the geographicalFor example, the floweringof Africa

than in anotherbecausesense, itdata for each geographicalof Vigna species

may be earlierof the relative

the predictive

for each region of Africa

or ecologicallatitudeorin oneand altitude

value of the survey if the floweringrather than for Africa as a whole.

The report
The reportimportantdiscusses

it must attempt

and most appropriate
shouldthecontentsof the database

to draw firm conclusions

conservation

strategy

and conspectus,

concerning

for the group.

but vitallythe conservation

If possible,

priorities

the following

pointsbe covered:•E Delimitationof the target taxon.

•E Classificationof the target•E Mode of selection

taxon that has been used, and why.

of representative

specimens.

•E Choice of hardware and software.
•E Ecogeographic

database•E Discussionof database•E Targettaxon

ecology.

file structurescontent.

•E Target taxon phytogeography

and distribution.

•E Interestingencounteredtaxonomic variants•E Current and potential•E Relationships•E Anyuses of the target taxon.
species

identification

problemsof identification•E Priorities

threats

for conservation

strategy.
•E The ecogeographic
appendixandfloral and fruiting

activities,

includingthe extent

to taxonomic

and genetic

diversity.

actionconspectus

or as a separate
of Conservation

The ecogeographicof the conservationencountered,possible
specimens.

of diversity

conserved.

•E Current and potential

Identificationand their wild relatives.

aids to vegetative,•E In situ and ex situ conservation
alreadyduring the survey.

between the cultivatedparticularpresentation

4.3

and inter-relationships.

priorities

and recommendations

for a conservation

may be included

the report

withinas anentity.
Priorities

survey or study should
and proposedconcludeconservation

with a clear, concise

strategystatement

for the target taxon. This87

can be achievedby establishingtarget taxon, and an evaluation
Withinthe patternof variationof potential

genetic

status.

which are of particularinteresteithere.g. areas with highconcentrations

the target area, zones may be identified

or endemic taxa, low rainfall,

By establishingresearcher

the occurrence

may decideof the target

has been found at only one location
the taxon may be restricted
genetic

that requiredifferent
furtherbut not at anotherthat the second location

habitatsampling.

shouldecogeographic

be searched.

that has a set of unique ecogeographic
and the conservationist

taxon, specificvariants

the

For instance,with similar

to that location

types,

If a taxon

conditions,

then

may wish to establishareserve at that site.
Withinconservation
pods,

taxon withinto select areas or habitats

then it may be suggestedofhigh frequency of saline soils or extremes of exposure.

if a taxon has been found at one location,
conditions,both the target area and theerosion and current conservation

because of the taxa found there or local conditions,diversewithinthe target
priority,e.g. restricted

populations

particularrare species,

features

The establishedwithinin the light

genetic

materialfrom a particularlyinteresting

conserved either

in situ or ex situ?

If not, shouldof theGalguduud,

Somalia,collectionof thisof herbariumis alreadytargetarea

For example, is sufficientniche

be expended

may indicate
activities

alreadyto actively

safelyconservethat there is a rare relativeon coastal

further

erosion.

dunes

indicates

collectionof Mudug andthat a large ex situ

would not be justified.

using in situ techniques,

you may suggest

theof a genetic reserve at that location.
levelsquestionsbe monitored

collecting

shouldto collect

Maxted (1995),

missions

and the establishment

speciesin Turkey as part of his ecogeographic
If the ecogeographic
data obtained

also be considered.

to assess the threat

team be directedconserve the taxa in situ?

ecologicalfrom genetic

taxa and the potential

activities.

growing

conserved,

is not conserved

A number of otherinternational

endangered

effortbut if a review of conservation

species

noted,

ecogeographic

material

monantha Thulin)

However, if the species

population

land races withof current conservation

Thus, analysis

cowpea (Vigna

establishment

not previously
the target

must be assessed

may warrantwith amphicarpic

or taxa that are particularly

level of variability

whichforage legume species

that may have crop potential

evaluation

this material?

can be identified

erosion?

the priority

targetdiscusses

Shouldtaxa?

shoulda nationalor

Is it possible

totwo specificcollection

reserve for rare and endangeredVicia

conclusions.

data has been gathered
may be insufficientof genetic

for example,of an in situ genetic

For instance,solelyfrom herbarium

to draw detailed

conclusions

specimens,

theon the targettaxon's

habitatadvisable

preferences.

to obtain

conservation

In this

case,

image

of appropriate

habitatto be proposed.

Once specificareas have been selected

a clearer

strategy

a survey

missionto the target
types,

area may beand so help clarify

for survey, aroute that covers the maximum number of sites in the minimum time- can be suggested.
phenological

data will indicate

to be collecting"collecting

duringwhen a collecting

the survey mission,team should

notably

visit the targetThe

area. If seed isfor the Leguminosae,

window" during which the team must find the target

the

there is a narrow

population;

too early andthe seed will not be ripe, too late and the seed will be shattered
5.0

Conclusion
Analysisfor assessingof a taxon's

ecology,geography

its conservation

status,and permits

which the taxon is likely
gene banks, botanic

to be found.

gardens,

for the effective

The ecogeographic

detailedconservationin situ and ex situ conservation

Acknowledgements
The authors

wish

to

the prediction

acknowledge

that

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Natural91

Questionsand answers:

Session1

Question to Dr. Egawa
Q. What is the origin

of Vigna glabrescensl

A. Vigna glabrescens

was selected from the wild species

erect growth habit
widelyQuestions(Debouck)and relatively

Vigna reflexo-pilosa

as a mutant with

big seeds in South Asia, where V. reflexo-pilosa

isdistributed.

to Dr. Maxted.

Q. Howcan eco-geographic
A. If a population

surveyingspot off-types

or outliers?

has been found in the past then we are likely

information

availableon that species,

ecogeographic

survey and we will be able to predict

However, if a population

the potential

information

available,

suffiecient

eco-geographic(typicalQ. What efforts
fashion,ecologyso the information
its location

will

be included

and habitat

to predictThough having said this various GIS programs canlocationof taxa, even where there is no direct population

if there is no direct population
informationand habitat)in thepreferences.

has not been found in the past it would be impossible

where it might be found as an outlier.
be used to predict

to have some ecogeographic

available

information

available,

if there is

to form an ecogeographic

envelopof the taxon.

are being made to collate

and distibutetaxonomic data in a more standardisedsuch as is done with genomic data? (Gepts)

A. This is a good question.

I believethat taxonomic data is largely

is an urgent need for clarity.

Althoughin legumes, notably the International
is attempting

to provide

baseline

there are various databaseorganised

and there

projects

and initiatives

Legume Database and Information

Service (which

biodiversity

Botanic Garden, US (which is databasing
botanistshave so much data available,

is unlikely

it will be made easily available

format in the near future.

chaoticallydata for all legume taxa) or the Missouri

all there herbarium specimen data. But generallythat has been gathered over the past 300 years, itto the user community in an easily

accessible

Session 2
Genetic diversity,

evolutionand conservation

95

Biodiversity,

ecologyand genetic

resourcesof Phaseolus

beans - Seven answered and unanswered questions.
D.G. Debouck
Genetic
Cali,

Resources Unit, Gentro International

COLOMBIA(e-mail:de Agricultura

Tropical(CIAT),

A.A. 67 1 3 ,d.debouck@cgiar.org)

Abstract
Any crop improvement effort if it is to make impact supposes an understanding
diversityof the crop. This paper reviews the genus Phaseolus, including

forms, their distribution
phyla, the evolution

and ecologies.

the wild ancestors of cultivatedIt presents evidence concerning the organizationof these phyla, and the history

There are at least two phyla, one

and one including

the Lima bean and several species

the commonbean with sister species,

distributedin Central and South America. The primary reason for the genetic distanceis geographic

isolation

duringtheirof the genus intoof domestication.

includingphylaof the genetic

early evolution.

between these twoThis paper also examines when bean

domestication

occurred and the reasons for it. Dates have been questioned,

domestication

was as ancient in Mesoamerica as in the Andes, and that beans are pre-ceramic. Beans areoften used as boiled

grains;muchreflect spacial

gradients,

implicationsof these findings

but it seems certain that

there are other uses as well and not all recent. Patterns of genetic diversityand domestication

has often resulted

in an important founder effect. The

for a improved conservation and use of these genetic resources are discussed.

1. Introduction
Yield

has long been recognizedbeans in the tropics,

subtropics

1 999;

Yield

Singh,

1 992).

against pests, diseases,
potential.

increasing

and abiotic

physiologicalgenetic

breeding,

stresses,

processes.

factor for expanding

regions(reviewed

can be obtainedse, for instanceredistribution

into harvested

by improving

One of the most powerful

and is thus based on the assumption

by Gepts,

1 998;

Kellyof the actual yieldof existing

bean plantparts. A third approach

photosynthetic

ways to achievethat there is genetic

et aL,of the yield potential

the full expressionis throughof metabolism

the acreage ofPhaseolus

through protectionthat is securing

for improvement

more products

the yield potential/^?*basic

and temperate

improvements

Another possibility

biomass, by allocating

as a limiting

efficiency

is byand othersuch improvements
variability

isfor any trait

96
involved.
Breeding
traits,for resistanceand knowledge

Harlan (Harlan,

to biotic

and abiotic

about inheritance1978),

stresses

and transfer

the point is in getting

implies

different

access to the genotype

of biomass or improved photosynthesis:means of allocating

efficiency,

respectively,

the products

sources of superior

into the target material.

with the superiorbyand

The same approach prevailsforone has to find genetic variants

withof photosynthesis

in order to start a breeding

As indicatedtrait,

this may imply the search through large germplasm collections.
a novel distribution

findingor with improved photosynthetic

program. Studies

of common wild bean

germplasm have shown that such variants do indeed exist (Lynch et aL, 1 992).

Genetic variability

shouldGenetic

thus be studied,and made available,

endeavours are thus the cornerstone

and appropriatelyof any sustainedbeans, as has been the case with several
achievementsin the understanding

conserved.

effort for increasingother crops (maize,of genetic

the productivity

potato,resources of Phaseolus

resources

tomato).

of

The majorbeans are presented,

byfamilyasanswering a series of questions.

2. How many beans are there?
The genus Phaseolus(subtribe

currently

understood

Neotropicalorigin, and has approximately

namely in relation
Sigmoidotropis(Marechal(Delgado

Salinas,to the American(Marechal

et aL, 1 978a),Phaseolinae,

tribe Phaseoleae,

1985;

1983;

Lackey,speciesof Vigna of the subgenera
1 970),

and recently Misanthus (Lewis

over 400 species

good type specimens.

of synonymy. Recently,

Phaseolus,

and confirmed the existence

and Delgado

DelgadoSalinas,Salinasclearlydemonstrated

as invalid,

Schmitet aL, 1996).

In the case of P. augusti

have been clearly

identified

Ramirezella

1 994).

While weand Urban (1928)(1 985)orhave contributed

has made an exhaustivetoreview ofof 36 species.

a somewhat expanded list of species.
as for P. glabellus

synonymy seems to be valid (Caicedo

andspecies exist and which are they.

On the basis of extensive herbarium surveys and field explorations,has presented

is ofhave been named, often with poor description

Reviews by Piper (1926)

clarification

Lasiospron

the genera Macroptilium,

know clearly what a bean is, it is less clear how manyPhaseolus

lackinget aL, 1978b)50 species. Generic limits have been clearly determined,

et aL, 1 98 1 ; Verdcourt,

Over the past two centuries

Marechal

Leguminosae)

for synonymy have been

(Hamann et aL, 1995;
Harms or P. bolivianus

et aL, 1999).

such as P. costaricensis

Some proposals

Debouck (1 99 1 , 1 999)

Llaca et aL, 1994;
Piper,

a proposal

for

There have been cases where new species
Freytag& Debouck (Freytag

and Debouck,

97
1996)

(Fig.

la).

several studies
et a/.,1995),

On the basis of species
including(reviewed

by Debouck,the use of molecular markers (Fofana et ah, 1 999; Jaaska,

an updated tentative

phyla in the section

cross compatibility

Phaseolus

The list proposed

list of Phaseolus(Tableexample, the collection

many questions,and we shall address

go together

with known species

as P. altimontanus)

as P. minimiflorus)

with P. neglectus,

sonorensis,of P. chiapasanus,and P. tuerckheimii

explorations

in Central

available,

P. macrolepis,P.

It seems also that we don't know yet
pendingon additional

germplasm

estimate.

origin, the majorityof the New World, between 37° latitude
available

specimens currently

thedistributed?

The genus has a Neotropicalof currently

phyla. For

will go with P. microcarpus,

P. leptophyllus,

50-60 species,

America, would be a reasonable

3. Where are beans naturally

and subtropics

P. glabellus,

species are there:

there are several

and DGD-233 1 will fall within

may remain monospecific.

exactly how many Phaseolus

while othersof already recognized

phylum of Lima bean. On the basis of germplasm and herbarium
it seems that the phyla

Jacobsome of them innow. Some phyla have indeed several species,

DGD-427 (proposed

DGD-1 525 (proposed

1 996;

as well as their likely

look as if they have only one. As a result of recent germplasm explorations,
unnamedtaxa that will likely

and

1).

in Table 1 raises

this paper. One is worth considering

species can be proposed

1999)

floristic

knowledge,of species

are distributed

North and 28° latitude

there are no Phaseolus

in the tropics

South. On the basis

species growing wild naturally

in other parts of the world; weedy and escaped forms of Lima bean have however been reported
in eastern Africa (Marechal
polystachyus

et al., 1978b).

to this general pattern.

occurs as far north as in southern Quebec, Canada (Rydberg,

USA (MacMillan,1892).

Argentina(Burkartand Briicher,Riverside

county in southern

DesertThere are however outliers(1 16°W).

P. vulgaris

would give an extension

1953).

Accordingto Isely(1993),

the extreme northwestern

to Piper (1926),
in longitudeand Minnesota,

as a wild form goes as far south as 32° in San Luis,

California,

According

1 965)

P. filiformis
extension

P. lunatus as wild form is present

up to 45°W. The majorityof species

enters theof the Sonoranin Brazil,

in Central

America in the western mountainous ranges of Mexico (Sierra

Occidental,

Eje Volcanico),arid in the northern and central Andes of South America.

Endemism is restricted

in Phaseolus,that is, species

be due to limited

work. Little

is known about variants

smilacifolius

and sinuatus),

field

restricted

to northern

Floridawith a very limited

Madre

range, whichof P. polystachyus

and South Carolina,

thatare howeverconcentrated

might

P.

(i.e.

USA (Debouck,

98

Fig. la.
Phaseolus
costaricensis
Freytag& Debouck. Population#3120 with plants 6m high, at pod
filling stage, in Costa Rica, provinceof Alajuela, districtof Zarcero, 2 km
N of Laguna to Tapezco, 1710 masl.
February 8, 1998. Stars mark cerise
flowers and pods. These plants wereseen visited by humming birds.

Fig. lb. Phaseolus
rosei Piper.
Population#2863 found in Equador,
province of Chimborazo, districtof
Alausi,on pad from Sibambe to
Huigra, 1550 masl. June 6, 1990.
Vines are shown climbing all over 4mhigh bushes. This species has beenshown since to be identicalwith thewild Lima bean, Phaseolus
lunatusL.? of the Andean genepool.

99

Table 1 - List of Phaseolus species organized by possible phyla (adapted
1 999). Species with cultivatedform are indicated with *.

from Debouck

Section Chiapasana(seraw Delgado 1 985)
chiapasanus Piper

Phylum of P. microcarpus
microcarpus Mart.
minimiflorus Norvell ex Freytag & Debouck

Section Phaseolus
Phylum of commonbean
costaricensis
Freytag & Debouck (Fig. l a)polyanthus Greenman*
vulgaris L.*

Phylum of P. neglectus
altimontanus Freytag & Debouckneglectus Hermann

Phylum of scarlet runnercoccineus L. *Phylum of tepary beanacutifolius Asa Gray*
var. acutifolius
var. latifolius
var. tenuifolius
parvifolius
FreytagPhylum of Lima bean
jaliscanus Piper
lunatus L.* (Fig. lb)
maculatus Scheele
marechalii Delgado
mollis Hook. & Arn.
pachyrrhizoides
Harms
polystachyus Britt., Stern & Pogg
ritensis Jones
salicifolius
Piperxolocotzii Delgado
Phylum of wild beans with rugose testa
angustissimus Asa Gray
carteri Freytag & Debouck
filiformis Bentham
Phylum of P. pedicellatus
esperanzae Seaton
grayanus Woot. & Standl.
laxiflorus Piperoaxacanus Rosepedicellatus
Benthampolymorphus S. Watson
purpusii Brandegee

Phylum with 2n=2x=20
leptostachyus Bentham
macvaughii Delgado
micranthus Hook. & Arn.
Isolated phyla
glabellus Piper
leptophyllus
G.Don
macrolepis Piperoligospermus Pipersonorensis Standley
talamancensis Deboucktuerckheimii
Donnell- Smith
Section Minkelersia(sensu Lackey, 1 983; including
Alepidocalyx M.M.S.)
amblyosepalus (Piper) Morton
nelsonii Marechal, Mascherpa & Stainier
parvulus Greene
pauciflorus Sesse & Mociiio
perplexus Delgado
plagio cylix Harms
pluriflorus Marechal, Mascherpa & Stainier
tenellus Piper
Section Xanthotricha{sensu Delgado 1 985)
esquicensis Freytag
gladiolatus
Freytag & Debouckhintonii Delgado
magnilobatus Freytag & Debouck
xanthotrichus
Piperzimapanensis (Delgado)
Jaaska100
1 999).

P. amblyosepalus

is restricted

is distributed

to central

Guerrero, P. purpusii

Nuevo Leon, Mexico (Delgado
from Solola

to Guatemala

Salinas,(Debouckknown only from the Galapagosthat P. chiapasanus

only in Durango, Mexico (Nabhan,

and P. xolocotzii

to western San Luis Potosi,

1 985).

P. macrolepis

and Libreros

Ferla,

Islands

1 990).

(Wiggins

and Porter,

can be considered

and P. plagiocylix

is present

1995).

P. leptophyllus

in central

to

Guatemala,In South America, P. mollis is
1 971).

From field work, it seems

as rare (growing

in stands with very few

plants).
On the other hand, some species

are relatively

widespread.

Mexico to San Luis, Argentina;

8,000

P. vulgaris

is found fromsouthern

Chihuahua,

Km (Toro Ch. et aL, 1990)

lunatusis found from Sinaloa,

1 997)

as wild forms. P. polystachyus

1 903).

P. maculatus is distributed

1 985).

P. microcarpus

is distributed

from Durango, Mexico to eastern Guatemala (Debouck,1999).

P. leptostachyus

is distributed

from Nuevo Leon, Mexico to Costa Rica (Debouck,

Mexico to Salta, Argentina;
is distributed

7,000 Km (Maquet and Baudoin,from Ontario, Canada to Florida,

is distributed

from Tamaulipas

tuerckheimii

is distributed

from Guatemala to Panama (Debouck,that none of these last mentioned
In Mexico, Phaseolus
Rzedowski,

California

and Sonora harbours

1978)

and Wiggins,

1 964).

P. maculatus(Gentry,

species:

to Chiapas,

from Loja, Ecuador to Tucuman, Argentina

defined:species

species(Caicedo

desert ephemerals

1957).

forests

and to the West the rare species

In the forest

openings

P. pedicellatus

et al, 1 996).

The semi deciduous

P. pachyrrizoidesis

It is worth noting

types(as currently
scrub of Baja
(Shreve

Mexico, one can find P. acutifolius

P. ritensis,

P. salicifolius

one finds P. microcarpus,

is distributed

P.

and P. acutifoliusof northwestern

P. parvulus,

pluriflorus

P. glabellus

1991).
1996).

2). The desert tropical

Mexico harbour P. coccineus,

Mexico harbour P. leptostachyus,

et a/.,

et aL, 1 999).

such as P. filiformisof northwestern

The oak-pineof central

1 999).

Salinas,has been domesticated.

with a marked dry season (TableIn the oak grasslandforests

Mexico (Schmit

can be found in almost all vegetation

P. grayanus, P. leptostachyus,

The oak-pine

USA (Small,from NewMexico, USA to Puebla, Mexico (Delgado

P. glabellus

distributedand P.

Mexico harbour

P. sonorensis(Nabhan,P. marechalii,

several
1 990).

P. pauciflorus,

and P. jaliscanus

P. vulgaris.

and

The oak-pine

and to the North P. neglectusP.

(Debouck,

1 999).

forestsof eastern(Debouck,

1 999).

around the Gulf of Mexico in montane humid or cloud forests (Schmit
montane forests of Oaxaca and Chiapas harbour the rare species

P. chiapasanus(Delgado

Salinas,forests,

subhumid

hot forests

In Central

America, certain

1985).

and tropical
speciesP. lunatus

is found in lowland

scrubs (Maquet

and Baudoin,such as P. oligospermus,

tropicaldeciduous

1 997).

P. tuerckheimii

and P.

101

Table 2 - Ecologicalhabitats

colonized

by some Phaseolus

species in the Neotropics.

G e o g rap h ic ar ea s ; v e g etatio n ty p e s

P h a s e o lu s sp e c ie s (a ll a s w ild )N o rth A m e ric a
P in e ju n ip e r d ry fo re st A rizo n a N ew M ex ic o

a n g u stiss im u s, r ite n s is

P in e fo re st an d th ick e ts A tlan tic C o astal p la inp o ly s ta c hy u s

M ex ic o
G u lf o f C a lifo rn ia a rid tro p ic a l sc ru b

a c u tif o liu s, fl lif o r m is

In lan d an d P ac ific co a sta l d ry th ick e ts

m icr o c a rp u s

O ak p in e fo re st E j e V o lc an ic oc o c c in e u s, m a re c h a lii

P in e m o n ta n e su b h u m id fo re sts S ierra M a d re O rien tal

p e d ic e lla tu s

O ak p in e d ry fo re sts ce n tra l M ex ic op o ly m o rp h u s

O ak a n d lo w e r m o n tan e d e cid u o u s fo re sts

v u lg a ris

P in e su b h u m id fo re st c en tral O ax ac ao ax a c a n u s

S e m id e cidu o u s m o n tan e fo re st O ax ac a C h iap as

c h iap a s a n u s

G u lf s h u m id m o n ta n e fo re st

g la b e llu s

L o w la n d tro p ic a l d e c id u o u s fo re sts an d sc ru b s

lu n a tu s

C en tra l A m erica
G u a te m a la h u m id m o n ta n e fo re st

p o ly a n th u s

C en tra l G u atem a la m o n tan e su b h u m id p in e o ak fo re stm a cr o lep isC o sta R ic a‑ P an am a h u m id m o n tan e fo re st

c o s ta r ice n s is

M o n ta n e m o ist fo re st

tu e rc ke im ii

L o w e r m o n tan e su b h u m id fore stv u lg a r is

L o w e r m o n tan e h u m id fo re stx a n th o tric h u s

L o w e r tro p ica l d ry an d su b h u m id fore sts

lu n a tu s

S o u th A m e ric a
C o lo m b ia P eru B o liv ia A rg e n tin a e a ste rn su b h u m id m o n tan e fo re stv u lg a r is

S o u th E cu a d o r P eru w e stern su b h u m id lo w er m o n tan e fo re stv u lg a r is

C o lo m b ia V e n e z u ela lo w er tro p ic a l d ry fo rest

lu n a tu s

S o u th E cu a d o r P eru w e stern su b h u m id low er m o n tan e fo re st

lu n a tu s

M o n ta n e su b h u m id fo re sts e astern C en tra l A n d e s

p a c hy rrh iz o id es

Notes:

Vegetation

Holdridge

vulgaris

according

1 967 (for Central

xanthotrichus(Deboucktypes

America),to Isely

1990

Cabrera & Willink

are found in Chiapas (Breedlove,
et al, 1989a).

(for USA),

Commonspecies

1 986)

Rzedowski(forMexico),1 973 (for South America).

and in Guatemala (Fig. 2) and Costa Ricain Mexico such P. leptostachyus,

have also been found in other parts of Central

1978

America (Debouck,

P. lunatus
1 991).

Speciesand P.
such

Fig. 2. Distributionof a few Phaseolus sensu stricto species In southern Guatemala (mainly fromthe author's and Dr. C. Azurdla's explorations).
Solid bar = 100 km. o: province capitals.
*:
Phaseolus macrolepis Piper.H: Phaseolus oligospermus Piper. •E: Phaseolus parvifolius Freytag.
A: wild Phaseolus polyanthus
Greenman. *: Phaseolus tuerckheimii
Donnell-Smlth.
V: wild
Phaseolus vulgaris L.

103

as P. microcarpus

and P. parvifolius

are found in seasonally

dry oak forests

Guatemala. P. coccineus as pure wild stands is found generally
forests,of Mexico andin montane pine-oak subhumid

and not found further east than central Guatemala and southern Honduras.
While Phaseolus

distributed

in regions

speciesare found in a variety

where the rainfall

temperature and photoperiod

patternand conservation

temperatures.

In this sense, the ecology

in the soil during

and Cicereae

it seems that they are always

coincides

for growth and reproduction;

seed dispersal

legumes of Viceae

of habitats,a dry season follows

the exceptionof the extreme distributionofthat allows forof under un- or less favourable

species

differs

fundamentallyto a Mediterranean

climateoccurring during the winter period. This would explain the absence of Phaseolus(withconditions

periodsof Phaseolus

with adaptation

with favourablefrom thewith rainfalls
from Californiaof the Sonoran Desert)

and Chile.

On the other

hand, survival is in the form of seeds, and in many cases with short-lived

perennials,

in the form

ofa tuberous root, sphericalin P. parvulus, long fusiform

to survive in summer and winter droughts.
season ensures distribution
grayanus, P. jaliscanus,in P. maculatus. This root allows plants

A periodic

but massive seed setting

and long-term survival of the species
P. maculatus, P. marechalii,

after the rainy(common in P. esperanzae, P.

P. polymorphus,

P. purpusii,

P. ritensis,

P. sonorensis).
A last consideration

deals with the wild forms of P. vulgarispresent in both Central and South America. Under reasonablespecies

as wild forms have experienced

evidence that they are both floristic

and P. lunatus. They are

disturbance,

it is possible

an expansion because of humans. There is, however, goodelements of climax forests:

subhumid montane forests in thecase of wild commonbean, and subhumid premontane and lower dry tropicalof wild

Lima bean. Contrary

to what has been claimed

domestication

does not seem responsible

Their ecologicalsuccess and adaptation

domestication.

Curiously,

elsewhere

in disturbed

habitats

relatively

while for others (e.g. P. oligospermus,

P. lunatus,

is scarce. It might thus be premature to conclude

Mexico are relatively

is also geographicallyunbalanced:

well surveyed for Phaseolus

in South America.
factor in theirsuch as P. leptostachyus

range of Phaseolus(e.g. P. leptostachyus,

knowledge1985),

perhaps because of their very small seeds.

known. For some species

Floristicin the case

Salinas,has been a positive

species with an important range of distribution

From the above, it is clear that the geographic

information

forests(Delgado

for the presence of these two species

and P. microcarpus have not been domesticated,

well investigated,that both

species,P. vulgaris),

speciesis unevenly

the range has been

P. sonorensis,

P. xolocotzii)

about the rarity of such species.

while, for example, Arizona and New
other areas (e.g. mountains of Coahuila

104

and eastern Guerrero in Mexico, Andes of Venezuela)
areas it might be too late: precise
City

floristic

information

have been poorly sampled. For certain
for the valleys

and San Jose of Costa Rica might never be gathered.

of Mexico City, Guatemala

Similarly,

the ecologicalunder which these species thrive are unevenly known. For those species
about numerous collection
ecologicalsites,

it is not too difficult

parameters at these sites (Jones

lookingfor genetic

deficiency,

variation

in relation

et ah, 1997).

to environmental

etc, while we know that variation

4. How many cultivatedFive Phaseolus

speciesthroughfactors:

light,

photoperiod,

and are cultivatedand P. vulgaris.

Debouck, 1991 ; Toro Ch. et al, 1990).
are basicallyto the domestication

1988;

Gepts and Debouck,indicatethat it is the same biologicalcultivatedform with all its variation

adaptations,is derived

1991).

and so may be consideredrelatively

form

the two forms

syndrome (Briicher,

Salinasinterfertility

species (Kornegay

1 988;

Delgadoand Cardona, 1991 ; Weiseth,in seed colors and patterns,

1954).

The

pod types, growth habits,

andprocess. Morphological

Velasquez and Gepts, 1 994; Gepts et ah, 1 986)

indeed for an additionalin Middle

more (Bannerot

and Debouck,

event in Colombia(Chaconin Mesoamerica is still unclear;it cannot

S. et al., l 996).

The precise location

be discarded

that there would have been more than one event, for instanceraces (Singhevidence

and South America, thus atdomesticationof the domestication

et al.,

between the two forms in crosses

events occurred, and possibly

Durango for each of the respective

late in the

between the wild and the cultivatedtook place independently

least two separate domestication
existsunderandthus from a wild bean through the domestication

has shown that domestication
The possibility

P. acutifolius,evidence, such as traits distinguishingComplete(Evans, 1 976) and molecular (Becerra

1 992).

today:

America and in South America (Gepts

The relationshipfirst based on morphological
all related

phosphorus

domestication.

wild forms have been found in Middle

wasestablishedin Jalisco

several parts of Central America (Standley(Mackie,

1 943).

and inet aL, 1991).

One type of wild Lima bean, P. lunatus, with small seeds has long been reported
all cultigroups

when

Other species might have been collected

The ancestry of the commonbean, P. vulgaris, has been establishedcentury:maps to infer

to exist (Lynch et ah, 1 992).

species have been domesticatedor unfinishedsoil and climate

This is of paramount importanceas emergency food such as P. maculatus (Nabhan et ah, 1980),
incipient

for which we have dataand how many cases of domestication?

P. coccineus, P. lunatus, P. polyanthus(past)

conditions

and Steyermark,1 946),

However, other authors (Heiser,

1 965;and proposedfromas ancestor of

Kaplan and Kaplan, 1 988)

105

have hypothesizedbiological

two separatebasis.

domestication

The discovery

seeds (Debouck

et al., 1987)

has since changed the picture.
from two different

and distinct

et al., 1997;(Fofana'Andean' wild form is relatively
Andes of Peru (Debouck

restricted

Salgadobased on poorly known
Peru with slightly

largerTwo separate domestication

eventswild forms, with different

Gutierrez

Salgadodistribution

et al., 1995).

ranges

The range of theto the southern Andes of Ecuador and the northwestern

et al.9 1989b;

assumption that the domestication
small-seeded

althoughof another wild form in northwestern

have since been demonstrated
ecologies

histories,

Maquet and Baudoin,1997);

this would lead to theevent occurred within that range. In contrast,

the range of thewild Lima bean is huge, from Sinaloa in Mexico to Chaco in Argentina

et al., 1995;

Maquet and Baudoin,1997),

and a precise

location(Gutierrez

for the domesticationof the small-seeded

cultivatedLima beans has still to be determined.

Interfertility

between wildforms and cultivatedderivates

has been demonstrated,

althoughspectacular

based on limited

genetic differencesand differences

which are all related

to the domesticationaresyndrome (Baudoin,1990).
The scarlet
economically,runner, P. coccineus, probably

has a particularly

Guatemala and Honduras (Delgado

variable

gene pool of wild

Salinas,1 988),

form. Hybrid swarms between cultivatedthe original
preciserange, indicating

ancestordomestication

the third most importantand likely

not all ancestralcurrently

only during
available

Mexico, around Sinaloa
southwestern
suffered1913;

there is fair evidence

to the cultivated

Pratt and Nabhan, 1988).
to support a single

form wasOn the basis ofdomestication

in western(Garvinand Weeden, 1994).

It migrated

in prehistoric

times to the

USA and Central

America (Guatemala,Nicaragua,

Costa Rica),

where it has

an historic

regression(Debouck,

1 992).

P. polyanthus

was thought to be the resultof a natural hybridization

between cultivated

Xolocotzi

Accordingforms of P. coccineus and P. vulgaris(Hernandez

authorsand Eguiarte,(Delgado

Salinas,1988;

Pifierovariant of P. coccineus. A wild form has been discovered1991),

where theand Smartt, 1 995).

the century (Freeman,material,

to the cultivatedunknown, as well as the place(s)Tepary was first known by its wild form and the relationshipestablishedin Mexico,

between these forms. Which wild form is therunner is still

took place (Debouckforms, distributed

species

and wild forms have been mentioned in several parts ofcross compatibilityof the scarlet

Phaseolus

proven to be compatible

with the cultivatedshown to be the ancestor of P. polyanthus.

1988),

et al., 1959).
it was thought

to be a particular

in Guatemala (Schmit

form (Schmit

From a domestication

to otherand Debouck,

and Baudoin, 1 993),

and thus

in western Central America, this106

cultigenhas migrated

habitats(Schmitto the northwestern

Andes, where migration

and Debouck, 1991).

Fromthe above, it seems well establishedthat five species

were indeed domesticatedwild forms, and that for the commonand Lima beans extending
America and South America, independent
least

goes on in humid montane

seven domestication

processes.

domestications

fromas wild forms in both Central

occurred in both places,

The time at which these processes

resultingin attook place is worth

considering.

5. When have beans been domesticated?
Archaeology

and linguistics

this question.

Archaeology

is also subject

to ecologicalinference.

Datingarchaeologists

providesand linguistics

have been recently

for New World crops (Kaplan,

1994;

record for commonbean [4300

Mass Spectrometry

disciplinesthat can give us some insightan answer, which may change with further,conditions,

methods

The oldest
Accelerator

are the scientificdirect dating]

may not be precise

the subject

years before present

revised,1 992).

Lima bean seems to be an ancient crop, although
Early records (3,000
although

1 994),

presented

Tamaulipas,

1997).
using

using radiocarbon

some dates have been revised to more

and more ancient in South America compared to Central America.

years BP; revised

dates) are from the central Andes of Peru (Kaplan,

a pod has been dated 5600 years BP from the coast of Peru (Kaplan
Scarlet1 994)and Lynch, 1999).

runner seemed to be known as wild form as early as 8,000 years BP from Ocampo,
Mexico, but this might

be incorrect(Kaplanand Lynch, 1999).

appeared some 400 years ago in Tehuacan, Puebla, Mexico (Kaplan,
1999;

(BP);

among

is from Ancash, Peru, and 2,200 years BP for

indirectrecent figures (Kaplan,of much discussion
Smith,

much earlier dates have been repeatedly(Pearsall,data andenough for time

Kaplan and Lynch, 1999;

Puebla, Mexico, althoughdatingearlier,on

Pearsall,1995)(reviseddate),

probably

introduced

As a cultigen,it

1 967; Kaplan and Lynch,from humid parts of that state or

Oaxaca.
The earliest

record for tepary is about 2,300 years ago (date revised)(Kaplanand Lynch, 1999).

This is slightly

possiblethe fully

cultigenrecords

report parchedgrains,P. polyanthus

is not known in the archaeologicalalthoughdeveloped

it is not certain

outsidewas introduced

so it is possiblethat archaeologists

in Tehuacan, Puebla

the present range of its wild forms, so it isinto this dry valleythat its domesticationof central Mexico. Someis preceramic.

record to the author's

are always distinguishingknowledge,

P. polyanthusfrom P.

107

coccineus,

althoughmorphologicaldifferencesin the seed hilum region are clear (Hernandez

Xolocotzi

et aL, 1 959).

Linguistic

evidence indicatesa long growing tradition

among Amerindsof the humid montane zones of central and southern Mexico and western Guatemala (Schmit
Debouck, 1991),

that could be assumed to be pre-Columbian.

Its migrationin CentralandAmericatowards the northern Andes could well be historic;its move in the Andes southwards from Cauca,

Colombia, would be during this century (Schmit

and Debouck, 1991).

A couple of archaeologicaldates for remains of Phaseolus

major sites of Tehuacan and Guitarrero1999),

but it is still a priority

Cave, have been criticallyspecies,

namely in the tworeviewed (Kaplan

for future research, as is the disclosureof new sites. However, twofacts are worth noting. First, beans appeared in all cases as fully domesticated,
seed sizes and colour patterns,
Even if we speculate
1 996),
Errorsarchaeologists

and Davies, 1990;

Koinange

et aL,

of earlierof many beans in preceramichorizons of almost all sites is striking.

and contaminations

are possible;

would not have noticed

beans, at least of the species
preceramic(Hillmanfrom the wild would be indicate

Second, the finding
in the datingthat is modern informs from the wild type have been observed.

about rapid domestication

the lack of transition

available.

and no transitionaland Lynch,

it is however unlikelythat so many

the presence of ceramics. One could thus conclude thatP. acutifolius,

times. Then the question

dates than those currently

P. lunatus

and P. vulgaris

is: why domesticate

were domesticatedinbeans if you don't have a pot to cookthem?

6. Why have beans been domesticated?
This question
been predominantly
cooking

obviously

relates to the uses humans have made of these plants. Uses haveas grain (dry beans),of fresh seed of P. vulgaris,

fresh seed (shell

P. lunatus

above 2,000 masl, given physicalnowadays regularly

cooked in water, but toasting(e.g. in tepary:aL, 1995).
hypothesizedthat beans were initially1990),

advantages. Dry beans are

seems to have been commonin preceramicAmerica (Tohme etContrary to a commonopinion, the use of beans as snap bean seems to be ancient as
in the pre-Columbian

et aL, 1 991).

it is likely

and nutritionalTheis commonin traditional

Kaplan, 1 956) and still is in some parts of traditional

it is reported(Singhand pod (snap beans).

and P. polyanthus

America, particularly
periods

beans)

Mexican books or codicesdomesticated(Estrada

Lugo, 1 989).

We havefor their very young immature green pods

Given the presence of many antinutritionalthat humans watching birds that eat seedlings

factors,

and the lack of pottery,and seeds of young pods (Debouck,

could use beans in the form of very young pods. The use of leaves, tuberous

roots and

108

flowers have been reported
have been limited(e.g. in forms of P. coccineus:1988),

but such uses

in time and space. However, the use of fresh leaves of P. vulgaris

observed in eastern Africa and Indonesia
bean domesticationis the aesthetic(Westphal,

selection

colours and patterns in traditional
were originally

Delgado Salinas,

dimensionto (early)

and would explainin part the diversityof grain(Debouck,

So if domesticationpressures

agricultures

1 974).

An important

has been

1 989).

more diverse and not oriented to a single use, it is likelythat some diversity

mighthave been saved during the process.

7. How is genetic
Wild

diversityorganized

forms of the five cultivateddomestication.

Cultivatedamblyosepalus,

P. leptophyllus

beans originallythrived,et al, 1988)

in wild and cultivatedbeans have all had a wide distributionbeans have not come from wild endemic bean species
or P. plagiocylix).

so that their original

commonand Lima beans were distributed
the two species

acutifolius

and P. polyanthus)

It is however possible

in climax habitats

physiologicalhas not been greatly

alteredof which the wild forms have the smallest

where wild
Salinasto show that wild

by domestication.

original

range (i.e.

are also the ones that today have experienced

the smallest

differences

and Brticher,

1953;

Gepts

with respect to photoperiod

and Debouck,differences
1991).

along theThere are also

and temperature requirements,

but definite

evidence came frommolecular markers, and first from seed storage protein polymorphism
and Bliss,

1986;

populations

with different

1994;

Khairallah

one locatedP.

America and elsewhere.

(Burkart

Gepts

Koenig and Gepts,

as P.

in South America long before humans

Taking the example of wild commonbean, there are morphologicalarc of distribution(suchrange expanded in Mesoamerica (Delgado

entered South America, and that their diversity
Curiously,

beforeHumans mayhave affected habitats

as well as in the Andes (Freyre et al, 1996).

expansion in traditional

forms?

et al,

1986;

geographic1989),

Koenigoriginet al,Such differences

between wild

were shown with isozymes (Debouck

RAPD (Freyre et al, 1996),

et al, 1992),

1990).

RFLP (Becerra

and AFLP (Tohme et al, 1996)(Geptset al, 1 993 ;Velasquezand Gepts,

markers. Two major gene pools -in Mesoamerica and another one in the Central Andes - were indeed confirmed withthese markers, but in addition
S. et al, 1996;

unique genetic variability

Tohme et al, 1996),

and for the region

northern Peru (Chacon Sanchez et al, 1999;

Khairallah

was also revealed for Colombia
encompassing
et al, 1992;(Chaconsouthern Ecuador andTohme et al, 1996).

The existence of two major gene pools already in the wild long before domestication
the question of their origin.

The two gene pools are distinct

at the morphological,

biochemicalraisesand109

molecular level. However the differences
an assemblage
Additionalof convergent legumes but a single species

evidence of that differentiation

between themselves
1992).

are small, and would indicate

and analysing

Two scenarios are possible:(or speciation)

fertilitythat commonbean is notin which differentiation
is obtained

and recombination

when crossing wild forms

problems (Koinange

either one gene pool is a direct derivative

the two gene pools come both from an ancestral

has started.
and Gepts,of the other one, or

stock. Along the former scenario, the diversity

found so far in both gene pools would suggest that the Andean gene pool is a derivative
Mesoamerican one rather than the opposite.
that the highest

diversity

aL, 1990).

one, given the factof species is found in Mesoamerica not in the Andes. However, a groupof wild commonbeans disclosed
unique genetic

The former scenario is a plausibleof thediversity.

in the western Andes of Ecuador and northern Peru showsIts phaseolin

Allozyme

diversity

T is a simple one in comparison to other types (Koenig

has also shown its uniqueness

mtDNAhas been shown to be more and uniquely variable
evidence to consider this form as close to the ancestral(Koenig(Khairallahetand Gepts, 1989).

Its

et aL, 1 992). There is some

genepool ofP. vulgaris (Kami et aL, 1 995;

Tohme et aL, 1996).
The other wild species
variationwith a very large range, P. lunatus, also displays

across its range (Debouck et aL, 1987).

based on lectin polymorphism(Lioi,

1996;

populations

Gutierrez

Salgado et aL, 1995),

RAPDs (Fofana

et aL, 1997;

Nienhuis

and AFLPs (Caicedo

et aL, 1999).

WithinMaquet et aL, 1994),

separated

Bi et aL, 1997).
bean prevails

by short distances

MutatisTwo major gene pools have also been described(Debouck et aL, 1989c;

RFLPs (Fofana et aL, 1999),

mutandis, one can wonder ifa situation

would favour the concept of a single

et aL, 1999)

Each cultivatedbefore the split into two different
mainly harvested parts (seeds,in time of such harvested

unnecessary or harmful (antinutritionaltimes, just a few populations
all populations

speciesThus it is likely

ratherpods)

and their nutritional
Perhaps

process. Not surprisingly,

over 8,000

wild
1 996).

quality.

because it was

factors ! ) to go back to the wild and re-domesticate

of wild common bean distributed

than twothrough the domestication

parts was also affected.

were involved in the domestication

et aL,

that an ancestralgene pools (Debouck,species came froma couple of wild populations

process. This process affectedfrom each

evidence accumulated so far (Caicedo

legumes converging into a single morphotype.

Lima bean genetic stock existed

The distribution

Molecular

PCR-

to the one of common

in Lima bean that both wild Lima bean genepools are not derive directly

1999;

evolutionary

et aL, 1995),

have been shown (Zorosimilargenepool.

isozymes

one gene pool, amongin central Costa Rica, differences

other but from an ancestral
Fofanamorphologicalseveralamong

Km, only a few have been

110

domesticated,having as a consequence a marked founder effect (Bannerot

and Debouck, 1 992;

Sonnante et al., 1994).

The same has been shown to have occurred in Lima bean (Gutierrez

Salgadothe year-bean (Schmit

et al., 1995),

and Debouck, 1991),

Weeden, 1994; Schinkel

and Gepts, 1 988).

lies

forms, given its historicin fact in its wild

consequence, much of the genetic
because of speciallynegativediversity

all wild populations.

the cultivatedcommonbean is provided

ofa storage protein

studiedacid- arcelin (Osbornby bruchid

resistance,
Ishimoto

domestication
noticed,1992).

As astocks,notleft out of the gene pool ofdue to a natural modification

this volume). This protein

at low frequency (Osborn
et al., 1 998),

et al., 1986),

where domestication

legumeswould be attempted(Liener,they would have been quickly

selectedand speciallybut have been little

factors (e.g. lectins,

trypsin inhibitor,

Nowacki,it seems unlikely

1980;

several times:

exists only

did not take place.

have been observed in the other cultigens,so far. Given the presence of antinutritionalin wild

variationa trait that surely would have been

is caused by antibiosis

et ah, 1 988;

et ah, 1983),

founder effects

glycoside)

as a crop (Debouck,

A good example of useful traits

in Guerrero, Mexico (Acosta-Gallegos
Similarregressionthat all basic genetic

has been left out of the domesticatedBruchid resistancein wild forms (Schoonhoven

it is likely

andtraits in the wild, but simply because people were not there in time

to domesticate

kept by early agriculturists.

In the latter,and the tepary (Garvin1980),

once palatable

and less toxic

prussic
thatvariants

were

for and passed on to the next generationsoffarmers. This would explain the extent of the founder effect and the reductionof diversity

upon

domestication.

Wild beans today are used as emergency food (Brucher,

Debouck,

1 990);

the recurrent

use of this resource could be a mechanism by which diversity

The end products
species,

an observationof such domestication

effortsare similar

1922).

explored,potentials

in terms of unrealized

There are thus similar1990).

comparative genome research across Phaseolus

species.

The practicalsimilarities

trends in the genus.

us back to the evolutionaryorganization

A question that capturedof. species

relationships,

practical

importance

Linnaeus considered

across the five cultivatedpatterns,(Smartt,

8. The phylogenetic

is recovered.

already made by Darwin and Vavilov. This served as the basis for the Law

of Homologous Series (Vavilov,

brings

1 988;

This

and possibilities

shouldto be

be kept in mind in

exploitationof such geneticof the genus: how many phyla?
attentionand theirby researchers

since Linnaeus was the understanding

phylogenetic

relationships.

Answeringfor effective

wide crossing

and looking

for increased

that cultivated

P. vulgaris

this question
variability.

and P. coccineus were so closely

related,is ofIndeed,that heIllconsidered

the latter a synonym of the former in the second edition

Recent works (Delgado
At the subgenerical (1999)

Salinaset al , 1993,1999)

have confirmed the monophyly

level, there might be several subclades:

and at least two accordingof Species Plantarum (1 763)!

nine according

to Fofana et al (1999).

1 which partly

reflect

Extensive
compatibility

hybridization
1 979),

donor parent (Debouck,presented

in Table

althoughpopulations

1999).

segregating

Both species

et a/., 1 992).

hybridization(Schmittaxa share sequences in common, supportinget al., 1994).

rapidly

can be consideredinterspecificrevert to the cytoplasm

as belonging

with P. coccineus,

to one another's

as revealed

by resultsofThe study of mtDNA has shown that the three

the view that these taxa share a commonancestor

Studiesof polymorphisms

using CPDNA have shown the very closed

between P. vulgaris

and P. polyanthus(Schmitmore distantly

DNA sequences
hypothesis

however inand P. coccineus has shown some level ofshows affinity

slightly

diverge

Conclusionsetbetween P. vulgaris

secondary gene pool. P. polyanthus

relationshipSalinasthese works are thus tentative.

(Smartt,(Hervieu

to Delgado

These studies

the number and kind of taxa examined and type of DNA studies.

of the genus.

related.

These results

and non-molecular

proposedby Delgado

while

P. coccineus iswere confirmed by a recent study using ITS/5.8S

data (Delgado(1 985)et al., 1 993),

Salinasthat evolutionary

et al,

1999).

If we accept the

recent taxa have epigeal

germinationand fibrous roots, P. coccineus would be somewhat more ancient and the vulgaris phylum would
be its more recently
vulgaris

migrated

derivedevolutionary

branch. In contrast

with companion species

in Central

to the situation

America, enriching

There is enough evidence(morphology:

et al., 1982;

Mercado Ruaro and Delgado

Salinaschiapasanus

karyology:

as a separate phylum;

these differences

separate

section in the genus (Delgado

includedin the Phaseolus

as a distinctive

section,Fofano et al , 1999)

1991),

the two other species,

Mexico (Delgado
recognizedSalinas,by Piper (1 926)

Salinas,1985;

Salinas,1985;

palynology:

Salinas,Lackey,1983).

1998)

to consider

and P. macvaughii

1 985; Mercado Ruaro and Delgado Salinas,(DelgadoP.

it even as awith 2n= 2x= 20 can be considered

would support this view. IfP. leptostachyus

forms a group of related species

DelgadoOn the other hand, whilefrom molecular marker studies

P. micranthus

its gene pool.

are strong enough to consider

a small group of species

phylum. Recent results

1999;

Delgadoin the Andes, P.

(Delgado

Salinaset al ,

is widespread(Debouck,seem restricted

to western

1 998).

P. pedicellatusaswith P. grayanus, P. oaxacanus, etc.,

and this has been confirmed by molecularstudies

Salinaset al , 1999;1999).

I am not ready to include P. glabellus

into this phylum as suggested

Salinasef a/., 1999).

Fofana et al ,elsewhere(Delgado112

Wide hybridization
knowthat P. flliformis
Baudoin, 1987b).

has been extended
and P. angustissimus

bySalinas et al , 1999),

characterized

to dry areas to allow a quick imbibition

vulgaris phylum (Schmit
1996).

et al., 1 993);

of the seed testa. As wehave seen, wildin Central

it is only known fromCosta Rica and Panama (Freytagof the P. vulgaris

around Central America. One is tempted to locate the origin

as ancestral

distributed

andphylum is centeredof this phylum there. The branch of

in western Ecuador and northwestern(Kami et al., 1995),

America and only there.

Freytag & Debouck, was shown to belong to the P.

It seems thus that most of the diversity

wild P. vulgaris

thought

by the presence of rugose testa - anare both distributed

P. costaricensis

as initiallySo, weI would however maintain them within the same group (a view supported

a novel species,

Debouck,are not as related

species.

andP. coccineus and wild P. polyanthus
Recently,

beyond the other cultivated(Katanga

recent results by Delgado
unique adaptation

littlePeru, althoughdemonstrated

would occupy there a relic zone, as a example of past migrationsof the flora between Central and South America.
The diversity(Gutierrez

Salgadoshows affinity(Katangaof Lima bean is equally
et al., 1995).

importantbetween Central and Andean America

There are several species

in Mexico with which the Lima bean

in wide crosses, perhaps to be qualified

and Baudoin,marker studies(Delgado

as belonging

to its tertiary

gene pool

1987a;

Katanga and Baudoin,1990).

Recent resultsfrom molecular

Salinaset al 9 1 999; Fofana et al. , 1999)

would indicate

the presence ofa group of (mainly

Mexican) species indeed related to the Lima bean, although

Andean species.

The existenceof a secondary

pachyrrhizoides

and P. bolivianus- 1 would be tempted to include P. mollis in this group as well -would indicate

a center of speciationin this region, more recent in comparison to the species

evolved in Central

America (Fofana

The phylogenetic

distancegene pool in the Andes with taxa such as P.

et a/., 1999;

that

Maquet et al, 1999).

between the commonand the Lima beans, demonstrated

cross compatibility

and molecular evidence

separateof their ancestral

evolution

not as close as the(reviewed

phyla in different

by Debouck, 1 999),
geographic

can be explainedbybyareas. While the P. vulgaris

phylum evolved in Central America, the phylum of P. lunatus evolved in the Andes. From thedifferentiation

expressedby the gene pools of Lima bean, it is possible

such genepools

is anterior

to that of commonbean. Chloroplast

Phaseolus

to infer that evolution

DNA analysis

has shown thatis one group as compared to other New World legumes such as Strophostyles,

subgen. Sigmoidotropis

and Macroptilium(Delgado

Salinaset al., 1 993).

the two major economic species (Delgado

et al., 1993;

different

reflecting

evolutionary

Vigna

It could however have

evolved into at least two phyla, each including
Fofana et al., 1999),

ofpathways.

SalinasMany aspects of

113

these working hypotheses

are still speculative.

be addressedby molecular

geneticists

9. Concluding

Remarks

Beans had a long evolution

However, they offer some interesting

compatibility

prior to domestication.

Phaseolus

by substantial

available

we are gaining

about the chronology

distance
throughby the separate early history

have epigeal

Andes).

explorations

and work with molecularmarkers is

lived

perennials

photosynthesis

success

specieswith tuberousof some wildroot systems, indicating

degradation.

Several wild species (e.g. P. chiapasanus,

exercise,

as we have seen with the bruchid
It is linked

to the breadth

for future bean breeding.

into the domesticatedstocks after originalexist to Phaseolus

the other hand manynovel opportunitiesof these genetic

resourcesof translocation

P. xolocotzii)

P. jaliscanus,
disappearall

is a process rather than aof these events is not an academic
base at domestication,inwhich in turnstill has to be addressed.

resources in their native habitats,are opened by progresses

in biotechnology,

and while onconservation

seems thus more than ever a priority.

Acknowledgements
This work has been made possibleforThe extent to which genes have been introgressed

domestication(s)genetic

are short-(See the papers by Gepts and Ishimotoof the genetic

suggests possibilities
While threatsof location

resistancewere

domesticatedof wild species

P. esperanzae, P. glabellus,

Crop domestication

event in space and time. The question

speciesand those with some level of ecologicalare destroyed.

definite

towards

P. coccineus, P. lunatus, andP. tuerckheimii,

when their forest habitats

element

aggressive

pattern

P. salicifolius,

togethervolume).

anotherAs

However, muchand P. vulgaris)

species (P. acutifolius,

do well in openings of their natural habitats

thisand ecologicallyand fibrous roots, while the majority

P. vulgaris)

P. polymorphus,

is a favourable

P. lunatus, P. polyanthus

products. Four domesticatedP. macrolepis,of these phyla.

remains to be elucidated.

germinationof each phyla,

America and northerna better idea about the definition

Four (P. acutifolius,

species

into phyla,(Centralbut not the only one, as some ubiquitous

not domesticated.

has early differentiatedof their formation

The ecologicaldomestication,

can be explained

geographic

germplasm is becoming
progressing,while being one group ineach the commonbean and the Lima bean. Formation of gene pools andin wide hybridization

separatedtoon the basis of new germplasm.

comparison to other New World legumes, such as Arachis,two of which include

questions

thanks to several grants to the author from the114

Centro International
Plant Genetic(Beltsville,de Agricultura

ResourcesUSA),

acknowledges(Rome, Italy),(Cali,

from national

Costa Rica, Colombia,

Colombia),

the United

and the World Conservation

the support

Guatemala,TropicalStatesthe International

Department

Union (Gland,

Switzerland).

Chile

of Mexico,and Argentina.

The author

his profoundgratitude

to Dr. G. F. Freytag

information

about Phaseolus

species,and to Prof. P. Gepts (UC-Davis)

The helpful

support(USDA-ARS,

Ing. R. Rios, Ing. R. Ricci,

acknowledged.

Comments on a former version of the manuscript

help in the preparation

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Ing. E. Velasco,

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125

Vida faba L. and related

species:

Genetic

diversity

andevolution
Elena Potokina

l , Duncan Vaughan 25 Norihiko

1 Dept.

of Plant Introductionof Plant

Industry(VIR),

Tomooka 23 Sergey Bulyntzev3

and Taxonomy 3 Dept. of Food Legumes, N.I. Vavilov
BolshayaMorskayaStreet,42, St. Petersburg,

Russia;Institute

2 NIAR, Japan

Abstract
Wereport results
geographicaldispersal

faba bean (Vicia
investigationsof the molecularanalysisof the most economically

faba

L.).

The wild

have not indicatedcomplex as had previously

to elucidate
importantancestora close relationshipbeen assumed from morphologicalhas been investigatedpopulations,collected

in 1916-1928

geographicalbegan replacing

origin, fulfilled

monograph of
populations

fromdifferent

purity,through

identityof populations

belonging2) although

the populationsof the same origin usually clusteredgeneticallyvery distantfrom each other, except for slightly

Afganistan

and clearly

separatedin the famousby RAPD.

to different

botanical

conditions

based on passport1929).

One of them, V. Pliniana

populations

data: on quartzite,

varieties

in terms of RAPD variation;
the populations

studieddifferencesin populations

from India andfrom the North-African coast (mountainsof Algeria andof the century.

These plants

among shrubs and in pastures(Trabut)Murat,

immediateconversion to the cultigen,differenceswere growing in wild
(Trabut,

191 1; Maire,by the author as V. faba var. Pliniana.

This plant was considered by Muratova (193 1) as the nearest wild relative
who explained

are not

was found in the wild state by the French botanist

Trabut (191 1) in Algeria at a height of 900 m and describedof Cubero (1974),

andtogether,were found in beginning

hypothesis

From thevarieties

The North Africa coast is the area where several plants, having some morphologicalV. faba,of different

origins we can conclude:

among V.faba botanicalfrom typicalof V. faba

the world, before

as describedat VIR, have been studied

1) there are no differences

Egypt).

expeditionsof genetic diversity

farming systems. Fourty six accessions

and depositedgeographicaldata. The enigma of the origin of V.faba

by N.I.Vavilov

all the criteria for originalcomparison of the RAPD banding patterns

remains unknown. Recent molecular

based on analysis

the traditional

Muratova (1931)

origin andbetween V.faba and species of the V. narbonensis-

dispersal

modern agriculture

relationships,

crop in the genus Vicia L. subgen. Vicia -of V. faba

and its geographicalmostlyphylogenetic

the actual

it is possibleof V.faba. If we accept theabsence of wild plants

the clear genetic

differenceof V. faba by theirof North African

126

populations

from other V.faba gene pool can be explainedby their possible

with former wild plants of V. faba, growing in this region.
ancestor of faba bean, having no major morphological
was distributedin ancient time in the East

recent close relationshipWe suggest a hypothesis,

differencesthat a wildfrom currently cultivatedV.faba,Mediterranean region and neighbouring areas, like mostother Vicia subgen. Vicia species. Thus, it seems likely that wild plants of faba bean could be found in
the beginning of the century in the North African coast.
Introduction
Faba bean {Viciafaba
Despitenumerous studies,

et al.,

1991).

RecentZimniak-Przybylska,
relationships

economicallyis known of the origin

and domestication

littlemolecular
1995;

betweensubgenus Vicia species

de Ven et al., 1996;

Raina and Ogihara,

1994;

Jaaska,V fabaand speciesof thedata. Our analysis

thatspeciesof phylogenetic

of V.faba populations

during their expeditions

objectives

and desribedof the present

genes (Potokina
relationscollected

These collections

traditional

byby N.I.
were madefarming systems. Most of

by Muratova (193 1) in her famous monograph. The

by Muratova, by RAPD analysis,

to find possible

from different

and try and elucidate

regions

we analysedin 1916-1928

work was to analyse the same faba bean populations

geographic

with somefrom any of them. To
dispersal,

worldwide.

prior to the time when modern farming methods replaced
are identified

chloroplast

directly

methods the genetic

Vavilov and his collaborator

among 29 Vicia

of Vicia, was not domesticatedmolecular

as previously

relationships

phylogeneticalthe enigma of the origin of V faba and its geographicalandhave not shown a close

V.faba while, having

diversity

Przybylska

V. narbonensis-complex

elucidate

the accessions

1997)

using RAPD and RFLP of PCR-amplified

existingof faba bean (Maxted(vanlead us to suggest,other currently

crop in the genus Vicia L.

investigations

assumed from morphologicalet al., 1999)

L.) is a most important

relationships

withinas thosestudiedthe V. faba gene pool

the routes of dispersalof faba bean as acrop.
Materialand Methods

Populations
Fourtywiththreenarbonensis

six accessions
accessionsof Vicia faba L. depositedof V johannis

L. of different

by Muratova was analysed

origin (Appendix
in the present

at VTR were analysedin comparison

Tamam. in Karyag.

and seven accessions

of V

1). Unfortunately,

only 6 % of accessions

studiedstudy.

A rather large part of Muratova's

collection,

127
includingauthenticalaccessionsof V.faba var. Pliniana

Trabut. and V.faba subsp. paucijuga
purityby reproducing

thecriteria

Mural, were lost since

1931.

Some other lost theirpollination

control.

theaccessions

established

before the work, were included

accessions

was made according

completeinformation

Only

subsp. paucijuga

originatedallin the analysis.

Muratova's

we also analysed

namely three accessions

fulfilledfrom India and Pakistan(1931)(Table

five accessions,(Indiafor original

purityTaxonomic identification

classificationfrom China (China

them without

received

weof V. faba

1).

To get more

by VIR between 1953-1963,

3, 4, 5) and two accessionsof V.faba

30, Pakis 3 1).

RAPD analysis
Materials
leaveswere grown in pots at NIAR, Tsukuba,from 8-12 individuals

were used to extract

standard

CTAB method for small-scale

extraction

primers,

which gave clear

bands

primer tested.

polymorphic

Japan (36°05'N;

140°05'E).

DNA from each population,of DNA (Williams

with six test

using

et al., 1993).

samples,

1. Primers used for amplification
P rim e r n o .

Amplification

P7

B a se seq u en ce
A G CA C TTC G G

P9A CT CC G CA G T

P 19
P 22

TA G A CA GT CG
A TG A G TC CA C

P 54
P 68

C GT A G CG C GAC AT CG G C CC T

P lO 5TG G TC G CTG A

P 12 0TG G TC A CA G A

P 15 2

G TTT CG CT CC

P 15 5
P 15 8
P 16 3

G G A CT G GA GT
G G TG A C G CA GC CTT GA CG CA

P 19 5
P 205
P 206

G TA G G C GT CGG C CG TG A A G T
C TC GA C TA G G

P 226
P 284
P 300

C CA G A C A A G C
TC CA T G CC GTC A G G CG G C GT

P 30 1G C TG G A CA TC

of DNA was performed as described

RAPDs were scored as dominant markers (presencetransformed

into a 1 (present)

data matrix

was subjected

for Windows.

The binary/ 0 (absent)
to principal

were chosen from 70

previously

matrix over all accessions

data matrix included

(PCA)

1).

by PCR(Potokina

et. al, 1999).

versus absence of specificcomponent analysis

theNineteen

The 10 base sequence of each primer used in this study is shown (Table
Table

Young

bands)

andstudied.

The

using software Statistica

5.0

of all species

only V. faba accessions

was also analysed

128

separately.

Pairwise

genetic

distances,

new independent

variabilityof the original

dimensional

continuous

distanceswere calculated

axial co-ordinates

using Jaccard's

were calculated

data. The V.faba accessions
space definedcoefficient. From thewhich represent

were then plottedby the first two co-ordinates.

done by the NTSYS-pc program (Applied

Biostatistic

Inc., version

most of theas points

in a two

These calculations

were1.70),

using the DOUBLECENTER and EIGENVECTOR options.

Results
Nineteen primers used to analyze 46 accessions
and 7 accessionsof V. narbonensis

Each primer produced

resultedin 246 unambiguously

between 6 and 25 bands per accession.

made by Statistica

5.0 package,recognized

analysed(Appendix

2). According

to those factors,

groups,corresponding

similarity

between an accessionof the group (Sokal

variability

intraspecificPrincipal

all the accessions

1963).

(Group

The first three factors

2) and V. johannis

diversity(loadings

Factor(GroupThe degreeofseparated

the accession

species3) respectively.

isof V. faba
Factors

4-6

is the principal

more than 0.7000),

and clearly

distinguishedthe

5 separated

accessions

fromaccessionsof V. faba (Group4). Factor

Afganistan,

Pakistanand India(Group6 distinguished5), factor(Group6). All the accessions

plottedfactors

1 and 4 are shown (Fig.

1). The plot shows the clear

as points

component foraccessions

in a two dimensionalfrom Russia

space defined by thedivergenceof the North Africanfrom others of the V. faba gene pool.

To provide

additional

insight

analysisof the genetic

into the structure

similarity

confirmed that the most of the populationsof V.faba by biochemical
No differences

Most of the accessions

analysisof the data, we used the principal

matrix of V. faba accessionsof V. faba studied

result agrees with those found by other researchers1993).

related.

into six

4,and Egyptian

ordinatevariability

can be divided

the more typical

Algeria

populations

component analysis,among RAPD patterns

The higher the factor loading

among V.faba accessions.

V. faba

RAPD bands.

and the average aspect of the group, which a factor represents,

and Sneath,1), V. narbonensis

described

six factorsof V. johannis

interpretable

to the factor to which they are most closely

is given by the factor loadings.

(Groupof V. faba, 3 accessions2). The analysis

are very close to each other. This

who have studied

the intraspecific

Polignanoet al., 1991;

were found among the botanicalvarieties

in terms of RAPD variation.

together,forming a core of faba accessions

Seradillavariation

1975;

clustered(Ladizinsky,

(Fig.

co-

et al.,

on PCA plot (FigFigure 1. Positionsof V.faba, V. narbonensis, and V.johannis populations
in two dimensional
space of factors 1 and 4. V.faba - Mediterranean and Europe (o); V. narbonensis (°); V. johannis(0 ); V.faba - Algeria and Egypt (a ); V.faba - India and Afghanistan (•E ); V.faba - Russia - (å );V.faba - Caucasus (æf ).

Figure 2. A plot of the first two principal coordinate axes computed from RAPD analysisof V.faba accessions. Designation of the taxa as in Appendix 1.

data

130

2, Group 1). The core group includes

mainlyaccessions

origin. Very close to the core places accessions(Groupla, Group lb). Considerable

accessions
calledof Indo-Afgan

Mediterranean

and European

from Caucasus, southern Russia and Near Eastdifferences

origin (Groupofare found between the core accessions

2, Group 3). Closeto these

accessions"black Russian beans", of Russia (Group 4). The main divergenceandare the so-

within the faba bean

gene pool was the populations

from the North African coast (Group 5). Based on the resultsof

RAPD analysis

some new informationofdifferent

we discuss

geographicalabout the links

between populations

regions :India
Accordingto Muratova's phylologicalstudy the cultivationof V.faba in India is not ofHowever, she indicated,that the indigenous

Indianvery ancient origin.
Indoreand Pusa were clearly

distinguishedendemism. They were singled
accessionsof this subspecies

some Afgan accessions

from the other forms of the collectionout as Vicia fabain our study ( India

clearly

reallyforms from Lyullpur,

subsp. paucijuga

by their

Murat. We had two

30 and Pakis 3 1). Both of them, together

separatedfrom the otherV. faba accessions

withanalysed(Group 2). The samples from Kashmir collectedby V. V. Markovich (India

found by Muratova to be very close to thosefrom Farakh and Ethiopia,

habit

was

particularon the Figure 2, both these accessions

wereplacedto these accessions
together

only. Nevertheless,with Mediterranean

28, India 29) was
theirand European forms, showing their close relation

to them.

Afganistan
The group of Afgan beans collecteddistinguishedin 1924,by its endemism and considerable

during the expedition

diversityof N.LVavilov,

of forms (Muratova,

were forms with medium and small seeds, the colour varied from reddish
purple.

Four different

botanical

Three of them f asiatica,
subsp. paucijuga

forms, describedf. farachica,

A little

Afgan 36, Group 3) from Bamian and the

distantly

Kabul valley,

placedwith accessions

f laterita

35 andwhere at the beginningof the(Muratova,

indicatesthat

relatedseparatedfrom the populations

on our RAPD analysis
Transcaucasia

and Afganistan

are closeof Europe, the Western Mediterranean

the botanicalvarietiesof(Afgancentury faba bean was second to wheat in importancefrom India

green andby Muratova, were present in our analysis.

twentieth

populations

193 1). There

to black,

f. nigra, showed clear relationshipfrom India and Pakistan.

was

193 1). Our data
to each otherand Near East.

and
Based

appear to have no meaning at the DNA level.

131

Six accessions
Two accessions

from Armenia, Daghestan

from Armenia, identified

megalosperma, clustered

togetherand Georgia

in our analysis.

by Muratova as var. major f. orbiculariswith Mediterranean

populations.

region these forms are common.One of two accessions
Daghestan was placed among populations

were present

and f.

In the Mediterraneanof var. minor f. daghestanica

from Italy and Syria. This supports

fromMuratova's

pont

of view concerning the similarityof the Caucasian local forms to the Mediterranean

ones. Onother hand, the second accessions

from Daghestan was very close to the accession

from theSouthern

Russia, identified

as var. minor f. agrorum. This form is widely

German forage faba beans, and, according
regionsof Russia

suggestions,distributed

to Muratova, has been introduced

from Europe several hundredyears ago. The resultsthat some European forms could have been introduced

among

into the Western
confirm Muratova'sby Russians

into the to

Caucasus.
Muratova reported,that the mountain regionsof Caucasus (Svanetia

have endemic forms of Vicia faba. One of them, describedas var. minor f. svanetica
habit similar

and Daghestan)by Muratova from Upper Svanetia

was found by the first author to be a rather peculiar,

with its general

to the German forage bean (f. agrorum), but much earlier maturing than the latter.

In our study we had two accessions

from Upper Svanetia.

One of them (Georg 46), was reallyvery close to the accession

off. agrorum fromRussia. The other one, identified(Georgplace

45)

has a special

populations.

on the PCA plot, beingThus, our RAPD analysis

Transcaucasianratherdistant

could confirm the hypothesecountry is characterized

as f. svanetica

from other Caucasianof Muratova that:

1) theby forms having come both from the West and fromthe East, and 2) that some mountainous regions(Svanetia)

have their endemic forms.

European part of Russia
Six accessions

from Russia are presented

identified

as f. agrorum originatedaccessions

representing

peculiarin the study.

from the Southan assortmentof Russia

to the Russian North and occur nowhere else. The latter

Altai,

in province

1931).

The black

pumilaand f. borealis,

distribution
northern(KurskIrkutskand in the region

Russian beans include

several botanicalthe otherseeds,

fivemostlyof Russia to the East, occuring

adjoining

the Baikal

varieties

that dark-colouredof the area, while in the Central

20)

are known as black Russianand Southerninlake (Muratova,

and forms: var. minor f.

var. equina f. opaca, var. major f. ianthina.

of the forms Muratova noted,
limitsreg.),

of forms with dark purple-colouredbeans and are grown from the northern and central regions
Southern

One of them (Russi

With regard

forms are spreadregions

to thealong theof Russia there

occur

132

mostlyforms with light-coloured

seeds.

Muratova clearly

orbicularisand f. megalosperma, as well as the small-seeded

western,southern

accessionfrom Kursk in our study (Russi

and central

regions

close to the Euro-Mediterrnean
suprisingly

that

the large-seededf.

f. agrorum, were introduced

intofrom Europe. The single

20, f. agrorum) is actuallyplacedlight-coloured

on PCA plot verycluster.

The black Russian beans (Russi

17, 18, 19, 21, 22)

showed a very close relationshipwith Indo-Afgan populations.

This fact contradicts

the Muratova's
gardenersof Russia

statedsuggestionthat the black

who in the Xl-XIIthRussian beans have been introduced

by Bulgarian

century began to settle in Russia and carry on their trade. Basedon our RAPD results we suggest that the black Russian beans may have an Asian origin.
China
Five accessions

belonging

Based on the PCA plot (Fig.

to three different2), populations

from the western Mediterranean,
supportbotanical

from China clustered

Ethiopiaand Central

the point of view that the faba crop reached

the silk route by the beginningforms were included
togetherin analysis.

with populations

Europe. The relationshipfound canChina from Mediterraneanof this century (Hanelt,

countriesvia1972).

Ethiopia
Accessionsof V.faba collected

at different

altitudes

accessionsof Mediterranean

Ethiopian

populationsas previously

biochemical(Seradilla

et al., 1993)

North-Africanfrom 1800

seeds

of N.I.Vavilov

in 1927 in Abyssinia

showed a close relationship

RAPD analysis

did not indicate

assumed from morphological

with

faba

core

essential

divergenceof(Muratova,

1931)

and

data.

coast
was identified

analysed

distinguishes

coast. All the accessions

the medium seeded accession

as f. mediterranea.

minor f aegyptiaca. According
flattened

to 2800m

origin.

Among three accessions
morphologicallyby the expedition

Two accessions

to Muratova (193 1) the latter
Egyptianaccessions

showed the clear divergencefrom Algeria(Tiaret)from Egypt belong to var.

form of short habit, with small,from other countriesof the North Africanfrom the other faba gene pools.

Discussion
The first pointis the divergenceof the North-African

regionhas a special

meaning in respectslightlydifferent

morphology

from real faba beans were found in the wild condition

Trabut in Algeria,

at Sersou de Bourbaraki

diversity.

of V. faba. The

Algerian

French botanist

to V. faba

populations

a Vialar,

In 1893

at an altitude

plantswithby theof 900 m

133(Trabut,

1911).

In thislocality,collected

the green pods of the wild

faba bean plants

preferred

to call the plants{Faba vulgaris Pliniana

as varietasof the plant first discovered
independent

species.

Trabut),thinking

for raising

women

The author
the characters

it to the rank of an

the geographicallocalization,

its deliminationfrom the cultivatedbean forms by singling

under the name V.Plinianain the northernmentions a form differingshow orange reddishof the seed coat),justifiedfinds of V. Pliniana

Africafrom V.Pliniana

keptedPliniana

sample, collectedby A. Battandier

locality,

where the accession

analysed

1926. If we accept the hypothesisof V. faba by theirdifferences

Battandierin Algeria,of Cubero (1974),
or very rapid

3 seeds each which(MPU, Montpellir)

growed in this region.

allow us to suggest

faba bean, having no major morphologicalrecentby N.I.Vavilov

was collecteddistributedin ancient times in the East Mediterranean

all other

species

species

concentrations

Vicia subgenus

Vicia.

as indicatedby isoflor

Europe and SouthwesternAsia.

we can explainfrom others of the V.

with wildforms of V. faba,a hypothesis,that a wild ancestor

offrom currently

cultivated

wasV. faba,region and in the Near East, like almost

Accordingto Maxted (1995),

maps, the centre of origin
Taxa of the subgenus

based

on relative

for subgenus

Vicia ishave subsequently

spreadwest and north, and to a lesser extent, east and south from this area. Thus, it seems likely
wild plants

of faba bean could really be found at the beginningof the North African coast.
(1974)

suggested

This hypothesisthat is logicalbegan in the Near East, following(Zohary& Hopf, 1973).

to suppose
the Neolitic

in the Mediterraneancountries.

(Greek,Hebrew, Arabic and Berbers)

thatof the century in the mountains

agrees well with that of Cubero (1974).

Cuberothat the use of Viciafaba

as a cultivatedspecies

across the inhabited

world

cultureas it spread

This means, however, that the plantparticularly

inthe actual absence of wildto the cultigen,relationshipdifferences

is a V.

37),

conversion

by their

thereThis is the same

who explained

faba gene pool may be explained

Amongat Sersou de Tiaret.

between the of North African populations

Our results

Maire (1929)

among shrubs and in pastures.

in this study (Alger

immediateof the century.

The form was found in the Western part ofon quartzite,

at the Herbierit

There were some moreby 45 mm long pods containing

in wild state,

specimens(Tabut)Murat.

at the beginningspots on a black background.

Morocco (Capar-Boulhaut)Southeastern

to use them for cooking.

by him not sufficiently important

speciesofAfricanand the wildof the plants,

the clear genetic(1911),

structure

condition

plantsof Trabutcharacteristics

anatomical

V. Pliniana

to evidence

Muratova (193 1), however, found that the morphological(arillus,out into a separate

accordingwas unknown in other regions,

Four names for V.faba in the different(Muratova,

1931,

Hanelt,1972)

languages

in a relatively

small

134

regionis evidence

that the plant

Near East (Cubero,natural1974).

It is possible

suggested

four different

confirmed by our results
the principal(Fig.

Asia, Afganistan

From the principal'a centre of third

centre Vicia faba spread to Ethiopia.

divergenceof Ethiopianwith Mediterraneanorigin.

forms are also very similar

Caucasianwere, probably,

introduced

Like other Vicia species,

importance'(cit.

core-accessions.

to Mediterranean(Sersouconclusion

de Boubaraki,

investigation
possible

we suggest

to get more precise
most closely

Chineserelatedforms

a group of recent
was introducedof the latter

diversity

in mountains

and Western Morocco need additional

information

any

to thethat these forms have an Asian origin.

that Vicia species

Sersou de Tiaret)

193 1).

ones. Some European forms

into Russia from Europe, except for the black Russian beans. The proximity
As a practical

Muratova,

RAPD analyse did not indicatedand European beans and represent

level led us to suggest,from

the common beans were

into the Caucasus by Russians. Faba bean evidently

Afganian forms at the molecularas

V. faba formed a secondary

regionforms from Mediterranean

are also relatedof V faba, had its

and the Near East regions

and India,

Asia. From the Mediterranean

to Europe and formed there

and thefrom the Near East (Fig. 3). They are3). We propose the Mediterranean

in Centralof Mediterraneanof subgenus Vicia originated.

centre where the forms of V.faba have originated.

centre of diversity

particularspeciesroutes radiatingarea Vicia faba spread to Central

distributed

peoplesthat at that time the wild ancestor

area in the same region where all other cultivatedCubero (1974)

thiswas known by different

about the phenomena of V. Pliniana,of Algeria
and detailed
as a form

to the wild ancestor of V.faba.

Acknowledgements
This research

was undertaken

while the first author was an Science and Technology

fellowof the Japanese

Goverment. We are grateful

analysisof data.

to Dr. Leonid Malyshev (VIR)

for help in

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Appendix

Id en tific a tion
nu m ber ofa cc e s sio n s

1. V.faba,

C a ta l.
num berV.johannis

C H IN A 1

51

1 9 2 3 /e x p e d .

C H IN A 2

10 9

1 9 2 3 /e x p e d .

C H IN A 3

14 5 0

1953/
A g ri cu ltu re
I n stitu te

C h in a , H a n ‑C h z h o u

C H IN A 4
c H IN A S

1 5 151529

1 9 5 5 /ex p ed .
1958/

C h in a , N or th ‑ W est
Cp rovh in ina ,ceC ,h me narkd u ,etS y ch u an

A g ricu ltu re
In stitu te

p rov ., a lt. 4 5 0 m

Y ear ofre ce iv in g /s ou rce

and V. narbonensis

accessions

O ri g in a n d (lo ca l n a m e) o f
a cc e ssio n s

C h in a , P e k in , K a lg a n
d istr. (D am u )M a n c h z h u ria (T z an D o u )included

C o llec to r's
nam e

P isa rev
P isa rev

Iv a n ov

in analysis

T axonV .f a b a v w .eq u in a , f.
r o se sc e n s
V .f a b a v a r . m aj or
f. u n d u lo s a
V .f a b a v ar .m aj o r ,f .p ro c u m b en s

V .f a b a v zr .m aj o r,fV .p.froa bc au m b en s

C H IN A 6697

1 9 2 7 /ex p ed .

W est C h in a, H an a iy k

D u litis

S P A IN 7802

1 9 2 7 /ex p ed .

S p ain , B ar ce lo n a

V av ilo v

S P A IN 8788

1 9 2 7 /ex p ed .

V av ilo v

S P A IN 9924

1 9 2 7 /e x p e d ..

S p ain , M a d ri d ,R o b u stia n o D ie z O b e sa
RSpo bainu stia, M na od Dridie, z O b esa

V av ilo v

V .f a b a v ar .eq u in a ,f .e q u in a
K f a b a v ai .eq u in a ,f.e q u in atV v.fiha db aia vn ai
a .eq u in a ,S P A IN l O

800

1 9 2 7 /e x p e d .

Sp ain , M a d rid

V av ilo v

V .f arob ar uvm ar . m in o r,f.ag

E T H IO l1

74 1

1 9 2 7 /e x p e d .

V .f a b a w ax . m in o r
f.a by ss in ic a

E T H IO 1 2

76 3

19 2 7 /e x p e d .

E T H IO 1 3

77 3

19 2 7 /e x p e d .

G E R M A 1488

F R A N C 15266

E N G L A 16

60 5

A fr ic a, E tM o p ia, D eb a re k , V av ilov
8 0 k m N E G o n d a r, a lt.
2 8 0 0 ‑3 0 0 0 m
A fric a , E th io p ia, D eb a re k , V av ilov
80 k m N E G o n d a r, a lt
26 00 mA f ric a , E th io p ia , H ara r
V av ilov
re g ., m n t. M u lla t, a lt.
26 00 mG e rm a n y (E c k e n d o rfer
F eld b o h n en )F ra n c e, P a ir s (F ev ero le d e
P ic a rd ie )E n g la n d (E a rly M az a g a n ,B ro ad B e an )R U S SI 1757

19 2 2 /S tru b efirm
19 2 5 /V ilm o rin
fi rm
1 9 2 7 /S u tto n 's
S ee d s e t B u lb s
fi rm
19 2 3 /A g r icu lt. R u ssia , K o stro m a reg .,C o lle g e ,K o lo g riv y d istr .
la n d ra ce

V .f a b a v ox .m in o r,f.a by ssin ic a
V .f a b a v a r.m in o r

V .f a b a v z r.m iri o r,t ag ro ru m
V .f a b a v ar .e q u in a ,t e q u in a
V .f a b a v ar .e q u in a ,t e q u in a
V .f a b a v zr .m in o r,t p u m ila

Appendix

1. cont

Appendix

1. cont.

Appendix

2. Principal Component Analysis of RAPD pattern variability
V. faba, V.johannis, V. narbonensis analysed

among accessionsofAppendix

2. cont.

143

The

genetic

structuresoybeans

revealed

mitochondrialof naturalbyisozymes

DNAs: possible

cross-pollination

populations
andinfluenceof wild
RFLPs

ofof seed dispersal,

and demography

JunAbeLaboratoryof Plant

SapporoGenetics

and Evolution,

Facultyof Agriculture,

Hokkaido

University,

060, Japan

Abstract
resolving

An understanding of genetic structures within and among populations
is important forevolutionary processes of genetic differentiation, developing strategies for in-situ conservationand sampling wild germplasm. In this paper, the spatial pattern and structure of genetic variation in wild
soybeans is assessed

based on genotypes at isozyme loci and RFLPs ofmitochondrial

The wild soybean is highly
mtDNAs. Populations

variable within and among populations

possibly

between populations

self-pollinating

nature of thisof genetic variation

caused by disturbanceof isozymes and RFLPs of mtDNAs reveals that populations

structured

populations

demographic

featurespopulation

genetic structure.

species.

has resulted from limited gene flow due to geographic

also from the reorganization

combined analysis

for both isozymes and RFLPs ofconsist mainly of a number of different homozygous plants. Heterozygous plants

are absent or rare because of the predominantly
differentiation

DNAs (mtDNAs).

to more randomly variable populations.
of wild soybean are discussed

Seed dispersal,

as possible

Genetic

isolation,andand migration.

Avary from highlycross-pollination

andfactorsthat influence

the intra

soja Sieb. and Zucc, is distributed

throughout

China,

Introduction
The wild soybean,

GlycineKorea, Japan, Taiwan and areas of Russia adjacentto China (Hymowitz

and Singh,

1987).

It is

144

a twining annual, and grows in sites that are subject
roadsides,

riverbanks

proteinprofile,and dry riverbeds.

and RFLPs of chloroplastrRNA and non-coding
the wild

ancestor

consistent

reproductive

regionssuch as wasteland,karyotype,seed

DNAs, and sequence analysis

genomes supports

the hypothesis

G max (L.) Merr. (Hymowitz

ofthat G. soja is

and Singh,

1987).

No

barriers exist between G maxand G. soja. Thus G. soja has been used
to furnish

The genetic

useful genes not in the cultigen

variation

characters

1971;

BroichFukui et al., 1978;

Kiang et al., 1992;

RFLPs of chloroplast(Sekizuka

and Yoshiyama,

and Palmer, 1980;
Yu and Kiang,

and mitochondrialand single

genepool.

of the wild soybean has been extensively

to morpho-physiological

Abe et al., 1999),

disturbance,from studies on morphology,

and mitochondrialof chloroplastof the soybean,

in soybean breedingal., 1992;

Evidence

to frequent

repeatsurveyed with respect

1960;

Kiang and Chiang,

1993;

Fukui
1989),

Dong et al., 1995;

DNAs (Tozuka et al., 1997;
sequence markers (Powell

and Kaizuma,
isozymes (Abe et

Fujitaet al., 1997),

Shimamotoet al., 1998;

et al., 1996).

These

studiesreveal that G. soja is geneticallymore diverse than G max.In this paper, the spatial

patternvariationand intra population

genetic

on bothisozymesand RFLPs of mitochondrial

cross-pollination,and demographic

Spatialof genetic

patternof wild

soybeanDNAs (mtDNAs).

Possiblefeaturesvariation

Yu and Kiang (1993)

structureon the structure

and intra

evaluated

six South Korean wild soybean populations.
the number of alleles

per locus (A), 0.372

Nei's

(H).

gene diversity

plant populations
systems (1.53,
thus

highlypollinating

(1.3,
0.342

0.200

variable

at isozyme

evolutionary

situ conservation
structure

population

mean values of 1.4 forof polymorphic

loci (P),and Godt, 1989).

lociwithinpopulations,despiteof diversity

withinand among populations

and 0. 134 forfor 1 13 self-fertilized

and 468 plant populations(Hamrickloci forof all mating

The wild soybean is

its predominantly

self-

nature.
The organization

resolvestructurethan those reported

respectively)

respectively)of seed dispersal,

at 35 isozyme and seed protein

for percentage

and 0.074,

and 0.113,

genetic

They reported

These values were higher

roles

based

are also discussed.

population

the variation

is assessedofof genetic

processes
and sampling
variation

Japan, were assayed individually
These populationsof genetic

differentiationof the wild germplasm.

but also to develop

forty-three

for genotypes

at 15 isozyme

populations

on riverbanks of different

not only to

strategies

In order to assess the spatialin the wild soybean,

were mostly collected

is importantin northern

for in-patternandHonshu,loci and RFLPs of mtDNAs.
river basins, which aretopologicallyseparatedwere characterizedfrom one another by mountain ranges (Fig.

1). The observed variations

as follows.

1) Isozymes
A totalof 36 alleles(2.4

corresponds

to 63% of the alleles

throughout

the distribution

homozygous plants,
self-pollinating

per locus)(57)

were observed

observedin the whole population

area. Most of the populations

and heterozygous

nature of this species.

in the 43 populations.

consisted

collected

This
from

of a number of different

plants were absent or rare because of the predominantly
The observed heterozygosityof isozyme loci at the seedlevel was, on average, 0.004 for the 43 populations,being the same as the value obtained

South Korean populations(0.004,

Yu and Kiang,

1993)

OmonoRiver populations(0.032,

Fujitaet al, 1997)

for six

but lower than the values for fourand four KitakamiRiver populations

(0.023,

Kiang et al., 1992).
The H value for all 43

populations

was 0.253.

Only threemonomorphic. These three populations

consistedof a single

multi-locus

the monomorphic populations,genotype.

were

Excluding

ranged from 0.017 to 0.261.

The

meanH value for each of the 43 populations

was 0. 132. The Gst value, which represents

theproportionof total

genetic

to differentiation

calculated

at 0.476,

indicating

between populations.
parts:

the H value in each population

populations

diversitythat

The genetic

one for the diversity

attributable

about half of the total
diversity

allocatedamong populations,variation

between populations

to the differences

was thus

arose from the difference

can further be divided

among regional

into twogroups of populations

and the other for the diversity

between populations

within the groups. The 43 populations

weredividedgroups, based on the river basins in which they were located.

The

into eleven regional

Gst value among the regionalgroups was 0.33. Sixty-nine

the populations

from the variation

thus resulted

The genetic

differentiation

confirmed by cluster
43 populations

analysisamong different

between populations

using Nei' s standard

were classified% of the variationof different

between populations.

analysiswith the UPGMA method

frequency

similarity(Fig.

2), and are plotted3). The populations

withinriver basinstend to have similar

one another, while those of different

river basinstend to differ

differentiationof different

gene flow among the river basinssame river basins,

the genetic

theirdistances

geographic

withinthe river basin,

et al.,

1992).

However, a difference

downstream populations
and Kitakami

rivers,the downstream populations
the extent

and frequency

populations

which resembledof disturbancedownstream region has often been subject
activities.
reorganization

Genetic

differentiationof genetic

allelic

sites on thetofrom one another. The genetic
be attributed

may be partly

et al., 1997)

was also foundThe

compositions

as was found in riverbank

even within the same river basins.
the upstream

collection

by mountain ranges (Fig.

between populations(Omono 6 to 9 in Fig. 3) of the Omono River (Fujita(Kiangat theirriver basins may largely

which are separated

compositions

was also

distanceby cluster

between populations

river basins

genetic

13 groups

map (Fig.

among

river basins.

intobased on the allelicobserved

to limited1). Withinthea functionofpopulations

and the KitakamiRiverbetween the upstreamandAs shown in Fig. 3, in the Omono

were similar

to one another but different

to one another. This may reflect

froma differenceinbetween the upstream and downstream regions.

The

to human interventions

between populations

variation caused by disturbancesuch as riverbank protection

would also occur as a resultand migration.

of theNei's standard

genetic

distance

Fig. 2. Classificationof natural populations of G. soja in northern Honshu, Japan, based on thesimilarityof allelic frequencies at 1 5 isozyme loci.
Cluster analysis was performed by the UPGMA method. The populations were classified into 1 3groups on the basis of genetic distance of 0. 1. Data for three populationsof the Omono river(Omono 7 to 9) are cited from Fujita et al (1997).

Fig. 3. Genetic differentiationin natural populationsof G. soja innorthen Honshu, Japan. The populations are marked with different
colors according to the groups classified
based on the results ofcluster analysis (Fig. 1). The data for three populationsof theOmonoriver (Omono 7 to 9) are cited from Fujita et al. (1997).

Fig. 4. Frequencies of three major mtDNA haplotypes
populationsof G. soja in northern Honshu, Japan.

in natural

1492) RFLPs of mtDNAs
The RFLPs of mtDNAs are useful for determining
plantsin natural

populations

and for tracing
genome is maternallyhave so far been identified

for the wild soybeans collectedbased on the RFLP profiles

obtained

et al., 1997,

Honshu population

Shimamoto(Tozukainherited.

et al., 1998).

differedamong theconsistingof plants which belonged

slightly.

The diversity

possessed

3) Intramaternal lineages.

analysis

intrapopulation
populations

genetic

within populations

accordingto theirof populationgraphswere obtained

polymorphic

of isozymes

are labeled
relatedness

by Hayashi'sloci for all of the plants
Populationsof mtDNA haplotypes
one another. However,different

size and by migrations

incompositionseven-River, which varied onlyby thefrom other populations.

in the extent of disturbance.

structure

structures

genetic

along themay also be influenced

and RFLPs of mtDNAs enables

in more detail.

us to assess

An example of the analysis(Omono 2 to 6) of the Omono River is presentedof the five populations

One population

the compositions

between river basins may also reflect differences

A combined

while the other populations

to those of the Kitakami

of mtDNA haplotypes

genetic

for mtDNA

five mtDNA haplotypes.

nearby, in contrast

population(Id, IVa

were monomorphic,

the same river basins tend to resemble

random drift due to the fluctuation
Differences

populations

along the Omono and Mogami rivers exhibitedthough they were locatedwere found in the4). The diversity

to the same maternal lineage,

As in the case of isozyme genotypes,
within(Fig.

Thirteenfrom different

Aka River, Yamagata Prefecture,

the populations

genes, cox2 and atp6, as probes

Of these, twelve haplotypes

populations.

derivedpopulations

most of its range,of the three most commonly observed mtDNA haplotypes

haplotypes

neighboringof 26 mtDNA haplotypes

from throughoutwith two mitochondrialfor each of the 43 populations

were a mixture of plants

A total

et al., 1997).

The frequencies
and Va) are presented

of individual

the movement of seeds between populations

because the mitochondrial(Tozuka

the maternal lineage

with their mtDNA haplotypes(Fig.

5). Individual
and are plottedforplantsfivein eachon the graphs

at isozyme loci. The horizontal

and vertical

Quantification

to the variation

at nineto more randomly

variable

Theory III appliedaxes on theof the five populations.

vary from highly-structured

populations(Fig.

5). Only two multilocus

genotypes

populations(Omono 2 and 3). In the population

populations

were observed in the two downstream
nearestthe coast(Omono 2), the twoFig. 5. Intra population genetic structure in riverbank populations of the Omono river revealed
by isozyme genotypes and mtDNA haplotypes.
The horizontal and vertical axes in the graphs were obtained by Hayashi' s Quantification
TheoryIII applied to the variation at nine polymorphic loci for all of the plants of the five populations.
Figures below the population code indicate Nei' s gene diversity for each population. Different
sizes of circles shows different numbers of plants.

Fig. 6. The expected dynamics of genetic diversity and structure that are influenced by migration,
outcrossing and disturbance in the wild soybean populations.
The model starts with migration from other sites into a less polymorphic population/ Circle andsquare indicates different mtDNA haplotypes, respectively.

151

genotypes

were different

disequilibrium.

from each other at five isozyme loci, showing a high gamete-phase

The H value for population

values of the two upstream populations
the isozyme genotypes
Furthermore,'Omono T was 0. 1 15, being slightly(0. 154 for Omono 4 and 0. 157 for Omono 5), in which

at each locus were more randomly combined with those at the other loci.

each of the two genotypes

corresponded

or IVa. One or the other of these two genotypes
artificial

disturbance.

to the different

each locus were only weakly associatedmtDNA haplotypes,

Id

a recent migrant by naturalor(Omono 4, 5 and 6), the genotypes

at

is probably

In the three upstream populations

with those at the other loci, and multilocus

were also only weakly associatedwith mtDNA haplotypes.

In addition,of these

populations

possessed

multilocus

thatregardlessof their

mtDNA haplotypes,

thoughthey belonged

between individuals

section.

but differed

were thus not associatedcross-pollinationand ecologicalreorganization
diversity

Populationsin less

genetic

disturbed

plantsstructured

influence

and would consist

population

genetic

is presented

populationin Fig. 6. The

and the migrants

populationbywith a high gamete-

may generate progenies
generations.

This

with

may causeand organelle

genomes. The consequence of genetic

is dependent

on their dynamics,

and fragmentation
may maintainmay lose genetic
of a limitedin the next

on the Omono River. Infrequent

genomes in succeeding

habitats

bottle-neck,a highwhich is influencedof populations
genetic

by disturbance.

diversity,

while

those

variability,

as a resultof random drift or anumber of related

genotypes.

Therefore,

the intraof wild soybean depends on the extent and frequency of migration,

cross-pollination,and disturbance.

Factors

intra

population

patternand possible

from other sites into a less polymorphic

may produce a highly

exposed to disturbanceThe spatial

structuresoybean which will be outlined

frequently

affectinggenotypesof wildsuch as mortality

structureof nuclear

characteristics

within populations

factorseven

by isozymes and RFLPs of mtDNAs and the

between nuclear genotypes

by demographic

populations

assessed

and organelle

and structure

to one another,structure

between the native

genotypes

in eachwith their maternal lineage.

such as the downstream populations

cross-pollinationdifferent

The similarities

plantson the structure

Such a migration

phase disequilibrium,

were similar

genotypes

and disturbance

The model starts with migration

disturbance.

individualfrom those of different

dynamics of the intra population

model is based on the genetic
biological

genotypes

to the same maternal lineage.

The expectedof migration,

lower than thegenetic

and structure

structuresof genetic

of wild soybean
variation within

populations

is influenced

152

by a species'
dispersal

biologicalcharacteristics

such as its breedingmechanism. Moreover, spatial

significantly

affectevolutionary

importantconsequences

and temporalfactors

structure

the spatial

pattern

Four factors

influencing

populationsof wild soybean are (1)disturbance,and (4) feeding

variations

such as selection

for the geneticseedsystem, pollinationin populationand genetic

of populations
and structure

dispersal,(2)

and grazing pressures.

system and seeddynamics also

drift and thus may have(Hamrick and Nason, 1996).
of genetic

variation

cross-pollination,in natural(3)

Each of these is discussed

mortalityandin the following

sections.

(1) Seed dispersal
Seed dispersal
intrapopulation

genetic

genetic

statistics

diversitythandevelopmentthat determine

structure

species.

in plant

the spatial

patternof variation

andHamrick and Nason (1996)

comparedtheamong plant groups with various seed dispersal

found that the species
valuesis one of the factors

with gravity-dispersedor explosively-dispersed

did wind-dispersed

species.

of spatial

heterogeneity

genetic

mating system or breeding

Localized

decreased

Gst

leads

to thebetween populations

regardlessof a species'

structure.

proportionallythe mother plant in an experimental
dispersed

seeds had higher
evidently

around the mother plant by pod dehiscence.

seeds dispersed

andseed dispersal

The wild soybean has no device for long-distanceare dispersed

characteristics

seed dispersal.
Oka (1983)

to the reciprocal
population

The seeds mainly

found that the number ofof the square of the distanceof wild soybean:

a maximumof 5 m from the mother plant. Besides

fromthe seeds were found to be

the dispersal

by pod dehiscence,the seeds are also taken to other places from around the mother plants by mice. Ohara et al.
(unpublisheddata)found that the seeds dispersedand partly

hoardedunder leaf litter

Apodemus speciosus, in a riverbank
seeds by mice and seed dispersal
between patchesof individuals

A long-distance
genetic

structure

many genetic

Clethrionomys

rufocanus andpopulation

The hoardingof wild soybeanof Hokkaido.

by pod dehiscencewithin populations,migration,

seed dispersal

floodsand human intervention.

may have importantand possibly

when it happens,from the native plants

distance

were mostly eaten

by two mouse species,of wild soybean populations

aspectsaround the mother plants

roles in the migration

also between populations.

would cause a large impact

because the migrants
in a population,

are expected

as is outlined

in wild soybean may result from natural and artificial
Genetic

similarities

associatedon theto differinin Fig. 6. Longdisturbanceswith the geographic

likedistances

153
between populations
et al., 1997).
caughthave often been found in riverbank populations

Kiang et al. (1992)

likelysimilarity

duringburiedbetween the riverbank

gene flow from the upstream populations

periods"

between populations.

distanceof flooding.

Artificial

disturbanceto a pre-existingis mostto the downstream

is also involved

seeds is often taken from place to place. Thus a new population
may be introduced

association

populations

During clearing of land, and levee and road construction,

newgenetic materials

Fujitapods and thoseon water for over 24 hours. The observed

and geographic

the resul of unidirectional

populations

et al., 1992;

found that the seeds of wild soybean in intact

in a dry pod wall can float

between genetic(Kiangin migrationsoil containing

may be establishedor

population.

(2) Cross-pollination
The pollination

system also determines

the genetic

and Godt, 1989;

Matsuo,1997).

Wild

and populations

consistmostlyof homozygous

heterozygousof2.3%

along the Kitakami

values (2.4%

to 3.0%)1994;

and Kiang, 1987).

Chiangsuch as honeybees

and carpenter

plantsreported

to flowers

has hardly

been disturbedal., 1997).

Large populations

genotypic

self-pollinating

nature,plants.

an attractive

estimated

proportionofan average outcrossing

rateRiver. The estimate

observations

bees frequently

visit

flowers

et al. (1997)

estimated

rate reportedby shore-protection

activities

food reward for pollinators

revealed

that pollinatorsof wild soybean
multi-locus

by Kiangin a relatively(Matsuo,by cross-pollination

combinations

in subsequent

generations.

OmonoRiver populations.

and Caviness,(Fujitaoutcrossing

et al.,
rates forwith an average of 13%,et al. (1992).

The highand other human intervention

seeds generated

index (F) at the maternal(Ahrent

The upstream region of the Omono River

with a high plant density,

Wright's

the same as theof the Omono River may be ascribed to frequentin these populations.

and organelle

is almost(Omono 6 to 9 in Fig. 3) of the Omono River.

populations

nuclear genotypes

on thesoybeanfield

Heterozygous

fixation

Based

However, detailed

than the outcrossing

rate in the riverbank

of pollinators

providehave predominantly

ranged from 9 % to 19 % among the populations

which is much higher
outcrossing(Hamrickin the cultivatedin four upstream populations

The mean estimatesvisitspreviously

Using isozyme markers, Fujita

individualof plant species

seeds at isozyme loci, Kiang et al. (1992)

in four populations

1997).

soybeansstructure(Fujitaetundisturbed

habitat,may

segregants

with different

1 997).
produceThis may cause reorganization

genomes, as modeled in Fig. 6. Fujita

et al. (1997)

parent level (not the seed level)

The F values ranged from zero to 0.383

betweenmeasured

for each of the fouramong the populations.

154

Heterozygous

plants were present in only two populations,althoughcompared with that expectedfrom a random mating population

were observed

two populations

questionplantsin the other

why there were no heterozygous
producedheterozygous

with smaller
plantsof wild soybean populations.

size. This

raisesare needed to understandthethe individual
rates ofthe actual role of out-

and disturbance

Plantsmay often be subject

to different

nearby. Ohara and Shimamoto (1994)
twining

population(mean outcrossing

crossing(3) Mortality

plantshigh frequency

Furtherstructure(H). No heterozygous
although

0. 122 and 0. 1 15, respectively).
in the genetic

were lowin these populations

seeds at a relatively
studiesthe frequencies

and branchingin an inland

branchingform was predominant
intensity

The twining form wascover and soil moisture

were high,from the seed poolsubsequent

The mortality

while

the

where plant cover and soil

was high. Different environmental

genotypes

owing to waterlogging

of Hokkaido.

in an open area of dry riverbed

emergence of different
development.

population

area where plant

moisture were low and light

schemes even though they growfound wild soybean plants with different growth forms,

forms, in a riverbank

predominantselection

stressesand in different

resultedsurvivalin therates

was higher in the open area than in the inlandby typhoon-caused

flooding

in

area,

in the late summer and autumn. A highmortality

may cause a rapid

decreaseof population

size,

and as a consequence,

resultstochastic

fixationof alleles

for neutral

markers

such as isozyme

and DNA

polymorphism.
genetic

or loss

In wild soybeans,the influencestructure when exposed to spatial
Habitatfragmentation

plant

populations

consistand their patchiness

The smallest

assemblage

adjacent

patch

which

are linked

and these local populations

more loosely

structured

more or less linked

withinpollenpopulationof genetic

variation.

In general,or the natural(Kawano and Kitamura, 1 997).
population'.

A group ofgene flow constitutea 'locala larger area constitutea 'metapopulation',and empty patch habitats

and/or seed dispersal.

may occur frequently

clarification.

dynamics, and ascommunities

a 'patchby a significant

of local populations

by gene flow throughand fusion of local populations

is called

schemes on population
still requires

in biotic

substructured

in a habitat

population',assemblage

and structuresof patchinessis hierarchicallyof individuals

populations

may also influence

patternsof various levels

landscape,selectionand temporal heterogeneity

by disturbancea consequence, play a role in the spatialof different

Habitatawhich areHowever, fragmentation

and as a consequence,flow may expand or contract (Kawano and Kitamura, 1 997).

inthe range of genefragmentation

causes therestrictionof gene flow and the development

between neighboring(4) Feeding

patchesof genetic

Ohara et al. (unpublished

20 to 200 seeds per individual

data)

found that wild soybean seeds were mostly

populationof Hokkaido.

in natural habitats

The number of seeds produced

the wild soybean populations(Oharaper individual

and possibly

Wild soybean plants produce

et al., 1989;

Ohara and Shimamoto,seems to be enough to at least maintain

increasethem. However, in this population,

few seeds were left around the mother plants that weren't eaten or hoarded
reductionthatin seed numbers appears to be compensatedmigrated

population

variationby animals

eaten or hoarded by mice in a riverbank
1994).

heterogeneityand populations.

and grazing pressure

Recently,of a spatial

downstream

from upstream

seems to remain constant.

for by buried

populations.

maypose a serious problem to isolated

populations

the

size

seeds
of thisin seed numbers due to feedingin which migration

would always be exposed to the danger of extinction

by mice. This

seeds and probably

Consequently,

However, the reduction

only ais rare. Such populationsin spite of the high reproductive

capacityof wild soybeans.
Another example of interferenceby animals is grazing

as is found in southern China (Yamaguchi
China is often limited

plantscommunity if the grazing

succession

pressurefrequency of disturbancevarious

populations

features,may finallysuch as a cliff. However, a moderate
to conserve such a populationinexclude the wild soybean from the plantis removed (Yamaguchi

et al., 1999).

of permanent study sites is necessary

and its influence

or individuals

withinnaturaltopographicalsizes within

1996;

Kawano and Kitamura, 1997).

habitatcan dramatically

disturbance,of wild soybean in southern

remarks

Long-term observationplant

and topologicalby animals and humans maybe a necessity

this area because secondary

Concluding

The habitat

animals,

to only a few safety sites from which the animals keep away because of

the presence of aculeatedlevel of disturbanceet al., 1999).

pressure by domesticon the population
populations

Isolatedof wild soybean. Local

areas or plant communities
or disappear

and new patches may become establishedthe case with wild soybeans.

dynamics

are often clustered,

Such local clusters

expand, contract,

to evaluate the extent andforming patches(Hamrickof& Nason,and patchesof individuals

withintheas a resultof natural

mortalityor(Kawano and Kitamura, 1997).

or fragmented populations,This issuch as those along roadsides

156

and in wastelands,

may be particularly

populations,migration

fragmented

and isolatedof extinction,

because,from other sites may be very rare without

the only sources of recruitment

are locally

produced

processes

shouldannually

be well understoodparticularly

focusing

on how the dynamics influencevariation.

A molecular-biological

study

understandthe dynamics

variation

samplingof the wild soybean.

of genetic

coupledunlikeriverbankhuman intervention.

populations,season are the seeds that
demographic

in danger

In suchfor the next growing

and buried

seeds.

The dynamics

ofand comparedamong various habitats,the spatialand structure

withpatternconservationand design

strategies

biologyof genetic

shouldfor in-situ

help

to

conservation

andfor this study.

Thisvariants observedinAcknowledgments
I would like to thank Mr. S. Kitajima
work includes

and Mr. H. Kuno for their assistancea part of Master thesis of Mr. Kitajima.

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Introgression
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In-situof wild azuki bean and wild soybean

conservationof wild genetic

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Res. l(suppl.

l):

1 93 (Japanese)Yu, H. and Kiang, Y. T. 1993.

Genetic

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Korean natural

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159

The Vigna angulariscomplex in Japan

Duncan Vaughan1 , Norihiko

Tomooka1 , Ru-Qiang

Doi1,

Kooichiand Akito^rop

Evolutionary

Agrobiological

Dynamics

Resources,

2 Currently:
0002,

Kashiwaba1

Genetic

Resources

2- 1-2, Tsukuba, Ibaraki,

JIRCAS

subtropicalstation,Instituteof Plant Protection,

Konarev3, Kooj i

Kaga1

Laboratory,

Kannondai

Xu1'2 , Alexander

2, National

InstituteofOkinawa

907-

Japan 305-8602

Maesato Kawarabaru 1 09 1 - 1 , Ishigaki,Japan

3All-Russian

Podbelskogo

3, St. Petersburg,

1 89620,

RussiaAbstract
The Leguminosae is the third most specious
ecologicallydiverse of all plant families.

the Papilionoidae

plant family but is perhaps

andVigna belongs to one of the two major groups of genera within

which contains manymajor cultigens.

of 17 species

the most morphologically

The subgenus Ceratotropis

within Vigna consistsof which 8 are cultivated.

The azuki bean [Vigna angularis (Willd.)
its distribution

Ohwi & Ohashi]

extends as far west as Nepal. On morphological

species whose diversity
occurs naturally

center is mainland Southeast

at a more extreme latitude

is a major cultigen

grounds it is closelyin East Asia but

relatedto a group of

Asia. The presumed wild progenitor

of azuki bean

than any other of the VignalPhaseolus

cultigenpresumed wildprogenitors.
Azuki bean is one of the few crops which forms a true crop complex in Japan. Wild and weedy
populations

can be found throughout

the main islands

Honshu. The current center of diversity
prefecturesof Japan except Hokkaido

in Japan based on a nationwide

around the Kansai (Osaka/Kyoto)

region.

This may reflect,

and the far north ofsurvey, appears to be thein part, the adaptationof thecomplex to the environment of this region and Japanese history.
With the azuki bean complex, four population

types can be clearlyrecognised:and genetic diversityof these four groups has been

wild, weedy and complex populations.

The relationships

revealed by molecularand are discussed.

Finallymarker studies

it will be argued that there are reasons to consider

domesticatedin Japan. These reasons include archaeo-ethnobotanical,

the distributionof members of the azuki bean crop complex.

the cultigen

andthat azuki beans could have been
cultural and historic

information

and

160

Introduction
The azuki bean (Vigna angularis
production

is about 80-90,000

of Hokkaido(Fig.

is an importantcrop in Japan and annual

metric tons. 65% of azuki bean production

is in the northern island1). Japan currently

var. angularis)

imports about 20,000

primarily

fromimportance of azuki bean in Japan is best summed up by its association

with

China. Azuki bean is the second most important
The cultural

success and good fortune. Thus to celebrate

metric tons annually,legume in Japan after soybean.

happy occasions,such as a births or weddings,

success in an exam,rice is mixed with azuki bean to produce 'sekihan'
an importantcomponent of traditionalor red rice. Azuki bean isJapanese foods such as steam "an" (azuki paste)azuki paste mixed with agar-agar to make ajelly

like dessert(yokan);ortaiyakibuns;

a waffel filledwithazuki paste.
Azuki beans is one of the few crops in Japan which form a true crop complex with its wild
and weedy relatives.

This crop complex in Japan can furnish germplasm to study morphological,

ecological,

and molecular

genetic

model for developing
relatively

changes during domesticationconservation

strategies,

both in-situ

and it is a crop which can be aand ex-situ. Since azuki bean has asmall genome size compared to Phaseolus vulgaris and Glycine maxit is a logicalto choose for genome studies of the phylogeneticallysynteny the azuki bean maybe helpful

relatedtropicalcrop

or grain legume group. By

for comparative genome studies among legumes and otherdicots.

The systematic

positionof Vigna angularis.

The Leguminosae is the third largest

plant family

There are 650 known genera and about 18,000
the broad outline of legume evolution
to which recently
Papilionoideae
species)

molecular
sub family

species

has resulted

in this family.

from basic analysis

data has been added (Chappill,(440 genera,

12000

species)and Vigna. The subtribe

Phaseolinae

cultivatedMacrotyloma, Phaseolus,

Lablab,Vigna is a large genus consistingworld (Marechal
domesticated.
groundnut,

et al., 1981).

and Asteraceae.

Current understandingofof taxonomic characters
Doyle,

Phaseoleae

1995).

Withinthe(84 genera,

1500

to man, Cajanus, Glycine, Phaseolus

has 23 genera of which the following
Psophocarpus

genera include

and Vigna (Lackey,

1 98 1 ).

of 7 subgenera which are found in the New and Old

Only in the subgenera

The subgenus Vigna cultigens
Vigna subterranea.

1995;

the tribe

has the main grain legume genera of importancespecies:

after the Orchidaceae

These speciesof Africaand Asia have species

are the cowpea, V. unguiculata
are in different

sections

been

and the bambara

with this subgenus. The

FigureFigure1. Azuki bean production

2. Distributionof Vigna angularis(by area) in Japan in 1 997.

var. nipponensis(basedon Tateishi,

1 996).

162

widelydistributed

species

forage but in Ethiopia
is remarkable
subgenus

V vexillata

is in the subgenus Plectotropis

and Sudan its roots are eaten (Duke,for the number of cultigens

8 are cultivated.

it contains.
is alsonomenclature having a total of 1 4 intra-specifictaxa.

cotyledonsthe subgenus Ceratotropis

and petiolate

and V. umbellata
2000;

notableused as a

The subgenus Ceratotropis

Out of the approximately

The subgenus

Within1981).

and is generally

20 species

for the complex

in theintra-specificazuki beans belong to the group having hypogenous

first and second leaves. Its closest

relatives

based on many studies (Kaga et al., 1996;

are V. hirtella,

V nepalensis

Konarev et al, 1999;

Konarev et al.,Tomooka and Egawa this volume).

The distributionof the componentsof the Vigna angulariscomplex in Asia.

The presumed wild ancestor of azuki bean is Vigna angularisform occurs from northern Honshu, Japan to Nepal (Fig.
var. nipponensis
prefecture,

var. nipponensis. This wild2). At southernoccurs in mountain areas. Thus while V angularis

Japan, it does occur in the mountains

Phaseolus/Vigna

cultigens,wild populations

latitudes

V angularis

is not present

in Okinawaof Taiwan. Among wild relativesof V. angularis

naturallyoccur

at the highestof thelatitude(Table1)
V angularisvar. nipponensis

is recorded

from China, India, Korea (North

littleis available

Myanmar and Taiwan. Unfortunately
nipponensis

in China and Tateishi

Korea V. angularis
1 999, personal

var. nipponensis-scommunication).

China the closely(1 985)

only records

Myanmar, Laos and southernand presently

or formerly sympatric populations

may

have occurred. This region is thus particularly

importantto gain an understandingof the V. angularislarge parts of this area have yet to be explored

distributions

complex. Unfortunately

var nipponensis,(Mun Sup Yoon,andhave overlapping

V. angularisand possiblyvar.

specimens from China. In

less commonthan Vigna nakashimae

In Nepal and Thailand

wild species

3 herbarium

on V angularis

V. nepalensis

V. umbellata

relatedinformationand South),V. hirtella,of the evolutionbygermplasm scientists.
The cultigen

Vigna angularisvar. angularis

South Korea and Taiwan. In China
northeastern

provinces

Outside(Lumpkin

is a major crop in China, Japan, North andazuki beans are grown in Hebei,

andand McClary, 1 994)

Japan the authors are not aware of information

form of V. angularis.

Henan, Shandongon the distributionof the weedy

Table 1. Wild relatives

Table

of Vigna/Phaseolus

cultigens

2. Summary data for three characters of field collected
weedv Donulations of the Vigna angularis comnlex.

The components of the Vigna angularis
Twovarieties
cultigenof Vigna angularisand V.angulariscomplex and their distribution

in Japan.

have been recognised.

var. angularisvar. nipponensis the putative

Hymowitz, 1985).

These two varietiesand pod shattering.

Another form of V. angularis,

calleddiffersuch as seed and pod size, that are intermediate

V. angulariswild progenitor

in such traits

the weedy type of azuki bean (Yamaguchi,

Based on field observations

samples of wild andas habit,of the cultigen(Smartttheandseed and pod size and color

which grows naturally

in Japan, and has been

1 992). Weedy azuki has several characteristics,
between the wild type and cultigen

the weedy type is sometimes found in distinctly(Table2).

Figure 3. Distribution
collection.

of Vigna angulariscomplex population

types in Japan based on direct

165
different

habitats

from wild azuki such as beside

uniform set of characteristics
are of various colors.

since it is sometimes determinate

In addition

and intermediate

plants.

and seeds or pods

some populations

consistSuch apparently

dynamic

we have called complex populations.

Between 1 996 and 1998. a series of missions(i.e.

It does not have aor indeterminate

to wild and weedy type populationsof a mixture of both wild and weedy plants
populations

streams or in wet habitats.

individual

was conducted to collectplant seed samples, bulk seed samples, herbarium

complete passportdata) germplasm of the Vigna angularis

The distributionof the wild, weedy and complex populations

shown (Fig. 3).

The production

These distribution

specimens, root nodules

andcomplex across Japan (Appendix

1 ).

collected

during these missions

centers in Japan for azuki bean is shown (Fig.

maps furnish the following

1. The northern limit

comprehensively

is1)

information:

at which wild V. angularisvar. nipponensis

grows is 39°N.

2. There are several lowland regions regions south of 39°N which lack wild and weedy azuki mostprobably

due to urban development

3. The area with the greatest

around the inland sea on Shikoku

abundance of different

populations- includes the prefectures

4. In partsof Honshu, particularly

southern

Shikokuwild

particularly

populationand Honshu.

types - wild, weedy, and complex

around the Osaka, Kyoto and Kobe urban conglomeration.
in Tottori,

Kihan tou (Mie and Wakayama prefectures)azuki bean populations

environment to which this crop relativeandare common.This area appears to have theis best adapted.

5. As with many crops the main production

center and area where wild and weedy relatives

grow

do not co-inside.
Vigna angulariscomplex diversity

studiesa Isozymes
Analysisof 4 isozymes(6-PGDH,populationsof Vigna angularisb Proteaseinhibitors

The intraspecificresults

inhibitor

collected

Due to high activity

collectedof trypsin

no variation

among 35

(Tomooka et al., 1 998).
inhibitorsfrom various locations

was investigatedin wildon Honshu, Japan. The

in the northern region of Honshu island are shown (Fig. 4).

and chymotrypsin

variation differences

analysedin trypsin and chymotrypsinof V. angularis

for five populations

Three types oftrypsin

var nipponensis

variability

and weedy populations

GOT, SDH and LAP) revealed

are visible
inhibitors

spectra were found among these populations.
among components with positionsin the conditions

used further

The trypsin

less than p6.0 (Fig 4A).
variation

could not be166
elucidated.
(only

A gelatin

the results

replica

2 and 3 (Fig.

inhibitors.

Population

1 (Fig

4B)

In generalsuggest that a careful
to reveal

had similar

inhibitor

chymotrypsin

4-1 1
spectra.

trypslnor chymotrypsin

4B) did not have bands p6 and p8 but possesseda band at aboutvariation

were observed for trypsln

survey of Vigna angularis

potentially

Is shown (Fig 4B). Populations

did not have a band at p6 for either

p l O. To check for intra-population

examined but no differences

Is likely

with chymotrypsin

for 4 and 5 shown)

Populations

positiondeveloped

useful

5 individual

seeds from 5 populations

or chymotrypsin

populations

inhibitors.

for variation

wereThese results

in proteaseinhibitors

variation.

Figure 4. Polymorphism
for trypsin (A) and chymotrypsln(B) Inhibitors
in Vigna angulariscomplex accessions from various parts of Honshu, Japan. Iso-electric
focussing of seed proteinsin Servalyt precotes pH 3-10 gel 50 mm long. Inhibitors
were detected In two consecutively
obtained gelatin replicas, a-e: populations
1 -5 from northern Honshu (a, b, c and e wild; d- weedy).
The proteins were extracted from 4-6 seeds from each population.

c DNA marker polymorphism and population
Twoseparate methodologies

have been used to study the population

component members of the Vigna angularis
2000a;

Xu et al., 2000b).

(Table(Tablecan be summarised as follows:

4 and 5). These results
has lower populationthat domestication

resultedb. Greatest intra-population
AFLP analysis

in other population(discussed

diversity

in a genetic

types

diversity

banding

observedmeasured based on both analyses

issuggestingneck.

was found in complex populations.

showed polymorphism

suggesting(Xu et al.,than either weedy or wild populations

bottlepolymorphismof complex populationsof3).
genetic

a. The cultigen

structurecomplex, RADP and AFLP analyses

The intra and inter population
presented

genetic

The materials used, primer number and polymorphic

using both methods is presented

these populations

Both RAPD andin complex populations

may be of evolutionary

not foundsignificancebelow).

c. Of the totalgroup genetic

variation(wild,

between populations

and 35% withinrecommendation,

based on other crop genetic

populations

weedy, complex groups)

populations(Marshalld. Between population

type diversity

i.e. wild-cultigen,wild-weedy

by RAPD analysis.

resources,

rather than a greater number of individuals

germplasm collectingthat sampling

within a population

about 65% was foundThis

confirms thea greater

number ofis the best strategy

forand Brown, 1975).
is very similar

when pairs of population

types are compared

and weedy-cultigen.

The results of these analyses

Tablegenetic parameters

shed particularlight

on weedy and complex populations.

3. Materials and methods used to estimate genetic diversity
population types (from Xu et al., 2000a and b).

for Vigna angularisTable 4. Intra population variation based on RAPD and AFLP polymorphism
in Vigna angularis complex population groups.

Table 5 Inter-group variation based on RAPD and AFLP polymorphism
Vigna angularis complex bean complex.

detecteddetectedin theWeedpopulations
Three hypotheses
Harlan, 1 975).
(a) Escapes

have been proposed

These hypotheses
from cultivation;

for the evolutionof weedy races (deWet,and

are that weedy races are:(b) Hybrids

between the wild type and the cultigen;(c) Directly

evolved from the wild type.
Of these three hypothesesof hybridization

our results suggest that weedy populations

and sometimes be directly

data supports weedy populations
cluster with cultivatedevolved from the wild type. Cluster analysis
from both processes

azuki and three populations

to support the weedy type resulting
a. Seed coat color variationb. Populations

resultingof AFLP(Fig. 5 ). Two weedy populations

cluster with two groups of wild azuki. Evidence

from hybridization

in some weedy populations

which appear to be hybrid

may sometimes be the result

includes:resembles

swarmpopulations

artificial

hybridpopulations;have been observed by the authors

169

Figure 5. Populations neighbor-joining
tree based on Jaccard' s dissimilarity
weedy •E ; wild O ) (modified
from Xu et al., 2000a).
w e ed y

fM ln

"k

coefficient (cultivated;蝣o**; cW o em e pd ley x‑l l *:

I一 ・
I一 ・
・
・

W ild

・
・
・
・
・
.

W ild

*
* C o m p le x*

Figure 6. UPGM dendrogram representing the associations among individual plants from threedifferent types of natural populations (wild •E; weedy O; complex *) belonging to the azuki bean
species complex from Tottori prefecture (modified
from Xu et al., 2000b).

170
and others (Vaughan and Kaga, 2000;

Yamaguchi,1992).

Evidence to support weedy azuki having evolved directlyfrom wild azuki includes:a. RAPD bands in the wild and weedy types but not in the cultigen

and no weedy specific

RAPD

bands;
b. Higher genetic

diversityc. Field observations

in wild populations

suggesting,than nearby wild populations.
flooding

than weedy populations;in some locations

that weedy azuki is adapted to wetter habitats

For example, stream sides are particularly

disturbed

due to seasonal

may be specifically

adaptedtoof azuki are the consequence of evolution

at aand weedy azuki, which can be found in such habitats,more highly

disturbed

ecologicalniches than the wild type.

Wecan conclude that weedy populations
local level. However, more informationis needed concerning

dominant markers, such as micro-satellite

markers, will be helpful

Such studies

may help clarify

how weedy populations

gene flow. The application
to identify

hybridof co-

individuals.

evolved at a local level.

Complex populations
Field data and results of RAPD and AFLP analyses have enabled characteristics
populations

to be measured. Complex populationsin the case of Phaseolus
1 997).

vulgaris

For the Vigna angularisof complex

have been reported

for other crop complexes,

they have been called inter-breeding

complexes (Beebe et al.,complex these populations

tend to be large and consist of different

plant types - wild, weedy and intermediate

plants. We have visitedin Tottori

years and on each occasion recorded changes. During

prefecture

over three consecutive

the first visit no area within the population(1973).

appeared similar

However, during the second and third visits

one large complex populations

to a hybrid

to the site plants

swarm as describedin different

to be like a hybrid

swarm having a high level of plant to plant variation.

AFLP methodology

to analyse individual

growing within

6 km of each other in Tottori

adjacent

to farmland where small fields

resultingfrom this analysis

which are similar

plants from three populations,prefecture.

of cultivatedHieserareas appeared

Consequently

we usedwild, weedy and complex,

All three populations

analysedwereazuki beans are grown. The dendrogramis shown (Fig. 6). This shows that the complex population

to plants from both the wild and weedy populations.

like plants

in the complex population

have considerably

populationand these plants

populations

leads us to recommend that complex populations

both in-situ

and ex-situ conservation.

It also shows that the wild

more genetic

form two groups. The high levelhas plantsof genetic

diversity

than the wild

diversity

in complex

should be a focus of attention

for

171

Where was V.angularis
Studying
possibility
againstdomesticated?

the genetic diversitythat Vigna angularisof the Vigna angularis

was domesticatedcomplex has lead us to consider thein Japan. Arguements which tend to be for andazuki beans being a Japanese domesticate

are listed

below:

Points which may support azuki bean as a Japanese domesticate.
1. Archaeologicalfinds of legume seeds from Torihama in Fukui prefecture,

Japan (Maeda, 1987)

are most likely

Vigna angularis.

These remains of legume seeds dated at 5000 B. P. are about

3 000 years older than the earliest

report the authors know of from China (Mawangdui of Changsha

fromthe Han Dynasty +-21 00 years ago).
2. Japan has all the evolutionary
domestication.
currently

In addition,

components which would be expected

Vigna angularis

exists in southern to central

appears to be best adapted
called

4. In Japanese culture the azuki bean has particular
sekihanto the climatethatazuki bean island (Shoodoshima).

association

with success and celebration(red rice - rice mixed with azuki beans).

5. While the Jomon people of Japan livingfrom about 12,000 to 2,000 years ago were essentiallyhunters and gatherers recently evidence that they cultivatedcultigensof crop

Honshu.

3. Close to the current center of Japan is an island
particularly

in a regiona few crops has emerged. Amongthese

are rice and chestnuts. Thus the date of early agriculture

in Japan is being pushed back.

6. V. angularis var. nipponensis is a legume which would lend itself
the pods are relatively

to easy hand harvestinglarge compared to other wild legumes of disturbed

Japan. For example, one pod of wild Vigna angularis

habitatssinceindigenous

towill yield about 3 times as much weight ofseeds as wild soybeans.
Points which may not support azuki bean as a Japanese domesticate.
1. Most cultigens
directlywere introduced

or via the Korean peninsula

2. The center of distribution
hirtella,(Kihara,studiesof close relativesof the cultigen

such as V. umbellata

1 996) and is the likely

area V angularis

have yet to clearly

identify

we found very little

area in Asia. Yee et al. (1999)

no conclusions

regardingevidence exists to suggest multiple

the center of diversity
domestication.

and V.

evolved.

a center of diversity
variationin cultivatedfound a greater diversity

germplasm based on AFLP and RAPD analyses. However, due to lack of pedigreeaccessions

China eitherof V. angularis,

Based on RAPD banding diversity

beans from a wide geographic

most notably

1 969).

occurs in mainland Asia (Tateishi,

3. Genetic diversity
angularis.

into Japan from other countries

of V.
azuki

in Chinese

informationonof azuki bean could be drawn. No

172

Final

comments
While many scientificJapanese

agriculture

studieshave been conducted on the relatives

there have been very few on the Vigna angularis
has some characteristics

to Glycine

similarHowever, G. soja can be found in areas where V. angularis

cannot such as along the Honshu sea board of the Japan inland
nipponensis
reflect

degree of disturbance

Whereas good estimates
outcrossing

estimates

Very little

which each speciesof outcrossingfor V angularis

is known about the distribution

since the V. angularis

Studiesof V. angularis

to try and identify

usefulplants to be identified

V. angularis

var.

These differencesmay

can tolerate.
for G soja (Fujita

carefulsurvey for V. angulariscomplex inin mountains

ofcomplex occurs as far west as Nepal.

at the genome level are few. Sequence data is only now being
micro-satellites.

The objectives

and provide additionalof this research

Vigna species,

studied,many be useful in understanding

gained by studying

V. angularis

are to enablegenome markers to improve the genetic

mapof azuki bean. Given the number of cultivatedinformation

et al., 1 997)

of the taxa in the Vigna angularis

Southeast

Asia may be revealing,var. nipponensis

have not been reported.

In addition,

heterozygous

sea. Similarly,

have been obtained

China and the Korean peninsula.

obtainedsoja, such as growing in

is more commonin some areas than G. soja, such as Tottori.

the relative

tocomplex. Vigna angularisvar. nipponensis
habitats.

which are similarof crops importantmany of which have been little
others within

the genus (Tomooka et al., 2000).

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and collecting

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and Introductionin Tottoriin Japan with a focus on
and Okayama prefectures.

of Plant Genetic

resources.

Vol. 1 6. (inThe Vigna angulariscomplex:press)Xu, R.-Q, N. Tomooka, D. A. Vaughan and K. Doi. 2000a.
genetic

variationand relationships

in-situconservation

revealedand domestication.

by RAPD analysis

and their implications

for

Genet. Res. Crop Evol. (in press)

Xu, R.-Q., N. Tomooka, and D. A. Vaughan. 2000b.

AFLP markers for characterizing

the azuki

bean complex. Crop Sci. 40 (3) (in press)
Yamaguchi,

H. 1992. Wild

and weed azuki beans in Japan. Econ. Bot. 46:384-394

Yee, E., K. K. Kidwell,

G. R. Sills,

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275.

(Azuki)among selected

Vigna

175

AppendixCollecting

trip reports for Vigna angularis

documents are all published
Tomooka, N. 1997.
angularis

complex and related species in Japan 1 996- 1 999. These

by NIAR and are available

Collectionof wild

var. nipponensis)

and Saga prefectures

soybeanin Ibaraki,

via the library{Glycinesoja)

of NIAR on request:

and wild azuki bean (Vigna

Fukushima, Niigata,Tochigi,

in Japan. Annual Report on Exploration

Kumamoto, Fukuoka

and Introductionof Plant

Genetic Resources 1 3 :41 -57
Tomooka, N., D. A. Vaughan, R. Q. Xu and K. Doi. 1998.

Wild relatives

in Japan with a focus on Vigna spp. Introduction.
Introductionof Plant Genetic

Tomooka, N., E. Potokina

Resources.

1 4:45-6

and M. Akiba. 1998.

Collection

Annual Report on Explorationin Tohoku

Exploration

regionof the wild relatives
genepoolof Plant

Genetic

of crops, 1997.

and soybean {Glycineof Honshu, Japan. 6-11th

and Introduction

and1

1. The azuki bean {Vigna angularis var. angularis)
genepoolof crops conservation

October.

Resources

max)

Annual Report(in Japaneseonwith English

summary). Vol. 14:63-70
Tomooka, N., D. A. Vaughan, A. Konarev and S. Tsukamoto. 1998.
relativesof crops, 1 997. 2. The azuki bean {Vigna angularis

soybean {Glycine

Japanese with English

and Introduction

genepool andin Kinki

October.

Resources 1 4:7 1 -83(insummary)

3. The azuki bean {Vigna angularis

Englishvar. angularis)of Plant Genetic

Vaughan, D.A., N. Kobayashi and R. Q. Xu. 1998. Collection

Explorationof the wildmax) genepool in the central region of Honshu, Japan. 1 5-24th

Annual Report on Exploration

genepoolCollection

regionvar. angularis)of Honshu,

and Introductionof the wild relatives
genepoolof crops, 1 997.

and soybean {Glycine

max)Japan. 4-1 1th November. Annual Reportof Plant Genetic

Resources

14:85-97(inJapaneseonwithsummary)

Tomooka, N., H. Kuwahara and S. Tsukamoto. 1998. Collection
1 997. 4. The azuki bean {Vigna angularismax) genepool in Tochigi,

IbarakiAnnual Report on Exploration
Japanese with English

var. angularis)

and Chiba prefectures,and Introductionof the wild relatives
genepoolof crops,

and soybean {Glycine

Japan. 6th and 1 1 -1 2th November.

of Plant Genetic Resources

1 4:99- 1 05(insummary)

Vaughan, D. A., N. Tomooka, K. Doi, R.Q. Xu and K. Kashiwaba. 1999. Wild relatives
conservationin Japan with a focus on Vigna spp. 1. Introduction.

Exploration

and Introductionof Plant Genetic

Resources

1 5 :53-6 1of crops

Annual Report on

176
Tomooka, N.? D. A. Vaughan, N. Nakagahra,

K. Hammer, R. Q. Xu and Y. Tsubokura.

Wild relativesof crops conservation in Japan with a focus on Vigna species.

and collecting

missionOctoberto Tochigi,

Ibaraki,Shiga

1 998 Annual Report on Exploration

and Tottori

2. Monitoring

prefectures.4th

and Introduction

1999.

and 6th -8thof Plant Genetic Resources.

15:63-66
Tomooka, N., K. Doi, Y. Tsubokura. 1999.
azuki bean {Vigna angularis)

genepoolregion of Honshu and northwestern
on Exploration

Collection

and Introductionof the wild relatives

and soyean (Glycineof crops, 1998. 3. Themax) genepool

in the San-inpart of Kyushu, Japan. 9^- 1 5th October

Annual Reportof Plant Genetic

Vaughan, D.A., T. Yokoyama and T. Komatsuzaki.

Resources

1 5 :69-77

1 999. Wild relatives

in Japan with a focus on Vigna species.

4. Collecting

Octoberand Introduction

1 998. Annual report on explorationof crops conservation

missionon Kyushu. 16th - 21stof Plant Genetic Resources.

15:79-84
Vaughan, D. A., T. Chibana, M. Akiba and S. Hattori.

1999. Wild relatives

in Japan with a focus on Vigna species.4.

missionOkayama, Aichi
Explorationand Shizuoka

prefectures.

and Introduction

Vaughan, D. A. and K. Kashiwaba.

1999.

Wild relatives

Tomooka, N., Y. Egawa, Y. Tateishi,
crops conservation
Okinawa, Ishigaki
Introductionof Plant Genetic

T. Yamashiro

of Plant Genetic

Japan.2b.

2a. Monitoring

Information

Kyoto University
Introduction

23^-30*Resources.

prefecture,
Coordinators

in East Asia.
Japan.

andpreparation)of crops conservationand collecting

missionin Japan with a focus on

in Tottori and Okayama prefectures,of wild legumes with emphasis

on Vigna in the

1 999. Annual Report on Exploration

andResources. Vol. 1 6 (in preparation)

1. Collecting

21st - 29th October

Meetingof

22nd -26th Feb. 1 999. Annual Report on Exploration
Resources. Vol. 1 6.(in1. Collecting

Kaga, A., M. S. Yoon, N. Tomooka and D. A. Vaughan 2000.Collaborative
Vigna species

1 5 :9 1 -98

and D. A. Vaughan 2000. Wild relatives

herbarium. 22-24* September
Genetic

October 1998. Annualonon the distributionof Plantin Japan with a

missionVaughan, D. A. and A. Kaga. 2000. Wild relatives
Vigna species.

1 5 :85-90

in Japan with a focus on Vigna species.
and Iriomote.

Hiroshima,

1 998. Annual Report onof crops conservation

mission on Shikoku.

and Introduction

in Yamaguchi,

22nd-27th Octoberof Plant Genetic Resources.

focus on Vigna species. 5. Collecting
Report on Exploration

Collectingof crops conservation

mission1999.

and IPGRI (in preparation).

on the islands

Reportresearchon theof southern

Okinawa

to East Asia Genetic

Resources

177

Questions
Questionsand answers: Session

2

to Dr. Debouch.

Q. Could you tell me about cyanide

compounds containedin Lima beans and their ecologicalmeaning (Fuj ii)
A.The presence of a glycoside
been reported

producingin the literature

contents of this glycosidewild forms (Bandoin

HCN in contact with chloridiic

acid of the stomach has

since the end of the last century. Thorough analysis(calledlinamorin)of thehas been done for Lima bean landraces

andand co-workers, 1995).

Wild forms all contain high levelsof the glycoside.

color of the testa, still have some significant

levelsSome landraces,

indistinctlyfrom theof the glycoside.

Most landracesaresafe and have no glycoside.
Usually

with pre-soaking

and cooking throwing

away wash waters, Lima bean food issafe. From my ownfield work in Central America in spite of the presence of linamarium,
wild forms of Lima bean host bruchids{Zabrotessp.) and pod weevil (Apios sp.).

C. There is a need to look more closely at the domestication
antinutritional

factors. What made humans domesticate

or plant products?
domestication
Questionsissue, especiallyand use for food such materials

There is a need to look for anthropologicaland not just genetic

similarietiesof species that haveand variation.

evidence

in relation

to(Ramantha Rao)to Dr. Potokina.

Q. I understandthat Viciafaba

Vicia faba

has a large genome. Does Vicia Plinianagroup you presented

chromosome morphology?
A. V.faba reallyin your paper have a different

or other forms of the
DNA content or(Vaughan)can be distinguishedby its highDNA nuclear

content. As far as I know, DNA nuclear content is a rather stable characteristic

and I havenoreferences reportingfrom the other Vicia species

intraspecificvariationwith stable chromosome number. V.Pliniana
all,

since

seed materialof thisspeciesin DNA nuclear content within Vicia species
has not been investigated

was lost and seems still

karyologicallyatnot to be available

anywhere.
Q. How does Viciafaba

subsp. paucijuga

fit into the pictureof the domesticationof the fababean? (Maxted)
A. Viciafaba

subsp. paucijuga

Cubero (1984)

reported

Murat. was considered
thatpaucijuga

as the most primitive

is so primitivein phenotype,form of faba bean.
that it is possibletothink that it is the wild ancestor of faba bean. As a logicalnorthern India, where paucijuga was collected,

conclusion,

Afghanistan

andhas been proposed as a possible

center oforigin of faba bean. This suggestionis also supported by the great morphologicaldiversityof K faba forms in the Indo-Afghan

region.

However, we suppose that
Mediterranean

faba bean originatedas a domesticate

in the East

region, and the primitive,

early domesticatedmaterialtransported

eastwards early in the historyof domestication,and may have resembled

paucijuga.

We agree with the hypothesisof Cubero (1984),

selection,

which would be favoured by topography

the genetic

characteristicsof the primitive

rather the secondary

domesticatedrepresents

origin'.

There are some arguments to support this:faba minor forms of Afghanistan.

material.

center of diversity,

paucijuga

thatgenetic

drift

Thus, Indo-Afghanrather than the 'center offorms showed no differences

They clusteredandof the Indo-Afghan region, would fix

region1 ) on the molecular level the primitive

could have been

togetherand slightly

from otherseparatedfrom theother faba gene pool.
2) paucijuga
indicating

forms combine primitive
obvious human selection,3) at present
vegetative

characters

originby means of separationof origin
Questionsdescribed

contain recessive

by Vavilov

increases in plant breeding

centers of botanicaland characters
genes for mostand dominant genes for most of the pod and seed characteristics.

to the situationof species

characteristics

for example, self compatibility.

it is known that the paucijuga-forms

This is similar
diversity

morphologicaldiversityof recessivethat in cross fertilized

plantsplots far from the geographicalforms, by artificialof a certain group of plants

inbreeding.

thecenters of

As a result, theare not always primary centersof forms.

to Dr. Abe.

Q. Environmental

conditions

genetic structuresuch as climate, photoperiodof wild soybean populations.

sea side and Pacific Ocean side have differentof snowfall.

seem to be important for determining

For example in northern Japan the Japan
topographic

characteristics,

like the amount

Could you comment on the effect of climate

on population

structure?

A. I think that, in some cases, environmental
wild soybean populations.

conditions

For example, populations

along the Amur River (Far East Russia),
soybeans, are insensitive

would influence

the genetic

structure

growing at high latitude

the northern limit

to long days and produce viableofsuch asfor the distributionof wild

seeds within the limited

frost

179
free season there. A high selection
affected

for this type of photoperiod

the genetic structure of the population

to the founder effect. Thereafter
population

structureconsequently

outcrossing

for markers linked

that had first colonizedthese plants

since migration
with plantsfrom low latitudes.

adaptivenature of this species.

behaviors

effect on
andThis seems to be true not only
independentlyof those

often caused by the predominantly

However, wehave not analyzed the differences

between populations

locatedon the Japan sea and Pacific

Honshu, Japan. Most of the wild soybeans have hard seededness,role in keeping

in a manner similar

would be minimal

but also those inheritedgenes, because of the high gamete-phase disequilibrium
self pollinating

would havewould have had a continuingfrom lower latitudes

with genes selected

insensitivity

seeds viable under adverse conditions

side ofthat has an importantsuch as freezing

conditions.

If there

are any differencesin the extent of hard seededness

between populationsin these regions,different

selection

schemes should be considered

as a factor influencing

the population

genetic

structure.

Further studies

Questionsinare needed on this topic.

to Dr. Vaughan

Q. Would it be possible

to have domesticationof soybean and azuki bean in the same place?

(Debouck)
A.I think this is possible

but may be unlikely.

the process of domestication
yield.

From the wild relatives

was different.

Pickingof these two crops I suspect

wild azuki bean can readily give a highHowever, for wild soybean I suspect the seeds for human consumption may initiallyhave been a by product of animal feed.

Session 3
Research, evaluation

and use

183

Evaluation

anduse of wild

Phaseolus

speciesinAgricultural

Experiment

Station,721-8514,Japan(e-mail:

breeding
M. Ishimoto
Laboratoryof Plant Biotechnology,

6-12-1

Nishifukatsu,

ishimoto@cgk.

Chugoku National

Fukuyama,

Hiroshimaafrrc. go.jp)

Abstract
Commonbean {Phaseolus vulgaris L.) is a staple food in many countries but, like othercrops, insect pests cause considerable

losses.

Extensive post-harvest

damage is caused by thelarvae of two bruchid pests, the bean weevil {Acanthoscelides
obetectus) and the Mexican bean
weevil {Zabrotes subfasciatus). High levels of resistance against the bruchids have been found inwild commonbeans collectedin Mexico. These wild accessions contain a novel protein, arcelin

which is divided

into at least seven electrophoretic

amylase inhibitor( Oi AI-2)

active against

variants. In addition,a second type of (X-OC-amylase of the Mexican bean weevil has been

observed in some accessions containing arcelin. Arcelin and <X-amylase inhibitorand structurally

closely

relatedto lectin(phytohemagglutinin).

develops different

defense proteins by duplication

are genetically

The commonbean presumably

and divergence of an ancestral gene.

Introduction
The bean weevil{Acanthoscelides{Zabrotes

subfasciatus

vulgarisL.). The female beetlelays^eggs

on the surface of pods or seeds in the field orSoon after hatching,larva eat the seed coat or pod, dig into thefacility.

cotyledons,

and develop

into adults.

and they

cause

storagegerminability,

becauselimitationof otherpotential

to eliminate

Internationalthede Agricultura

The life cycle

extensive

adult insects

control

Mexican bean weevil

Say) and the Mexican bean weevil

Boheman) are major insect pests of the commonbean {Phaseolus

the storage

seed

obtectus

lossesof thesebruchids

anddamagecan mate and oviposit

without

methods,use of bruchid

resistantlossesduringA research

Tropicalin a few wild

storage.

(CIAT)

found highcontinues

to

seed

common bean accessions

qualityandany feed. Due to thecultivars

levelsduringhas considerable

groupin the Centroof resistancecollected

to thein Mexico184(Schoonhovene/ a/., 1983).

Arcelin
The resistantarcelin.

Arcelinaccessions

accessions

are characterized

was named after Arcelia,

were found. Arcelinbeans and arcelincomprisesby the presence of a novel seed protein,
the town in Mexico where the resistantis the major seed proteinin the resistantmore than 30% of the total protein.

containa major seed storage

protein,phaseolin

protein.

Some wild commonbeans including

Seven allelicsulfate-polyacrylamideshow different

degrees of resistancethe arcelinvariant (Cardona

et al, 1990).

and arcelin-5

are highly
arcelin-2

shows a intermediate

presenceof arcelin,

the accessions

Bruchidresistancein the accessions

progeny from a cross together

The accessions
resistancecontaining

arcelin-1,

is highly

resistantthe unavailabilityof arcelin

may cause the malnutrition

Table 1. Effectof five arcelin

to proteolysis

A rc elin v ari an t

E m erge nc e
(% )
9 .3
2 0 .5
7 9 .58 .4
4 .0
9 4 .01). The accessions

or arcelin-5

is inherited

Feeding

theon the Mexican beanof arcelindigestiveof thetests show thathas yet to be

enzymes, therefore(Minney et al, 1 990).

on the development

arcelin-4

to the bruchid.

by larvalvariantsonare susceptible

But the mechanism of action

1
2
3
4
5
‑

The wildof over 10% has toxic effects

clarified.

or v ariety
G 12 8 82
G 12 8 8 6
G 12 9 22
G 12 9 5 2
G O2771'C a lim a 'et al,

1998).

1).

In spiteof arcelin.

et al,A cc essio n(Fig.

to the bruchid.

arcelin-1

weevil (Osborn
Arcelin(SDS-PAGE)

containing

arcelin-3

with the presence1988).

on a gelto the Mexican bean weevil depending

containing

in the high concentration

have a few

variation

to the Mexican bean weevil (Table

containing

arcelin-1

show genetic

so far (Acosta-Gallegos

accessions

resistantaccessions

gel electrophoresis

variants have been reported

commonbeans

36 to 46% of total

resistantintense bands around 30-40 kDa, and these polypeptidesof sodium dodecyl

Mostwhich comprises

the bruchid

wild commonof Z subfasciatus

D ays toad u lt em erg en ce
5 1 .0
4 8.8
42 .0
70 .0
64 .6
32 .9
(C ard o n a et q l. 19 90 )

Fig. 1. Seed
concanavalin
arcelin-1
(2),
1995)

glycoproteinsdetectedby Western blottingwith peroxidase
coupled to
A (Con A). Lanes: 1, Taisho-kintoki;
2-6 wild accessions
containing
arcelin-2(3), arcelin-3(4), arcelin-4(5) or arcelin-5 (6). (Suzuki
et al.

Fig. 2. Seed glycoproteinsdetected by Western blotting with peroxidase coupled to Con
A. Electrophoretic
banding patterns around 14-20 kDa correspond to each OCAI variants.
Lanes: 1-5 and 10, cultivars;
6-93 wild accessions. (Ishimoto et al 1995)

186

Some wild common bean accessions
bean weevil.

The resistanceindependentof the presenceof the resistanceis stillcontaining

is inheritedof arcelin

arcelinare also resistantto theas two recessive

complementary

genes,(Kornegay

and Cardona,

unknown. The resistancepresence of a high molecular1991).

may be at least

weight polysaccharides(Gatehouse

The mechanism
partlydue to the

et al., 1 989).

a -Amylase Inhibitor
A novel type of a-amylase
amylaseactivity

accessions
inhibitor

inhibitor,of the Mexican

containing

arcelin-4.

bean weevil

CDNA encoding

aAI-1

angularis

Ohwi and Ohashi)

effects

comprehensively

to the pea (Pisumthe(Callosobruchus

maculatus F.) which can not develop
the Mexican bean weevil toleratebeans and transgenic

comprehensively

in wild

and cultivatedupon the specificinhibitionbeans

are divided

intoinhibitor

(Fig.

2).

seed

four types,

a -amylasea AI-1a AI-1,

those of plants,

bacteria,

and two bruchid

inhibitor

and the cowpea weevil

containing

arcelin-3

pests

or arcelin-4,

of Z subjasciatus(C

is insufficient

containing

inhibitory

activity.

has

been

analyzed

et al., 1995).

Basedsources,commonarcelin-1

Therefore,

andis dividedof theinto

to preventor arcelin-5

four
but notof the common bean, the Mexican beana AI-2, was only observed

and shows inhibitory(Minney et al, 1990;

have shown that a AI-2 inhibits

accessions

inhibitora -amylases of mammals and some insects,

weevil and the bean weevil. The second inhibitor,

in seeds

such asa AI-2, a AI-3, and the absence

is a well-known

It inactivates

content

property.

to some bean weevils,of a -amylase enzymes from different

variants.

tests

bean (Vigna

its insecticidal

commonbeans (Ishimotoelectrophoretic

feedingL.)of thelegumes.
of thea-amylase

Transfera high seed content (0.5 to 1 %) of a AI-1 in common

variation

larvalresistanceon commonbean seeds. But the bean weevil andGenetic

accessions

1 997).

clearly

resistantchinensis

a-of common bean a -amylase(Chrispeels,

has demonstrated

legumes are completely

bean weevil

the larval

sativum L.) and the azuki

Seeds of these transgenic
azuki

a AI-2, inactivating

was found in the bruchid

The insecticidal( a AI- 1) have been studied(Willd.)termedactivityin the wild

specific

Suzuki e/ al, 1993).

to theAlthoughthe growth of the Mexican bean weevil, itsbruchid

infestation.

are classified

The bruchidin nulla AI-2 is not a main factor for the bruchid

type

resistantlackinganyresistance.

The

187
third

a -amylase inhibitor,a AI-3, was found in some wild and cultivatedcommonbeans, and showed a combined spectrum of a AI-1 and a AI-2. These commonbeans
contained

simultaneously

controlled

by tightlythat at least

both

linkedanalogous

proteins

genes (Suzuki

two a-amylase inhibitorof a AI-1 and a Al-2 which are

and Ishimoto,variants,

1999).

These results

indicatea AI-1 and a AI-2 are present

in thecommonbean.
The wild accessions

collected

a AI types and electrophoretic
centerof genetic

information

diversity

variants were observed
forof the original

in Mexico show high genetic

on a AI. All

in Mexico. Like arcelin,a AI seems to be Mexico. Although

distributionof the storage

bean weevil, the commonbean may have developed
bruchids

diversity

insectthewe have nopests such as the Mexican

the defensive

mechanisms againstin this region as a result of natural selection.

Evolutionary

relationshipArcelinvacuolesbetween areclin

and a AI are seed glycoproteinsduring seed development.

phytohemagglutinin
including(PHA)

PHA, arcelin,biochemical
proteins

which accumulate

Their primary structures(Chrispeels

and Raikhel,1991).

a AI, and a -amylase inhibitor-likeand defense properties
and the single

homologous

and a -amylase inhibitor

locus(Tablein proteinshow high homology

withThe proteinsof this family,

proteinhave different(AIL),

2). The high degree of homology among thewhere theirencoding

genes are locatedgenes may have evolved by duplicationand divergenceplant

defensetheseof a common

gene resulting

relationshipof amino acid sequences deduced from sequenced genes is shown (Fig. 3).
of two homologous polypeptides,

properties.

indicate

ancestral

PHA consists

in different

storagea AI is a single

gene product,

evolutionary

PHA-E and PHA-L, in the sameseed. PHA is a lectin that binds glycan on the glycoproteins
mammals and birds.

Theand shows toxic effects

followedonby posttranscriptional

processing

to make up an active tetramer complex composed of 2 subunits,a and (3(Morenoand Chrispeels,of signal

polypeptide,the proprotein1989).

After

glycosylationand theis maturated

in 2 steps;

endoproteolytic

removalcleavageat thecarboxyl

side ofAsn residue and trimming a few amino acids from the carboxyl

resultingin heterogeneity

shortest

amino acid sequence which has three gaps compared to PHA. The third gap isof the two subunits.

Of this protein

family,terminus,

a AI has theTable 2. Function

and structureof a family of defense proteinsin the commonbean

Fig 3. Evolutionary

relationships

of a family of defense proteinsin the commonbean.

189

also found in all the arcelins.

Arcelinis composed

and arcelin-5)

protein(arcelin-3

or tetramer

component of total

seed protein

mechanism of the toxicity
putative

precursor

otheragainstpolypeptide.

bruchids

is themajor

by bruchidlarvae,but theremains obscure. AIL resembles

AIL is also insecticidal

its mechanism of action is unknown (Ishimoto

in aAI,
againstof the PHA, arcelin,

speciesto the common bean as well as in the wildand Delgado-Salinas,

1997).

to our understandingof their precise

and AIL family

The study of this proteinfunctionabut it is dimer

the cowpea weevil,

et ah, 1 999).

The proteins
relatedarcelin-2

Arcleingap observedaAI,(arcelin-1,

arcelin-4).

to digestionof a AI and has the firstcomposed of a single
althoughandand is resistantof arcelinof dimer protein

have been observedincommon bean (Pueyo

family in different

species

may leadand their use in crop protection.

References
Acosta-Gallegos,(1998).

J. A., Quintero,
A new variant

Q., Vargas, I, Toro, O., Tohme, I, and Cardona,of arcelinin wild

common bean, Phaseolus

from southern Mexico. Genetic Resources
Cardona,C, Kornegay,
Comparative
resistant

Chrispeels,

Posso,

Chrispeels,

Transfer

Gatehouse,

Ishimoto,of bruchidgrains legumes by geneticeds.),

control:

pp. 139-156.

M. J., and Raikhel,defense.

variants

in the development

resistanceengineeringL.

H. (1990).

of dry bean lines
197-206.

from the commonbean to otherwith the a -amylase inhibitor

the role of transgenic

plants"(N. Carozzi

gene.
and M.

Taylor & Francis Ltd., London.
N. V. (1991).

Lectins,

lectingenes, and their role in plant

Plant Cell 3: 1-9.

A. M. R., Shackley,

S. I, Fenton, K. A., and Bryden, J. (1989).

of seed lectin

toleranceby a major insect

Acanthoscelides

obtectus.

J. Sci. Food Agric. 47: 269-280.

M., Suzuki, K., Iwanaga, M., Kikuchi,
of Seedvulgaris

45 : 23 5-242.

F., and Ramirez,

to the Mexican bean weevil. Entomol. Exp. Appl. 56:

In "Advances in insect
Koziel,C. E., Morales,

value of four arcelin

M. J. (1997).
starchyJ,

and Crop Evolution

C.

a-Amylase

Inhibitors

storagepestof PhaseolusF., and Kitamura,

K. (1995).

in the Common Bean. Theor. Appl.

Mechanism
vulgaris,
Variation
Genet. 90:

425-429.
Ishimoto,

M., Yamada, T., and Kaga, A. (1999).
inhibitor-likeproteinresemblinga putative

Insecticidal

activityof an a-amylase

precursor

of a -amylase inhibitor

in

190
the common bean, Phaseolus
Kornegay,

Minney,I L., and Cardona,vulgaris

C. (1991).

h. Biochim.
Inheritanceobtectus

in a wild common bean accession

Euphytica

52: 103-ll

Acta 1432:

of resistancecrossed104-1 12.

to Acanthoscelides

to commercial

bean cultivars.

1.

B. H. P., Gatehouse,

A. M. R., Dobie,

P., Dendy,I,of seedresistanceto Zabrotes(common bean);a mechanism

Gatehouse,

J. A. (1990).

Biochemicalbasis

subfasciatus(bean

weevil)in Phaseolus

vulgarisfor arcelin

toxicity.

J. Insect

Physiol.

Moreno, J., and Chrispeels,

Biophys.

M. J. (1989).

Cardona,C,and

36: 757-767.
A lectingene encodes the a -amylase inhibitorof the common bean. Proc. Natl. Acad. Sci. USA 86: 7885-7889.
Osborn,T. C, Alexander,

D. C, Sun, S. S. M., Cardona,C, and Bliss,

F. A. (1988).

Insecticidal

and lectinseed protein.

Scienceactivityhomologyof arcelin

240:207-210.
Pueyo, J. J., and Delgado-Salinas,
members of the subtribe

A. (1997).
Phaseolinae

Presenceof a-amylase

inhibitor

in some(Phaseoleae:

Fabaceae).

AmericanJournalJ. (1983).

Resistanceto the bean weevilof Botany 84: 79-84.
Schoonhoven,

A. V., Cardona,C, and Valor,

and the Mexican bean weevil
bean accessions.
Suzuki,K., and Ishimoto,aAI-3,
280.

(Coleoptera:

J. Econ. Entomol.
M. (1999).

Bruchidae)

in noncultivatedcommon76: 1255-1259.

Characterizationin the common bean {Phaseolus

vulgarisof the thirdL.). Breedinga -amylase inhibitor,
Science49: 275-

Vicia(Mucuna191

Allelopathyin

Mucunaand related

leguminous

species

Yoshiharu Fujii
National

Instituteyfujii(@,niaes.

of Agro-Environmental

Sciences,

Tsukuba, Ibaraki,305-8604,

Japan

afflrc.go jp

Abstract
Screening of allelopathic

plants by the bioassay called the "Plant Box Method" and "Sandwich Method"

showed that hairy vetch (Vicia villosa) and mucuna

pruriens) are the most promising allelopathic

ground cover plants. Unusual amino acids and chemicals with the catecol group in these leguminous plants
might play important roles as allelochemicals. Vigna species, including azuki bean also showed relatively
strong allelopathic

activity.

Direct application of hairy vetch to farmlands showed that these plants could suppress weed without
using herbicides. The most promising ways were:1) as cover crop in orchards;
2) as cover plants for weed-protection in abandoned fields;
3) as living mulch plants which can be used instead of plastic-film-mulch.
By using these plants, sustainable agriculture without or with reduced artificial chemicals might be possible

invegetable and fruit production.
Introduction
In Japan abandoned
farmers is increasing

and industrialization.

Government regulations
species.

paddy fields

are increasing

in number because the average age ofToday almost 5% of paddy fields are not used because of

to prevent over production

To maintain this land it is usuallydifficultof rice.

These fields

and requiresconsistof many weedy

much investment to restore paddy

fields to their original condition.
Wehave been engaged in the search for allelopathic
and understandin order to identify

its mechanism. In the course of this study, we developed

identify

allelopathy(Fujiiet al, 1991).

al. 1992, 1995),

plantsand discriminate

from other competitive

We have reported allelopathy

and medicinal

plants (Fujii

of allelopathy,

called1991),

hairy vetch (Fujii

among other species.

In the course of this research, we have developed
and assesses the possibility

some new methods tofactors such as nutrients, light and waterin velvet bean (Fujii,

et al., 1991)

allelopathy

new bioassay

systems that demonstrates

the "plant box method, " and "sandwich method".

et

192

These methods involve mixed planting

using agar medium, and exudation of alleloehemicals

leaves and/or roots. These methods can show allelopathic
However,it is difficult to use allelopathic
competitive

factors such as lightaction of root exudates and leaf leachates.

plantsdirectly

way to use allelopathy.

in agriculture,

because of otherand nutrients. Thus we started to use the allelopathic

combined with other factors to enhance the effect of allelopathy.
probablefromIf other competitive

factorIn this sense, cover crops are afactors are the same, allelopathy

can play animportant role in weed suppression in combination with other factors. Some farmers have suggested
from theirexperiencespossibility

that some cover crops could be used for practicalscreen for allelopathic
primary selections,that some ground cover plants
activity

suppress

weeds drastically.

weed suppression.

There is a

Thus we began toof cover plants by the plant box and sandwich methods. From theseand experiences in the field, we have conducted field experiments to select themost promising cover crops for weed control in abandoned fields.
Materials

and Methods

Plant materials and definitionof hairy vetch. Most of the seed used in these experiments were from

"Kaneko Seed Co.", "The Takii Seed Co.", "The Yukijirushi

Seed Co ", and "Sakata Seed Co ". The

term 'hairy vetch' was used in Japan both for hairy vetch and woolly pod vetch. All seeds of hairy
vetch were imported from the USA (although

one seed company cannot tell us the origin and we

presume this is the origin), we call both species hairy vetch. There are, however, differences between
Vicia villosa and Vicia dasycarpa (=syn. V.villosa subsp. varia) but the allelopathic
inhibitory

activity

Cover Crops by the Plant Box Test. Primary selection

allelopathy

was done using the "Plant Box Method", developed(Fujii,

Young plants were cultivatedcontaining

andof weeds on the field of both is almost the same.

Screening of Allelopathic
1991).

activitya nutrient solution.

for the assessment of allelopathy

for one to two months by sand culture, in standing water

The receiver plant used for bioassay was lettuce (Great Lakes 366),

because it has a high sensitivity to bioactive substances.
Extraction of Inhibitory Chemicals from CoverCrops. Plant growth inhibitory
cover crops were tested by water and methanol extraction.

activities

of some

Dried leaves and shoots (60°C, overnight, forced air dry) were extracted with water and methanol in a ratio of 150:40.
were mixed with agar (0.5%)of cover crops for

These extracts

to make a bed to support lettuce seeds. After three days in the dark at

20°C, radicle and hypocotyl length were measured.
Screening of Cover Crops for Weed Control in the Experimental Fields
ability

To determine thepractical

aspects of weed suppressionin cover crops, field tests for weed suppression were

designed.

All field trials for screening were done in the experimental field of the Shikoku National

193
Agricultural

Experimental

Station. For the first screeningof spring seeded cover crops, 46 plants

were sownon lm x lm quadrates on May 23, 1992, then after three months of no weeding, weed
densityand dry weights were measured on August 25, 1992. For the preliminary

seeded plants, 39 candidates

screeningof fall

were sown on 2 x 2m quadrates on November 5, 1992, and after noweeding for 6 months, weed dry weight of each plot was measured on April 20, 1993. For 21
selectedfall sown cover crops of fall sown, 4 replications

design were designed

using the complete randomized blocand seeded in 2 x 2m quadrates onNovember 5, 1992, and weed and crop dry

weights were measured on April 20, 1993.
Application of Hairy Vetch to Abandoned Field in the Experimental Station

From the results ofscreening of cover crops, we focussed on hairy vetch, and the fourth set of trials

were conducted on

an experimental field at Shikoku National
1) A larger scale application

Agricultural

Experiment Station.

test of hairy vetch was conducted in comparison with Chinesemilkvetch on the uniform paddy field of 1000 m2.This field was divided into 16 blocks and 4 replicationsof4 different

cover crop trial were designed.

and oat, (c) Chinese

Cover crop design was; (a) hairy vetch, (b) hairy vetch

milk vetch, (d) no treatment as control. This experiment was started in 1992 andcontinued. Data from 1992 to 1994 is reported in this paper.
2) Increasing

the seeding ratio from standard seeding ratio to double, triple and quadruple

wasexamined using a field of 500 m2, and arranged by the split-split-plot

seeding

test using 4 replications.

This test started on October 25, 1993 and weed and crop yield were measured in May 20, 1994.
3) Changing

the seedingdate from October to February was tested using the same field with tree

replications.

This test was started in 1 992 and ended in 1994.

4) Effect of soil water content on the growth of hairy vetch was tested in a lysimeter.
was designed to know the suitablewater ratio for crops, and it is possible

This lysimeterto change the water levelby a stair step system of over flow.Each block was about 10 m2and there were four stages with two
replications.

Soil typeof above four tests was sandy loam, paddy field converted

to upland

conditions.
5) Mixed
experimental

plantingsof leguminous

field at Shikoku

National

cover crops with grasses
Agriculturalslope angle of this field was 9 degrees. Seeding
yieldswere examined on a sloping

Experiment Station on Oo-asa mountain. The
date was November 6, 1993, and crop and weed

were measured on May30. The total area of this field was about 800 m2.

Direct Application Test of Hairy Vetch to Farmer's Field. Direct applicationand exhibition

trial ofhairy vetch was conducted using the field of cooperative farmers by the courtesy of each districtcounselors for farmers (in Japan, there are counselors for farmers in each county. These counselors

194

are public

service officers belonging

to the Ministryof Agriculture.)Six different farming systems

werechosen.
1) Paddy

field in Man'nou town. The owner is Mr. Tsune-kane. The area is about 800 m2. This

paddy field is in a slightly

mountainous area, and slightly

dry condition.

This field had just stopped

rice production.
2) Paddy field in Marugame City. The owner is Mr. Hirata. The area is about 1000 m2.This field is
close to the road and in humid conditions.
3) Paddy

field in Zentsuji

This field was abandoned two years ago.

City. The owner is Mr. Kawada. The area is about 900 m2.This field wasabandoned for 4 or 5 years and they used Chinese milk vetch as green cover crop, but suffered fromserious weed infestation.
4) Grassland
grassland,

in Chu-nan town. The owneris Mr. Morichika. The area is about 600m2. This is sloping
and used for exhibition.

5) A persimmon orchard in Kounan town, Oka village.

The owner is Mr. Oka. The area used for thecover crop trial is about 5000 m2in first year, 1992, and extended to 10000m2 in the following

year

and has continued until now. Oka family is a pioneer in this orchard area, and produces the sweet
type of Kaki of high quality.
6) Pear Orchard in Toyohama town. The owner is Mr. Oohiro. The area used for the trial for hairy
vetch is about 80 ha.
Results
Screenings of Allelopathic

Cover Crops The results of screening for aJllelopathic

leguminousand gramineous species

these tables,

radicleby Plant Box Method are shown (Table 1 and Table 2).

percentage means the percentages

lettuceplants present

length

within the root zone controlled

cover crops fromof the root radicle

in the root zone of each donor plant.
by donor plants.

by length of the young

Based on the calculation(Tableof radicle

Leguminous cover crops such as velvet

bean, hairy vetch, yellow sweet clover and white sweet clover have strong allelopathic
activityIn

inhibitory1). Of these legumes, velvet bean, Crotaralia, Canavalia, Cajams, Cicer, Vigna andGfycine are summercover crops, and most of others are winter cover crops. Melilotus and Pueraria
are perennial

crops. It is important

to know the characteristics

of each cover crop to use for thesuch as oat, wheat, millet (Setaria),

rye also showed strong inhibitory

farmland.
Gramineous species,
activity,

but most of Compositae family such as Helianthus showed only medium inhibitory

by this method (Table 2).

activity

Table 1. Assessment of Allelopathic
by the Plant Box Method

Activityof Leguminous Cover Crops

Table 2. Assessment of Allelopathic
Activity of Gramineous Cover Crops
and some Other Family by the Plant Box Method

196

Extraction of Inhibitory

Chemicals from Cover Crops

allelochemicals

in plants, we checked the extraction

The inhibitory

activities

To determineof chemicals
activity

the results

concerning the allelochemicals(Fujii1994).

VelvetFujii,

bean contains
activityofcover crops.
Hairy

from both water and methanol extract.

Wehave already published1990,

for some principalof water and methanol extracts of cover crops is shown (Table 3).

vetch and velvet bean showed the strongest inhibitory
et al.,

the presence

extracted

a very highfrom velvet beanamount of L-3, 4-

dihydroxyphenylalanine

and the inhibitoryof this compound is rather high in acidic

non-oxidative

conditions.

However, in the case of hairy vetch, there may be other more inhibitory

allelochemicals

than velvet bean. The results of solvent extraction

are consistentandwith the results

fromPlant Box test for root exudates in these experiments.
Screening of Cover Crops for WeedControl in Experimental Fields

To assess the activity

these cover crops, field experiments were started using the small-scale

field test in Experimentalof

Station, and it was found that spring-sown cover crops were not promising (Table 4). In Japan there
is a rainy season in June, and soon after the beginning
Some cover crops such as Helianthus,
activityof this season, weed growth is very vigorous.

Celosia and Panicum showed relatively

strong inhibitory

to weeds, and Muama,Vignaand Cassia were somewhat less strong. Competition

for light

and nutrients are the most important factors in the field, and all of these plants have vigorous growthrate and huge leaves. Most of these cover crops were reported as allelopathic,
strong inhibitory

and have relatively

activityby the Plant Box test (Table 1 and 2). For example, there are many reports

concerning the allelopathyof Helianthus, and Celosia (Rice, 1 984). However a field with more than

20% of the field looking

weedy is disagreeablesuccessful

for farmers. Spring-sown cover crops have not been

in Japanese conditions.

The results from the trial for autumn seeding are shown (Table 5, 6). If cover crops are sown inautumn,they tend to grow slowly but steadily
could eliminate

in winter to make enough biomass in spring and thisvigorous weeds. Table 5 shows a part of the results of autumn-sowncover crops.

Since this was a preliminary

trial, we had no replicationof this test. It is obvious that the dry weightof each cover crop is most important to reduce the growth of weeds. It is true for rye, oat, wheat,
woolly pod vetch and Italian ryegrass. Due to the overwhelming canopy of these crops, weed haveno space. But if compared with the relationshipbetween dry weight and weed suppression

activity,

Brassica, Vicia, and Medicago do not have enough biomass but show improved weed suppression
than estimatedfromtheir biomass. Allelopathy

The result of a replicatedmay play a role in these cases.

study is shown (Table 6). Without weeding, hairy vetch, oat, barley,rye, wheat showed strong inhibitory

activity

to weeds, but Chinese milk vetch, which is a traditional

green manurein China and Japan, showed little weed suppression.

Table 3.

Effect of Water and Methanol Extracts of Cover Crops

Table 4.

Effect of Spring Sown Cover Crops on the Weed Growth in
the Field; A Primary Selection

Table 5. Effect of Fall sown Cover Crops on Weed Control in
the Field; A Primary Selection

Table 6.

Effect of Selected

Fall Sown^Cover Crops on the Weed control

of

199

In conclusion, spring sowncover crops are not promising in Japanese conditions,but autumn

sowncover crops such as hairy vetch, rye, oat, wheat, and barley have sufficient inhibitory
for weed from spring to early summer.The inhibitory
traditional

methods of controlling

Application

activities

were almost the same as that ofweeds such as the rice straw mulch or herbicide

application.

of Hairy Vetch to Abandoned Field in the Experimental Station

heavy amount of seed application

activity

The results of aof hairy vetch, Chinese milk vetch and mixed plantingof vetchand oats is shown (Tables 7 and 8). These data were taken between 1992 to 1994, and in each year
four replications

were made. Hairy vetch almost completely

On the other hand, Chinese milk vetch, traditionallycould inhibit

inhibits

the growth of weeds in spring.

used in Japanese paddy field as green manure,

the weed biomass only up to 80%. If 20% of weeds grow this will make the fieldweedy the following

year, and will be abandoned in two or three years without weeding. These

results coincide with the observation^ of farmers that continuous use of Chinese milk vetch will cause
a serious infestation
The addition
years, additionof weeds.
oats to hairy vetch was aimed at increasingof oats decreases the population

the weed suppression ability.

of weeds nearly perfectly

until

In bothautumn. Mixed

planting of vetch and oat is complementary and yield of both per acre increased. However,if no careis taken, the outlook of the field from spring to summerof mixed cover field is not beautiful

becauseof the remaining oat stems. On the other hand, hairy vetch keeps a stand height of 50 cm(maximum)
and the appearance of the field is uniform and flat, and free fromweeds. Hairy vetch dies when thetemperature reaches to 30°C. In our experimental field, hairy vetch made a straw like mulch without
any work and this mulch protect the field from weed invasion after the death of the mother plant. We
concluded that using hairy vetch alone is better to recommend to farmers because of the simplicityofsowing and reduced the labor.
Some abandoned paddy fields in Japan are sufferingfrom water logging.

Resistance of hairy

vetch to water logging in paddy fields was measured using the water-logged lysimeter. Hairy vetch is
resistant

to heavy water logging. Water content of 40% covers most of the water logged paddy fieldand vetch showed no growth retardation in these conditions.
of water is possible,

But, in this lysimeter,

so if there is heavy water logging condition

continuous supply

and no movementof water in thefield this might disturb the growth of vetch.
Standard seeding rate was sufficient for the weed control, and increasing the seed volume madeno difference for biomass to weed suppression.

The cost per 1000m2 (Japanesestandard

farming) is about 2500 yen or about 20 US dollars on the basis of Japanese market price.

unit ofTable 7. Effect of Cover Crops on Weed Control in AbandonedFirst Trial on the Experimental Station (1992-1993)

Paddy Field;*1 Percentage of weed control. 100 % means complete control. Dry weight ofweeds in the control plot are 281 (First)
and 155 (Second)g nr2.
*2 The same letter means not significantly
differentby Duncan' s Multiple Range
Test(P>0. 05).
*3 Dominant weeds species in this filed are, Alopecurus aequalis and Lamium
amplexicaule.
*4 Seeding
date: October 28, 1992, Sampling
date: May 7(First),
JunelO(Second),
1993.
*5 Four replications,each plot is 5 x 10 m (500 m2).
*6 Place: SNAES Experimental Field P8 (Zentsuji,
Kagawa), abandoned paddy field.

Table 8. Effect of Cover Crops on Weed Control in Abandoned Paddy Field;
Second Trial on the Experimental Station (1993-1994)* 1 Percentage
of weed control 100 % means complete control. Dry weight ofweeds in the control plot are 156 (First) and 247 (Second)g nr2.
*2 The same letter means not significantly
different by Duncan's Multiple Range
Test(P>0.05).
*3 Dominant weeds species in this filed are, Alopecurus aeqitalis and Lamium
amplexicaide.
*4 Seeding date: November 10, 1993.
*5 Four replications,each plot is 5 x 10 m (500 m2).
*6 Place: SNAES Experimental
Field P8 (Zentsuji,
Kagawa), abandoned paddy field.

201

Since late seeding tends to result in growth of more weeds and it was concluded that the
optimum seeding date for weed suppression in Japan is no later than the 1 st week of November.
Direct Applicationof Hairy Vetch to Farmer's Field. Direct applicationand exhibition

trialsofhairy vetch were done using the fields of cooperative

farmers.

suppress weed almost completely,of farmers were agreeable. Before this trial,

and the impression

In most cases, hairy vetch could

there was no custom to use hairy vetch as a cover crop to control the weeds. Many farmers cameto
our Experimental Station or exhibition

fields of cooperative

farmers, and most of them started to try

hairy vetch. Now,in central Japan, the use of hairy vetch under persimmon orchards has spread toabout 600 ha, and is still expanding.
Incorporation of velvet bean leaves into soil and weedprevalence in the fields of velvet bean
standsAn experiment was carried out to examine the effects of velvet bean on the growth of otherplants in a mixed culture (Fujii,1990).

The treatment also included

leaves into the soil. Fresh leaves incorporated
reduced the emergence of succeeding

into soil (1.0%

commonbean (Phaseolus

an incorporationof velvet bean

WAV in dry weight-equivalent)vulgaris) by 60%, and the plant

growth to 30% of the control. This effect ran out two weeks after the incorporation.
incorporated

Dried leaves

into the field showed no inhibition.

Weed populationsin spring in the continuous cropping velvet bean plots showed a lower populationof weeds dominated by sticky duckweed (Cerastiun glomeratum), than the other plots of egg plant,
tomato plant, upland rice and fallow.
Attelopathic
restraining

compound in velvet bean

Analysisof the effective compound of velvet bean in

the growth of companion plants confirmed its associationthat velvet bean seeds contain a high concentration

with L-DOPA It is well knownof L-DOPA (6 to 9%) , which plays a role as achemical barrier to insect attack. In the mammalian brain, L-DOPA is the precursor of dopamine, aneurotransmitter,and also an important intermediate

and insect cuticle, L-DOPA is oxidized
intermediateof alkaloids.

In animal skin, hair, feathers,

through dopaquinone to produce melanin. As L-DOPAis ancompound and rapidly metabolized,

normal tissues have little L-DOPA.

Fresh velvet bean leaves contain as much as 1% of L-DOPA. It actuallyroots, and its concentrationroots. This concentration
inhibitionrurreaches lppm in water-culture solution,

exudes from theand 50ppm in the vicinity

is high enough to reduce the growth of neighboringofplants and the growth

in a mixed culture is shown in Agar-medium culture. It also leaches out from leaves withrain drops or fog dew. Since velvet bean produces 20 to 30 tons of fresh leaves and stems per hectare,approximately

200 to 300kg of L-DOPA maybe added to soils a year (Fujii,1994).

202

Discussion
Haiiy vetch is thought to have originated between western Asia and the eastern Mediterranean
coast. It was cultivatedin England and Germany in the early 1 9* century then introduced to the USA

in the middle of 19th century. It was recommended by the USDA, and nowis widely distributedinthe southern part of the USA Hairy vetch was introduced to Japan in the early 20th century and goodresults

were obtainedat the Agricultural

Experimental

Station, but has not been distributed

until

recently.
There are some reports of allelopathy
Balinevichiute(1972)

tested the inhibitory

vetch showed the strongest activity.

and weed control using hairy vetch. Lazauskas andactivityof extract of seeds to barley, and found that hairy

White et al. (1989)

reported that the incorporationof residue of

hairy vetch and crimson clover reduced the emergence of Solanaceae weeds by 60 to 80%, andwater extract of hairy vetch had stronger inhibitory(1 993) reported

than crimson clover. Johnson et al.

that the mulch made fromhairy vetch or rye completely

systems by killingthat the living

activity

them by herbicide
and Teasdalecompensate for vinyl plastic

weeds in non-tillage

or mechanical cutting. Teasdale and Daughtry (1 993) reported

mulch of hairy vetch showed better inhibitory

one. Abdul-Bakiinhibit(1993)

activity

to weeds than the desiccatedreported a unique system using hairy vetch mulch tofilm mulch in tomato production.

There are so many reports and fieldobservation about the weed suppression of hairy vetch, but still the contributionof allelopathy

allelochemicals

fromhairy vetch.

are unknown.Weare nowplanning to separate allelochemicals

Hairy vetch has much merit in addition
1 ) Nitrogen

fixationand its

to weed control in the field. The merits are:to reduce chemical fertilizer;

2) Organic materials to reduce chemical fertilizer

or soil conditioner;

3) Prevents soil erosion by surface cover;4) Deep root systems help to increase the soil porosity;5) Thickground coverage ameliorates

the microclimate

to reduce maximumtemperature andincrease minimum temperature;6) Induce carnivorous insects which reduces the population

harmful insects.

After these series of experiments, it was concluded that hairy vetch is the most promisingcover crop for the control of weeds in abandoned fields, grasslandand orchard in the central andsouthern part of Japan.
Velvet bean smothers weeds due to its rapid and thick covering effect with leaves. L-DOPAor
its associateallelochemicalcompounds, accumulated in an extremely high concentrationin plants, function

as anand reduces weeds. The role of L-DOPAin velvet bean seeds was earlier regarded asa chemical barrier to insect attacks. It is now confirmed, however, that it plays another role with, its

203
ailelopathic

activity,

in weed control.

Since velvet bean has special abilitiesof nematode population,such as weed smothering, tolerance to pests, suppression

and soil improvement in its physicalto reduce applicationsof artificial

chemicals.

structure,

Velvet bean seed yields

it could be more widely used
are very high in the tropics,

the seed contains a high level of protein with a useful protein score. If detrimental
DOPA and trypsine
to alleviation

inhibitors

could be eliminatedthroughof the food problems in some tropicalcountries.

andfactors such as L-

proper cooking, it would also contribute

References
Abdul-Baki,

A. A and J. R Teasdale. 1993. A no-tillage

and subterranean clover mulches. HortScience,
Fujii,

Y, T. Shibuya,candidate

Fujii,

Fujii,and T. Yasuda. 1990. L-3, 4-dihydroxyphenylalaninefrom litter leachate

Y and T. Shibuya.

1991.

ailelopathic

activity

36 (Suppl)

152-153.

Y, T. Shibuya,

A new bioassay

for allelopathy

1992. Establishment

properties.

of a new bioassay

Assessment

152- 153.
for allelopathy;

Assessment ofplants for the detectionof

Weed Res., Japan 36: 36-42.
1992. Allelopathy

Y 1994. Screeningof ailelopathic

methods for allelopathy,

Fujii,

1 50-15 1.

from root exudates by using Plant Box and agar medium. Weed Res., Japan

Y and T. Shibuya.

Sciences,

Assessment ofwith agar medium, n.

and T. Yasuda. 1991. Survey of Japanese medicinal

the most promising

8.

by sandwich method. Weed Res., Japan 36 (Suppl)

of hairy vetch. Assessment of ailelopathic

weed control in the field. Weed Res., Japan 37 (Suppl)
Fujii,as an allelochemicalwith agar medium I.

from root exudates. Weed Res., Japan 36 (Suppl)

Y and T. Shibuya.

ailelopathic
Fujii,

28: 1 06- 1 08

Y and T. Shibuya. 1991. A newbioassay for allelopathyof allelopathy
Fujii,

system using hairy vetch

fromMucunapuriens (L.) DC. var. utilis. Agric. Biol. Chem. 55 : 617-61

allelopathy
Fujii,

tomato production

activityand160-161.

candidates

by new specific discrimination

and assessment

and the identificationof L-DOPA as the allelopathic

substancevelvet bean. The Bulletinof the National

Institutefromof Agro-Environmental

10: 115-218.

Y S. Ono. and K. Sato. 1994. Weed suppression

by winter cover crops such as hairy vetch,oats, rye, barley and wheat, and its relation to Plant Box Test. Weed Res., Japan 39 (Suppl)

258-

259.
Johnson, G. A, M. S. Defelice,
Corn.
Lazauskas,

Weed Technology,

and Z. R Helsel.

1993.

Cover crop management and weed control in

7: 425-430.

P., and Z. Balinevichiute.

1972.

Influenceof the excretions

from Vicia villosa seeds on

204

germinationand primary growth of some crops and weeds. In "Physiological-Biochemical

Basis of Plant Interactions
Rice, E. L. 1984.
Allelopathy,
Teasdale,Manipulated
2nd editionin Phytocenoses"

Vol.3 pp.76-79

ecosystems:

Roles of allelopathy

in Agriculture,

pp. 8-12,inAcademic Press. London.

J. K and C. S. T. Daughtry. 1993. Weed suppression

by live and desiccatedhairy vetch.

Weed Science, 41 :207-212.
White, K H., A D. Worsham, and U. Blum. 1989. Allelopathic
aqueous extracts, Weed Science 3 7:674-679

potentialof legume debris and205

The Use of Wild

Relatives

Mungbean and Blackgram
PeerasakSrinives1,

Ngampongsai

Napaporn

department

Pathom 73 140, Thailand;2

and Gamma RadiationinBreeding

Hual-alai1,

Supavineeof Agronomy,

Chai Nat Field

Saengchot1

andSaen, Nakhon

KasetsartUniv.,

Kamphaeng

Crops Research

Station,Chai Nat 17000,

SumanaThailand

Abstract
Interspecific
its related

hybridization

Vigna spp. Pod setting

cross V. radiatax V, radiata

was performed between mungbean (Vigna radiata (L.) Wilczek)

andpercentage was high when V. radiata was the female parent.

The(wild)

gave the highest

combined with embryo rescue were successful
mungo (wild),

V. umbellata,

from the cross involving

pod settingof 26.1%.

Interspecificwhen male parents were V. radiata (wild),

V. umbellata(wild),

V. angularis

moth bean (V. aconitifolia)

was unsuccessful.

staining

percentage

V. mungo, V.
Embryo rescue

Most of the Fi plants

was the highest(20.6%)

set

flowers profusely

without pod setting.

interspecificprogeny of V. radiata x V. mungo.Attempts have been made to obtain F2 seeds

hybridPollenand V. glabrescens.

crossesin thefromthese crosses.
F2 seeds from the crosses V. radiatax V. radiata (wild),

mungox V.mungo(wild) and V. mungo(wild)of gammarays. Quantitative
Quantitative
included

traitsincludedx V. mungowere irradiatedresponses to irradiation

fromthe black gram crosses.

x V. radiata,V.

with 10, 20, 50 and 70 kradin both quantitativeand large-seededleaf mutants (dark green, waxy, multiple,(wild)

were only found in 1000 seed weight of Mi plants

Mutation was detected

early maturity

V. radiataand lobed),

and qualitative

traits.

lines were extracted.

Qualitative

traitspod mutants (large

and top podding),and semi-dwarf plants, were identified.
Introduction
The advantages
scientists

including

improvement,
cultivars.
(L.)

Rick (1967),

Hepper],have been well documented(1970),

cultigensspeciesof thewith potential

by several

Knoblock(1972),and Harlan (1976).

is to transfer

desirable

traitsfor using wide hybridizationand other

Ceratotropis[V mungo (L.)

Barberthe main use of wide crossing

The potential

Wilczek]subgenusof wide hybridizationin breeding

subgenus

Ceratotropis(Thunb.)In plantfrom wild relatives

toof mungbean [Vigna radiata
is high.

for use in mungbean improvement

rice bean [V umbellata

plant

Ohwi & Ohashi],Among Vigna
are blackgram
azuki bean [V.

206
angularis(Willd.)Ohwi & Ohashi],moth bean [V. aconitifolia

legume V. glabrescens.

These species

except V. glabrescens

which is a tetraploid(Jacq.)

Marechal],and a wild

have the same chromosome number (2n = 2x = 22),(2n = 4x = 44).

Attempts

have been made to produce hybrids

among Vigna spp., aiming to incorporate

certaincharactersfrom wild to cultivated

For example,

transfer

mungbean yellow mosaic virus resistanceand insect resistancefrom V. umbellata

performance of the lines
obtained

to V. radiata.

derived from these crosses.

any seeds.

AVRDC (1990)

radiata and V. glabrescens,
Chen et al. (1983)
concluded

and obtained

made crosses

that poor development

this problem,

reported

differentInterspecificfrom radiata
genotypes.

a successful

degrees

on parental

withinto V. radiata,

such as resistanceto diseases

non-shattering,and high methionine.
involving

cultivatedtraits(V. radiata

silvestris)

can be crossed

and insects,

cultigen.

Since transgressivefrom interspecific(GP1).

and blackgramwith similar

before selection.

In plant breeding,depending

The Fion parental
from V. mungo

are normally

For example, mungbean and its wild
relativeusing varieties(V. mungo varof the respective

may not be as high as that obtainedwith a mutagen to enhance their

genetic

gamma rays have been widely used mutagen andhave been developed1993).

Gupta (1 996)

extensively

reviewed mutation breedingradiation

mutagens.

He groupedmutant traits

intoattributes,

pest resistanceof 13 mutant mungbean varieties

is difficult.

of the same species

80% of mutant cultivars

to yieldand Chen et al

environments,about

mutants which were related(1989),

to adverse

and its wildof the progenies

crosses the F2 seed can be treatedsterility

tolerance

success rate as crosses

segregation

To overcomethat could be tranferred

and wild varieties

var. sublobatd),

andembryos from various Vigna crosses.

of pollensince they belong to the same genepool

relative

Chen et al

V. radiata-mungo complex itself

usefulto thrips.

and umbellata,

genotypes.

V.

them in modified MS media, combined(1983),

degrees

betweenmoderate resistanceof Fi seeds depended

x mungo showed varying

Crosseshybridization

mungo, angularisof success in rescuing

hybridizationonhowever, the Fi plants

among radiata,

listedvariation

successfully

4 pure lines carrying

Tomooka et al (1991)

successful

Ahn and Hartmann (1978)
and V. angularis,toand diseasereports

reported

Gosal and Bajaj

tried

There have been no further

the Fi embryos were rescued by culturingwith some other techniques.

AVRDC (1974)

from V. mungo to V. radiata,

Fi seeds from the cross between V. radiata

did not produce(1990)

species.

officially released

after gamma rays irradiation
chlorophyllin India,(Wang,

in mungbean using chemical

andmutants and morphologicaland higher nodulation.

The list

Pakistanduringand Bangladesh

207
1982

to 1992

maturity.

revealed

that major attributes

So far, there

before performing

Materialsof the mutants were disease

was no report on combining

wide crossing

resistanceand early

with mutation

breeding

pure line selection.

and Methods

Plant materials
The Vigna spp.
were 9 parental
germination.
1.

used in thisexperimententrieswhich have two distinct

Their scientificnames, accession

The mungbean group (epigeal

were of the subgenus

Ceratotropis.

germination

epigealgermination)

V. radiata var. radiata (cultivated

1.2

V. radiata

1.3

V. mungo var. mungo [cultivated

1.4

V. radiata var. silvestrisvar. sublobata

mungbean, cv. CN 36)

(wild

(wild

2. The azuki bean group (hypogeal

mungbean, ace. TC 1966)
blackgram,
blackgram,

V. umbellata(cultivated

2.2

V. umbellata

(wild

2.3

V. aconitifolia(moth bean, ace. 80053)

2.4

V. angularis var. angularis

2.5

V. glabrescens(wild variety, ace. V 1 160)

in all possible

were cultured

1963)

agar (White,
Rooting

2 mg/1 DBA added.

some qualitative

traitsotherand divided

Fi plants

to observe

to embryo rescue. Dissectedsupplementedof the Fi plants

combinations

embryoswith 200 mg/1 yeast

extract

was done in B5 medium (Gamborg

were grown and compared with theirparents

toet ah,
andwere recorded.

F2 seed from individual
harvested

ace. TC 221 1)(azuki bean, ace. Fukuoka)

were subjected

with

2)]

rice bean, ace. 80021)

Fi seed set. Fi pods that fell prematurely

1968)

2 (PSLrice bean, ace. 80015)

were made between the above species

enhance shoot production.

cv. Phitsanulok

germination)

2.1

in White's

and hypogealnames and numbers are as follows:

1.1

Crosses

habits,There

Fi plants

into 5 equal

sets were irradiatedfrom cultivatedsets of seeds.

x wild mungbean and black gram were

One set was used as the control

with gamma rays at 10, 20, 50 and 70 kilorads

gammator (J. L. Shepherd& Associates,

CS-137.

It is installedat the Department

of Applied

University(KU),

The irradiatedseed (Mi seed)

Bangkok.

Model MK 1-30) was loaded
Radiation

while the(krad).

with 4,500

The

Curies ofand Isotope,

Kasetsart

were sown in the field

and bulk

208
harvested
plantsin each family to obtainseparately

harvested

M2 (F3) seed.

Mutant M2 plant traits

for pure line selection.

Remaining

Weightwere recordedandplants were bulk harvested

toobtain M3 (F4) seeds for further

observation.

of 1000 seed was recorded

while number of pods per plant,

number of seeds per pod, and plant heightin Mi to M3,

were observed

onlyfor M3plants.
Hybridization,
Nat Field

Crops

embryo rescue, and plantingof the Fi plants

were performed at Chai

Reseach Centre of the Departmentof Agriculture.

Irradiatedsown at the fieldof the Asian Regional

Centre,AVRDC, locatedF2 seed werein KU, Kamphaeng Saen

Campus.
Data analyses
The observed

quantitative

data were analysed

accordingto Steeland Torrie (1980).

To test for differences

between 1000 seed weight among the lines in each generation,

analysisin completely

randomized

designcombining

the residuals

from direct

and reciprocal

crosses,

F-testsof all sourcesof variation.

While variation

due to irradiation

Bartlett's X2 (Chi-square)

Results

and Discussion

Results

from interspecificcrossesA reasonableprecentage

the female parent,(CRD)

to test for homogeneity

was used. A pooled

male parent.

However, seed set was low in all crosses.

crosses between V. radiata
generallyretained(femaleFi pods until

embryos gave a higher percentage
thus more delicate
techniqueembryos.

was not as highrescue disturbs

parent)A relatively

higherdays after pollination)
was higher

as naturalin olderembryos.

was theof Fi

by Chen et alplantsDissecting

the embryos while natural seed and pod development

their rather maturethan from younger andobtainedin these

For example,

V. mungo, and V. mungo

and thus more seedlings

pod development

earliersettingof the above crosses

with V. radiata (wild),

The number of adultpercentageasand grew until flowering.

15 days after flowering.

germination1). Podglabrescens

when V.radiata was the female parent, as reported

for embryo rescue, germination

having V. radiata(Tableeven when the tetraploid

When embryos from young pods (2-15(wild)

in all crosses

Only Fi seeds from 5 out of8 crosses germinatedwere dissected

rates was compared by

was the pollinator

was satisfactory(1983).

in theof variances.

percentage

seeds were obtainedin most crosses,

error was formed by

and used as the denominatorof pod set was observed

except when V. aconitifolia

nestedfrom embryo rescue

3 crosses

becauseembryowas almost normal. This

Table1. Number of flowers pollinated,
pod set, Fi seed sown, and adult Fi plants obtained
from crosses havine V. radiata as female Darent usine without embrvo rescue.

Table

2.

Number of embryos cultured,plants obtained from crosses
rescue.

embryos germinated,seedlings
obtained,and adult
having V. radiata as female parent, using embryo

Table 3. Analysis of variance of 1000 seed weight from Mi plants
and TC 1966xCN36.

of CN 36 x TC 1966

210

was not the case in the othernumber of adult plants.

5 crosses

Embryo rescue helped

from crosses using V. umbellata
radiatax V. angularis

was not successful.
1).

Morphologicalcharactersand it can be concluded
is dominant

epigealspot and powdery mildew diseases
This
relatedVigna species.

seed set. Pollen

Althoughstaining

revealed

Plantscan be vegetatively

bridgecrossing

Results

hybrids

that some Fi plants

available(Gosal1983).

and Bajaj,

to use desirable
profusely,

are being

doubling

Photoperiod

V. radiata cultivar

their

leaf

to date.
traitsfromthere has been no

do not produce viable

and attempts(1997)

to Cercospera

flowered(AVRDC, 1990),

flowers with growth regulators

by Ngampongsai

by Tomooka et al (1991)

propagated,

to V. radiata

in detailgermination.

from someare more resistant

than the most resistantinterspecific2). The cross V.

Fi plants

to hypogeal

All Fi plants

confirmed the recommendation

adult Fi-plantsof the adult

were studied

is dominant

to non-sensitivity.

gave only a low

as male parent (Table

Morphologyof all Fi plants

thatseed development

increase the chance of obtaining(w) and V. glabrescenscrosses is shown (Figure

sensitivity

in which naturalpollen(Fig.

2).

made to use them by doing
chromosomes, and treating

thefrom gammairradiation
Fi plantsfrom V. radiata

mungox V.mungovar. silvestrisindividual

Fi plants

var. radiatax V. radiataand their reciprocal

were harvested.

Finally

var. sublobata,crosses set seeds normally. F2 seed from19, 37, 32 and 36 F2 seed families

from the cross CN 36 x TC 1966, TC 1966 x CN36, PSL 2x TC 2211,
respectively.

Thus

variation

betweenrecombination

as well as irradiation.

V. mungo var.

Mi plants

withineachwere obtained

and TC 2211 x PSL 2,
F2 familycame froma. Variation in metric characters
Analyses
36 x TC 1966)
3).

There

of variancesin 1000 seed weight

revealed

that variationand (TC 1966 x CN 36) is high among F2 families

was no dosage effectfrom gammaradiationfrom the same cross (Table

upon this trait,

plantsfrom (PSL 2 x TC 2211)

and (TC 221 1 x PSL 2) gave slightly

4).

In these

in addition

effectof irradiation

backgramcrosses,

was also detected

for this

is not clear.

variability.

Response

to further

populations

with higher

genetic

to variationin (TC 2211

However, it helped

variation

however.
differentin eachThe Mrresults(Tableamong Mi lines found, the dosage

x PSL 2) (reciprocal)

identifying

selection

in seed size in (CNpopulations
cross

will

cross.

The reason

that

showed highgenetic

help

determinewhethergive improved response to selection.

21]

Fig. 1 Fi plants from interspecifichybridization.
(a) V. radiata x V. radiata (w) (b) V. radiata x V. umbellata (c) V. radiata x V. mungo(d) V. radiata (w) x V. mungo (w) (e) V. radiata (w) x V. glabrescens(f) V. glabrescens
V. radiataFig. 2 Pollen staining of Fi plants(a) V. radiata x V. radiata (w)(c) V. radiata x V. mungo (w)(e) V. radiata (w) x V. glabrescensfrom interspecifichybridization.
(b) V. radiata x V. mungo(d) V. radiata x V. umbellata(f) V. glabrescens
x V. radiataxTable4.

Analysis of variance of 1000 seed weightand TC 2211 x PSL 2.

Significant

Table

Table

at P =.05

of PSL 2 x TC 2211

and P =.01, respectively

5.

Analysis of variance of 1000 seed weightand TC 1966 x CN 36.

**

Significant

6.

Analysis of variance of 1000 seed weightand TC 2211 x PSL 2.

Significant

from Mi plantsfrom M2plants of CN 36 x TC 1966

at P=.01

at P =.05

from M2 plants ofPSL

2 x TC 221 1

Seed size showed less variationin M2. In V. radiata x V. radiata (w), only variation between

M2lines of TC 1966 x CN 36 was significant
gene loci and thus could be easily(Table5). This trait

is governed

fixed after one or two generations.

by a few major

Similar

responsewas

also detected

in V. mungo x V.mungo(w) crosses.

Variation between M2 lines was reduced

from highly

significant

at P =.05

detectedlines(Table4) to justin M3 lines in all 4 crosses

by whichhomozygous)

found among the selected
reciprocal)

9).

crosses (Table
Effectdue to differences
cross progenies

The reason is unclear.

wasis equivalent

to F4

The M3 generation

As a result,M3 lines.

6). No variation

Variationin overall

had larger

This variationof a genotype
significantly

becoming

less variation

between crosses(direct

vsaverage seed size between bothseed than those from direct crosshas not been detectedof the gamma rays can also be measured using Bartlett'sof variances.

High variation

gammadoses would give a significant
weight of the cross (TC 2211

between populations

X2. Just significant

variation

x PSL 2), but not in the other crosses.

weight in M2 and a few agronomic traits

in V. mungo

was not determinedin the M3 were not significant

in the blackgram

X2 (Chi-square)
treatedwas detectedin Mi seed

Variances of 1000 seed
(Tablecrosses due to twining

10). Variation

growth habit

7. Analysis of variance of 1000 seed weight from M3plants ofCN 36 x TC 1966
and TC 1966x CN 36.

Significant

at P =.O1

testwith different

progenies.
Table

was

8 and 9).

for homogeneity

plant height(Tablemay be fixed (probability

large-seeded

could still be detected

7 and

7 and 8).

= 7/s or 87.5%.

groups in which the reciprocal(Table(Tabletime some gene lociequal to l-(y)3significant

inof theTable8. Analysis of variance of 1000 seed weightand TC 2211 x PSL 2.

Table 9.
generations.

Average1000

seed weightMeans of direct and reciprocal
different
at P =.01 by F-test
Table 10.

incrossesfrom M3plants of PSL 2 x TC 221 1

4 mungbean

followedcrosses

at 3 different

by different

letters

are significantly

Bartlett's
X2 test for homogeneity of variances of some quantitative
mungbean crosses, measured at Mi to M3 generations.

b. Variationin qualitative

Since Mi plants

showed multiple

leaflets,charactersin 4from gamma irradiation,nocharactersshowed mainly physiologicalmutants were found in this generation.
and M3 generations.

mutationdisorders

A number of mutant chararacters

In the M2, 20 leaf mutants were detected
the rests

were semi-dwarf plants,

leaflets.

A plant from the cross (TC 1966 x CN 36) treatedmultipleleafletsand semi-dwarfhess.

(Tablewere observed in M2
ll).

dark green leaves

Fifteenplantsand narrow

with 50 krad gamma rays had both

No mutant was observed in the cross TC 221 1 x PSL 2.

More mutants showed up in the M3 generationfrom which 36 mutants from all crosses

were

11.

Table

MutantA ='multiple

charactersfound in M2 and M3 of the mungbean+ semi-dwarf

B = profuse pod + large leaflet

and blackgram

crossespod

E = profuse pod + erect stem

C = extruded pod + profuse

D = erect stem + large leaflet

irradiatedwith different

doses

of gamma rays.

216

mungbean crosses and gamma doses in M2 and M3 generations
Pictureswere summarized (Table

12).

of certain mutants are shown (Fig. 3).

Table 12.

Summary of mutant plants found in M2 and M3of mungbean and blackgram
crosses irradiatedwith different doses of gammarays.

G en eratio n

C ro ss

0
0
0
0
0
0
0
0
1
0
0
1

C N 3 6 x T C 19 6 6
T C 19 6 6 x C N 3 6
P SL 2 x T C 22 11
T C 2 21 1 x P SL 2
C N 36
T o talC N 3 6 x T C 1 9 66
T C 1966 x C N 36
P SL 2 x T C 2 211
T C 2 2 11 x P SL 2
T o tal

M 2

M 3

T o talD o se
20
3
0
1
0
2
6
3
5
0
0
8

10
1
1
0
0
1
3
2
4
0
0
6

50
0
5
2
0
1
8
3
1
0
0
4

70
2
3
0
0
1
6
7
1
3
1
12

6
9
3
0
5
23
15
12
3
1
31

Conclusion
Althoughwild relatives

offer a high potential

as genetic

resources

blackgram

breeding,

it is difficult

to obtain

F2 seeds

interspecificcrosses.

An alternative

is to obtainF2 seeds from the cross between cultivatedand wild varieties
increaseof the same species.

genetic

variationas pure lines for further

Subsequentprior to selection.
testingfrom Fi plants,

for mungbean anduse of mutagenesis

Mutants carrying

and use in a breeding

especiallyfromon F2 seeds cannovel traits

can be establishedof Agriculture,

ARC-AVRDC,

programs.

Acknowledgement
The above work was supported
Kasetsart

University

and the Thailand

by Thai Department

Research Fund.

References
Ahn, C. S. and R.W. Hartmann. 1978.
radiata(L.) Wilczek]

1974.

AVRDC
Centre,.

AVRDC

Development

Centre,(Wild.)

between mungbean [Vigna
Ohwi & Ohashi].

J. Amer.

: 3-6.

Development
1990.

hybridization

and azuki bean [V. angularis

Soc. Hort. Sci. 103(1)
AVRDC.

Interspecific

Progress

Shanhua,
Progress

Shanhua,

ReportTainan.

AsianVegetable

Research

and

1989.

Asian

Vegetable

Research

and

86p.

ReportTainan.

1973.

350p.

217

Fig. 3 Mutants found in V. radiata x V. radiata (w)(a) albino seedling(b) dark green leaves and early maturity (c) semi-dwarf plant(d) extruding pods (e) waxy leaves (f) narrow leaves (g) variegatedleaves (h) wrinkled

Barber,H.N. 1970.

Chen,

H.K., M.C. Mok and D.W.S. Mok.

Chen,

Hybridizationand evolution

organogenesis

from interspecificPlant

9 : 77-79.

Cell Rep.

hybridof plants.

Taxon.

19 : 154-160.

1990.

Somatic

embryogenesis

embryos

H.K., M.C. Mok, S. Shanmugasundaram
hybridization

between

Vigna radiata

of Vigna glabrescens

and D.W.S. Mok.

and V. glabrescens.

Theor.

leavesand shootand V. radiata.

1989.

Interspecific

Appl.

Genet.

78 :

641-647.
Chen, N.C., L.R. Baker and S. Honma. 1983.
Vigna food legumes.
Gamborg,

Euphytica

O.L., R.A. Miller

culturesof soybean

crossability

among four species

32 : 925-937.

and K. Ojima.

root cells.

Interspecific

Exp.

1968.

Nutrient

Cell Res.

50 : 152-158.

requirementsof suspension

of

218
Gosal,

S.S. and Y.P.S. Bajaj.

1983.

Vigna radiata

embryo culture.

through

Gupta, P.K. 1996.
Kim (eds)

Mutation

Breeding

Recent Advances

IIPR, Kanpur 208024,
Harlan,J.R. 1976.

Knoblock,

Genetic

I.W. 1972.

Ngampongsai,

S.

Rick,
Steel,

hybridization

3 : 217-229.

Bangkok,

specieshybrids

1980.

McGraw-Hill,

633p.

Inc., USA

N., C. Lairungreang,

The Subgenus
Present

Seminaron Plant

Beijing,

China.

R.P. 1963.

of Pulses

Crop Sci.

16 : 325-333.

Thailand.

of Slides

Mutation

Research,

Taxon.

21 : 97-103.
(L.)

Wilczek]MS thesis,

Graduated

Thai with English

abstract)

inprovement.

and Procedures

Proc.

XVIII

of Stalistics

Int.

(2nd ed.).

Y. Egawa and C. Thararasook.

1991.

Show, pp 1-24. In Mungbean and the Genetic
Chai Nat FCRC. Dept. of Agric,of crop mutation
Breeding

1 04 p.(infor vegetable

Principles

Ceratotropis.

statusThe Cultivation

plants.

of Mungbean [V. radiata

P. Nakeeraks,

Summary in the Style

1993.

Society

Using Embryo Rescue Techniques.

R.G.D. and J.H. Torrie.

Wang, L.Q.

In AN. Asthana and D.H.

Indianof crops.

in flowering

Hybridization

University,

Hort Congr.

Resources:

Vigna mungo and

32 : 129-137.

pp. 124-136.

in wild relatives

Interspecific

Exploiting

Research

White,

in Mungbean,

resources

CM. 1967.

Tomooka,Euphytica

betweenIndia.

Vigna species

Kasetsart

hybridizationin Mungbean Research.

Intergeneric

1997.

and Related
School,InterspecificThailand.

breeding

in China,pp.

ll-19.

In Proc.

for Regional

NuclearCooperationin Asia,of Animal and Plant Cells.

Ronal Press, New York. 228p.

219

Questions
Questionsand answers: Session

3

to Dr. Ishimoto

Q. Given that the genes for a amylase inhibitor,
expressed, how do you eliminate

arcelin and lectin are closely

linkedand often co-confounding effects of the action of individual

proteins?

(Gepts)A. The functionof the proteins

method. Bruchids
purified

are usually

analysed

are feed on the artificial

by the bioassay

using the artificial

beans made of susceptible

bean

bean flour and theproteins.

Q. Do any arcelinvariants

represent

an evolutionary

advantagein the wild commonbean?

(Debouck)
A. There is a selection
bringspressure by bruchids

in some areas. In these places the presence of arcelinadvantage to the wild commonbean, a Amylase inhibitor

is present

in mostcommonbeans. Wedo not know the full story of how the a amylase inhibitor-lectingenefamily evolved.
Q. You mentioned co-evolution

between Phaseolus

evidence showing that distributionof allelicvariants

of arcelin

A. Weand other researchers

or density

distribution

Do you have anyof weevils is associatedfound the insecticidalof these

and weevil.

with the frequency

etc.(Morishima)

amylase against Zabrotes subfasciatus.
development

species

defenseof Z subfasciatus.

effect of arcelin and the inhibitory
The pressure by Z subfasciatus

proteins.

migratedof

may result in the

But we have no information

The bruchids

activityof primarywith humans around the world.

Questions to Dr. Fujii
Q. Is the character for differencesin allelopathic

A. As I am not a genetic scientist,

I do not know.

Q. Could you tell me the biosynthetic
A. Bio-transformation

activity

inherited?(Ishimoto)pathway for the secondary metabolites?

of L-DOPA is the most interestingand easy way. L-DOPA is converted

fromtyrosin by tyrosin monooxygenase. By introducing
the dream of making an L-DOPA (allelopathic)
Q. Does hairy vetch have a toxic effect on all species

this enzyme to other crops I haverich plant.
in the fruit orchard?

strong, is there any inhibitory

influenceon fruit tree growth?

A.Hairy vetch shows species specificactivity.

It suppresses

but has little(Ishimoto)If the toxic effect is(Shimamoto)the growth of Compositae

species

effect on Graminae. So, in the field, graminaceous weeds will dominate after

hairy vetch application.

But in combination

with the effect for light, fall seeded hairy vetch

220

can suppress

springto early summer weeds almost completely.

Late summer weeds

emerge again, so at least one weeding is necessary. After removal of hairy vetch there is
no residual

effect.

There is no measurable

differentroot systems.

Q. Which is more allelopathic

cultigensor their wild relatives?(Morishima)

A. I have not so much experience with cultivars.
allelopathic

activity

cultivars

hi my experience wild relatives

compared to cultivatedshow a wide range of diverse

types show strong activity

affect on the fruit trees because of there

species.

In the case of azuki beans, traditional

activities.

This means some wild or traditional

but some show very low inhibitory

Q. Have you looked at L-DOPA allelopathic

have a stronger

activity.

effect in Lathyrus sativus, as it has high levels

of L-

DOPA? (Maxted)
A. I have checked the concentrationacid,

P-cyanoalanine,

Lathyrus

varietyof L-DOPA in Lathyrus. It contains

which causes lathyrism.

with little

another unusual amino

In India they have successfully

bred aof this unusual amino acid.

Q. What is the toxin in Viciafaba

surely it is not L-DOPA as it would then cause favism inhumans? (Maxted)
A. Favism is also caused by an unusual amino acid cc-cyanoalanine
Q. Why did the pears not die from the allelopathic

and their relatives.(Maxted)

effect of Vicia villosa

being grown underthem?
A. Usually

there is no inhibitory

activity

because of the differences

reportunder Vicia villosa,in a strong drought,lightdrought

Vicia villosa

villosa

is said to reduce the soil temperature

conditions,in root systems. There is a

Vicia stands show water shortage,conserves water. But the coveringbut

effect2 to 3°C. So sometimes floweringof V.

is delayed

5 to 7 days in the case of pear or grape.
Questionsto Professor Peerasak

Q. In addition

to bruchids,

bean pod borers (Maruca spp.), commoncutworm etc., are importantpests for mungbean cultivation.

Did you detect mutant(s)you plan to screen your mutant(s)A. I have noticed
insects

to those pests?

are complex (they included

Q. Is there any possibility

beetles

and worms). I cannot identify

insects.

of using interspecifichave identified

hybridizationin disease

Since thethe resistanceor broad spectrum. We are planning further studies

and his colleagues

Or dofor those pests?(Egawa)the waxy mutant that had less damage from leaf feeding

to whether it is specific

A. Dr. Miyazaki

resistantason this topic.

and insect breeding.

a wild mungbean (V. radiatavar. sublobata)?(Vaughan)

221
ace. No. TC I 966 resistant
had a backcrossprogram in Thailand

gene was involved)
from rat feeding

to bruchids

to our cultivated(Callosobruchus
to incorporate

maculatus and C. chinensis).
this resistantWegene (luckily

only one

mungbean. However, weare still waiting

for results

tests to be more sure that the resistant

substance(s)are not harmful

tomammals.
Q. What are the main causes for incompatibility

among azuki bean group species

in the subgenus

Ceratotropis
A. There is no definite

answer to thisquestion.

Even within

the Vigna group(epigeal

germination)

if we cross between V. radiata X V. mungo, wecan obtain seed only whenV.radiata

X V. mungo, we can obtain seed only when V. radiata is the female parent but

not when it is the male. When V.mungo is the female parent the pollen
cannot germinate

from V.radiata

to reach the egg cell of V. mungo. Thus there seem to be several

mechanisms involved.

Closing

Remarks

225

Closing

remarks

Makoto Kumamoto
Director,

Liason and Coordination

Council Secretariat,

Ministry

Division,of Agriculture,

Agriculture,

Forestryand Fisheries

Forestryand Fisheries,

Japan

Research

Ladies and gentlemen, at the close of this workshop I would like to say a few words onbehalfof the Ministryof Agriculture,

Firestryand Fisheriesof preserving

genetic

resources

The importanceattention

as traditionalvarieties

The Ministryof Agriculture,

the National

Instituteare replaced

research related to biodiversity.

sustainability
priorityand bioscienceon the conservationof conservation
collaboration

support joint

researchunderstanding.

including

The Japanese

resources is that it is of basic

importanceto agriculture,Thus the Japaneseand utilizationof genetic

Government has placed a highresources.

technology

transfer and capability

will improveof genetic resources. The Japanese Government has initiatedThis International

here today. We also

germplasm using state of the art technology.

on international

workshops

severalwith this in mind. One example is in-situ conservation

with Chile and Vietnam which has been presented
to characterise

andwas started in 1 985. Since then adevelopment.

Weplace high priority
have been holding

research institutes,

and studied.

Overseas Development Assistance projects
research

international

have been accumulated

It is our hope that, through collaboration,in the field

increasing

and rain forests are lost.

(MAFF)The MAFF genebank projecton genetic

Secretariat.

Resources, have been active in both conservation

great number of germplasm accessions
Governments position

is receivingby modern varieties

Forestry and Fisheriesof AgrobiologicalResearch Council

like thiscollaborationin the field of genetic resources. We

as a part of our effort

to improve

international

Genetic Resources Workshop is the 7th in the series.

It is my duty to close this the indoor part of the workshop. The efforts of all participants
is greatly

appreciated.
Tomorrowa field trip has been planned

for the last day of the workshop. I hope that in

the more relaxed atmosphere outside the seminar room, linkages
countries

represented

here will be strengthened.

The indoor part of the workshop is closed.

among the peoples

Please enjoy.
Thank you very much.

of the many

226

Photographof workshop

Front row left to rightparticipants:

HirokoMorishima,

DanielDebouck,

Shimamoto, Paul Gepts, Ramanatha Rao, Peerasak Srinives,
Kalyani Wasala, Faithi

Devra Jarvis,

Hassan, Maria Teresa Federici Rodoriguez,

Oo, Maria del Rosario Josefina

Fumio Kikuchi,

Castro Munoz;

Duncan Vaughan, Kiyoharu Oono, Shinichi

Takanori

Kitamura,

HitoshiNaoki Katsura, Indra

Hakim Kumiawan, Than Htay

Middle row left to rightSato, Keisuke

YoshiyaNakagawa, Shoji Miyazaki,

Masao Ishimoto,
Yoshiharu Fujii,

Yoshinobu

Sakamoto,

H. P. Bimb,

Egawa, Kouichi

Kadawaki,

Kasuto Masuo, MunSup Yoon, Kooji Doi,

Akito Saito;

Back rowleft to right

Norihiko

Tomooka, KunihikoAbe, Nigel Maxted, Tomotaro Nishikawa,
Kaga, Kazuto Shirata,

Kato, Akihiko

Michiko

Yoshino, Jun

Hironobu Jinno, Toshio Koj ima, Shintaro

Iguchi, Akito

Tsukasa Nagamine, Nobuya Kobayashi.

Fujii,

MAFF International
Workshop on Genetic
Resources
13th-15th
October
1999 Tsukuba,
Japan
Theme 1 Wild legumes
Theme 2 In-situ
conservation

research

List of ParticipantsList of ParticipantsAbe, Jun

Hokkaido

University,

Asanuma, Shuichi

JIRCAS,Japan

Bimb, H. P.

Nepal Agric. Research

CastroUniv. Nac. Agaria, la Molina,

Munos, Maria del Rosaria

Chibana,

Takashi

CIAT, Cali, Colombia

Ebana, Kaoru

NIAR, Japan

Egawa, Yoshinobu

JIRCAS,

Rodriguez,

Maria Teresa

INIA, Las Brujas, Uruguay

Akihiko

Hokkaido

Fujii,

Yoshiharu

NIAES, Japan

Shuuichi

University,

University

Harada, Kyuuya

Chiba University,

Hautea, Desiree

University

Hisada,Migihiko

NIAR, Japan

Hoshino,TsuguoJIRCAS,

Japan

Iguchi,Shuntaro

Ibaraki

University,

Jarvis,

Masao
Devra

Japan

NIAR, Japan

Gepts, Paul

Ishimoto,

Lima, PeruJapan

Fujii,

Fukuoka,

Center, Nepal

NIAR, Japan

Debouck, DanielFederici

Japan

of California,

Davis, USAJapanof the Philippines,

Chugoku Agricultural

Los Banos, Philippines

Japan

Experiment

Station,

IPGRI,

Rome, Italy

National

Plant Gene Bank of Iran, Iran

Jinno, Hironobu

Hokkaido

Prefectural

Kadowaki,

NIAR, Japan

Javardy,

Firouzeh

KouichiKaga, Akito

NIAR, Japan

Kato, Kunihiko

NIAR, Japan

Katsura, Naoki

NIAR, Japan

Katsuta,

Masumi

NARC, Japan

Kawabe, Susumu

NIAES, JapanPGR Center, JapanJapan

Kikuchi,

Fumio

Tokyo Agricultural

Kitamura,

Keisuke

NARC, Japan

Kobayashi,

Hiromi

NIAR, Japan

Kojima,ToshioKokubun,Makiei

Ibaraki

University,

JIRCAS,Japan

University,

Japan

Japan

Kumamoto, Makoto

AFFRC Secretariat,

MAFF, Japan

Kumiawan, Hakim

Research. Institutefor Food Crop Biotech.,

Matsuo, Kazuto

NIAES, Japan

Maxted, Nigel

Birmingham

Miura, Kiyoyuki

MAR, Japan

Miyazaki,

Shoj i

NIAR, Japan

Morishima,

Hirokoformerly NIG, Japan

University,

Nagamine, Tsukasa

NIAR, Japan

Nakayama, Hiroki

NIAR, Japan

Nakayama, Shigeki

NIAR, Japan

Nishikawa,

NIAR, Japan

Obata,

Tomotaro

Taro

JIRCAS,

Okuno, Kazutoshi

NIAR, Japan

Oo, Than Htay

Seed Bank, Myanmar

Ookawa, Masao

NIAR, Japan

Oono, Kiyoharu

NIAR, Japan

Potokina,

N. I. Vavilov

Institute

IPGRI-APO,

Malaysia

S

aito,

Elena
Rao, V.

Japan

Akito

JICA,

Sakamoto, Shinichi

Hokkaido

Sato, Takanori

NIVOT, Japan

Shiina,

NIAR, Japan

TsugioShimamoto.

UK

NIAR, Japan

Okada, Kensuke

RamanathaIndonesia

Yoshiyafor Plant Industry,Japan

Hokkaido

University,

University,

Japan

Japan

RussiaShirata,

Kazuto

NIAR, Japan

Srinives,

Peersak

Kasetsart

University,

Tomooka, Norihiko

NIAR, Japan

Vaughan,

NIAR, Japan

DuncanThailand

Wasala, Indra Kalyani

Plant Genetic Resources Center, Sri Lanka

Yamada, Toshiaki

NARC, Japan

Yamamoto, Akio

Agricultural

Structure

Yoshino,Hokkaido

University,

Michiko

Improvement Bureau, Japan
Japan

Acronyms
AFFRC Secretariat

Agriculture,
Forestry
Secretariat(Japan)and Fisheries

Research Council

CIAT

Centro International(Colombia)

de Agricultura

Tropical,

INIA

Instituto Nacional(Uruguay)

IPGRI

International(headquartersIPGRJ-APO

International
Plant Genetic
Pacific Office (Malaysia)

JIRCAS

Japan International
Sciences (Japan)

MAFF

Ministry(Japan)of Agriculture,

NARC

National

Agriculture

NIAES

National(Japan)Instituteof Agro-Environmental

NIAR

National(Japan)Instituteof Agrobiological

NIG

National

Instituteof Genetics

NIVOT

National Research Institute of Vegetables,
Ornamental Plants and Tea (Japan)

de Investigacion

Agropecuaria

Plant Genetic Resources Institute
Italy)
Resources Institute- AsiaResearch Center for Agricultural
Forestryand Fisheries

Research Center (Japan)
Sciences

Resources(Japan)MAFF International

Workshop on Genetic Resources Series

1. Root and Tuber Crops
2. Soil Microorganisms
3. Animal Genetic Resources:

Efficientconservationand effectiveuse

4. Plant Genetic Resources: Characterizationand Evaluation:
NewApproaches for Improved Use of Plant Genetic Resources
5. Diversity

and Use of Agricultural

Microorganisms

6. Genetic Diversity and Conservationof Animal Genetic Resources:
1. Improvement and conservation of genetic diversity
in livestock.
2. Current status and perspectivesof effective use of sperms.
7. Part 1. Wild Legumes*
Part 2. In-situ conservation*Publishedseparately

research*

Editors
Editorin Chief:

Kiyoharu

OonoManaging editors : Duncan Vaughan
Norihiko

Tomooka

Akito Kaga
Consulting

Published

Editor:Shoji

MiyazakiMarch 2000

Research Council Secretariatof MAFF and National InstituteofAgrobiological
Resources Kannondai 2- 1 -2, Tsukuba, Ibaraki 305-8602,
ISBN4-931511-03-1

Japan

The Seventh
Ministryof Agriculture,
International

Forestryand Fisheries(MAFF),

Workshop on Genetic Resources

Part2

In-situ Conservation

National

Institute

Researchof Agrobiological

Resources

Tsukuba, Ibaraki,Japan

13th-15th

1999

OctoberSponsored by

Research Council Secretariatof MAFFandNational

Institutein cooperationof Agrobiologicalwith other MAFF institutes

Resourcesin TsukubaJapan

In situ Conservation

Proceedingsof an Internationalwith additional

PrintedPublished

Research

invitedConference held from 13th to 15th Octoberpapers.

in Japan by Asahi Printing
by National

ISBN 4-93151

1-05-8December 2000

1999

InstituteCo., Ltd., Tsukuba

of Agrobiological

Resources,

Tsukuba, Japan

Preface
On the second day of the 7th MAFF International
NIAR, Tsukuba, Japan. In-situ conservation
Six papers were presented
several internationally
University,

research was the topic of papers and discussions.

and Dr. Morishimabroaden and deepen the proceedings
respected

Genetic Resources Workshop held atgave a summary of the six papers.

of the workshop on this topic,

genetic resources scientists.

Hammerof the Universityof Kassel and his colleagues

As a consequence we have organised
papers into three sections.
second part consists

globalof papers that deal with theory and practical

The third part provides

diverse case studiesof particular

the topic of in-situ conservation,
literature

will provide fresh perspectives

on the topic of in-situ conservation.

and invited

perspectives.

The

aspects of in-situ conservation.

crops and wild relatives

It is hoped that these papers, several from scientists

Karla paper.

at the workshop
and regional

fromof Reading

Japan and Professor

all agreed to contributethe papers presented

The first two papers provide

papers were invited

Dr. Barbara Pickersgill

U. K., Dr. Katsuei Yonezawa of Kyoto Sangyo University,

In order toof crops.

that have not previously
and add usefully

writtenon

to the body of

CONTENTS
Preface
Globaland Regional

In situ conservation

Perspectives

Perspectivesof diversity

withinabout in situ conservation

field

crops: is this necessary
B. Pickersgill

and/or feasible?
3of wild relatives of crops in Latin America
D. Debouck

19

Theory and Methodology
In-situ conservation strategies
in population
genetics theory

for plant species:

some comments based on the recent advances
K. Yonezawa

A methodology

Checklists

for genetic

and in-situ

43

reserve conservation:
Case study for Vicia
N. Maxted

conservation- case report from Italy
K. Hammer,T. Gladis,
and P. Perrino

83

H. Kniipffer,

G. Laghetti
11 1Case studies
Research on Vigna subgenus
conservation

Ceratotropis

genetic

resourcesthat provides

insights

into in-situ

strategies.
D. Vaughan, N. Tomooka, A. Kaga, K. Doi and
M.S. Yoon

In-situconservation

researchon buckwheat(Fagopyrum

129

spp.)

in NepalH. P. Bimb

In-situconservation

between Chile

researchon wild relativesof tomatoes

151

in Chile:

Collaborative

research

and Japan.
T. SatoSpacialand temporal

aspectsof genetic

the Red River Delta, Vietnam, revealed

variationin landraces

165

of aromatic

rice populations

inby RAPD markers
S. Fukuoka, T. D. Suu, L. N. Thrinh,
T. Nagamine and K. Okuno

187

Closing

remarks
H. Morishima

Photographsof participants

List of participants

and paper authors

203

206
209

Global and Regional

Perspectives

In situ conservationof diversity

withinfield crops:is thisThe Universityof Reading,necessary and/or feasible?
Barbara Pickersgill
Departmentof Agricultural

Whiteknights,

Botany,School

P.O. Box 221, Readingof Plant

Sciences,

RG6 6AS, UK

Abstract
Diversity
regional

in crop plants occurs on regional,

scale involves

adaptation

effectively

adaptedare to be utilisedby the many distinct

agricultural

systems.

and new agricultural

populations

environment. It is

in field

and their effects

but poses problems

ex situ.

types of particularlosses,techniqueschanges also affect diversity
field

or conserved

Changes, and possibly

and natural selection,

components of the regional

conserved by local cultivators,

represented
materials

Diversity

to biotic and abiotic

valued and usuallyaccessions

community and field scales.

Diversityon the community scale is
to fit distinct

niches in local

in such diversity

are inevitable

as new planting

and as market forces change.

The actions

both on existing

have not been adequately

when regionallycrops maintained

become available

populations.

on thestudied.

of, and interactions

between, humandiversityand on generating novel diversity

Material

which growers no longer wish to

maintain may need conservation

ex situ.

maintain them most appropriately

for those purposes, need re-examination if ex situ collections

be morereadily

utilised

These

However, the purposes of ex situ collections,inand how to
are toby plant breeders.

Introduction
Duringboth national

the 1960s

and international,

eggs out of whichIndia

a feeling

thesewere in danger of killing

successes

to become self-sufficientgrew that the successes

had hatched.

the more variable,

subsistence

farmers,

which had proved valuable

Epidemics

such as that of southern

to confirm

that diversity

low, at precisely
were disappearing.

corn leaf blight

among modern cultivars

the time when traditional

cultivars,

and Indonesia

lower-yielding

breeding

programmes,

the goose that had laid the golden

High-yieldingin wheat production,

said to be displacingof plantin the Unitedof small-scalefor use by plant
StatesUrgent programmes were therefore

with their

set up to collectattendant

were
and

breeders.

in the 1970sof major crops was becoming

crop populations,

had helpedin rice production,

crop populations

sources of alleles

which

seemeddangerously
diversity,

extant diversity

fromfarmers and conserve it off-farm,
providein ex situ gene banks.

a resource for plant breeders,content of these collections,of maintaining

The original

but there are now increasingraison d'etre was to

doubts about whether theand the extent to which breeders are using them, justify

the costs

them.

The United Nations Conference on the Environment and Development, held in Rio deJaneiro in 1992, directed
Signatories

attention

to the Convention

to in situ conservation
on Biologicalfor crops as well as for wild species.

Diversitycommitted

situ conservation

before there was any clear idea of what should

shouldAs a result, basic

be done.

research,

manage programmes for in situ
implementation
1997).

conservation,of management decisions,

The lack of basic knowledgeof diversity

in fieldprogrammeof variations

foreseen or monitored,

forresources

is present

disadvantages

in environment and/or in agricultural

to growingof particular(for

generallyin situ conservation

allelesand possible.

in general,suggestthreats

to and needs for conservation

whether conservation
Diversitytherequiresin the firstpractice

Decisionsanplace.

may then be

have to be madebe sited and how many sites arearticulated,

especially

farmers asked to allocate

crops by traditional(for use by plant breeders)toscarcemethods).

Is theor conservationof co-them)?

The advantages

and(listedin Table1), and the literatureonconsiderable

confusionon this point.

widespread,with different

scale, the community scale and the fieldon each of these scales,tactictraditionalcited for in situ conservation

diversity

feasibleshoulduse by the farmers who developed

Many crops are geographicallyat the regional

withand Hodgkin,of diversityof such programmes need to be clearlyof land and labour
genotypes

on how tothem (Hawtin

and maintained

to pay for them in cash or kind (for example,

purpose conservation
adaptedconcurrently

on-farm conservation

and action taken if necessary

The purposes

those expected

place

in

acute with regard to on-farm conservation

about where programmes for on-farm conservationnecessary.

or how thisdecisionsrather than precedingof the reasons why the diversity

The consequences

be conserved

underpinis takingis particularly

to support

crops.

An effective
understanding

which should

themselves

insofarscale.

as it is possibleof diversity

ex situ, in situ or a combination

types and levels of diversity

This paper attempts
to separate

at each of theseto consider

them; to examine thelevels;and to assessof both is the most appropriate

or mostfor each.

on a regional
Particularly

scaleclear examples of diversity

on a regional

scale occur in the belt of Africa

Table 1. Commonly cited advantages
conservation of field crops

2 .

and corresponding

Disadvantages
Enforced preservation of primitive
agricultural
systems is unacceptable
and impractical(Ingram and Williams,
1984); diversity
conserved on farm isoften uncharacterised
so requires
evaluation before it can be utilised.

In situ conservation is dynamic andpermits continued evolution (Qualset

Evolution involves change, which
includes loss of diversity(Smithson

et al., 1997; Bellon et al., 1997; Lenne
and Wood, 1999), including
evolutionand Lenne, 1996;

W

itcombe,Lenne and Wood,

1 999), hence an inherent contradiction
with the objectivesof conservation.

1999).

Pressure

Larger populations
can be conservedin situ than ex situ. These are lessliable to genetic drift or loss of genetic
integrity
through hybridisationorhuman error (Brush,al.,1997).

4.

of in situ

Advantages
Samples from heterogeneous field
populations
are difficult
to maintain exsitu but more easily maintained under
traditional
management (Vaughan andChang).

of locally adapted types of immediate
value to local farmers (Qualset et al.,
1997; Wood and Lenne, 1997;

3 .

disadvantages

1991;

Qualseteton farmers' land and/or timemeans they may no longer be able tomaintain all the variants previously
grown but are bulking them into
composites (Vaughan and Chang,1992; Cleveland et al., 1994);
preventing access by farmers to newhigh yielding
cultivars is unacceptable(Witcombe, 1999), but introduction
may result in hybridisation
with local
types (Bellon and Brush, 1994; Wrightand Turner, 1999).

Fanners and/or national programmes
control access to their genetic

Access by plant breeders at national
international
institutes
to genetic

resources (Qualset
et al., 1997).

resources
physicallyet al., 1997;

Bellon(Brush,maintained in situ may be
and/or politicallydifficult1991).

orbounded by the Sahara desert to the north and the tropicalforests

Bunting

the start and duration

defineand Pickersgill,
the length

shortest

1996).

Throughoutof the growing

average growing

this region,season, hence the potential

mean lengths

the linesof latitude.

This range of environments

maturity

in localunpredictable.

yieldssouth.

forms predominate

the region,

the datetimes toin the north, longer-of onset of the rains

In the north, the date of the end of the rains is also unpredictable,

wetter south the rains end, in most years, withinso thatto each other and tois matched by a range of inherent

Short-season

Throughoutof the rainsin the south,of growing season run roughly parallel

crop populations.

1975;

from the crops grown. The

seasons occur in the north, and the longest

zones with different

season forms further

to the south (Bunting,the same 1-2 week period.

isbut in theThis occurs when

the days are shortening.
Throughout
the rains

the region,

have begun.

photoperiodic

farmers start to sow their cereal crops as soon as they feel thatIn the wetter

south,

the first

forms, which respond to the shortening

date in every year.

This date coincides

sowings,

days by floweringwith the periodof the available

water stored during

are of

at much the same

when the rains usuallywhenever these forms are sown, they will use the full length
will mature after the end of the rains (using

in most years,

end. Thus,

growing season,

the rains in the soil profile),

and will avoid damage to the grain by the fungi and insectsof the wet season.

In the morearid

farmersa range ofnorth,

where the end of the rains

is not predictable,

photoneutral

forms, each of which flowers

at a fixed (but different)

therefore

earlier,i.e. adapted

node number. Some areto a shorter growing season, than others.

the farmers choose, from the range of planting

material

maintain

which

When the rains begin,

they maintain,

those forms

which they feel will make the best use of the season to come.
In addition

to theselarge-scaleregionaldifferencesin average

season, there are potentiallysubstantial

variations

betweenseasons and groups of seasons.

The farmers'

againststoragemain defenceof surpluses

producedthe faminesthat may follow

in more favourable

years.

fill

until

in a single

good season, and so survive

production

systems

maximising

output in all years.

resistant"
years.

cultivars,

are geared to securing

but locallyIn the wetter regions,bumper harvests

So they grow, not specially
adaptedpopulations,these are usuallyfor 3 years.

isThey canthe next big harvest.

in favourable

Their

years, rather thanbred non-photoperiodic

which can give large yields
photoperiodic,

seasons

So the Dogon of Mali maintainby a family

storesof growing

unfavourable

stores which are large enough to hold all the grain requiredtheselength"droughtin favourable

because such populations

cantake full advantage
Implicationsof long seasons in which the rains begin early.

for conservation.

These traditionaltypes

of sorghum and pearl

environments of their particularthem. In the so-called

regions,

are certainly

and are valued for this adaptation"conversion programme", breeders

from sorghums for the southern belt of West Africa.
cultivars

milletcould not produce heavy yields

adaptedby those who growremoved photoperiodic

The resulting

to theadaptation

short-season

photoneutralin the longer growing seasons of favourable

years,

because the plants flowered,and their grain matured, too early.

Moreover, the grain matured

under unfavourable

conditions,and insect

damage was more severe.

surpluseson which the local people(wet)

so that fungal

These cultivars

not only failed

to provide

rely for survival,

but also often produced little

There are many similarintendedexampleshave found such cultivars

the periodic
usefulgrain and so were rejected.

where those for whom improved cultivars

inappropriate.

The International

tend to release elite breedinglines rather than finishedcultivars,

added

types

to regionallyadaptedin national

breeding

backgroundgenotypesthat confer regional

adaptation.

are alreadyeffectively

conserving

parts of theirregional

value, without

thoseInstitutes

thereforenow

so that desirable

traitscan beprogrammes without
crop genotypes

the need of formal on-farm conservation

concerned with adaptation
In a global

theirbackgroundbreeders'

lines.

context,

utilisation

genotypes

conditionsof regionallyadaptedthat are of direct

programmes.

genotypes

in whichthey occur.

growing

conditions

different

difficult.

One may therefore

the genetic

integrity

is often more importantat the location

ask whether the emphasis
is misdirected,

for adaptation

usefuloften requires
alleles

For breeders

Moreover, conservationfrom thoseof accessions
diversity

alleles

As long asuseful elements ofaccessions

be broken up and only the generallyof usefulthe

do not appear to be threatened.

This often imposes delays and difficulties.

germplasm, conservation

ex situ collections,

to regional

losingThis is turn means that local farmers

farmers are allowed their own choice of what to grow, the agronomicallydiversity

were

that

transferredtoworking with exotic

than conservation

ex situ of accessionsof theadaptedtoof the gene bank is notoriously

in gene bank practiceand whether attempts

on maintaining

to maintain,

to a diverse

range of exotic

types

a crop being maintainedin global

environmentsaremisplaced.
Diversityon the community scale
There are many examples of diverse

withingiven community: up to 12 types of maize per farmer in Chiapas,

Mexico (Bellon

withinand Brush,a1994),

50 distinct

types

of potato

in a single

community in the Andes (Brush,

than 100 types of cassava among the Jivaro of Amazonian Peru (Boster,Some of this diversity
for labourat planting

is preserved

and harvestingin order to spread potentiallyover a greater"hungry gap" which occurs when productsof earlier

1995),

more1983).
limiting

requirements

periodof time and to minimise

theharvests

have been consumed but the

new harvest has not commenced. For example, the Mende of Sierra Leone in West Africa
have maintained distinct types of rice with short, medium and long times to maturity since atleast the 17th century (Longley and Richards, 1993).
Short-season rices are planted at the startof the rainy season and are the first to be harvested.

However, the bulk of the harvest comesfrom mid-season rices,which are planted

after the rainy season is well-established.

long-season

rices are plantedin the valley

and floating

growing season is longer, and are harvested
A surprisingly

similarsystembottoms

Finally,

and watercourses,

where theafter the main crop.

to that of the Mende is developing

in a different

hemisphereand over a much shorter time span. The Tawahka, who live along the Rio Patuca

in southern

Honduras, acquired

importantcrops,

althoughlabouramount that can be grown.
yielding

This has drastic

agroecologicalon its known large yieldniche.

Instead,the upland fields
blocks.

years andare mostly sown

These mature at different

farmers in the Andes of South America, at altitudes
householdet aL, 1990).

may maintainas much food as possible1997),of different

with potatoes

to the diet (Quiros
If one planting

Farmers plantis destroyedby frost,

in different

areas.

1997).

Similarly,

a mixture of types with different
so that some yieldflavours,

et aL, 1990).

fieldswhere potato

as many as 26 morphologicallyThis is done to obtainuses adding1997),

in favourable

dwarf rice

the labour of harvest over a 2 month period.

(Ortega,(Ortega,withinshallow.

the year as possible

field

are maintainedin dry years, because its roots are relatively

an individualinteresthigh-fields,Amongsubsistence(Quiros

includingIn upland

types of upland rice, grown in separate

times, thus spreading

types,

is higher.

sowed it in an inappropriate

is a staple,

ll different

Rice Research Institute,

where the water-tablefor farmers who gambled

with 3-4 distinct

that

the

Most farmers have learned that dwarf rice does best in lowland

along the river margin,consequences

It has now become one of their mostland and sowing the crop often limits

records

dwarf rice bred by the International

may fail completely

types

for clearing

House (1997)

the community as a whole.
fields,rice only in the 1950s.

for as much of

textures,tubersdistinctcolours

andof each type inanother may survive (Ortega,levelsof frost resistanceis grown in eachwill be produced

from every field.

Modern cultivars

have been introduced,
storage properties,
Rice

but the traditional

hence are preferred

is inbreeding
selectionfor "truenessdistinct.

This selection

is imposedmaterialin the case of potato,
from their tubers

basic

principle

with

seed

characteristics

to type" will

betterandso in thesecrops1997).

vegetatively,

sufficeflavour

to maintainthe different

reports

that farmers can identify

In outbreeding

crops such as maize, the same
integrity

takenfrom the largest

ears

conform to the ideal

thatof each type,

and Brush, 1994).

selection,even when stringent,

does not prevent a continual

In Mexico, an improved

shorter-statured

Tuxpeno, the race of maize dominating

commercial productionin Chiapas,

1976.

height

The farmers say that it is gaining

situation

adaptedhas been describedmaize grown higher
slopes (Wrightto localconditions(Bellonsides

to findmaize typesofwas introducedinwith local

1994).

A similar

among offspringof crosses

maize which they grow on the valley

on the valley

cultivar

hybridisation

and Brush,flowbottom lands and local

suitablefor the intermediate

and Turner, 1999).

In potato,

the smaller

tubersZimmerer and Douches (1991)
sexually

each year and, throughin Honduras, where farmers selected

between a commercial hybrid

different

the genetic

into the system.

betterasis used to maintainfor that type (Bellon

maize, becoming

types

selection
being

However stabilisingas havingat harvest in the case of rice, and in the choice of planting

with great accuracy.

sowingof new variation

are propagated

where Ortega (1997)of stabilisingforare regarded

for home consumption (Ortega,and potatoes

stabilising

types

types

producedfrom outcrossing,are chosen for planting,suggestthat these may include

true seed. These will contributeadditionalsegregation

as in maize.

and recombination,and both Ortega(1997)

first-generation

tubers fromvariation

to the fieldandpopulation

In rice, the Mende operate a system which allows some gene flow between short, midand long season types.

According

mid-season rice are harvested
are segregatedto Longley

as possible

in maize, permits

early off-types

new short-season

which remain after the bulk of the fieldsource of seed for sowing on the bottom lands.
via pollen(1993),

in fieldsofas they ripen and, if not used as extra food in the "hungry gap",

and tested in small trial plots

The late off-types,

and Richards

allelesis harvested,

planting

material.

are gleaned

as aThis gene flow via seed, like the gene flow

to reach the populations

in which they will be favoured by

selection.
Implications

for conservation.

These and other studies

of different

crops, with very different

reproductive

biologies,

10

fanned by peoples
settings,

of different

cultures,all show that subsistenceabout matchingon different

order to make best use of their limited

niche for it, but they do not abandon their tried

lost through(Brush,1991,

Louettesuch replacement

farmers or to plant breeders.
to demonstrateagriculturaland trusted

et al., 1997).

of planting

material

witha presumably

introductions

and completely

irrevocableof faba bean (Vicia faba)

declinesin the cultivation,pulse

crops (tarhui(Lupinusconservation.

if they perceive

until

somethingmutabilis)

assumed that any diversity

or quantitative

studies

displacement

was introduced

replacedtheir

to localhave been doneof a crop species

fromto the Tawahka of southernsinak bean (P. lunatus)(House,1997),

Similarly,

to the Andes and commonbean to Africa have led tothe diversity,and cowpea (Vignamaize seems largely

cannot always be foreseen

Rather than attemptaclearly

to be of value either

vulgaris)and presumably

much of the wetter part of Africa,
Such situations

in observing

loss of that part of the gene pool of P. lunatus.

drastic

millets.

to them inwould not be easy to conduct.

Commonbean (Phaseolus

Honduras in the 1950smaterials

is unlikely

A greater cause for concern may be the complete
a culture.

systems

It is implicitly

However, no qualitative

this, and such studiesof the land available

Most farmers are interested

into theirecologicalbecome very knowledgeable

attributes

resources.

and will take such material

better is offeredand in different

farmers have, of necessity,

crops, labour and the agroecologicalnew material,

continentsof the locallyunguiculata)

domesticatedrespectively).

to have displacedsorghum and theand are not always appropriate

to compel or induce farmers to continue

which they no longer value, it may often be betterOver

for in situ

to grow crops

to conserve such replacedor displaced

resources ex situ.
Diversityon the field scale
Many collectors

populations

grown by subsistence

many intermediates,
1973);

have been impressedin single

farmers:

fieldsmore than 10 morphologicallyand Chang, 1992);

mixturesof Africa1992);(Voss,

Honduras (House 1997);

by the diversity

10-12 differentof tetraploid
distinctthat occurs within

inflorescence

wheat in Ethiopia

types,

together(Porceddu

types of rice grown together

the field
withand Perrino,

in Sabah (Vaughanof 6-29 types of common bean grown in the Great Lakes region

an average of 9 typesof cassava per fieldamong the Tawahka of

5-30 morphologicallydistinctin a single

types of potato

fieldin theAndes (Brush et al., 1981).
Theory and experiment
the target population

both suggestthat if selection

continues

in the same direction,

should become more uniform as the less fit components

are eliminated.

ll

In artificial

mixturesof equal proportions

for 90% of the surviving
1968).

In synthetic

plants after 3 generations

mixturesbecame dominant within
as rapidly

same genotypes

consistently

counter the effects

of natural

subsistencefields,eitherof natural

selection

selection,

as Smithson

the dynamics of these situations(Jennings

accounted
and de Jesus,

competitive

types similarly1992).

Since diversity

does not disappear

naturalselectionmust fluctuate,

not favouring

theor gene flow must intervene

toout, most studies

inor both.
and Lenne (1996)

point

taken over a limited

periodare lacking

or 2-row and 6-row barley,

grown in the highlandsof Eritrea.

Accordingpopulationof time.

and even anecdotalof wheat and barley,of the field

cultivar(Voss,

over time, or human selection

systems are snapshotscompositionof rice, a single

of beans, one or a few particularly

a few generations

as this in farmers'

Unfortunately,of 5 cultivars

Sustainedstudies

evidence is limited.

Mixturesor other barley-barley

to Adhanom (1995),

has shiftedof diversityofcombinations

after

are

about 5 years, thetoo far from what the farmer considers

desirable

and the mixture is recomposed,

often using seed stocks brought in from areas which

specialise

in maintaining

seed of distinct

Lakes region of Africa,and supplying
homogenising

effects

greater extent by human intervention.
crop is sown.

These

mixturesnamed types.

of natural

A speciallycontainselection

In beans in the Greatare counteracted

composed mixture is createddifferent

components

to an eveneach time thein different

proportions,dependingon whether the beans are to be grown during the long rains or the short rains, or onless fertile

or more fertile

soils (Voss,

1992).

Newvariants may be incorporated
after trial

and evaluation

variants(Trutmannin isolated

et aL, 1993;

into these diverseplots

to determine

Thurstonfield populations
the appropriate

et aL, 1999

for beans).

but usuallyplacesonly

for the new

In one indigenous

community in the Upper Amazon of Peru, the shaman collected

types of cassava from outside

the community and maintainedwith volunteer

giftsof the ancestors")(Salickand evaluated

Bellonet aL, 1997;

involves

selection,others.

It is therefore

than ex situ.

Qualsetof in situ conservation
et aL, 1997,

both human and natural,also recognised

is that it permits

Lenne and Wood, 1999).
which willthat diversity

Moreover, as Lenne and Wood (1999)

of useful characters

togetherseedlings("theet aL, 1997).

A much-cited advantage(e.g.

these,

continued

evolution

But such evolution

favour some alleles

may be less securely

and disfavour

conserved

in situpoint out, the rate or extent of evolution

on-farm is unknown.

Some guidance

as to what may occur in the heterogeneous

populationsof subsistence

12

farmers

comes from studies

1990).

Composite

on barley

Cross H (CCII)

composite

cross populations

in California(Allard,was developed

from intercrosses

among 28 cultivars,

which differ

in resistanceto Rhynchosporium

secalis(leaf

blotchor scald).

However, thepopulationof the pathogen

on CCII contains

allelesfor virulence

matching

each of thesealleles

for resistance.

Five races of the pathogen

damaging.

Most of the 28 parents

generations

without

resistantconscious

to all three races.

of CCII

predominate,

three of which are moderately

are susceptible

to theseselection,

the most frequent

Resistanceto a fourthparent, and has remained rare in the population.
years, and alleles
disadvantage
alleles

for resistancegenotype

to the fifth

present

This race is prevalenteffects

maintaining

race has also not become fixed,

in some but not all

selection

withinapparently

for similarreasons. Only

accessions

wild barleyplants,are resistantto all 5 of these races (Allard,resistancecould easily

be found among plants of the late generations

new alleles

have evolvedin this population,changedand they have recombined

variable

population

co-evolving

resource for plant breeders
is stored

and reassorted

than traditional

However, lines withof CCII.

No

alleleshaveof the existing

into novelwith a pathogen

and less than 0.1% of1990).

but the frequencies

genotypes.

thus often constitutes

gene bank collectionsthese

the population.

1.5% of the 18,000
multipleso are at athus favours

polymorphismof barley in the USDA global collection,

wasin only one

on seed production,in years when this race is uncommon. Natural

Resistancein the population

race was initiallyhave strong negativein some years but not in others,

races but after 45

A geneticallya more attractivein which genetic

variability

in a more static state.
Diversity

in a crop is likely

to select

Zhu et al. (2000)

containing

a single

type of rice contained

only one or a few races of the pathogen,populationsof blast

isolatedfrom fields

containing

non-glutinous

hybridrice (lesssusceptible)

among rows of hybrid

the second year of the project,
its yield

in monoculture

project,

interest

townships

yield

that

populations

mixturesrice, the severity

Mixturesor multilines

decreased

rice in the mixtures

were no longer needed.

had expanded to an area of more than 40,000
and Lenne, 1996;

from fields
whereas(susceptible)

and
rice

so much that, by

was 89% greater than

By the thirdyear of theas an experiment on fields

of 5

ha.

of other crops have been grown under mechanisedin other parts of the world (Smithson

in theWhen rows of glutinousof blastfrom farmers was such that what had started

isolatedof glutinous

were more diverse.

of the glutinousand fungicidesof rice blast

diversity

pathogen.

were interplanted

foundfor or conserve corresponding

Zhu et al, 2000)

agriculture

but seldom on such a

13

scale.

Interestingly,

glutinous

cultivars

Zhu et ah (2000)
produced little

that, if the developed
have to reacquire

foundthat some mixturesof glutinous

or no control of blast under local conditions.

world is to revert to growing heterogeneous

knowledge

fieldof how to compose mixtures for optimal

and non-This suggests

populations,

it will

results

under differing

calledthe dynamics

conditions.
Implications

for conservation.

More studies

are needed of what Smithson

diversity,

to establish

what

responsible

for maintaining

aspects of traditional

selective

diversityand Lenne (1996)

pressures,in field

by farmers

populations.

and/orSuch studies

environments,

whichmanagement need to be conserved in order to conserve diversity

in situ,

We do not know whether re-creation

annually,

regionas in the

Great

Lakes

every 5 years, as in Eritrea,

components of a mixture.
evolution

as on-going evolutionof Africa,or re-creation

imposes a significant

If it does, then diversityof new and possibly

better-adaptedbarrierof bean mixturesof cereal

mixturesto gene flow between

may be conserved in these mixtures,types may be hindered,in the much-criticised

ex situ gene banks.

are also

neededto document

whetherthedemonstrated

in the composite

crosses of barleyof charactersof novel(Allard,1990)

describe,with implicit

separately

the 10 or more components of the rice mixture grown by her parents,

so wasmaintaining

insteadsuggestthat thetypesof bluea bulked composite.

desirable

Similarly,

Cleveland
togetherof these cases is it clear that ceasing

lead to a loss of alleles.

(1992)

a rice farmer in Sabah who was no longer able to maintainof North America are combining

maize. In neither

Vaughan and Changison-farm in heterogeneous

Hopi Indiansof crops.

evolution

duplicatedregret,populations

but

in much the same way

is said to be hindered

More studies
combinations

aremay establish

and which may be less important.
approximatelyofOn the contrary,

new genotypes by reassortment

et al. (1994)

previously

distinct

to conserve genotypes

composite

populations

will necessarily

may lead to evolutionofand recombination.

Conclusions
The adaptations
environments
thosetherefore

to obtain

farmers.

internationalalreadyclimatic

regimesthatenablefarmers

in marginal

some harvest from their crops in most years are vitally

Local

breeding

local value.

to regional
farmers

evaluateand often

programmes as inappropriate

effectively

conserving

reject"improved"

for their particular

to locallyimportantcultivars

conditions.

those parts of their crop genotypes

This should apply to resistances

importanttofrom

They arethat are of direct

pests and diseases

as well as

14

to adaptation

to climatic

variables.

view "exotic"

accessions

as among the least

farmers are best met by in situ conservation
breeders

by ex situ conservation
The ideal

may involve

scenario,collectionof alleles

allelesvarieties,

interest.
its

unquestionably
too long.
likelyadaptedgenotypes;

needs of plant

fieldsprogrammes are tending,programmes in any relevantin localparts ofraces of the crop wherever

improved local types which in the future may be re-collected
and thence national

breeding

programmes.

such as occurs when landraces

and

This

would

are replacedby highto the extent that they themselvesof plant breeders.
of the community or field,

there

may be conflicts

and the conservationist's

existing

current

maintenanceimportantfor both in situ and ex situ conservation,

Most studies

suggestdiversity

diversity

between the farmer's

perception

Studying

to maintain

often

for use in ex situ "pre-breeding"

could then be incorporatedof what is in his interest,

promotingof locallyand would also allow farmers to benefit,

At the levelglobalNeeds of local

of elite lines to national

local genotypes,

wish, from the activities
perception

materials.

from farmers'back into international

yielding

starting

breeding

the world.

prevent the loss of entire

desirable

to which many multinational

by release

they are of value, producing

locationsgenotypes).

of material

Desirable

working in different(not necessarily

programmes, followed

re-cycled

However, breeders

at the community or field

and its possible

futureof what is in thelevel,

replacement

and the factors
or erosion,isand has been neglected

for

that, given freedom of choice in what they plant,that they regard

as valuable,

unless or until

farmers are

somethingthat they

regard as "better" becomes available.
Changesin traditionalfarmingtechniquesfarm. The Tawahka of Honduras are not linked
neither
agriculture

draught

animals nor tractors

eventually

may cease to maintainto cultivate

may affect the diversity

by road to any other communities
or harvest their rice fields.

comes to this part of Honduras,
all the types

harvest will no longer be limiting,of uplandso it will

maintained

no longer be necessary

and useWhen mechanised

the Tawahka (if theyrice that they currentlyon-possess.

have survived)
Labour forto spread the harvest

over

several weeks.
Changes in market forces
countries,

rural populations

may also affectare drifting

to the cities

on-farm diversity.

In many developing

and urban markets are becoming valuable

sources of income for farmers who have surplus production

for sale.

Althoughfarmers of theGreat Lakes region of Africa prefer to grow mixtures of beans with seeds of diverse
samplesof a single

colour fetch

a higherprice in the market (Voss,

1992).

colours,

Voss (1992)15
therefore

advocates

introducingof a single

colour.

Diversity

even though the highlyand currently

into the farmers' planting

material

more components of seed

for traits

such as disease

resistance

may thereby

be maintained,

diversity

in seed colour, noticeable

to collectorsof germplasm

visibleused by farmers

is, in effect,

advocatedfor various crops in developed

countries

characters

subject

selectionby consumers(qualitytime to maturity,etc.) but diverse

in other characters.

germination,

Human-aesthetic
They are reported

in maintaining

human aesthetic

choicecultivars

and Lenne, 1996):

uniformcharacters)inor growers
information.

in sorghum in West Africa and inwhere breedingis the basis

existing

akin to thosehere for lack of adequate
diversity

the Nuba mountains of the Sudan. In livestock,controlled,(Smithson

reasons are not considered

to be significant

of multilinein their

may be lost.

to strongdevelopment

components

mixtures,(synchronousThis

as markers for the different

can usuallyof much, perhaps

be more precisely

most, of the diversity

between breeds and land races.
Changesshouldin traditional

agricultural

systems

are inevitable,

but whether

be to turn certain farms into museums to conserve in situ the diversity

with or without

incentives

to the farmers involved,

accessby subsistence

farmers to improved

lucrative

markets

to justify

is difficult

improved

Furthermore,

introductionsin Mexico and Honduras show, attempts

evolutionin much the same way as occurs when accessions
Ex situ conservationconservationnecessarilyof genotypes

to prevent

may need to emphasiseor accessions.

need to be in perpetuity:or potentiallyas the examplesof maizechange may retard further

are conserved ex situ.
of diversity

conservationof alleles

ex situconservation

Furthermore,

re-collection

found,

Preventing

techniques,

may compensate to some extent for losses

However, ex situ conservation

currently

is much more questionable.

material,

morally.

the response

may be more useful

on-farm.
ratherthan

may notthan long-term

maintenance of any accession.
Some of these conclusions
to further

work. A significant

both crops in the field
crops, cultures

may appear heretical,

body of data, obtained

and farmers'

and climates,

conservation

to be clearly

implementing

it.

practices

is urgently
establishedbut heresy is often a usefulover several years from observations

as they affect those crops, and including
required.

and satisfactorystimulantThis would enable

diversethe need for in situ

recommendations

put forward

Acknowledgement
I am gratefulto Professor

A.H. Bunting for extensive

discussionofand criticalcommentfor16

on successivedraftsof this paper.

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19

Perspectives

about in situ conservationof wild relativesofcrops in Latin America
D.G. Debouck
Genetic Resources Unit, Centro Internacional
COLOMBIA e-mail: d. debouck@cgiar.

de Agricultura

Tropical,A.A. 671 3, Cali,org

Abstract
In situ conservation is often perceived as the methodology of choice for the conservation of genetic
resources of wild relatives
well as of inadequateof crops. This paper reviews specificof Neotropicalmaize, and tomato. This review indicatesdemography and genetics are often overlooked,

these populations,conservation in situ, as

documentation, poor coverage and current threats for wild populations

crops with examples in beans, cotton, cucurbits,
population

cases of effective

perspectives

inand their consequences for long-term survival ofand their use in crop improvement programs. This review suggests what can be done in

the short-term, namely as priorities
waste land, such as roadsides.

in inventories,

Technical

small-scale

refuges, agronomical practices

and uses ofand socioeconomic issues of these choices are discussed.

1. Introduction
Plant
agendas,conservation

mostly since

was being eroded

is a relatively

1950.

that increased

topic

to review, as records
learned.

Neotropics,

earlier

were sounded (Harlan,

breeding

lessons

Slightly

recentfood supply

and that thisin this1975).

thatis precisely

Mesoamerica, Species 2000,

This

and population

are being collated,

It is indicative

issue

on the public

species

Conservation

the first alarms that plant variability

erosion

was a result of the success of plant

growth. Plant conservation
methods

is conserved,

thus is not an easy

experiments

carriedout andwe don't know yet the extent of plant

heritagein thesuch as Flora Neotropica,Flora

the purpose

developed,

of projects

etc. This means that the objectsof plant

and international

century

A clear goal is however often agreed upon, when considering
perspective.

national

genetic

resources

for conservationplant

are still il defined.

conservationmeans that the diversity

from a geneticof genes in plantso that they can continue to exist and evolve. From a resource perspective,

it

20

meansthat conservationof all options

about a plant biological

heritagethat has been, is and could

be used in the future.
Conservation

can be achieved

evolve under abiotic

stressesin originaland selective

conservation

is known as in situ conservation

plant species

and their populations.

most important
botanical

quality,gardens,pressures

where natural populationsof diseases,

and aims at preserving

For some plant species,

conservation

genebanks,

habitats

can also be achievedarboreta,

livingcollections.

conservation.

The fact that these

two groups of methodologies

been widely

discussed

1991;

conservation

strategy

shouldconsiderboth methodologies

change or new market opportunities
one methodology
Table

open up, agencies

etc. Thispotential

facilities

In this case access is desired

evolution
1991)

the evolutionary

ex situ, in special

thus continuing
Shands,predators,

toofnamely crops, for which access is the

germplasm with fixed properties;(Hawkes,pests,

continue

is not particularly(Tableare indeed
1).

desiredsuch asto known
in ex situ

complementary

A comprehensive

has
plant

and in a dynamic way. As land uses
may orient conservation

efforts towards

or another.

1 - Conservation

methods

for plant

biodiversity

Item sP u rp o se

ex situ m eth o d o lo g ies
C o n serv ation an d access to k n ow n
g en oty p e s

in situ m eth o d olo gie s
P res erv ation o f ev o lu tion a ry p o ten tial

O bjec t"crop s"; o rth o d o x see d g e rm p lasm

all m aterials; an y k in d o f p ro p ag u les

K e y k n o w led g e
fie ld s

P h y sio lo g y ; g en e tics

E c olo gy ; p o p u lation g en etics

W ay sG en eb an k s, field an d in v itroco llectio n s

P ro tected areas, farm e rs ' field

L im it ation s

S m all am ou n ts; lim ited n u m b e rs
ev olu tion froz en

A v a ilab le lan d , ch an g in g lan d u ses
low co n trol o v er m aterial/ d ata

A d v an ta g es

A ccess to germ p lasm , ev alu atio n s

R o b u stn e ss, n o v el v a riatio n

H u m an re s ou rce s

S p ecially train e d

S p e cially train e d

C o sts : le v el/ ty p e

H ig h ; d ire ct

H ig h ; in d irect

C om m itm en t

co n tin u in g

co n tin u in g

21
Commitments
nature,towards

as recognizedplantbiodiversity

by signatories

conservation

would

to the Convention

howeveron Biologicalbe of continuing

Diversity(Anonymous,

1992).

2. Purpose

and some implications

The preservationof in situ conservationof evolutionary

potential

presumes a few basic assumptions.

one hand, plant populationsof a given species

have size and multiplication

rates high enough for their long-term survival

dimension)(Nunney and Campbell,

which viable

propagules

are spread periodically.

the creationof variablity,

genetic

load(Ledig,1986;

so that populations

can maintainsome levelof heterozygosity,and fitness

for long-term

survival(perspectiveof plant

Nevo, 1998).

Such levelsof genetic

diversity

Yet it is unclear

1994;

Soule and Simberloff,

Hamrick and Godt, 1996;
On the otherenvironments

hand,

are in place.

mutationsselective

are eliminated,outbreaks

pressures

imply the maintenance of original
the presence

As microorganisms

diversity

key parameters

for survival

also

selection

important(Attenborough,existence

1995;

processes

pressures
Darwin,wipinghabitatsof plant

by boththespeciesbioticworking,(Avise,and abiotic

so that lethal

out the evolutionaryoras mayplant

species,thesewith enough space (Gilpin,

1987;

Gilpin

For wild

sometimes the presence of symbionts.

have often a much higher

multiplication

rate comparedto match virulence

genes by pathogens

are

1989;

Pollinating

agentsarein plant

populations

as they favour
Jain,

authors

processand diseases,

and Jarosz,

is needed;survivalof synecology).

1984).

Keen, 1992).
certainvariants

A time dimension

is a condition

for theof most directuse (Plucknett

et al.,

landraces

selection

as the sources of variabilityof crop species

do not escape

may differ from those of wild plants.

by rules of plant demography
all steps

1858;

to address

be effectivelyof populations(Burdonallow plant populations

and work of all these mechanisms.
Often considered1987),of pests

causing diseases

size and genetic

should(perspective

genetics)

1986).
causedbut withoutwith disease1986),

to long-term

These pressurespopulation

how much diversity

pressures

happento plants,

demographysystems that allow

is detrimental

and Soule,(plantThey also have breeding

agree however that inbreeding

deleteriousthat is, theymechanisms byenvironment.

They have effective

themselves,

multiplication

any change in their1993).

are able to maintain

On theof the selection

and population

processes(Harlan,selection

processes

but these

The survivalof landraces

is also conditioned

genetics.
1992).

Humans however interfere

Additional

space throughat almostploughing

and

22

planting

might be offeredWith relay cropping
which specificsubmitted

to specificor differing

landraces

to biotic

with growing

landraces,

planting

as living

and abiotic

conditions:thus expanding

patterns,

plantsstresses

humans also change periods

are subjectedin differing

unfavourable

to selective

seriesenvironments

Plantssuch as droughtor early or late frostsorselectionby keeping'useful'variants,

others.

Another action by humans is in bringing

genes, while discarding

speciesthatotherwise

creationof novel diversity.

favour a particularetc. Humans also take an activethat is allowing

would not be in contact,
This

fruit

process

are thusof space and time. Humans interfere

by irrigation

theirof time during

pressures.

can be mitigatedprocesses

heating,

the size of their populations.

part into thethem to reproduce

sometimes resulting

and spread

togetherplant

in hybridizations

andcan be conscious

when farmers or their families

colouror flavour(Heiser,1988).

It can also be unconscious

humans unintentionallyintroduce

plantsor pestsduringtheirlead to the maintenanceof thousands

otherwise

have most likely

variants

are crucial for their availability.
In traditional

responsibility

disappeared.

for maintaining

for the conservation

systems,

farmers

have, apartthousands

of useful variantsof farmers as keepersof the global

Merrick,

1987;

1995;

perhaps

unwanted situation

diversity

and Alcorn,in theirin the Andes.

While

culturalvalues(e.g.

plantsraisedagriculture?

Apartdemography

and population

implications

that

Ford-Lloyd

and Jackson,

long-term stability
traitsand contributecontinue

genetics,

1986).

pressuresof landraces

farmers often deal

exemplified

by the verticalindustries,

farmingwasneeds, extending

to socialandIs that role recognizeddealing

with

there

are several

social,economic

in the society(Bellon,1996;

one aspect that should

is not merely the plantingof plant

and cultural
Brush,

1995;

be consideredis thewhat modern breedingof quantitative
may be selecting,

However, these biologicalof time make a relevant

in modernissuesthat may result in novel combinations

to future crop evolution.

andin part from amainlyAmong others,

need to act over a long period
conservation

purposes).

In that sense, farmers anticipate

significantlyrole resulted

and society's

taken(Altieri

implications,

to be debatedof selection

and polygenes.

1987).

In the Neotropics

food

That unique role

been highlighted

farms were long 'primary'for ritualfrom technicalof crop species.

This

has

that wouldfrom producing

farm environments,

almost the only way to cope with all farmers'

processover time of all thesehas rightlyof a premium on diversity.

with an broad ecologicalland exploitation

agrobiodiversity

OldfieldThis

for a couple of crop species

Conditions

farmingBrush,of plant variantsmigrations.

whencontribution.

of some diverse plant items.

mechanisms
Surely,in situ23

In plant genetic
case (Harlan,

1978;

resources

for food and agriculture,

Hawkes, 1977).

On the one hand, duration

systems might still

be too short to isolate

cultivatedderivates.

Althoughbreadwheat, rapeseed),biologicallyor when the crop is itself

This means that many wild species

derivedcrop relatives,

and that a relatively

both groupsof plant

wild relatives

may represent

material(Beebestill

under cropping

wild relatives

in the cases of ploidy
an artificial

the most extensive

Quirosbasic

et al, 1992).

variation

construct(i.e.

systemswith their

possible

betweenfree exchange of genes is still

et al., 1997;change (e.g.

breeding

form open biologicalfrom their

On the other hand,ever accumulated

in a cropand be needed in future crop improvement programmes as well for the evolution

such specieson their own. Several of the benefits

have been documented (Old field,

1989;

currently

many possibilities

for extended(Tanksley

and McCouch, 1997)

exist

it is not worth conserving
commerciallines.

requirementsof wild

difficult(Ingramwide hybridization

sources of variability

speciesgenomesand ecologicallythere are specific

Given this context,

mechanisms of genetic
it is worth asking

in relation

protection

schemesand theconservation

problems they present.

insteadhighofspecific

ex situ conservation

reasons,
diversity

namely climate
at work for these

how far wild relativesof crops area few examples from Neotropicalcrops, andto existing

pertinent

one wonders if

in form of wild relatives

1984).

the generator

distribution

and re-buildingof crops often make their

In addition,of crops
In addition,and relatives

and Williams,of1983).

of genes through transformation,conserved in situ. We will consider

examine theirand Prescott-Allen,

The enormous amount of variationof materials.

effectively

to be drawn from wild relatives

Prescott-Allenand transfer

the widest

change, to try to maintainkinds

alreadyof crops are a specialof evolution

and geneticallythis has happened

triticale).

species,

wild relatives

protected

documentation.

areas,

We will

their
next

inclusion

into

examinea few

3. Examples from the Neotropics
Beans
The genus Phaseolus
domesticatedspeciesfrom still

are short-lived

understory

livingmay have 50-55
wild forms (Debouck,perennialsof pine or oak forests

with tuberousof Central

eastern USA, and the Andes. There is anotherwith epigeal

germination

wild

and fibrous

species,
1999;

five

Debouck,of which
2000).

have been

The majorityofor woody roots and spread or climb in theAmerica extending

to the southwesterngroup of wild species

root systems, that generally

andthat are annual plants,

colonize

open habitats.

The

24

following
latterthree examples

are about the first group of species,

and the following

group.
P. maculatus is a component of the grasslands

Sierra Madre Occidental
1983;

Gentry,varietiesof northwestern

1957).

This short-lived

in these grasslands.
that1978).

P. mallis,

drought

Populations

distributed

from the foothills

has a tuberous

taprootare now being quickly

ploughedsuch as bean, barley,

and triticale

outsidelava bed land frequentan endemic of the Galapagos

archipelago

and covers several m2
to crops having modern(Debouck,in this region(islands

Wigginsand Porter, 1971)

and is possibly

considered

as protectedin this nationalpark of Ecuador. Goat eradication

be a priority

as wild beans are particularly

is a floristic

western Panama (Delgado

some populations(Guatemala),Parque Nacional

continuing

threatforest(Davis1985).

From our own fieldwork,of thisspecies:(Costato establish

et al., 1997).

is a desert

Mexico (Delgado

Salinas,Arizona to Baja California

Thisde las Minasmaize fields

and vegetablespeciesneed to befrom Chiapas

does not survive

toprotected

Biosphere

Reserve

La Amistad(Costacropping

systems is athe destructionof itsephemeralof the Sonoran Desert and Baja California

Shreveand Wiggins,1964).

Its range

Sonora (Buhrow,1983).

A few populations

1985;

and coastal

includedin the following

National

Park (Texas),

California

Sur).

protected

Vizcaino

Free grazingof native habitats

Argentina

areas:

cattleas a wild species,(Debouck,

Resistance2000).

have beenof this species

SaguaroNational

Park (Arizona),

Big Bend

Reserve (limit

Baja California

Norte andand goats are a potential
flowering

cropping

is widespread
The wild

is from southern

Populations

and opportunistic

by irrigatedof1959).

Desert Biosphere

may escape by its earlyP. vulgaris,

attributes.

SierraParque Nacionalfrom the trans Pecos in Texas (Turner,

Luis,plantsthe followingRica),

reportedreplacement

invasive

habitats.

P. filiformis

species

In addition,in the park should

et ai, 1995).

Chirripo

Rica and Panama). Deforestation

Fernandina,of the Lima bean, could becomponent of the pine-oak cloud forests

Salinas,areas harbour

original

vulnerable.

such as feral guayava (DavisP. tuerckheimii

a relative

personal

are at risk.

of Isabela,

San Salvador;

controlledof theMexico, to Puebla and up to New Mexico (Buhrow,perennial

These grasslands

withstandobservations,Baja

two are of thehazard,and pod setting.

systems is a problem

althoughinsects

is a useful trait present

only in populations

Mexico and at low frequency(Osborn

et al., 1986).

for improving

TraitsthisSonora.

Mexico to San

form of common bean has several

to bruchidbe

The completein coastal

from southern Chihuahua,

might

positiveof western

photosynthesis25
might be present

as well (Lynch et al., 1992).

already

at risk, namely in the surroundings

Rica),

Bogotapractices.

(Colombia),

Introduction

all detrimental(Beebebut

alsoof herbicides,

From our own fieldwork,of Latin American cities

due to the modifications

et al., 1997).

A few populations

Manantlan (Jalisco,aresuch as San Jose (Costaof traditionalchange of corn varieties

to wild and weedy populations,many populations

agronomical

and abandonment of fallows

areas they sometimes grow in the cultivatedare included

Mexico)(Benz et al, 1990).

The American speciesof cotton include

in protected

plots

areas such as the SierradeCottongenome)and 12 diploid

distributedin the Galapagos

in La Libertad,
deciduousPeru;

forests,wild

speciesIslands,

all otherthickets

4 tetraploid

(DD

wild

genome)(Fryxell,

and weedy forms (AADD1984).

G. mustelinum in Pernambuco,

American wild species

and dry scrub (Fryxell,

G. klotzchianum

Brazilisand G. raimondii

are in Mexico, mainly in subtropical1979).

There are several endemic species

in Mexico with limited

and/or a poorly known range: G. armourianum on San Marcos Island,

Baja California

1982),(Parry,G. gossypioides

around Guaymas, Sonora (Fryxell,

1979).

in northern

Oaxaca (Fryxell,

None are currently

included1993),

in protected

G. raimondii(DD; 2n = 26) is distributed

in a few westward flowing

Libertadin northern

Peru (Simpson

It offers(Follin,1986).

projects,

and by weeding1988).

It could

It is not currently
G. thurberi

Mexico (White,

et al., 1988).

1948).

areas.
valleystolerance

to Verticillium
withbe threatenedby encroaching

agriculture

practicesagainstwild Malvaceae

and feral cottons

includedin any protected(DD; 2n = 26) is distributed

G. turneri

of La
fungusmajor

irrigation(Simpsonet al.,

Arizona and northern

Sonora,area.
in southernIts seed has no lint;

however, it confers

resistancetopart of its range in Sonora could be affectedbyuplandcotton (Harlan,1976).

A significant

irrigation

agriculture

and ranching

projects(West,

fibre

breakage

1 993).

Cucurbits
Cucurbita
have a cultivatedC. ficifolia

in deciduous

and indehiscent;forests

to Lakeare short-lived

by rodents,

it seems that Okeechobee

Okeechobee

in Florida(Andres(Nee,1990),

five of whichhave not yet been establishedperennial

or in arid zones. Fruits

seeds are dispersed

bees. Among all wild gourds,
restricted

13 species

form. True wild forms and crop ancestry

and C. moschata. Cucurbits

open habitats
skinnedis an American genus with perhaps

vines, generally

sprawling

are relatively

large to large,

and flowers

pollinatedgourd is the rarest:
and Nabhan,

1989;

for
in
hard-

by solitary

its habitat
Walters

is nowand26

Decker-Walters,1993).

There has been some discussion

US Endangered

Species

Act because of its relationship1989;

and Decker-Walters,

Walters

1992).

as to deny its protection
to C. martinezii

C. radicans

under the(Andresand Nabhan,

seems to be another elusive

endemic

from the Eje Volcanicoof Mexico, and C. galeottii

if a valid

speciesseems to be an endemic

from Oaxaca (Nee, 1990).

None of these species

is currently

includedin protected

species,C. foetidissima,

has been proposed

doing

well in Aridoamerica,

althoughnew agricultural
Chihuahua.
mightdevelopments

as a new crop (Gathmansome of its populations(plantingsof barley,

In South America, C. ecuadorensis

triticale,areas. One

and Bemis, 1990),

have alreadybeen affected

fruitsbycommon bean) in Durango andseems to be a true wild species,

have been used at some time (non bitterisin some populations!)

althoughit1990).

It(Nee,

seems to be related

to the complex C. andreana- C. maxima complex (Wilson

et al., 1992).

Given its multiple

resistance(Provvidenti,1990),

than presently

revealedvery valuablegenetic

to viruses

severelysquashes

resource. Its range might be more extensive

herbarium records(Cutlerand Whitaker,

highly(Dodsonand Gentry, 1991).

endangered

affecting1969).

Much of the vegetation

it is abyof western Ecuador isMaize
The genus Zea has, apart from Indian(subdivided

into 3 subspecies)

not been much exploited
1982;

Reeves,and 3 wild species

commercially1950).

has been carriedof teosinte

are at risk or becoming
and Valle

documentationout recentlyTheteosinte

populationsof changes

Guatemala,are also

at risk

becauseplantations)(Wilkes,

1988;

Wilkes,coccineus

and teosinte(Benz

populationsof teostinte

extinction

has occurredal,1986).

Tomatoand P. vulgaris;
et al.,

1990).
duringWild forms of maize havepotential

is recognised(Naultof locationand statusof all teosinte(becauseof changes

in croppingof suburbs

systems)

Hondurasof(Sanchezand Jutiapa,
cane

The Biosphere

Reserve Sierrade Manantlan

inof Zea diploperennis

harbours

Sanchezchecks;in landSome populationsof the extensionof southern

et al.,and sugar

its buffer

field

teosinte

uses (pastures1972).

for the conservation

Phaseolus

Chalco(because

1998).

1990).

progenitor

Gonzalez et al., 1998).

del Guadiana

et al.,

established(Sanchez

extinct:

Gonzalez

Jalisco(Doebley,so far, but theirAn accurate

populations

Mexico City)

corn Zea mays, its wildzone also includes
Gonzalezgiven the soil seed bank, but carefulofprimitive

races of maizereportsome missing

et al. (1998)

it might

wild populations

be prematuremonitoring

to conclude
is necessary

their(Iltiset

27

The genus Lycopersicon,
al.9 1993),
1995).

has nine speciesThe

cultivatedcerasiforme,

although(Warnock,tomatosometimes considered1988).

has

They are all of Southone immediate

wild

the range of which extends from Peru throughRick and Holle,

Solarium (Spooner

American origin (Rick,relative,

L esculentum

var.

the Andes to Mexico (Rick,1995;

is an endemic

speciesfrom the dry tropicalscrub

and tropical

thicketsof the Galapagos

Islands,

well known for its toleranceand Epstein,

1976)

and its pedicels

lacking

jointsare of interest

to the processing(Rick,

It has highsolids,a traitof economic

importance1967).

solubletomatoes(Rick

et al.9 1987).

differences

exist

between

Lycopersicon

speciesdeclarednational

animals(Davisthis volume).
tolerance1993).

(Rickindustry

area, genetic

It seems to be the soledue to the invasive

et al,of Galapagos
alien plants

wasand feralChilein driedof the northern

Atacamiandesert1987).

because of its habitat,

This species
it requires

irrigation

projects

this species

is not presently

pollensolubleincluded

from other plants.

solids,of its value to the tomato industryof alien invasive

as the original

in any protected
Some populations

has expanded1990),

and Bolivia(Rick

Mexico.

The cherry

tomato has many uniqueanthracnose,

leaf spot, verticillium

a colonizer,

it could

populations

are however threatened(Iltis,1988),

and from there,by uses of herbicides

could behabitatin Peru to

into Central

America andresistanceto leafwilt and potato Y virus (Rick et at., 1987).

be seen as saved and not deserving

in anyendangered.

from its original

attributes:

andtomatoes

it is not included

and massive reforestation

are not particularly

var. cerasiforme

Peru (Brakoa key trait for processing

plant species

rocky habitats

and Holle,

is a source ofare initiated.

Colombia

weeds.

Peru and northern

is known from Ayacucho and Apurimac in central

area. Introduction

L. esculentum

southern

and xeric vegetation

It is a source of increased

In spitethe sole hazards,

tolerating

limited(Rushfor processing

landschemes as the archipelago

Not surprisingly

L. chmielewskii

protected

range.

in coastal

it is self-incompatible,

1982).

over theirYet, there are threats,of river channels

might be at risk if large-scale(Rick,

has a limited

in protection

is distributed(seeZarucchi,the archipelago

to salinity

et al, 1995).

slopesarea. Since

populations

park in 1959.

streambeds,

drought

Althoughincluded

L. chilense

Sato,

et

1990).

L. cheesmanii
deciduous

withinspecialprotection

and agricultural

mold,

fruit

As a weed andschemes. Many
practices

not

28

L. hirsutum is distributed

from southern Ecuador to central

valleysand on the coast (Esquinas-Alcazar,

wild

tomato has several

contributeto substantially

1997).

Some populations

northwestern

resistances
increasein montane humid forests.

to pests
yield

may exist

and nematodesof the central

is a sisterinter-andean

Apurimac (Rickvalleys

et al, 1976).

fruit color in tomato (Rick,
Yanachaga-Chemillen

species(Rick,

This odoriferous1982);

1979).

in

Peruanobiosphere

reservein current inventories(Daviset al, 1997).

theNoroesteof L. chmielewskii,

and McCouch,are green;

Park in Pasco,access problems

that would protect

however, it may contributesuch as eucalypt

would represent

but inventories

this species,ofalso found in montane dry scrubs

Due to its range, some populations

National

also

tomato (Tanksleyof Peru, from Amazonas to Cuzco throughIts fruits

it canin the cultivated

Peru; however, they are not reported

L. parviflorum

4. Future

1981)

Peru, in the inter Andean

Huanuco andto intensity

could be included

are lacking.

ofin theIts habitat

have

however, reforestation

with exotic

speciescan be drawn, startingwith the amount ofa hazard.

needs and prospects

From the above examples,some lessons

protection:
a. A full survey of populations

for wild relatives

inventoryof all wild

flora in the Neotropics

surveys

only exist

for teosinte,acaule, S. demissum).
Ch. et al,of crops is badly

is unlikely

and perhaps

to be achieved

a few wild relatives

Some surveys can be considered

many wild species{Capsicum,

Cucurbita,soon. Such exhaustiveof potato(e.g. Solarium

as advanced (e.g. commonbean: Toro1989),

Gossypium,

Ipomoea, Lycopersicon,

Manihot,remains a lot of work to be done. Herbaria

more than genebanks

are the repository

information,

sometimes herbarium

documenting

geographic

herbaria,

this information

1966;

cacao: Cuatrecasas,

distribution1964),

relativesof crops is lacking.

throughelectronic

limitedHeliconia

information

is publishedof populations

is not always readilyb. For many if not all protected(e.g.

of potato:

since a complete

Hawkes and Hjerting,species1990;

neededand orchids).

species,areas in Latin America,
How many protected

ornamentals,

When existing

Persea)monographs.

thereof thisBecausetask of taxonomists(e.g. avocado:

in

Kopp,

often needs updating.

means? In many cases, floristic

to timberis not the primaryavailable.

this information

as genericbut for the otheran inventoryof populationsof wild

areas have that list updated

and available

inventories

or fragmentary"flagship"

speciesare non existentsuch as Podocarpus,

Puya,

29

c. Can the number of protected
For many populations
protection,areas for populations(many species?),no documentation),the present

and protectionof wild relativesof crops be increased?

situationthat of res nullius

is basicallyhumans and introduced

livestock).

A quick analysis

relativesof Neotropicalcrops shows that their

existing

protected

relativesin toto and that of those included

areas. The matching- that of populations

bufferaspecttheexistenceor economic

surroundings.
incentivesof vernacularchanneled

Local village

future but mightof wild

relatives

for agrobiodiversity"ib choh":pepper,by the Chiriguano

Indians

radicansby the Otomi Indiansof Chuquisaca,

Bolivia;

Mexico).

Manantlan'

is perhaps

the onlyestablishedto preservea wild relative

mentioned

above, there might be many more opportunities,

in relation

to centers of plantshouldnot always

possibility

for Latin

be of a large

today very large acreages
Parque Nacional
several wild

perhapsof land is becomingof cassava (Killeenof Neotropical

protected

anymore because it is almost

vegetationin the northern Andes, will be protected'scenic'

reason to set aside

that has been initiallycoverage

et al., 1997).

inLand set aside

with the exceptionof Brazil,

to protect

a recent noteworthy
that holds

exception

theis theamong other crop relatives

1998).

In many cases, such as

and highlands

vegetation

cannot begone. Rather, we would like to see a dense network of

Due to water shortages,of potatodeIndeed,in lowlands

reserves.

to save wild relatives

Reserve 'Sierraof crops.

and Schulenberg,

dry foreststherelative

remote;small-scale

help

often recognize

many of which are discussed

area in the case of wild

variants

studiesof C. baccatum chili

The Biosphere

endemism of IUCN (Davisdifferent

immediateof crop - maize. Given the insufficient

Noel Kempff Mercado in Bolivia

relativesin theirand "yutmu" for Cucurbita

area in the Neotropics

American countries,

may

rat Lima bean, by the Mayas offor Capsicum microcarpum, a wild relative

detailcommunities

in situ. Linguistic

Yucatan, Mexico; "quii-rai"

protected

where to createof crops could help to raise economic

show however that rural communities

of Hidalgo,of wild

not always be aware ofof crops

conservationof crops (examples:in currently

ones and/or adopt other schemes inbecause wild relatives

names of local floras

of wild relatives

areas - will indicate

communities.

in the immediate

importanceThis is unfortunate
if properly

presence

partners

tocases of wild

inventories

in protected(noinaccessible

are not consideredof bothconcerns local(i.e. place

populations

and where to expand existing

be the best conservationpassiveof the above mentioned

new parks and reserves,
zones. This later

is simply

it is likelythat more paramos, the high altitudein the future. These protectedareas might

as well(Debouckand Libreros

Ferla,

1995).

Everyland for conservation

shouldalso integrate

aspects

dealingwith30

economicplants,communities

as thiswill

reinforcereasons

for conservation

towards

authorities,

can be designed

from different

perspectives:

throughcharacteristics(Jonesand the general public.

This

increasein protection

fieldwork,

interactionsal., 1997),

and life zones mapping. Actual visits

when defining

with local communities,

borders

of protectedmodels of ecologicalto the fieldseem unavoidable,

areas and buffer zones (Shafer,1990),

etparticularly

but also in order toraise awareness in local communities

in view of the future management of such areas. Once

local

the choicecommunities

important.
crops;are convinced,

Re-forestation

projects

shouldof species

also integrate

in many cases in the Andes re-forestation

impact in terms of conservationof understory

What type of protection
in contact

withmodificationin landsurprisingly,

many wild relatives

use and in the patternonce vegetation

might thus be required,or Pinus speciesof crops is needed?

Relatives

disturbance,and compositionof original

vegetation.

and populationsof cleared

returns to climax vegetation.

Periodic

et al., 1997).

Takinga cautious

quicklyaround large and mid size cities

of Latin

America should

there is likely
protected

to become more and more expensive,

areas by force (withfor ex situ conservation.
relatives

attitude,

the benefitof crops when maintained

to save 'green' areas in cities

withinthe cropping

types

land. They

disturbances

expandingfor the future!)

or to gois a threat

to somesystems. A 'win-win' approach wouldwith herbicides,

opportunity

for preserving

wild

in Latin America could be on roadsides

1997),

road networks

are increasing

would be to abandon frequent
practices
betterof roadsides
soil protection,

examples expanding

fires,

total

in almost
weeding,that would allow the survival(Arayaevery country.

continuing

grazing,of wild relatives

fewer landslides,

and higher

slightly

zones and/or raising

the buffer

et al., 2000).

or to introduce

and fallows

since

As landbut to installof management of wastelands
relativesforurban areas

annually.

prospectuse of herbicides

be to leave some edge rows, unusable corners, etc, untreated
other

Not

to determine

be monitored

there is thus little

On the other hand, intensiveaof many of them mightis very difficult(Beebeof crops havean ecologicalof which and periodicity

most speciesof

has a grim

is firstly

environments,

the intensity

is alsoof wild relativesof crops are weeds or early colonizers

succession

purposesnative plants.

humans. Agriculture

would thus survive best in disturbed
even regress

the dimension

with Eucalyptus

for wild relatives

long evolved

for recovery

Anothergreat(Allem,A win-win approachor other managementof crops and forages,

economic returns. Finally,

from the above

awareness of local communities

Iltis31

withinthem can make a difference

protected

The perspectiveof crops growingof protectedareas for wild relatives

and population

genetics,

in designing

Simberloff,1986).

of plant

populations,
harvestMolecular

is the sustainabilityof population

overlooked,

demography

and managing protected

Plant demographyof mature plants,

mortality

in or close todiversity

withinof alleles

per locus,

loci (Avise,

1994;

best

Gepts,

to establish

1987;

Soule andto assess survival

prospects(e.g.

species

relationships

measured by the presenceof heterozygosity

at certainloci,With these statistics,

populations

shouldbe protected,

shouldbe oriented,

etc. An objective

increaseof sites/

populations

and to lower the long-term
are not diverseofthe level ofthe number

conservationists

management practices

populations

and structure

the number of heterozygous

which

years of in situ conservation

et at., 1997).

of rare alleles,

about

the relevancenumber of juvenile

They can also be used to indicate

Hamrick and Godt, 1996).

decisions

Soule,

have been determined1993).

and among populations,the level

1990;

of crops.

are often completely

rates, seed bank in soil, etc) (Degreef

markers have been developed
1989;

areas (Shafer,models are being developed

once key parameterscrop gene pools (Doebley,thewild relatives

areas.
A further question

plants,for several

behindtheir

can makepriority,

these studies

howwould be tocosts (i.e. what if after

enough for conservation

or utilization

purposes?).

5. Concluding
Wildremarks
relatives

are known for most Neotropical

exceptions(e.g.

distributionof all populations

conservation

status (i.e. saved/endangered).

conservation

are often known, but information

missingsweet

potato).

However,

accurateof these relatives

is stillin many cases. The genetic

Fast progressin molecularapplications.

Awareness and training
for conservation

educationof local communities

obviously

an importantby(1988)

planning,

withknowledge
lacking,a few noteworthy

concerning

as is documentation

Basic life cycle characteristics

structurefrom the fieldis likelyof conservation
designaboutof most genepools

marker techniques

techniques

crops,

plant

has stillto leadof theirfor in situ

demographyisto be determined.

to lower costs

and widerconcerning

the use of theseand management is just starting.

Awareness andclose to protectedagencies

requiredgeographic

areas or in managing rural environments

task. There is urgency (Leakeyand Lewin, 1995),

there may also need to be an order in priorities.

isand as mentioned

While other planets- perhaps

32with no biodiversity- will stillbe there for a while,

our planet

is losingits biologicalcapital,and part of this is key to our food supply.

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andTheory and Methodology

43

In situ conservation
strategies
for plant species:
somecomments based on the recent advances in population
genetic theories
K. Yonezawa
Departmentof Biotechnology,

81-75-705-1914,

E-mailKyoto

SangyoUniversity,

Kyoto

603-8555,

Japan.

Fax :: yonezaw@cc.kyoto-su.ac.jp

Abstract
Recent advances in the theoriesof the effective

population

size and some related

topicsarereviewed to derive some general points for the optimizationof long-term in situ conservationof plant

species.

The minimum census population

for retaining

genetic

diversity

for a long-term persistence

species,and of the order

Homogenization

size necessaryis predictedof 103 in highlyin the reproductive

accelerated

effective

for maintaining

survivalselfing

contribution

or partiallyof plants

diversity.

reproducing

species.

via control of mating patternor progeny

plants

The soil

Restrictedgene flow within

subdivision

or isolation

to increases

the genetic

reproductive

local extinction

decreases

population(SeveralSmall)

strategy

with management to avoid local

Large)

strategy.

subpopulations
size.

When inbreeding
should be conducted

When a subpopulation

subpopulations.

success of subpopulations
genetic

diversity.

difficulties

a population

diversityor localsof a population

From a genetic perspective,
extinction

is preferredoccur in a subpopulation,

it should

be recolonized

will bethe effectiveby either

due to asymmetricalat a rate lower than the reciprocal

goes extinct,

with overlapping

seed bank acts to increase

size of growing plants.

whereas variable

In a population

with removal of old plants

population

by distanceclonallyof populations.

and growth of juvenile

the genetichigh

to be of the order of 102 in randomly outcrossing

number per plant helps to retain the genetic diversity
generations,a sufficiently

population
as a whole,migration

therefore,or

the SS

to the SL (Single

migrationfrom otherof the effective

subpopulation

using propagules

from multiple

Without these management measures, there will be no point adopting

the SS strategy.

44

Introduction
The importanceincreasinglyof in situ conservation

recognized.

There are limits

and ex situ conservation
Brown 2000).

freezes

The in situ
of genetic

the adaptive

resources

biodiversity

in nature, an essential

In situ conservation

diversity

evolutionof plant

for futureof crop plants

speciesplant

is important

breeding
stablenot only for the

but also

globalto retain

speciesstrategic

parameterssuch as the size, number, and positionsof conservation

regionare decided.

Whilegeographic

targetspecies

about the current

as well as the assessmentof economic, political,

for decision

theoriesand principles

designingin the second step.

Theoriesforlong-term

biologyconservation(cf.,

i.e., demographic

and genetic

Campbell1993).

Franklinhereafter,

establishedsufficiently

high

In genetics,

genetic

variability

strategies

to an ideal

population

importantworks).

biologicaltherefore

an essential

part

1988,

Soule

of animal species

conservation.

impactsofon thein the strategy

1987,

referredtheof thebasedbeen developed

and Soule (1980),

for long-term

under

two

Nunney andto as F-S (1980)of maintaining

This rule was originallybut has also been acceptedfor planningplant(e.g., Hawkes et al. 1997).

The parameter Ne is the effective

which(1980)

step, predictions

have

Simberloff

distribution

the well known, rule Ne=500 with the perspective

for the conservationconservationtakebiologicaldisciplines,

proposedwill

sites withinand socio-ethicalmaking in the firstin populationin theare chosen, and in the second, some

information

are importanta highenvironment.

first step, the target

the project(e.g.,

whether on farm or in nature, will occur in two steps;

and region for conservationisthat can be conserved ex-situ

of genes and gene complexes

factor for a healthy,

projects,

and wild relatives

to the genetic

conservationconservationof landraces

population

size, i.e., the size of population(Wright

1969;that incorporates

The genetic

diversityand structure

most of his pioneering,of organism populations

has been used as the key parameterstandardized
historicallydepend on Ne,

in developing

genetic

theoriesforsize or controllingsome reproductive

conservation.

Necan be increased
or demographic

variablesby eitherenlargingmentionedlater.

population
For strategicplanning

or decisions,

it must be45
known how management of these variables
be formulated
the species

explicitly

in terms of the variables

concerned.

of F-S (1980),
population

Ne theories

to increase

Ne. To this end, Ne must

characterizing

the reproductive

had not been well developed

developments(cf.,

Caballeroin theories,

1994,

Wang and Caballero

a new spectrum of molecularsystem ofat the time of establishment

but since then, have been extended to a wide range of breeding
structures

variability

contributes

1999).

systemsandBesidesthemethods for measuring

the genetic(Haig

et al. 2000).

and structureof populations

has been explored

These methods facilitate

the practicaluse of existing

theories

theories

for biologicalconservation.

Our files

will be enrichedby incorporatingthese new developments.

and strategies

conservation

strategies

Here, I present
relatedsubjects,

biologicaland give

population

Plant conservationin situ,some comments about theirshouldpresented

in Table 1 (Shaffer

may be classified

rejuvenatedinto three categories,

in Table 1.

time scales,

Possible
1981,

i.e., ex situ, on farm

Conservation

projects

and/orfor evolutionary

in different

demographic
categories

in a small populationin different

strategies

for the three conservation

categories

occurring in quite differentof the initial

genetic

diversity

project

differareadaptationis the major concern.

over generations

can be minimized

variables.

of conservation.

The variables

is to avoidofof a sufficiently

by the management of

to be managed

In ex situ conservation,

contexts

or characteristics

in nature, on the other hand, maintenancediversity

strategies

and the best conservation

time scalesofFrankham 1999).

In conservation

The decay of genetic

differenton the optimization

are commonin that the major concern of the conservation

large genetic potential

reproductive

in designingex situ and on farm in situ, although

populations.

as well as someconservation

or minimize the decay through generations
targetimpactof newfor designingsize as a key parameteroccur in different

with the time scale.

and strategies,

and tools

for plant species.

projects

largely

Conservationand the development

This paper will focus on the theories

and in nature in situ as characterizedcategories

Pritcharda short review of the recent advances in the Ne theories

conservation.

The effective

1998,

shouldbeeach entry must be

size because large numbers of entries must be maintainedTable1.

Categoriesof plant conservationand some characterizing

parameters.

47
with a limited

capacity

reproductiveof facilities.

patternof plants

control.

Projects

in bothconducted

in a wide region.

the number and spatial

In thisrathersituation,

where possible,

than population

categories

of in situ

size willconservation,

In this case, the population

arrangement

matingsystem andbe the major parameters

toon the other

be

hand,

will

size and, when it is to be subdivided,of the subpopulations

will be the major parameterstodecide.
The magnitude
using

variousparameters.

Table 2.

Declinein turn

reliesor change throughactualvariables

diversity

generations(census)as presented

Some typicaldeveloped
contributionsof populationsof these parameters

populationin Tablein recentsize

are presented

depends

and various

inon Ne, which

reproductive

and

3.

Formulationof Ne in terms of thesesubjects

in population

genetic

years (Caballero

1994,

to show recent trends in this research

There are two concepts

have been measured

and commonly used parameters

has been one of the core researchbeen significantly

1970,

of genetic

Some typicalon thedemographic
variablesand structureof effective

population

theoriesand hasWang and Caballero1999).

subject

3).

are listed

size (Wright1969,

denotedNej) thatwas definedof identity

by descent(TableCrow and Kimura

Wang 1996):
a. The inbreeding
progress

effectiveof inbreedingb. The variance

size(often(the probability

effective

size (Nev) that was defined

to predict

theof genes);

to describe

the effectof random

drift in gene frequency.
Nei and Nev are different
specificpatterns

under changing

of reproduction

Yonezawa 1997),

in the sense thatcoancestry

coefficientsare different

in early generations

smaller

1969)

1993,

after initiation,and increases
Wang 1997).

sizes

and/or(Chessor

1991,

Wang 1997,

A newly foundedor delimited

population

the increasingof the inbreeding

structurerateschange with generations.

value, termed asymptotic

than NeVinitiallyvalue (Chesser(Wrightover generations

and population

but are the same otherwise.

may not be at equilibriuma commonstable

population

andNei and NeV

but converge with the lapse of generation
effective

gradually,This

In this situation,some

difference

size (Ewens

1979).

whereas NeVdecreases,in the changing

pattern

Generally,to

Nei isto the asymptotic
of Nei and NeV

48
Table

2.

Descriptors

Genetic

diversity

Genetic

structure
SingleSof the genetic

diversity

1)

Either2)

The frequencyof populations.

Number1) of alleles per locus,
heterozygosity2),
gene diversity3),
proportion of polymorphic
loci, andgenetic variance

population

ubdividedand structure

Inbreeding
coefficient(fixationindex), andindex of deviation from Hardy-Weinberg
segregation
Wright's ^-statistics4): F^ FSTand FIS ,Nei's coefficientsof gene diversity :HvDsr iGsrf, and //,, anddistributionof genetic variance within andamong subpopulations

population

the actual or effective

number (Kimura 1971).

of heterozygotes

per locus.

Averagedover loci when multipleloci are considered.
3)

Definedas 1- ^pf

when multipleloci, where pi is the frequencyof allele/.

Averagedover lociare considered

Equal

to heterozygosity

in a

(Nei

1987).

randomly mating population.
4)

See, e. g., Haiti and Clark (1989).

5)

GST was named coefficient of gene differentiation(Nei 1973).

Table 3.

Determinantsof the effective population

size NeThe list is not comprehensive but will be sufficient to consider recent trends in this research subject.
Listing is confined to the definitionof Ne. The pioneering works of the late Professor Wright and other authorities
are not listed because these are well known and notdifficult to access. Netheories under the presence of selection have significantly
advanced in recent years, but are omitted here because
the effect of selection on Ne is not important unless the selection works directionallywith a high intensity.
Priority is given to the theories
with plant-specificor-commonbreeding systems, when available.
Outcrossing
versus selling, monoecious versus dioecious, ploidy, involvement of asexual reproduction,and so on.
Annual versus perennial (overlapping
generations),
structured in age versus demographic stages and so on.
Island versus stepping stone models, and so on.
Colonized from neighbouring
versus all subpopulations,
immediate versus gradual recovery, and so on.

50suggeststhat the optimum strategies

for long-term

conservation.

the asymptotic

effective

may be different

from thoseIn the present paper, Nei and Nev are not distinguishedand only

size

for short-term

is discussed

conservation

to focuson the optimizationof long-termcons ervation.
The appropriatenessmagnitudeof Ne.

of conservation
Contribution

strategies

and managements can be evaluatedto Ne of the control

in matingsystempattern

in ex situ conservation

has been examined by some researchers

1984,

Crossaand Vencovsky

1994,

the reproductive

success of plants

homogenizing

rule Ne=50 for a short-term
conservation.

These

conservation

projects.

regardingcalledconservationin captivity

However, as will1988

as N=Ne/10'1=500 x 10=5000,
The concept
extinction,

years with a probability(Shaffer

1981,

incorporated
survival

Simberloffof 99%) against
1988,(Harperdifferent.

with plant-specific1977,

Richardin situhave been disputes

Frankham and Franklin

of

1998,

the rule Ne=500 has often been
to census size ratio, Ne/N,
Ne=500 is estimatedof demographic

survival(e.g.,

1986).

offor one thousand

and environmental

change1993).

No genetic parameters

werein the so-called

genetic

If necessary

to define

extinction

theories

mentioned

breeding

systems

breeding

systems

Ne theories

theories

stochasticity

concept of MVP, whereas no parameters

There are many plant specificspecies

proposed

for a long-term

MVP satisfying

for a long-term

Nunney and Campbell

in the context of genetic

Ne for species

the

there

in the context

demographic

were incorporated

The two MVPs are completely
be definedoriginallysize requiredin the demographicof populations

F-S (1980)

i.e., of the order of 103.

of MVP was definedas the populationoflater,

and 1995,

1995a),and Lawrencein the planningBecause the effective(Frankham(Gale

as standards

size TV that satisfies

MVP (Minimum Viable Population).

is of the order of 10"1 in plant species

accepted

and reproductive

and the effectivenessand A^=500

be mentionedof the numbers (Lande
The census population

1996),was elucidated.

numbers have been widely

the validity

Frankham 1999).

Yonezawa et al.

by theMVP mentioned
the genetic

theabove.

MVP, it should

later.

and life cycles

for plant specificfor predicting

in the world
breedingof plantsystems except

51
for selfing

remain to be developed,

while theories

for animal specificbeen fairly

well developed

ChessorNunney 1993,(e.g.,The recent advances in this subject

1991,

are outlined

definedspecies

undergoeffectivesize)

Ne (inbreeding

life cycle models she investigated.

2LN

t2

2LN

V

both sexual

life cycle stages

and clonal

reproduction.

Olive(1993)

Fig. 1 shows one of the

Ne for this model was formulated

as

NJI/

ij*t

?,•E=expected

to a common ancestor allele,number)J; Nt= size of stage / ; L= generationi.e., coalescence

for two allelesof a stage / plant; tu= that for two alleles

stage

that

for two alleleschosen from a stage / planttimes

F's are calculated

using

The applicationof this equation,

however, is more or less limited,coalescence1994).

for some of these plant species.

size (plant/ ; ^=and Chessor

and multiple

where N=total population
time (years)

Sugg

systems have

below.

Species with mixed sexual and clonal reproduction
Many wild herb

breeding

the projection

lengthtime (Tajima

chosen from different1983),
plantsand a stage j plant.

matrix;ofThe

between two consecutive

years.

interpreted
holdin terms of the reproductive

unlessboth

sexualandPoisson-distributed(Yonezawa 1997).

coalescencetheory(Tajimapopulations

that undergo only clonal

Yonezawa (1997)
The models investigatedunder thesevariables
independent

models1983,

gave equations

and demographic
clonalreproduction

contributions

per

this

equationcannotand does not
plant

are

based on thebe applied

to thereproduction.
(variance

explicitly

and summarized in Table 4.
of clonalof the species,

because of having been defined1991),

by Yonezawa (1997)
are formulated

reproductive

Further,
Slatkinpatterns

because it cannot beeffective
are illustrated

size)

unaffected
(Fig.

2).

by these limitations.
The equationsin terms of reproductive

and demographic

It is known from these equations(S) and the same as Ne for fully

derivedthat Ne becomessexually

reproducing

Fig. 1.

Life cycle graph of a populationu}i = the rate of plantsof progeny

model used in the Ne formulationof stage / moving to stage;of stage j supplied

season. (Some modifications

eithersexuallyof Olive (1993).

in the next season, Ffi = the number

or clonallywere made for uniformity).

by a plant of stage i in each(a).

Discrete

generation

with mixed sexual and clonal reproduction(b).

Overlapping(c).

Structured

in two demographic

Fig. 2.

Life

generations

cycle

with mixed sexual and clonal reproduction

graphsreproduction.

stagesfor threetypicalpatterns

The census size population
In model (a),all the N plants

each year.

In model (b),a fraction

In model (c),plantsthe population(D1+D2=l).

move to adultstage,

Fractionsare renewed sexually

u of the entire

to the next year, the remaining

sexualfractionis composed

population,

1-w beingof ND2 juvenile

plantsun, u21 and 1-Uj of juveniles

renewed each year, respectively

remainadult,

returnto juvenile{u.f^un-\-u21, u 2-u12+u22).

inor clonallyi.e., Nu plants,

renewed each year.

and are renewed each year, respectively,u22, u12 and l-u2 of adults

and clonal

size N is assumed to be constant

all models.
surviveof mixed

and ND2 adultremain juvenile,
while

fractionsstage,

and areTable 4.

Formulationof Ne for the three population

models with mixed sexual and clonal reproduction

S= (1 - a) + (1 + a) vjk, (k and Vk = mean and variance

A= 2(1 + a)Vjc

a) - (l +a)Vk/k,

L= generation- (llength,whichequals3/(1~of sexuallyprogenyof clonallyu) and~ "22)}(l ~ "11^22 )/{(l~U= A(Wll+W2l)+A(W12+W22)>"2 =A(^l+W2l)2+A(W12

P= selfing

rate.

of Fig. 2 for the other variables.

See, the legend

produced(c and Vc = mean and variance+^'Mii)(ldefinednumber per plant,

produced

for tne models

progeny

in Fig. 2.

respectively).

number per plant,(b) and (c),respectively.

respectively).

55
species

if a condition

A=0, i.e.,the cases

3) and 4) of Table

variationin the sexualeffectof clonal

reproduction
or larger(largeVcjc - tyk/k +(l - a)/(l+

a)jj2

4.

The terms

Vk/kand clonal

reproductive

contribution

reproduction

depends

works to either

increaseand Vc/con the relative
or decrease

as exemplifiedinmeasure the magnitudeofof plants,

magnitude

Ne depending

than the term Vdc= fyklk +(l-a)/(l+a)]/2.

L) tends to decreaseis satisfied,Ne, in other words, accelerated

respectively.

of theseon whetherTheterms;

clonalVjc

is smaller

The long

persistenceof plants

recruitmentof plants

will help

plantsof some herbtoincrease Ne.
Some wild plant
species

speciessuch as Arisaema

have an even more complex life-history;
serratum and Fritillaria

camtschatcensis

persistdistinguishable

demographic

moving up and down among several unambiguously
producingsexualfor these specieseVand clonal

progeny

has been formulated

each year (Kinoshita

1987,

by Yonezawa et al. (2000)

Shimizu

for many years,
stagesandet al. 1998).

Ne

as,

V

where

V equals2(1+a)\t-u2yr(l-u){Avr(S)+Avr(AS)},extended

to the case of multiple

stagesfor the average over the stages(weighted

with(cf., Table

4).

u and

The terms Avr(S)by the reproductive

contributions

u2 now being
and Avr(AS)standof the stages)

of S

and AS, respectively.
In the case where the census population
modifiedyear by year, the equation

is

to,

'

V

[I]'

where Nh is the harmonic
constant

size N changes

rate X, the equation

mean of the census sizes
is presentedas,over [L] years.

When Nchanges

with a56

N=

2Nn %{X-\)V l-(l/lf]

where Nois the initial
The equation
camtschatcensis,
estimated
2000),

population
was applied

to estimate

the Ne values for two populations

the life cycle

data of which are shown in Fig. 3.

which fall in the range of estimates
The numericalthe effectivenesspredictedof controllingto increasesurvival

size (Yonezawa et al.

in plant

speciesgave some usefuland survivorship(Frankham

predictions

parameters;

and growth of the first stage plantsof the second and third plants

in many plant species

consistof females

and cytoplasmic

about
Ne was(juveniles)(adults).

such as Saxifraga

and hermaphrodites.

genes (note that cytoplasmic

and kinship

Newas formulated

granulata(Stevens

and Richards

Laporte et al. (2000)

defined

Ne for nuclear

genes as well as nuclear

with the lapse of generations).

under two types of gynodioecy,

where the determination
cytoplasmic

some fecundity

were

speciesPopulations

frequency

previously

using the equation

with accelerated

with reduced survival

Gynodioecious

calculations

obtainedin Fritillaria

These populations

to have an Ne of about 20 to 30% of the census population

1995a).

1985)

size.

Each of the nuclear

and cytoplasmic

i.e., nuclear and cytoplasmicof sex, female versus hermaphrodites,

genes, respectively.

genes change theiris controlled

In the case when the hermaphrodites

gynodioecy,
by nuclear andundergorandommating, Newas derived as

N e =Nh(\-h}(2 - y/f - y/h)h(l-iyhf+(l-h)([-y,fy'

where h= relative

frequency(ratio)in the familiesof femalesNor l+o-2/(2-^-^J',of hermaphrodites

in the populationof female maternal

parents; y/f= averagefrequency; y/h= average frequencyofFig. 3.

Life cycle graph of two populations
for the two populations

investigated.

of Fritillaria

camtschatcensis.

The two numbers intervened

by / stand for the measurements

58
hermaphrodites

in the families

reproductive
dependingsuccess

betweenthe sexes.

is modified

parents; andare formulatedor cytoplasmic(cf., Table 1 of Laporte

When the hermaphrodites

maternal

These variables

on whether the Ne is for nuclear

nuclear or cytoplasmic

Nof hermaphrodite

c?= variancein different

of

ways

genes and the gynodioecyiset al. (2000)).

undergo partial

selfing,yjhf-A)(l-V/f

the equation

for nuclear Ne gynodioecytoe =Nh(l-h)(l +F¥)h(l-where Fisf and Fish standfor the heterozygote

Weinberg proportion)

for females

Ne = \l(\+(l-h)Fisf+ hFtsh)cytoplasmic

Ne, although+ (1 +iL,Xland hermaphrodites,

under

nuclearvariables

via numericalet al. (2000)

showedgynodioecy,

in comparison

to hermaphrodism,of different

magnitudes

to hermaphrodites,

the genetic

Selfingin hermaphrodites

works generallywith a high reproductive

success of females.

from the HardyThe equation

No modification

should take different
calculations

is neededof theseon the reproductive

for

Ne, althoughequationsthatand cytoplasmic
success of females

system of the sex determination,
to decrease

becomesvalues accordingly.

reduces Ne for both nuclear

dependingrelative(deviation

respectively.

gynodioecy.

the constituent

Laportegenes, although

deficiency

and the rate of selling.
this effectis mitigatedSpecies with soil seed bank
Many wild plant species
soil

have a soil seed bank (Harper

1977,

Leek et al. 1989).

can persist

for a few to more than one hundredyears dependingand Legg 1995),

and can have a profound influenceon the genetic

the species
inferred
contributes(Templetonand Levin 1979,

that the seed bank buffers
to increase

Many et al. 1999).

Evans and Cabin 1995,

populations

Ne of the growing

It is crucially

importantstructure
Cabinfrom the loss of genetic
plant

populations

to formulate

on species(McCueSeeds in(Cumming

and evolutionof1996).

It has been

diversity,and then,and Holtsford

1998,

Ne in terms of some measurable

59
biologicalparameters

the seed bank.
not yet been
population

to estimate

To my knowledge,
given.

will be sufficient

Poisson-distributed

reproductive

reproduction,

equation

confirms(equilibrium)

survivorship(or monoecy),and germinabilityof growing

andin the number of growing

plants;and randomof soil seeds;

and absenceaof clonal

as

contributes

to increase

unless the term s2Np/Nsor sf takes a sufficiently

large value.

thatplantstheseed bank

but alsosoil

thus, both of them may be regarded
demographic

stages.

seeds

determine

Ne, although

the evolutionary

as members of the population

Now, Nefor the entire population

not

fate

of a

belongingtoof census size Nj{=Ns+Np)

asUi
Ds(2-R)+Dp(l-sf)/R'N

ewhere DS=N/NT,Dp=Np/NTand R is annual recruitment
Thus, the contribution
mentioned
valuesbut\-sf

Not only growing

is formulated

models,as used by Yonezawa et al. (2000),

hermaphrodism

contribution

possibleon theNl-s2Np/N/

different

annuality,

age-independentN

was obtainedof the seed bank.

procedures

Nefor the surface plants is derived

xr

population,

basedsuch as the constancy

plants;substantially

here, which

effectmating of growingThis

is presented

the essential

seeds (Ns);ofwith soil seed bank havethe same mathematical(Np) and live soil

contributionof Ne for species

This is just one of the simplest

some simplifications

plantsthe evolutionary

in Fig. 4.

to consider

Following

quantitatively

the equations

An equation

model described

adopting

and interpretabove;of seed bank is not uniquely

the seed bank works eitherof reproductive

rate of seeds, r+f.

and demographic

positively

variables.

determined

as in the firstcaseor negatively

dependingon thebased

on moreDetailed

discussion

Fig. 4.

Life cycle graph of a hypotheticalseed bank.

annual plant population/= entrance rate of newly produced

r= death rate of soil seeds, and s= contribution
plants.

havinga soilseeds to soil seed bank,rate of soil seeds to surface

61
general and realistic

Nefor sudivided
Plant

populations

populations

subpopulations
them.

models will be undertaken elsewhere.

may be conservedwith an appropriateThis invokes

eithermagnitude

the so-called

as a single
and pattern

SLOSS (Single

population

or in a number ofof gene flow (migration)

Large Or Several Small)

betweendisputes.

The

concept SLOSS as well as MVP mentioned above were coined in the context of demographic
theoriesof biologicalconservation(Higgsandsubpopulations,

Ne for the entire

subpopulations

as well as the rate and patterneffective

size of a subpopulation

population,

population

is definedUsher

in terms of effectiveof migration

among subpopulations.

in the same way as that of a single

with the census size of population

being substituted

diversity

retainedin a subdivided

by Ne. The SL (Single

Large)

strategy

will be preferredwith the total census size unchanged,

better

if Ne increases.

variousmodels of population

SLOSS issues.
decideRecently,

the number and spatial

terms of the magnitude

pattern

population

population

The
undivideddue to population

Small)

strategy

will behas been definedunder

the genetic

in theof populationof subpopulations,becauseaspects

subdivisionand toit is formulatedin

success of subpopulations
is eitheras,

In thispositive

In conservationand the rate and pattern

practically.

is expressed

subdivisionand migration.

number of subpopulations
or keep constantofand pattern of gene flow among subpopulations.

of populationof reproduction

size

as a whole is determined

if Ne decreases

the advantage

arrangement

Equal and constant size and reproductive
The effectpopulation

which will help to settle

The Ne allows us to discuss

inby that of a subpopulation.

and the SS (Several

Ne for subdivided

structure,

When conserved

is formulated

The amount of genetic

subdivision1980).

situation,

projects

or negative

dependingon thein situ on farm, the size andof migration

will not be difficult

the effective

population

to control,

size of the entire

N =-where Nej= the effective
Fsj^ thefixationdifferentiation

size of subpopulation;index

of Wright

among subpopulations

The value of Fst dependsof migrationand decreasinglarger(> 2) ; andwhichof genetic(1969),; e.g., FSt equals

respectively.

that Ne for the entire

withsignificant

unless the rate of migration

in a subpopulation\INes (large

Fst).

rate as highWitha migration

indicating

variability

size unchanged,
possible

Local extinction

due to inbreeding

than that of a single

advantage,(smallhowever, is not

population.

in n works to increase

lower thanFSt), Ne equals

has no effect,nNes

i.e., the populationWith

the total

Ne, but negligibly

censusslightly

may occur with a high value of Fst, which

in Ne. This adverse effectvia either

criticalupper limit value of FSt, denotedmn

subdivision

depression

avoidedincreasing

will be obtainedshows that Ne is always

is low, i.e., sufficiently

as m>llNes

as an undivided

the increase

0 and 1 with increasingis largerThis

under an island

values of m and n (cf., the data in Wang 1997a,b).

cause a large reduction

condition

census size.

that the population

the same genetic

under practicallythe same total

population

population

willll(\ +4Nesmn2/(n-l)2)

The above equation

undivided

populationand the rate (m) and pattern

Fst takes a value between 0 and 1, approaching

than nNes, indicating

retains

the degree

1 999).

on the number of subpopulations

rates of migration,

approximately,

measures(Wang and Caballero

among subpopulations

model migration.

n= the number of subpopulations

m or decreasingof populationn (increasing

FC9is imposed,

subdivision

could

be

the size of subpopulations).

If a

the values of m and n satisfying

thisusing the relation,

\-Fc

{n-lf

where Neois the effective
The theories

populationof the neutral

size without

additivegenetic

subdivisionvariancethat equals nNes.
under the balanceof mutation

and

63
random drift (Landevariances within

1992,

Whitlock

1999)

give the prediction(Vw) and among (V*) subpopulations

that the equilibrium

and for the entire population

genetic(VT) areVw=2nNesVm9
Vb = 4nNesFSTVm/(lVT =2nNes(lrespectively,
neutral+ FST)vm/(l- Fst)>

where Vmis the mutational

mutations.

decreasing

These equations

rates of migration

as that of a single
migration- FsT),

helps to increase

Variable size and reproductive
In conservation
subpopulations

success (asymmetricor excessive(1-FST)(1+V)+

diversityand recolonization

Population

subdivision

For monoecious,

diploid

species,

subpopulations

withinsuccess ofby the propagulesin this situation

causes quite

Ne under variablereproductivewith a constant size for subpopulations

is presentedas2&

2NesVFSTn/(n-l)'case (Whitlock

as in the preceding

V<]/^lNes-l+\lFST)<l/(2Nes-l)

meaning that population

homogenizationon the other hand, the size and reproductive

being

equation,with a minimum

success of subpopulations

success

definedwithof the entire population.

where V is the variance in the reproductive
similarly

conspicuously

Vwon the average is as large

genetic

may occur.

migration)

via

that the SS strategy

should be variable, and, local extinction

complex influences.

e

It follows

the genetic

each generationthat Vb and VT increasedifficulties

of wild plants,from other subpopulations

Nvariance suppliedFst), while surprisingly

population.

rate to avoid inbreeding

subpopulations,indicate(increasing

undivided

genetic

must holdsubdivision

is less variableof subpopulations,is advantageousthan expected

the other variables

and Barton1997).

for Ne to be larger

only when the reproductive

when Poisson-distributed

By this
than

nNes,

successof(Wang and Caballero

641999).

Ne approaches

nNe/(l+V)(n-l)/(2V)

when the migration

migration

therefore

acts

increase

Ne unlessmigration

when the migrationis severely

to reduce

rate

restricted

Ne.

Population

is high(largeFSt)-

subdivisionoccurs at a low rate (large(smallFst),

Asymmetrydoes

andin thenot contributeFSt) and symmetricallyto(smallV)among subpopulations.
The effectof population

subdivision

is even more complicatedin the presenceof local

subpopulations

1985,

McCauley1991).

subpopulations

byextinction

and recolonizationfrom other

Assumingmonoecy, diploidy

andcolonization,

Ne is presentedNe

immediate(Slatkinrecoveryof extinct

as

= 2^
4Njm+e)Fs/

where e is the extinction
depends

rate of subpopulations

on e, the number of founder

per generation.

The fixationindex FSt now

gametes (2k) and the mode (# the probabilityof eachpair of the 2k gametes coming from the same source subpopulation)of colonization,

formulated

an islandas (l+eNeJk)/(L+4Nesm+2Nese(l-0(l-ll(2k))))migration

under(Whitlockand McCauley1990,

the above equationthat a frequent

local

recolonization(j>) can cause a large reduction

be either(largepositiveor negative

dependingthe decay of genetic

diversity,colonization

i.e.,

type

a migrant-poolcolonization

Whitlockextinction

and Barton 1997).
(largee) together

being
model

It is predictedwith a highly

by plantsfrom multiple

in the terminology(m) can

To minimize

subpopulationsof Slatkinfromlocalized

in Ne. The effect of migrationon the values of the other variables.

of(1977),(small0),is crucially

important.
Theories

for the distributionof genetic

Whitlock1999)

genetic

variances,

Vwand Vt, and consequently

the total genetic

extinction

localized,

recolonization.

local

with

that

a highly

both

1992,

1999,

a frequent

have revealed

variance (Landeof the within-

Pannelland Charlesworth

and among-subpopulation
variance,a so-called

VT, decrease under
propagule-pool

type

65
Prediction

for completely

subdivided

populations

In the case where a population

is divided

asymptotic

variance effective

genetically

fixed,

subpopulation

while

subdivision

Now focusing
additive

effects,

reproductive

not exist

beforethe asymptotic

inbreeding

effective(Wang

before genetic
on the genetic

and assuming

1997,

after subdivision

where Vois the initialallcan be predicted

the homogeneity

the

the

subpopulations

size

is givenby thatof a1999).

The effect

of

trait

and constancy

the distribution

subpopulations,becomebased on Nes.

variance of a quantitative

is expected

genetic

isolated

Wang and Caballero

fixation

success of subpopulations,generation

definedsize does

Nes used before

population

to completely

governedby genes ofin the census

of the genetic

size

and

variance in the rth

to be,

variance of the entire population,

as before (Grow and Kimura 1970).

Vw,V&, and VTbeing similarly
index Fsr(t) in generationt relieson the effective
subpopulation
size Nes.
(Fsnor®)* FsT(t) is given by 1-£*/?(-/(2A^))

If the initial
population
is randomlyand (l-exp(-(2v+]/(2JV<w))f))/(4iV«v+

matingl) inthe absence and presence of neutral mutations,

respectively.

Obviously,
decreases

Vt in the above equation

with the lapse of time, indicating

total

genetic

trendof change

trajectory

diversity

by facilitating

The fixation

increases

as a resultthat population

genetic

holds

also

when nonadditiveof changein thevariance

subdivisiondifferentiationdiffers

in V&, whileVwworks to increase

theamong subpopulations.

gene effects

components

of increase

are involved,
dependingThis

althoughon the

the

kind

of

nonadditivity.
The progress
basedof genetic

on the concept

differentiationof the genetic

in the population
identity

by descentcan be more explicitlyof genes(Wright

described
1969).

The

66
probabilityof two genes randomly

sampled(Jw) and that for two genes sampled4i) - ^") +(^)

jj'from different

being

subpopulations(Jb)

identical

by descentare presented

as- /r)>"(2v+V(2^)>,-jiV2-,

where the superscript
initialfrom a subpopulation

values(t) stands(Nei1987).

for the generations

As generation

after subdivision,

proceeds,j^j£asymptotes

an(jto l/(4A^v+l)

diminishes,i.e., the subpopulations

become totally/^on only the mutation

rate v, not the size of each subpopulation.

depends

under the presenceof migration

increasing

the genetic

unlessNes is sufficiently

difficulties.

discussed

diversity

The equilibrium

to avoid
genetic

smaller than that under the presence

distributed

populations

The distanceof dispersionof pollen

genetically,

although

2nNesVm(Lande 1992,

and seeds may be relatively
A populationso-calledneighbourhood

size,

individuals

in an area from which the parents

distributedof gene dispersion
denotedCrawfordin thisi\fc,

Ne is definedfor a real, distinct

populationdelimited

population

as a whole cannot be obtained

Nefor distinctly

subdivided

plot within

populations

case should

is definedas theindividuals

may be treated

as if

the neighbourhood

size

Inappropriately,

for an imaginary,a large continuous

before.

be structured

in terms of thesize.

merely by substituting
mentioned

small compared to thehas been interpreted
whichof a finite

1999).

The geneticof central

1984).

Whitlockgeographically.

has often been taken as the same concept as Ne. Nb is definedor an arbitrarily

this effect may be mitigatedis now 2NesVm,muchdistance

subpopulationofsubpopulations

due to a restricted

1969,

has the effectvariance within

and uniformly

often

as

due to inbreeding

structuredrawn at random (Wrightalthough

Similarlyof subpopulations

as a whole.

continuously

subdivisionofextinctionof migration,

Nefor continuously

and /bt]

The rate of decline

populationof the entire population,

large

area size of the populationdifferentiated.

before,/b°> being thenumber ofnonexistent

population,

Ne for a continuous,

whereasfiniteA^ for Nes in the equationof

67
Ne for a continuous
random mating,approximately

population

diploidy

andof a finite
monoecy,total

censussize N, under the assumption

was formulatedbyMaruyama(1972,

of
1977)

as,

JAr/(z)cr2)---when

Do2 < X[N--whenD^->X

where D and a1 are the density
respectively.

The equationsof populationand the varianceshow that N£>~Nwhen Da2<\of gene flow distance,and Ne-N otherwise.

Thevariance a2 tends to be small in the absence of distantgene flow.

Then, a qualitatively

similarprediction

as with Ne for a subdivided

population

is derived;

Ne and therefore

genetic

diversityof the population

are enlarged

with a restricted

as a single

panmictic

gene flow, the population
theories

for continuous

behavespopulations

gene flow, while, with a highpopulation.

remains to be developed

theGeneralizationof Ne

under more general

breeding

system models.
Environmental
different
local

conditions

genotypes
adaptation

habitats

will be favoured in different
to different

restricted(Slatkinfacilitated

if the population

diversityof a population

1993,

Barton1973,

andsubpopulationsin natural

may be highly
subpopulations

Nagylaki1975,

is subdivided,

Levin 1988).

as a whole unless local
Whitlock1997,

the genetic

reproductive

In this case,

Bartonto increase

the genetic
and Rouhani

1999).

Occasionalmigration

due to inbreedingof a population

depression

or isolation

by distanceas a whole, whereas imhomogeneity
or local extinction

imhomogeneity

The migration

betweenbut

potential.

diversity

the reproductive

will be much

(Bartongene flow by either subdivisionof a population.

adaptation

is frequent

success due to either asymmetric migration

variability

plots.

extinction

restricted

Managements to minimize
genetic

The local

which in turn will contributealso to generate new genotypes with a high evolutionary

increaseor habitat

and then,genes can occur whether or not the movement of genes iswill help not only to avoid local extinction

In conclusion,

heterogeneous,

will

greatly

among subpopulations

works toin the local

acts to decrease

it.

help

retainthe(delimited

plots

in

68
the case of a continuous
avoid inbreeding

Disputes

population)difficulties

shouldon the generality

shouldwas proposedthat

the increase

be lower than 1%.

experienceof animal breeders

Drosophila.

The progress

inbreeding,trend,

plantsspeciesare more tolerant

1987,

Husbandof inbreeding

selfing

1996).

may not be serious in the planningon theotherbased on the perspective

variance for quantitative

but has also been accepted

traits.

insteadof 500.

between neutral mutations
theorybe saidthatis more comprehensive

stabilizing

Lande's

problems

projects.
forlong-termhighevolutionary

amount of additive

for animal conservation,and 1995)

proposed

to use the number

between the theories
the theoryof genetic(intermediateof F-S was establishedvariance)

optimum)
becauseof F-S (1980)

and random driftof neutral

selection

it considered(loss).

mutations
It maythe effectofselection.

Both theories
the generality

theoryInbreeding

was based on the balanceand stabilizing

and Charlesworth

The number 500,

(supply

random drift (loss),

selfing-inclined

species.

While(supply),As aof plantthat distinguishbased on the balancetonot be an important

plants.

a sufficiently

planning

in Table 5.

the tolerance

and population.

will

was proposed

Lande (1988Some parameters

Lande'swords,

This rule also was proposedfor conservation

variance),

in genus(Charlesworth

propagatinghand,

are summarizedof genetic

data of some speciesof plant conservationand Lande (1995)(loss

on thedepressionof maintaining

however, has faced some criticisms.
5000

clonally

i.e.,

based

and among plants,

speciesInbreeding

thereforeA^=500,or mainly

per generation,with the species

than outbreeding-inclinedof large animals

1%, was decided

in otherthan animals,

in highly

genetic

value,

different

conservation

coefficient

depression,

are more tolerant

problemconservationof inbreeding

as well as some experimental

and Schemske

The rule

for short-term

The criticalhowever, is known to be largely

generaltoof the rules Ne =50 and 500

on the perspectivel/(2Ne),at a minimum rate (m<l/Nes)in the subpopulations.

The rule Ne=50 of F-S (1980)
based

be conductedadoptedsome assumptions

in deciding

the criticalor validityof which remains

to be substantiatednumbers 500 and 5000,
with a wider spectrum ofTable5.

Parameters

distinguishing

VG=additive genetic

between the rules of F-S (1980)

variance for a quantitativeand Lande (1995).

trait,

VE=environmental variance,Vm=genetic variance newly supplied per generation
h2 = narrow-sense heritability,
Vy^+Vg),andp = proportion

relativeto the genetic

variance

via mutation,of an infinitely

large population.

70
experimental

data.

The assumption

Vm=10'3VEof F-S (see the legends

Ve) seems to have been based only on the data of some species
V^lO'Vtf

be ratherevolutionarily

contributory

an oversimplification.

mutations).

Heritability

The assumption

differs

greatly

fallingin the range of 0.2 to 0.8 (Mousseau

and Roff 1987).

suggestthat the heritabilities

traitsthose of length

0.8.

in Drosophila

because he assumed that only about 10% of new mutational

nondeleterious,

Hallauerof Table 5 for Vmand

traits1988).

of fitness-related

such as plantheightwithto 125 if h2 is assumed

p=0.5 is used, Lande's theory(1980)(cf.,

Table 1 in Lande 1995).

Lande's

theory,(1995)

also

employed.

However, Franklin

experimental

data support

susceptible

to the value

or logicalnotgives almost the same number as obtainedby F-S

Unlessevidence

arbitrarily,

If an

the F-S theory

this arbitrarinessis overcome, the superiority

than the theoryof F-S, is not proved.

gives

if the assumption

5000,

and Frankham (1998)

the relation

With such uncertainties

size is highly

explanation.

more sophisticatedout that

to be 0.2, and to 2000 if

he used seems to have been chosen rather

assumption

pointedeffectiveby any experimental

althoughin crop plants

than 0.8 (see, e.g., the data inthe criticalbeen substantiatedand populations,can be even higher

By the theory of Lande (1995),

havingtraitscan be even lower than 0.2, andis not a magic number to adhere to.

/?=0.67

is due toh2=0.5 of F-S would

Experiments

The number 500, therefore,

The assumption

varianceusedof plants

The number 500 changesof p.

(Landeand Frankham (1999)

of
Lande

Vm-104VE
ismaintained

that

Vm=10"3Ve rather than Vm=l0~4VE.

and simplifications

Lande, there would be no point specifying

being
the critical

population

size for conservation

can leniently

It can safely

be said that the census population

includedin the theories

size as 500 and 5000.

be regardedof F-S and
The effective

to be of the order of 102 or higher.

size for long-term in situ conservation

is of theorder of 102 or more for randomly

outcrossing

species,and of the order of 103 or more forhighlyreproducing

species,

because the effective

selfingand partiallyratio NJN is relatively(Caballero

1994,

close to 1 in the former group, while tending

to census size

to be small in the latterYonezawa 1997).

The magnitude
determining

clonallyof genetic

the criticalvaluevariancefor quantitativeof Ne for biologicaltraitsis not the only

conservation.

Usingsource forrecentDNA

71
techniques(Amos and Hoelzel

average number of alleles
the proportion
size.

1992,

of Ne that is necessarylevelsassessedthey

can be quickly

conveniently

data;

may be regarded

used as a tool for monitoring

A time-consuming

and speciallyof quantitativethese parameterstraits(e.g.,

to retain

effective

the genetic

traitsalone.

In addition,markers,

and can bemolecular

to estimate

1981).

Determinationof the criticalvariance for quantitative

above were defined

traitsbe maintained

however, gives no guarantee for a long-term persistence
There have been some population
extinction.

showed that finite

melt-down, i.e., synergistic

extinction

populations

interaction

Ne using

based on the perspective

by U/(roT)in populations.

ofThis condition,of populations.

theoriesthat

addressthe determinantsof Lynch and Gabriel

can become extinctof random drift

analyticallymagnitudeand(1990)and Gabrielby the so-called

mutationaland mildly

deleterious

mutations.

by Lande (1994,

1995),

who formulated

in terms of Ne and some other variables

relatedto the rate andof the mutations.

t scaled

genetic

The simulations

was developed

the mean time to extinction
effectthe genetic

theoriesshouldgenetic

projects.

elsewhere.

genetic

This line of discussion

Use ofThey represent

or about 70% of the maximum possible

et al. (1991)

commonlyis necessary

Falconercan besize.

that a commonly observed magnitude,of population

Nei 1973),

experiment

The rules such as A^=500 or 5000 discussed

process

effective

and managing the process of conservation

designed

will be discussed

The role of extinction

as the criticalusing

the criticallarge

has some advantages.

estimatedover loci, andvalue of these parameters

over the whole genome, not of quantitativeand precisely

such as theKimura 1971,

Ne that is sufficiently

as the source parameters

variability

variance

to retain the desiredof these parametersthese parametersor heterozygosity

depend on Ne (Maruyama 1970,

using the observational

observed

parametersloci can be used as the basis for defining

Because these parameters

determinedsome other genetic

per locus, average gene diversityof polymorphic

the magnitude

Haig 1998),

His theory(t/=themaximumgrowth rate of population

total

lead to the conclusionthat the mean time to

genomic rate of mutations

per unit time, and ^generation

per generation,

length)is asymptoticallyro=the72
equal

to e4N2/Ne when all mutations

condition

4Nes>l, and is equalwith a variation
deviationcoefficient

to N* i when the mutationsc = as/sof the selectionthat the t differs

have the same selection

satisfying

4Nes>lmuch depending(c=0)

with Ne of around 100 have almost no risk of extinction,

mutational

effects,

simulationof Schultz

the presence
populationsof beneficial

mutationseffects

Lynch and Lande (1993)
under the forces

mutations.

Assuming

explored

showedunder c=0,

can go extinct.

that the risk of extinction

and synergistic

are constanteffects

whereas, under variable

epistaticThe

is much reduced ininteractionsof mutations;with Ne of around a few hundred are immune from extinction

whether the mutational

occursor variable;with Ne as large as a few thousand
added

a

and as =meanand standard

populationsand Lynch (1997)

genetic

The numerical calculations

on whether s is constanteven populationss satisfying

have different(Jcoefficients, respectively).

coefficientirrespectivelyofor variable.
another line of extinctionof selection

underchanginga modelof directionallytheories

environments,changing

where extinction
random drift

environment

andwhere theenvironment changes in such a way that the optimum phenotypicvalue at time t equals

kt+ e(£=thetemporalvariation),theychange (kc) that is determined

by Ne,

rateof environmental

change,analyticallyderived the criticalrate of environmental

genetic

variance and some variables

The population
Extending
genetic

is expectedMills(geneticload)

the strength

if the environment

traitsor unpredictably

and Smouse (1994)

investigatedinteractionof inbreeding

anddemographic

andsimulations

based

on a stochastic

discrete-time

Leslierelation(lessbetween inbreeding
than 100),

extinction

coefficient

newly foundedpredicted

is not always advantageous;under constantpopulations,of selection.

it works to increase

environmentalofthe riskenvironments.

of extinction

that

occur

stochasticity,model incorporating

depression.

than kc.

that a high magnitudechanging

the process

and fitness

and patternchanges with a rate higher

Lande and Shannon (1996)

variance of quantitativeof extinction

e =stochastic

characterizing

to go extinct

this line of research,

and

They predicted

inbreeding

depression

prediction

can be perfect.

undermakinga hypotheticalthat, in small

can be a criticalcause ofin the first few generations.

Inevitably,no theoryforlong-termEachof the73
above-mentioned

theorieshas

been constructedadopting

particularassumptionsof theirown. The theory

sophisticatedtheories,

was based on a model that the environment

constant before
deleterious

extinction

mutationsand that the population

have been accumulated

environment and populationof Lynch and Lande (1993),

constant

population

size before

The prediction

extinction

criticalwere assumed,

In my opinion,

concept of genetic

for measuring the degreeof fitness

body prepares,

actually

whether

genetic

diversityused or not, many kinds

for long-term

Mills

and Smouse (1994)

survival

to changing

inbreeding

coefficientand fitness

demographic

and environmental

stochasticity,

theor experimental

have shown that inbreeding
systems of inbreeding

the environmentalofof lymph

although

it is usefulcells

Just like ouragainsta population

depression

depression,

the breeding

the attack

shouldkeep aThe predictionofmodels regardingtheand the patternofof whichPrevious

occurs in totallyremainsexperiments

different

to bein related

ways depending

systems of populationson

as well as

conditions.
extinction

theories

demographic

extinction

theories

have been constructed,

groups,deterministici.e.,

potential

persistence,

depression

genetic

categories,of the evolutionary

generality

evidences.

Besides

i.e.,

the

environments.

betweenthe genetic

unless(antigens),

relationshiptopicsis invalidated

was made based on a number of hypotheticalexamined with observational

are not realistic

in its current environment.

from a wide, unknown spectrum of alien substanceshighchange andload (cf., Crow and Kimura 1970)

of long-termof a populationIn

which again(1996)

populations.

the potential

extinction.

environmentalload, is a good indicatorfor evaluatingin thedirectional

they used, i.e., the genetic

not be usefulConstancyof long-term

criterionwilllevel.

size arewhen mildlyprocessof Lande and Schannon

Haldane'sand population

occurs suddenly

to a certain

a long-term

andone of the most advanced andextinction

size will not fit the actual

the theory

assumptions.

of Lande (1994),

simplifications

such

as mentioned

the latter

demographic

stochasticity,

environmental

1978,

1987,

1988).

(Caswell

genetic

parameters.

Soule

Shaffer

Some researchers(e.g.,

variouswhich can be classfied

versus stochastic,

theories

above,

being

further

into

three

and catastrophy

None of these theories

Nunney and Campbell

ofinto two large

classfiedstochasticity

types

1993,

incorporated

the

Lande

1995)

74
proposed

that the demographic

theories

whereasthe genetic

for long-termMills

theories,and Smouse (1994)

suggestedthatdemographic

early

mentioned

factorsand Lande (1997)

ratherfactors

situation,

the best strategy

state

small

allowedto construct

in reality

planning,

However, the prediction

made bypopulations

but due to genetic

and demographic

extinction

conservation

the prediction

proceeds

a grand,with particularshouldof the populations,

theories

Further,the simulationthat environmental

stochasticity

extinction.

Harmonization

theorywould not be fruitful.

of causes and effects

be to keep monitoring

incorporating

the current genetic

some cases. The role of extinction

theories

watch and the efficientmeasures to control

some importantthen should

be to define

Eachof its own.

and, when they are found to be deteriorating,management measures. Simulationsorremains to be developed.

universal
links

can occur not due tocauses.

can be a major cause of long-termof the genetic

However, seeking

planning.

of relatively

made by Schultz

unification

be used for short-term

above as well as some works cited by Frankham (1995b)

extinction

or environmental

than genetic

shouldIn thisand demographic
to take appropriate

parameters

may help

inthe core parameters

tothese parameters.

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54 (6)(in

83

A Methodology

for Genetic

Reserve Conservation:

Case Study for Vicia.
Nigel

MaxtedSchoolof Biological

Sciences,

The Universityof Birmingham,

Edgbaston,of plant genetic

diversity

Birmingham

B15 2TT, U.K.

Abstract
A methodology

for in situ conservationin genetic

reserves has

recently been proposed. To demonstrate this methodology, this paper attempts to provide both atheoretical and practical overview of the implicit questions and techniques associated with the location,planning,

establishment,

management and use of the genetic

reserve. The methodology is illustrated

resources conserved within a genetic

using the temperate forage legume genus Vicia L. It is hopedthat the paper will demonstrate the effectiveness of the general methodology proposed and provide an
exemplar that could be adapted to conserve other plant genetic resources groups.
IntroductionThe

fundamental

importanceof biodiversity

exploitation

by humankind

is central

to the Convention

on Biological

Diversityand this point

is underlined

in Article

2, which state the conventions

objectives:"The objectives
diversity,
sharing

Diversity

arisingalso

thestresses

where necessary,to conserve biological

1980;

(CBD,

1992)

of biologicaland the fair and equitableof genetic

resources...

needefficiency

of conservation

for increased
Article8 of the Convention

and Williams,

guidelines

for the selection,areas or areas where special"
effort,on Biologicalestablishment

andmeasures need to be taken

diversity."much research(see for example:
Holdensustainable

out of the utilisationof in situ conservation.

management of protected

diversitytoon nations to:"Develop,

Althoughtied

are the conservation

use of its componentsof the benefits

in the fieldcalls

convention...

the sustainable

The convention
particularlyof thisconservation

Frankel1984;

has focusedand Bennett,

on the ex situ conservationof plant

genetic
Hawkes,1970;

Frankeland Hawkes, 1975;

Brown et al.9 1989;

Guarinoet al,

1995;

Hawkes et al,

84
2000)

there has been relatively

situ conservation
appropriate

littleof plant genetic

and effectiveplant diversity.

progressdiversity.

protocols

Recently,

with the development
There is therefore

and methodologies

Maxted et al. (1997a)

conservation,

for the inan urgent requirement

to develop

to assistattempted

includingof methodologies

in the in situ conservation

to address

the application

all the diverse

aspects

genetic

farm projects

as specificconservation

techniques.

plant genetic

conservation

in a genetic

reserve.

methodology

for theestablishment

and management of genetic reserves using as a case study of the temperate

forage

It illustrates

region.

The In Situ Conservation

Diversity

various techniques,
Articleconservation

strategies,

1 (Maxted

et al., 1997b),see Figure

2 of the Convention

on Biological"Ex situ conservation
diversity

outsideDiversitymeans the conservationof componentsbyof biologicaltheir natural habitats"and recovery

natural surroundings

and, in the case of domesticatestransfer

involves

differenceof ecosystems and natural habitatsof viable

where they have developed

There is an obvious fundamental
situ conservation

are defined(CBD, 1992):

and the maintenancethe sampling,oneach composed ofand the two strategies

means the conservation

involves

the proposed

ex situ and in situ,"In situ conservation

surroundings

and onHowever, this paper focuses explicitly

legume genus Vicia L. in the Mediterranean

There are two basic

reservesofin situ plantof Plant Genetic

of both geneticof

populationsof species

or cultivatedtheir distinctivein theirspecies,

properties.

between these two strategies.

in the"Ex situ conservationand storage of target taxa away from the target area, whereas in

the designation,

management and monitoringof target

taxa where

they are encountered.
Definitionsin the literatureof in situ conservation

because they confuse two quite distinct
In part, this lack of clarity
in their native
other.

in their

genetic

is due to in situ conservation

nativereserves and on-farm conservation.

being appliedon the one hand and to crops within

Also use of genetic

conserved
genetic

habitats

techniques,

are often unclear and imprecise,reserves

as a term to describe

habitats

is not universallyto wild species

traditionalfarming systems on thethe placewhere wild species

followed,

reservations,as well as many other terms. However, the underlying

conservationis that plant

diversity

is conserved

aresynonymous terms include

reserve management units, gene management zones, gene or genetic
genetic

conserved

sanctuaries,

crop

concept of in situ genetic

in the locality

where it is currently

Figure1. A Model for Plant

Genetic

Conservation(Takenfrom Maxted et at., 1997b).

86

found, either

where it is naturally

cultivation.

It is important,these two distinct

locatedor where it has developed

however, if we are to avoid confusion

in situ techniques.

Maxted et al., (1997c)

distinctive

traitsthat we distinguishprovidethe following

underbetweendefinitionsofthe two techniques:"Genetic

Reserve Conservation

genetic

diversity- the location,in natural wild populations

management and monitoring
withindefinedareas designatedofforactive, long-term conservation.
On-Farm Conservation
locallydeveloped

speciesor forms by farmers

agri-silvicultural
Of these

methodology

doing

withinwith associatedtraditional

diversity

wild

agricultural,ofand weedyhorticulturalorsystems. "techniques,

there

under the scientific

the conservation,by the conservationists

is anothercontrolsignificant

distinction,of the conservationist;

on-farm

it is the farmer

and for this reason

the developmentof a general

In contrast

reserve conservationis largely

genetic

and as such is easier

to study,

as socio-economic

andfactors are of lesser importance.

Severalauthors

methodologies
Safriel,(1997),

independently

Maxtedshown in Figure

reserve is perhaps

et al.

conservation

discussing

thisfollowedslavishly;

proposedmethodology,(1997c)issues that requireof wild

speciesmanagement and monitoring

it is applicable

et al (1996),

Safrielet al(1998).

the overall

It is important

proposedin a genetic
for the bulk

for orthodox
reserveto stress

beforebeof wild species,
inexpensive,

will almost invariably
the location,

in a particular,formodel for plant

where it is to be applied.
naturalwhetherclosely

designation,
location.

Thisor distantly

when the management regime isand non-orthodox seeded species,
and allows

The latteror schema should

methodology

reserve involves

diversity

et alis meant to act as a guide to some ofThis generalbecause it can be relatively

in a single

or illustrated

and management of a plant genetic

taxon and each situationof genetic

is the most appropriate

minimal,and Maxted

and can be seen to fit within

discussion.

Conservation

to crop plants,

Gadgilproposed

however, that no model, methodology

for each particularconservation

most notably

in this case the methodology

relatedspecificallyby Maxted et al. (1997b).

adaptation

techniquehave

2, for the establishment

the most detailed

genetic

the importantworkingfor genetic reserve conservation,

methodology,

requiremanagement of genetic

crop varieties

has proved more difficult.

ethnographic(1997),

in situ

is not directlyis actually

directed

traditional

cultivation

two basic

conservationthat- the sustainable

continued

permits

evolution.

multipletaxon

However, theFigure2. A Model for Genetic

Reserve

Conservation

Phase 1

Reserve Planning

Phase 2

Reserve

Phase 3

(TakenFrom Maxted et aL9 1997b).

and Establishment

Management and Monitoring

Reserve

Utilisation

88

disadvantagesof conservation

immediately

available

for plant breeding

regime is minimal,

littlelatterthe reserve

case,

often

composition

in a genetic

reserveare that the conservedor other form of utilisation

germplasm characterisation

or evaluation

To illustrate

In themanager may even be unaware of the complete

specific

Diversity

the methodology

of Vicia L.
for establishingand managing plant diversityof Vicia taxa will be discussed.

ecogeographyof Vicia have been recently

and conservation

The genus Vicia comprises

approximately

Europe, Asia and North America, extending(Kupicha,

1981;

Maxted,1995).

to temperate

Perhaps

2.3. million

surrounding

ha were planted

the Mediterranean

with thissea (FAO 1999).

The diversity,

taxonomy,

by van de Wouwet al.,

(ILDIS

1999),

chiefly

locatedSouth America and tropicalthe most well known species
crop,

in a genetic

reviewed

190 species

the faba bean or broad bean, Vicia faba L., an important
1996

and, if the management

data may be available.

reserve, the in situ conservation

Africais notof the reserve.

The Taxonomic and Genetic(2000).

material

pulse, fodderof whichEastof the genus iscrop and vegetable.

570,000

Many other speciesinIn

ha in the countries

of Vicia are used as minor

seed, forage or fodder crops, these include

V. ervilia

Willd.,

V. narbonensis

L., members of theV. sativa

V. benghalensis

L, V. articulata

Hornem. and V.

L. aggregate,

V. villosa

Roth,monantha Retz. The straw and seed of these species
voluminous feeds for ruminants(Ennekingis also traditionallyet al , 1995),

in mixtures with cereal tutor crops, such as barley
Lathyrusboundaries

L.; Lens Mill.;Pisum

and oats.

major classifications

bean. The most comprehensive

subgenus

the four natural
Vicillahas proved

The precise

revision

a populargroup to study,1995).

into two subgenera,(1976),

Vicia.

The distinction

between the

Her subgenus

Vicilla,which includes

forage species

is further

into

V. ervilia,

V. benghalensis

Kupicha'sconceptionof subgenus

and V. hirsuta,

who"Ervum" and "Cracca" in herof the inflorescence

V. villosa,20

for the faba

lengthspots on the stipules.

andOne reason why so muchbased on relative

nectarifous

generic

there beingof the genus was undertaken by Kupichaand "Vicia" and "Faba" in her subgenus

alongwhich has led to an abundant

to Vicia is the lack of an obvious progenitor

groups of species

two subgenera being primarilyof the Papilionoideae

A. Fedorov.

of the genus since Linnaeus (Maxted,

taxonomic study has been devoted
dividedL. and Vavilovia

between these genera have been much debated,complex synonymy. The genus its-selftowhile the forage is often grown for hay

The genus Vicia L. is a member of the legume tribe Vicieae
withused as an additive

Vicia is more coherent, containing

and the presence ofdivided38 species

such as17 sections.
dividedinto 5sections.

Maxted (1993)

following

her suggestionsuggestions,splittingmany of Kupicha,(1976)

faba monospecificsectional

acceptedsativastatus.

Plitmann,

1970;

Schafer,

althoughsecondarySouthern

Siberia

1973;

Maxted,1995;

of diversity(Kupicha,1981;

the Mediterranean

Temperatezones and at higher

cultivatedonly known in cultivation
sites

basin.

altitudes

1988;

occupiedby Vicia species

roadsides,

as well as more pristine

et aL, 1988;

habitats

those

regions.
(Bond,

disturbed

from species
fieldsgroups of Vicia species.

Althoughthere have been several investigations

Link et at., 1995;
studiesVillax,andconsidered

open communities.

The

found in disturbed

habitats,(VavilovEhrman and Maxted

1926).

publisheddiversity

Kernick,

important
1978;

they wereecogeographic

withinand between
speciesof theAbd El Moneim etaL,

Potokinaof diversity

throughout

the genus.

withinofregimes

margins

and steppes. The species

et aL, 1993;

reserve conservation

by patterns

field

Robertson

The objective

This

It is

like

have recently

1963;

of Vicia Diversity

et aL, 1997c).

FAO 1999).

The range of habitats

habitatsof genetic

Reserve Conservationis represented

mostand climatic1995).

most of them have however focused on the agriculturally
1954;

and Southern

It is currently

1995;

soil types

Maxted,like woodlandsof various

reserves (Maxtedin areas

for which only very few

studies

diversity

Viciafaba)

endemics,

and Bennett and Maxted (1997)

range of genetic

3 shows

many restricted

Maxted (1995)

of genetic

Figureinfluenced(1989),

there have been no comprehensive

America andis entirely

the wild and weedy flora of agricultural

ICARDA, 1994;

for Vicia,

North(excluding

found in the more disturbed,

Foury, 1950,

Davis &

its distribution

originally

Genetic

Vicia species

and includes

have evolved in generally

1926;

1967;

Hanelt & Mettin, 1989).

and the Middle-East

species

1990;(Plitmann,

V

Bennett & Maxted 1997).

in some sub-tropicalcultivatedgenus (Tupikova,

Vicia

complexes,

centre of current diversity

Maxted et aL, 1989;

is diverse

more advanced are generally

Vicia species,

to two species

1997;

or which are bound by specific(Maxted,and giving

The faba bean is grown across the Northern

and therefore

have been documented

1988;
usefulThe genus contains
Khattabrelating

Potokina,

Nikiforova,

in China, North Africa

human distribution.

debate,are found in South America,of the economicallyadjoiningand enacted

The current taxonomy of the genus has been generally

Basin is the most importantcentresthe natural distribution

into nine sections

and the rank and number of taxa included

The Mediterranean

widelythe species

with only a few areas of continuing

and V narbonensis

further reviewed this subgenus

et al,

1999),

andis to ensure that the maximum possible

the minimum number and size of in situ genetic

is, however, a complex goal to achieve

because detailed

Figure3. Distributionof Economically

UsefulVicia Species(Takenfrom van de Womv,2000).

91

informationon the amount of genetic variation,

population

requirements

and the geographicalof the target taxon is required

commonly unavailable
finding

distribution

for even the most well studied

optimum locations

structure,requirements

Maxted et al. (1997c)

reserve conservation
It should
Figuregenetic

methodology

conservation

reserve conservation

for genetic

developed

fits within

the overall

by Maxted et al (1997b)

been undertaken:

the decision

shown inundertaken,

and the particularconservation

point must address

the issue as to whether

The latter

successfully,

for the target

conservation,

taken that the target

to warrant active

reserve is appropriate

establisheda general

However, bearing ininterest

outlined.

genetic

resource

must have alreadytaxon is of sufficient
strategies

proposed

to meetbefore a target taxon is conserved in a genetic reserve, several steps in theprocess

survey mission

Therefore(Fig. 2).

1. Therefore,

conservation

complicated,

are often a compromise attempting

be noted that this model for genetic

model for plant

The process ofa single species.

of the target taxa to be conserved in the reserve.

mind these constraints,

and these data areand management regimes for genetic reserve is further

and management regimes employed

the diverse

system, habitatgroups of crop relatives.

because reserves will seldom be set up with the aim of conserving
the locations

breeding

taxon.

an ecogeographic

If thisobjectives

survey or a

and appropriate
conservationin ais the case and the reserve

then a scheme that makes the conserved diversity

isavailable

for current

a review of existing

conserved

and future use must also be devised.
Part of the processof selecting

Vicia material

and therefore

situ collections

exist of cultivatedearliestcollections(Vavilov1996).

we must now briefly

Althoughpotential

collected

have not been systematicallyseriousgenetic(IBPGR,largely

as a result

permanent pastureserosionof intensificationand disappearance

and those species
particularlyof agriculture,of wild climax

of less immediate

are often

associatedovergrazing,

community species.

For example, in

There has been, however, been a systematic

attempt

to conserve Vicia

in the EasternSouthampton,

Weedy Vicia

are disappearing

diversityand latterly

Mediterranean.
University

farmingofwhich

the world.

traditional

declinesystems

throughout

withtrue thatin the Mediterranean

Albania all land races of Vicia sativa have already been lost (Hammer et al 1996).
speciesThe

over the years is considerable,

conserved ex situ. It is undoubtedlyof Vicia diversity,

large exand co-workers in the twenties

collected

for the South American species

Relatively(van de Wouw et al., 2000).

by Vavilov

the number of accessions

there is currently
1985),

review what is conserved.

and wild Vicia species

are the accessions

gaps still exist, particularly
utilisation

target taxa will involve

Maxted and co-workers at theof Birmingham,

UK, in conjunction

University
with

IBPGR,of92

ICARDA and CLIMA have undertaken
and various republics
been duplicatedcollection.

annual collecting

expeditionsof the former Soviet Union since 1985.

at the Genetic

to Turkey, Syria, Greece

All of the material

be noted that the ex situ conserved material

does not provide

found within the genus, either in terms of species

Table 1. Number of Vicia Accessions
de Wouw, 2000).

Conserved

Sp ecies
V.articulata
V.bengalensis
V.ervilia
V.fabaV.m onantha
V.narbonensis
V.p annonica
V.p eregrina
V.sativa
V.villosaO ther Vicia
species*
5ta1*Numbers in brackets

indicate

has

Resources Unit at ICARDA, as well as the in the country of

A summary of current Vicia gene bank germplasm is provided

AARI
1
0
147
77
0
22
4
12
370
35
2 14
(87)
882

collected

A TFC
26
58
254
14 10
15
267
35
4
724
110
276
(39)
3T79

B ari
19
49
65
1461
19
71
130
29
1726
275
306
(57)
4150

the number of otheror genetic

ExSituconservationof the full diversity

coverage.

in Major

Collections

Institu te
W ‑6
1C A R D A
14
21
34
19
168
324
564
5497
8
197
48
224
7
136
29
259
1178
2840
177
342
357
1105
(35)
(52)
2584
11064

Vicia species

in Table 1. It should

liM A
83
0
232
1064
0
17
5
13
863
143
15
(7)
2435

(Takenfrom van

IP K
14
9
137
1507
10
69
27
9
772
23
226
(49)
2SO3

V IR
ll
44
548
1261
54
54
21
2252
263
168
(37)
4679

conserved.

AARI = Aegean Agricultural
Research Institute,
Menemen, Turkey
ATFC = Australian Temperate Field Crop Collection,
Horsham, Australia
Bari = Istitutodel Germoplasma, Ban, Italy
W-6 =Regional Plant Introduction
Station, Washington,
USA
ICARDA = International
Centre for Agricultural
Research in the Dry Areas, Syria
INIA = Instituto Nacional de Investigaciones
Agrarias, Madrid, Spain
IPK = Institut fur Pflanzengenetik
und Kulturpflanzenforschung(IPK), Gatersleben,
VIR =Vavilov Research Instituteof Plant Industry,
St. Petersburg,
RussiaGermany

93

There has been no attempt to systematicallyconserve Vicia diversity

genetic

reserves or on farm. Undoubtedly,

genetic

Galilee,

Israel;Kaz Dag, Aegean Region, Ceylanpinar

Mersin in Turkey)
vetch species,
monitored

existingand other forms of protected

but here the conservationand managed) and therefore

Further attention
and the potential

it is susceptible

and Amanos,and genetic diversity
unobservedconservation

because their habitats

Turkey,the range of the genus containto further

is not being

genetic

of Vicia diversity

are threatened

the genus has for novel exploitation(a) Reserve Planningof South-east(species

needs be focused on the systematic

ex situ and in situ techniques,

reserves (e.g. Ammiad in Easternareas throughout

is 'passive'in situ in either

erosion.

using bothby anthropogenic

changes(van de Wouw, 2000).

and Establishment

Site Assessments
Althoughconjunction

ecogeographic

techniques(see Maxted et aL , 1995),

especiallywith geographicalinformation

systems,broad

genetic

diversity

location

for individual

reserves.

by surveyingtaxa). Increasingly
withinspecies,

The ecogeographic

targetpossiblyusing molecular

the conservation

1995;

Bennett

the precise
broad areasof taxa and variationsome form of assessmentof genetic

This allows assessment

sites. To facilitate

diversity.

be purposely

selected

withindiversityof the variationthat more than one reserve is establishedreserve sites should

security
for any

to complement

This will permit the conservationof diverseof the gene pool.

There have been several(Maxted,to provide

will need to be compared for(numbers

techniques.

into the future it is advisableeach other in terms of ecogeographic
withinsites

site, as well as comparison between alternative

taxon and that these multiple

ecotypes

however, potential

will involve

areas of potential

survey may suggest relatively

them for taxonomic diversity

this surveying

present at a particularof conservation

identify

where genetic reserves could be sited, they are unlikely

where the reserve might be established,suitability

willwhen used inrecent ecogeographicand Maxted, 1997;

surveysof various

Maxted, and Kell, 1998).

groupsAlthoughof Vicianone of thesecovers the entire genus, they focus on the subgenus Vicia taxa, there is still remains a need for astudy of the taxonomicallywere to determine

largerthe geographicalspecies,

assesstheir conservation

provideclear guidance

subgenus

Vicilla.

and ecologicalThe objectives
distribution

status using IUCN categories

on the most appropriate

conservationof the publishedof variousof threats
strategy.

groups(IUCN,

studiesof Vicia
1994)

and

Of the 190 speciesofVicia (ILDIS,
Threatened1999),
Plants22 species(Walters

and 1 subspecies

and Gillett,1998)

are list by the 1997

in the following

Red List Category

The species listed
taxa are entirely

in Spain,are not included

kalakhensis

Khattab,Syria;
that

descriptionof levels

for a thoroughecogeographic

V. hyaeniscyamus

Mout and V tigridis

Mout. frombased

monographerscase, the IUCN categories

forestablishingsurveys with extensive

genetic

field experience

to be drawn about where to site genetic
diversity.

seriously

threatened

highestconcentration

analysisconcepts

Maxted (1995)
of subgenus

ratherthan

the taxa

an accurate

There is therefore

a needMediterranean

1989)

for vetch diversity

the locationof rare and diverse

concentrationof taxa ofthat a similar

taxa. Maxted also concluded

that the most

Lebanon, Turkey and Israel,of gapto the Vicia gene pool has allowed the identificationof keyMaxted thereforeof fivein Syria:

in Syria.

and the

The application

recommended the establishment

Ain Dinar, Al Hasakah(37

15 N, 42 20 E),

Al Hosn, Horns (34 46 N, 36 18 E) and Mimas,

Druze (32 36 N, 36 43 E), as well as in the Olimpos

Turkey (36 21N, 30 25E),of theand it is believed

taxa are located(Margules,

combinationinreserves to conserve the maximum possibleof potentiallythreatened

theshowed that the highestVicillaare not very helpful

by Maxted and co-workers has allowed

to Syria,

of Vicia in the region.

reserves

populations

Ten. & Guss. and V.

of threats

reserves,

taxa are those restricted

Kessab, Kessab (35 54 N, 35 56 E), Qal'atDjebeland V. barbazitaeof threat for the taxa of the genus internationally.

is seen for the majority

genetic

Vicia

Gomb. V.

Vicia was found in the North-eastern

populations

subgenus

Mout., V. qatmensis

by nationallyareas

range of genetic
pictureendangered

it can be assumed the that the current list reflects

in this particularpriority

conclusions

to note that theV. dionysiensis

Warb. & Eig from Israel;studiedof threatened

However, it is interestingand several critically(e.g.

concentration

review of the threat status of the Vicia as a whole.

Although,
identifying

and Turkey.

in the list

Maxted from Turkey),

have been wellsubgenus

PortugalMaxted & Bisby,V. esdraelonensis

eristalioides

Number of Taxa

drawn from subgenus Vicillaspecies

Red List of

categories:

are all mostly narrow endemics, with the highest

taxa being indicated

IUCN

Beydaglari

National

Park, Cavus,of these sites are shown (Fig. 4). Each site has extensive

Vicia species

found in a range of habitatfrom climax maquis

95vegetation

throughplantationand cropped

diversity,

along with numerous other Mediterranean

privategarigue to deciduous
land.

and state ownership,unsustainablecultivation

they are appropriately

and pine forest,

In these sites

plantation,

species.

The potentialsites are in bothare currently

threatenedby over-grazing,

making these populations

When considering
relative

potential

conservation

low. It may be advisable

final

site selection.

particularof vetch diversity

geneticsuch as local National
will also affect selection

reasons reserves are often sited in forestry

are relatively
reservesites for establishinginterests,

cost of reserve establishmentthat for practicalvulnerable

unlessmanaged.

Fig. 4. Location of sites recommended for establishment

in mind existing

as well as pine

there is also a wealth of other forage legume

but the populations

and forestry

and steppe grassland,

to undertake

If the ecogeographic

area, it may be that this area or a closelyarea, whose management plan could be adapted
the wider reserve, therefore

genetic

reserves.

reserves it is advisable
Parks or Protectedof alternative

to bear

Areas. The

sites, which mean

areas where the establishment

costsome form of cost-benefit

analysisdata suggests

the reserveadjacent

locating

one contains an existingto permit genetic

saving some of the costs of genetic

conservation

prior toin aprotected

in a portion

reserve establishment

of
and

96
maintenance. For instance,
Beydaglari

National

one of the high diversity

Park at Cavus in Antalya,

reserve would be low as the land is alreadysites for the Vicia is locatedTurkey and the cost of establishingprotected.

The site has recently

which are unlikely

habitatestablisheda suitable

there would need to carefullyof Vicia eristalioides

Maxted and Lathyrus

the site would likely

become extinct

belinensis

Assessment of Local Socio-economic and Political

Factors

protected.

privatelyThis is particularly

pertinent

owned land. Government policies

to mismanage the environment,
promoted

to thedetriment

especiallyof broader

Therefore,

if the reserve wereand appropriating

on a voluntarywhen biodiversity

basis,

broad reservewhile

others are

hot spot areas are located on

may often need adjustmentin the agriculture
conservation

found atonly know from this one site).

in delineatingareas, and many reserve area networks are establishedlegally

with Pinus nigra L.,

Maxted and Goyder currently

is stillmajor costs involved

the local

and management or the large populations(L. belinensis

There are clearly

been planted

for Vicia species.

monitoringa genetic

However, in this particularmanagement plan would need revision.
to provide

in the Olimpos

to ensure that incentives

and forestry

objectives.

sectors,

are not

Soundly-basedin situconservationof gene pools with intensive

management may well show only slow progress,

two actions

will be clearlyin terms of economics,

helpful:firstlyfunding

for the initialreserve planning,

donors,and secondly,in terms of organisation,

blueprints

and summaries of organisational

could form part of the national

biodiversity

whetherfrom national,

the availability
international

the widespreadneeds to nationalon private

or even privateof standard

governments, such information

action plan.

land or on areas managed for activities

usage of the proposedof outside

knowledge

In our case study, some of the sites chosen for the establishment
situatedof genetic

reserves areother than conservation.

The current

valleyeither sidesite at Kessab, Syria is a cultivatedthat are used for grazing and the site at Mimas, Syria is alreadywith rocky slopes

heavilygrazed.

As over-grazing

is currently

a serious threat to taxonomic diversity

at both sites there is an obvious conflict

conservation

objectives.

case attempts

Ministryof Agriculture

However, in the latterand Agrarian

to declaresustainable

manner. The current lack of a management policy

considering
basaltic

local

part of the area a reserve and manage the remaining

led to a rapid

loss of biologicaldiversity,

for grazing
which

shepherds

andarea in a morein the Djebel

is particularly

the high number of endemic legume and grass taxa found on this relatively

enclave.

withare being made by the Syrian

Reform, Douma, to persuadeland-owners

has undoubtedly

but

Druze
alarming
small

97
Sustainability

is fundamental

long-term political
Appropriate

plans

example, any building,effectfor the region

industrialisation

trainedmaterials

in situ. Appropriately

unavailable

in many countries.
resultingshouldthere is a need for adequate

objectives

specialists

be checked

or hydro-electric

the conservationinternationallybenefits

conservation,and economic commitment to the reserve to ensure its long-term

development

adversely

to in situ

projectsof the reserve.

who can adequately
qualified

Lack of trained

from conservation

are, forthat might

There is alsoa need forand manage the genetic

staff remain in short supply

staff undoubtedly

to the general public

to see if there
in the region

conserve

and experienced

security.

limitsor areuse and the flow of

may be constrained.

Reserve Design
The previous
selected,stagesof the methodology

but now having identified

reserve for the target
factors,

the potential

taxon at the selected

such as reserve structure,

for the target
potentialsite(s),how the reserve site

This involves

consideration

size, whether a single large or multipleof various

smaller sites are bestreserve shape, environmental

see Hawkes, et al (1997)

Each of these factors is associatedis

we must design the most appropriate

location.

taxon, the use of corridors,

user communities,

have highlighted

heterogeneityandfor detailed

discussionof these issues.

with a copious and controversial

literature,but it is possible

to draw some overall conclusions.
The current consensus view of reserve structure
the Biosphere

program (Batisse,1986).

by a bufferThis

is that based on the UNESCO Man andestablishes

habitat,surroundedshielding

the reserve core from general areas of human exploitation.

core area is sufficiently

zone and outside

a centrallarge to accumulate

target taxon (Lawrence and Marshall,
it is more appropriate
the effective
diversity1997).

size (EPS)

for an indefinite

The reserve design

debatedistributed

but the vast majority
and therefore
diverseappropriate

In practise,

option.

segments

individualsof theconservationand thusof geneticof single

large

to have one large reserve of 15,000

ha orBoth large and small reserves have advantages

andof the wild

several small reserves,ecogeographicallybreeding

period.

is often centred

ha?

zone

This plan assumes that thethat form a viable population

ensure the effectiveon the relative

versus several small reserves, for example, is it bettera network of five each of 3,000

a transition

Rather than focus on the actual size of the reserve,that willin the target species

where possible,

1000 - 5000 potentiallyto target the numbers of individuals

population

disadvantages,

this,core area with a stable

relatives

advantageof crop plants

are rather patchilyrather than a single large reserve, establishedof the distributional

range

would provide

however, often there is little

choicebut to have several

inthe most
small

98

reserves because of extensive
fertilevalleybasin

naturalin Western

habitatSyria,

fragmentation.

there

and there are no undisturbed

combinationof a relatively

high rainfall

of endemic legume species

that thrive

so the target populations

this location
habitatin the disturbed

on the middle

human adaptation

or natural

combined with basaltic

soils

habitatsaof theareas.

However thehas resulted

in a number

present.

would require monitoringfor Vicia it would be wise to establish

corridors

Al Hosn, for instance,

has been extensive

environment for millennia

problem,

In Qal'at

Over grazing is again ato avoid populationa network of genetic

depletion.

reservesAt

linkedbyslopes above the valley basin, away from the towns and villages

and not as exposed as the mountain tops.
If multiple

small reserves

the reserves using habitat

are selected,

corridors,

between the component reserves.

where possible,

temperature,

deleterious

wind, the incidence

anthropogenic

effects
pipelines,

activities

diminishand may leave populations

can be increased

thus facilitatingby linking

gene flow and migration

small reserves do have an increased

micro-environmentalof fire, introduction

associatedreserve by roads, fences,
will necessarily

effectivenessHowever, relatively

to area ratio and so are prone to deleterious
light,

theireffects

includingof alien species,

dams, agriculture,and limit the effective

intensivechanges ingrazing,with the edge of reserves.

edge

as well as

Fragmentation

forestryof theand other humanreserve size, multiply

the edge effects

in each fragment unsustainable.

Taxon and Reserve Sustainability
Sustainability
conservation
reserve

is a fundamental

is not an inexpensive

once designatedconcept

and establishedhabitatnecessitate

the commitment of substantial

periodreserveconservation.

option compared to ex situ conservation.

monitoring,

would therefore

for genetic

will

requiremanagement and site security

be a mistake to consider

levelsactivefor a substantialof resources

it a 'cheap'

Nevertheless,population

periodThis willof time.

for a similar

time period

for conservationin an in situ genetic

reserve.

or a weed associatedsuch as many crop relatives,and itoption.

means that the taxon is suitabledisturbance,

theand consistent

Taxon sustainability

A highly

In situmobile species

may not be suitablefor genetic

over an extended
with humanreserve conservation.

These may be better conserved as part of an on-farm conservation

regime or using an ex situ

technique.

large populations

initially

Taxon sustainability

may be limiting

because althoughfound in the reserve site, over time the population

associatedplant

and animal

especiallyproblematic

populations

in reserves

may migrate,or environmentalof relatively

small

may bedue to changes in theconditions.

area. ConcomitantThis

may prove
with

this

the

99

management plan can probably
therefore

population

unforeseen

stability

be to maintaindefineda viablepotential

for evolutionary

fluctuations,in the short

can never be fullyor unavoidable

shouldonly be reviewed

long-terma viable

population

guaranteed

influences,

population

environmental

withinbecausethe reserve of the target
its genetic

and (c) is at minimal

variationsindefinitely

Sites

and

of possible

even if that is the goal. However, the goal

as one which (a) maintains
adaptation

or medium term.

and potential

taxon.

diversity,

risk of extinction

catastrophe,

IUCN has(b) maintains

itsfrom demographic

includingover-use (IUCN,

1993).

There is little

value in establishingby any form of human development
settlements

may be encroacheda genetic
project

reserve unless it is unlikely(Ingramon for building,and Williams,

amenities,

1984).

It may be difficult

use, but checks for development
however, impossible
time. Legislation
not developed

plans for potentialsites shouldthat once conservationfor other uses may assist

however, that environmental

legislationstrong.

It seems likely

providethe maximum protectionof national

to assess the short term requirementswith the security

that this will always be the case, but legislation

reserve sites are established,

the destruction

undertaken.

Experience

if the political

It shouldor
It is,

they are maintainedof the site.

can be circumvented

militaryin one or two hundred year'ssites are designated

for reserve sites.

nearfor human land

be routinely

to assess the long term human requirements
ensuring

Reserves

etc. and even remote sites may be

used in time for dumping human waste, or for example, the sighting
nuclear installations.

to be effected

andhas show,will is sufficiently

can be strengthenedtoalso be noted that if multipleof any one reserve will obviously

have less overall

impact.
To return to the example of establishing
might

be felt as the area is so dominateda reserve at Qal'at Al Hosn, in Syria,
by human intervention

may be exactly the factor that aids sustainability.

valleybasin was a major staging

post for the crusaders

built

one of their largest

at the head of the valley,

virtually

completely

associatedvisitprovidethe site anyway it could

Krak des Chavaliers.
attractionsin Syria.

with a reserve near the castle
a sustainable

The fertileon their way to the Holy Land and theyin tack and is one of the major tourist

scope to encourage ecotourism

it

it would be unsustainable.

However, that human intervention

castles

initially

The castle

isThere would be

and as many tourists

source of income for the conservation

project.
Formulationof the Management Plan

The reserve site will have been selected
geneticallydiversepopulations

becauseit containsof the target taxon. Therefore,

abundant

and hopefully

the first step in formulating

the

100

management plan is to observe the anthropogenic,
shouldbe surveyed

interactions
decidedwithinbioticso that the species

present

the reserve

be understood,

shouldand a means of implementation

and abiotic

in the ecosystemagreed.

dynamics

a clear conservation

Vicia are weedy species

vulnerable

to changes in human activity,

such as changes in agricultural

decreased

stockingof herbicide.

allowing

the site to reach a climax community, as the majority

climax communities.

application

A detailed

assessment

goal should

and certainlyof subgenus

levels,land.

This by definition

Therefore,of the management plan will require

extensive

appropriate

environment for the target taxa is supplied.

makes them verypractice,organisation,

staff, etc. that are involved.

of Vicia species

experimentation

includedare:description,site

history,prescription(what work needs to be carried out and precisely

and genetic

survey and monitoring

establishing

the genetic reserve will be to conserve a specificwill

requiredetails

associatedpreference,

breeding

system,populations

at the site (e.g.

publicobjectives,interest,

the target

are not found in

how and when to do it), ecologicaltarget taxon, the management plantaxon (e.g.

taxonomy, phenology,

autecology

fire and epidemics),

are a natural

as well as cyclicalcomponent of communityand successional

throughpractice,whichthat could seriously

threatenthe levels

incipient

for example.

enough to accommodate superficial(severechanges as long as

which take into account potential
management action

the natural changes seen in plant populations,on communities,

due to stochasticof the target taxon. The management plan should

taxon populations,but beyond

the site,

fauna and flora).

and density

for the target

and forestry

habitatwithinThe management plan must allow for natural fluctuations

effectfocus ofand density

contain actual limits

agricultural

management, management

and manpower. As the specificof populations

the long-term viability

dramatic

and abiotic

mappingthey do not threaten

emphasised

bioticof the target

levelssize,

format, but items

and description

in populationin populationon the location,size, etc.)

Changes

weather, floods,

and theminimum population
with associatedchangesanthropogenic,

influencing

budgetthe reserve, synecology

dynamics.

flexible

factorsschedule,
with

site

or

to ensure the most

There is no standard

generally

withinconservation

increased

it is not a case of simply

The actual content or style of a management plan will vary depending
target species,

bethe majority

the current management regime is required

applicationIt

are known, the ecologicalMany Vicia speciesof disturbedof the site.

humans undoubtedly

urbanisation
Therefore,

anthropogenicof the target population.

is triggered.

and pollution,

naturalHavinghave the mostor changes in

the management plan must be
factors,

but recognise

those factors

101(b) Reserve Management and Monitoring
The initial
experimentation,

managementof any genetic

and it is unlikelythat the ideal

reservewill

involvean elementofmanagement regime will be known when

initially

establishing

the reserve.

carefulintroduction

combinedwith evaluation,

revisionpractical

application.

Therefore,

the initialof management will be high, with intensive

and extensive monitoring
particularly

importantThus the initiation

procedures

levelof the management plan will
and refinement

and the plan will need to be applied

requirein the lightflexibly.

when the target taxon is a component of a pre-climax

of itsThis may be

community, suchas many Vicia species.
Changes in the structurewill

obviously

affecttarget species

the conservation
appropriate

process is likely

sample quadrat

locations,of the reserve.

regularmonitoring

selectionand timing

productionof recommendations

detailed

discussion

shouldincludespeciesof quadrat

to identify

defining

objectives,of data for quadrat
recording,identifying
recording,

accumulation

key associateddeterminationon the management plan.

and

See Maxted, et at. (1997d)

for

The management plansize and the population

reached.

This would allow management changes to be made, thus ensuring

mechanism indicating

to record

reserve, so the conservationist
the overall

pictureand monitor every species

in the reserve as a whole.

for monitoring

the establishmentof both fixed and random quadrats

taxa are likely

animal species,

on a regular basis.

to include

such as those without

primary herbivores

the target

quadratarethe secure

size should

withintheselectedwill

taxa and sites within

the

plant

obviouslyor transects

in a particular
be kept

withinusually

the reserve.

involves
The key

the other plant andof target taxa would decline,

such asetc.

or absence linked

or fruiting)This form of monitoringtaxon, but may also include

There are numerous methods of assessing
commonly used is presence

Key indicatorwhich the populationor necessary pollinators,

floweringor individualis forced to take samples of data that if effectively

reserve are selected(vegetative,

when these levelsof the target taxa.

It is impossible

indicator

taxa andanalysisa minimum and maximumnumber for the population
act as a feedbackTheof desirableof data sets, statisticalprocessreflectof theany actual or incipient

monitoring
conservation

the reserve

Thus, the populationsof these points in the context of in situ conservation.
shouldwithinmanagement review and amendment will be required.

to involve:

frequencyof the target

integrityin the reserve will requirechange and if detected,
monitoringor size of populations

constant

speciesabundance or diversity.

The most

to some estimate

of densityof a species

quadrat.

Havingbegun monitoring,to permit

easy comparison

theof data from

102

subsequent

surveys.

The actual number of quadrats

between the resources
recordingrequired

extra quadrats.

growth, though

Monitoring

for a long-lived

importantto monitor at similar

to record

comparable

diversity,

the conservationist

monitoring

species

Increasingly,
will require

the monitoring

rathermeasured using

a population
agents,than estimating

molecular

effectabout any significant

changes

vegetational

species

It isabundancein the future.

1995;

the habitat

or

which

The species

plant or animal species

canMaxted, et ai, 1997d),of your target taxon andstate or by acting

as pollinators,

seedof the data sets will indicate

population

densityfrom the previous

Experiencein conserving
was establishedprescription

reserve establishedforestindicates

for the maize relativeof population

change interm whether a trend is

between the natural ranges forthe importancein the tropicalforestof SierraAdethat if

was not amended successionalchanges would alter the balanceof theplantsthe wild maize eliminating

would out-competechanges1991).

sizes withinof monitoring.

the reserve indicatedto conserve it (Shands,mechanism, triggering

Routineby management or other intervention.

Zea diploperennis

Manantlan, Mexico. Routine monitoring
and larger

surveys and over the longer

and those induced

want topopulations.

whether there has been significant

However, care must be taken to distinguishcharacteristics

will

surveys to draw conclusions

that may have occurred in the target

analysis

becoming apparent.

survey the conservationistof the survey with previous

statisticalthe original

once in a season'son genetic diversity,

methods

by grazing

the data for a particularcharacteristics

reserve

from

etc.

compare the population

population

gainedmay be less frequent.

(see Cronk and Fuller,

at a pre-climax

Having collected

reserveto occur annually,

more precise information

or some animals may have a beneficial

genetic

and the information

may focus on other taxa than the target taxon; invasive

maintaining

will be a compromise

stages in the target species life cycle on each occasion to be ablepose a serious threat to target populations

dispersal

the quadrats

is most likely

perennial

results.

may be more routinely

to record

to be recorded

The monitoring

it from theprocess acted as a feedbackin the management of the reserveand ensuringthat genetic

and management of the reserve is not an end in itself.

There is an

resources were safely conserved.
(c) Reserve Utilisation
The establishment
explicit

link

between genetic

reserve must facilitatesustainable

conservation
utilisation,and use, therefore
eitherconservationin the genetic

now or in the future.

The use of the

103material

conserved in the reserve may be divided

among traditional,

generaland professional

users.
Reserves are very rarely establishedin an anthropogenic

vacuum. There are likely

local farmers, land-owners and other members of the local population
proposedreserve

traditionaluse made by local people

These local

peoplesimplysite

and who are likely

may have historicallyhave enjoyedvisiting

harvestedcommunities.

by the establishment
or collected

project

from, hunted

could succeed in the absence of local

definitely

fail if the local population

opposes the establishment

traditional

utilisation

with conservationbufferor transition

is compatible

zones by local,

However, to avoid negation

traditional

It is unlikely

supportand it will

Therefore

where

sustainable

exploitationof theobjectives,
objectives

over or mayof the reserve.

user communitiesof the conservationTheof the reserve.

the site on which the reserve has been established.

that any in situ conservation

hunting,

who may wish to use theto remain as neighbouring

may be disrupted

to be

their

shouldbe encouraged.

access and any harvesting,etc. may need to be regulated.
To illustrate

see the pertinent

how reserve managers can work together

with local

example of the Dana Nature Reserve in Jordan.

communities

we canOne of the objectivesof thereserve is to maintaina semi arid-zone ecosystem.

certain level of grazing

was required,

too much and it would encourage shrub and tree growth at

the expense of some herbs species.

Local farmers wished to graze as many sheep and goats as

possibleand for some time there was a conflict

was proposedthat a vet be employed

the animals

allowing

for some reductionlevels(M. Appleton,

a diverseby the reserve to ensure the healthwere healthierecosystem

between the farmers and reserve staff.

in return the size of the flocks would be reduced.
as a result,

To help maintain

Both the shepherds

so provided

Then itof the local flocks,

butand reserve staff benefited

the shepherds

with an increased

incomein the size of the flocks and so permitting

more sustainablegrazing

Pers. Comm.)

The second user group is the population

at large,

and whether

local,

national

orofinternational,

their support may be essential

to the long-term politicaland financial

viability

the reserve.

As discussed

and aesthetic

for species

conservation

earlier,

is of increasing

importancewill

ultimately

finance

Visitsby the general publicawareness exercise.

the ethicalto professional

the establishment

justificationconservationists,and commonly the general publicand continuationof the reserveto the reserve should be encouraged

If local people

who traditionallyreserve, they may be converted to supporters,
community resultingafrom eco-tourists,

through

as an educational

taxation.
and public

use the reserve site are opposed

if they can see direct

financialschool and other groups of visitors

benefits

to theto theirto the reserve.

The

104
reserve design

shouldtrails,halls,

lecturethroughtake into account the needs of visitors,
etc.

They are also likely

to bring

by way of visitor

additionalby the ecotourism

project

proposednatureincome to the reserve itself

guided tours and the sale of various media reserve information

be illustrated

centres,

packs.

Such usage canfor the genetic reserve near Qal'at Al Hosn, in

Syria
Professionalbasis

use of germplasm conserved in the genetic

to professionaluse of ex situ conserved

characterise,

evaluateand publicise

botanical

gardens publish

catalogues

and descriptions

germplasm.

For thisreason it is wisetothe germplasm held in the reserve. Just as gene banks andcatalogues

level of documentation

reserve need to be on a similarof their collections,so the reserve manager should publishof the germplasm held in the reserve to inform potentialof passport,

characterisation

as extensive

for in situ as for ex situ conserved

The quantity

and levelof documentationand evaluation

data recorded

germplasm (see Ford-Lloyd
has a direct

users. The

relationshipshould be justand Maxted, 1997).
withthe potentialfor

exploitation.
In many cases, the work of professional
linkedthroughpartnershipwithinusers, the general public

Non Governmental Organisations,and local people can beespeciallythose involvedin sustainable

rural development,

conservation

volunteers

or use of resources

with traditional

culturalpractices.

All partners

will therefore

share the goals of sustainableof biologicalresources

takinginto accountfactors,

which are likely

social,to form a cornerstone

in accordance

economic, environmental

to the nations'

proposalsuseand scientific

to implement

Agenda21.

Finally

it is important

to stress

that ex situ techniquesshouldalways be used to act as a

backup for in situ conserved taxa. The two strategies

are complementarybank manager will duplicate

in other gene banks to promote safety,

his or her collection

and just as a good genethereserve manager should also duplicate

his or her collection

in ex situ collections.

series of complementarystrategies

both in situ and ex situ conservation

are currently

being

conservation

enactedinvolving(van de Wouw, 2000).

conserved ex situ as seed accessions

Traditionally,in gene banks and it is importantto act as a back up for any in situ genetic

Vicia

For Vicia, a

specieshave beenthat this method continues

reserve conservation.

Conclusion
There has been a growing
conservationagri-ecosystemsof plant

genetic

threatenedinterest

resources,

among genetic

conservationists

because of the urgent

in the in situ

need to protectwith imminent change and the need to decrease

naturalandthe plant genetic

105

resource conservationists
conservationist
applying

technique,when formulating

an overall

conservation

techniques,

includingof different

methodologies

been focusedresources,

seed conservation.

complement each other.

on developing

techniquesof other biologicaldisciplines,

strategy,

The contemporary

shouldthinkof research

for the ex situ conservationand it is hoped

beginsin terms ofboth in situ as well as ex situ, where

With the majority

there is now a need to redress the balance

the experience
genetic

on a single

a combination

the different
havingreliance

to provide

in the pastof plantthat this paper,

a firm scientificgenetic

buildingonbase for in situ

reserve conservation.

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the in situ approachJ.G.).

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144-159.

Management and monitoring.
(eds.

Ecogeographic

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Maxted,

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E., 1997.
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Villax,

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occidentale,

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Algerie-

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Centre,pp.

1-862.

and Cambridge,

UK.

Plants.
IUCN

Compiled
- The

by

World

Ill

Checklists

and in situ conservation- a case report from

Italy
Karl Hammer1, Thomas Gladis1,
PietroHelmut Kniipffer2,

Gaetano Laghetti3

andPerrino3

1 Instituteof Crop Science,

Universityof Kasselof Plant Genetics

and Crop Plant(INK),

D-37213

Witzenhausen,

Steinstr.

19,

Germany
2 Institute3, Germany
3 Istituto del Germoplasma(C.N.R.),

Research

1-70126(IPK),

D-06466

Gatersleben,

Bari, Via G. Amendola 165/a,Corrensstr.

Italy

Abstract
The methodology

and use of checklists

for plant

genetic

resources

cussed. Italy is taken as an example to demonstrate the usefulness
the Italian

territory

has been covered. Two larger checklists

Germplasm in South Italy and Sicily"
Italy".

A catalogue comprising
Checklists

and a "Catalogue

small islands.

resources,a genetic interaction
This

conservation,

dis-

Meanwhile most of
into a "Catalogueofof Crop Germplasm in Central and Northern

wheat is discussed

The situationof specificof Linosa and Ustica is discussed.

and on-farm conservationpreconditionforcrops with respect to on-farm

here. In the same direction,is proposed

between wild or weedy and cultivatedis an excellent

are briefly

all of Italy is envisaged.

management. The example of hulledin plant genetic

of checklists.

have been developed

have also been useful for the investigation

useful for studying

studiesa dynamic,

and can be only achieved by an integrated

checklists

Both islands

areare rich

for their maintenance. There is often

species.

Introgressions

evolutionary

approachare quite common.
towards

germplasmin situ and on-farm management.

Introduction

largely

Checklists

are usefultools

neglected

in studies

on plantrino (1985),

theywhere results

had to be obtained

atic studies
lists

in all kinds
genetic

can be used in surveysof larger

has been describedwithinareas, e.g., Libyaof botanicalresearch.

They have, however, been

resources.

As demonstrated

by Hammer and Per-of remote areas, such as the Ghat oases of Libya,
a rather limited

time. They are also useful(Hammer et al. 1988).

by Hammer (1990).

The generalfor system-

work with check-

112

Meanwhile
comprehensive

our school

database,

has usedchecklists

in several

countries,

and createdathe contents of which is summarised in Table 1).

Table 1. Summary of the checklistsof cultivatedplants database. The years refer to therespective publications
for Cuba (Esquivel
et al. 1992) and Korea (Hoang et al. 1997). Figures
for areas in preparation
are still incomplete(after Kniipfferand Hammer 1999, partly updated). Data for Italy are provided in Table 2.
C o u n tr y o r r e g io n
T ax aS p e c ie s
G e n eraF a m ilie s
S yn on y m s
V e rn a c u la r n a m e s

C u b a (19 9 2 )
1 ,0 4 4
1 ,0 2 9
531
117
729
1 ,6 6 9

K o r e a (1 9 9 7 )
605
578
378
Ill
497
7 14

E A s ia

(in p r e p .)
996
940
52 9
14 7
6 86
2 ,8 8 9A lb a n ia (in p r e p .)
433
4 18
255
82
225
2 64

1 The database under construction for East Asia contains information on China (at present 552
species and 694 folk names), Japan (448 and 497, respectively)
and Korea (875 and 986, respectively).
Relevant information has also been collected during a mission in Japan(Tomooka et al. 1999).
Checklists

designed•E Nomenclature
original

by us (see Kniipffer(accepted

name: genus, species,

publication/original

information

family,containinfraspecificquotation),includingimportantabbreviatedlanguageor region,•E Vernacular names, including•E Geographicand Hammer 1999)(origin,distribution,

name, author, place ofand regional

synonyms,

own collections/observations),•E Usages and plant parts used,•E Narrative

text of any length,

wild relatives,•E Editorialand ethnobotanicalremarks concerning

specimens•E Literature
Checklists

e.g. for information

breeding,

aspects,a particularspecies,e.g. literaturereferences

and herbarium

references.
for Italianplant

genetic

resources

have been used intensively

flora of Italy.

For this purpose

This checklist

was used in the fieldwork

vated plant species

starting

diversity,

yet to be checked,

Checklists

fraspecificabout the history,a preliminary

and their wild relatives.

taxon found in cultivation
point

for floras,

catalogues

in our work on the agricultural
checklist

was designed

and horticultural(Hammer et al. 1990b).

for asking

the farmers for rare and obsolete

culti-It allows

to quickly

or in-

check whether a species

was known from that area before. It also forms a goodof cultivatedplants,or monographs.

The inclusion

of

113

widelyused synonyms and those found in local literature

the checklists

usefulalso for studying

elder literature

Based on the first checklist,
and Sicily

as well as vernacularsources (for example, see Table 4).

an early catalogue

which turned out to be extremely

was publishedrich in plant

genetic

comprisingresources

see also Table 2). This

catalogue

was mainly used for the further

resultedin another catalogue

for Central

and North Italy (Hammer et al. 1999a)checklistsof two smaller

islandof Sardinia

is still(Hammer et al. 1997,

in preparation

1999b).

A largerSouth Italy(Hammer et al.

1992;

islands

names, makes

explorations

andas well as inchecklist

for the(Hammer et al., in prep.).

Table 2. Summary as of mid 1998 of the checklistsof cultivatedplants database. The years
refer to the respective publicationsfor South Italy and Sicily (Hammer et al. 1992) andCentral and North Italy (Hammer et al. 1999a) (after Knupffer and Hammer 1999). The database for Sardinia is in preparation(Hammer et al., in prep.); the figures include some utilised
wild plants. The summary values for Italy take into account the overlap of plant species
between the regions.
R e g io n
T axa
S p e c ie s
G e n eraF a m ilie s
S yn o n y m s
V e r n a c u la r n a m e s

S I ta ly (1 9 9 2 )
540
52 1
298
86
348
2 ,9 8 1

Additionally,findings

C & N I ta ly (1 9 9 9 )
568
550
327
92
3 44
1 0 ,8 0 2

the data on Sicily

and literature,and the smaller islands

so that the figures

general comparison,S a r d in ia (in p r e p .)
375
37 1
247
81
250
2 ,4 2 0

in Table 2 are still

have to be updated
incomplete

I ta ly (to ta l)
687
665
380
10 5
459
1 5 ,6 2 1

from recent

and do not allow ae.g. between South Italy and the more northern part. A better preliminary

estimation

can be drawn from the number of accessions

Sicily1622

accessions

have been collected(Hammer et al. 1992),

the respective

result fromCentraland Northern Italy is 486 accessions(Hammer et al. 1999a).

Comparing

the groups ofin the central

and northern

uses (Table3), the large

number of medicinal

parts of Italyis evident,

whereas species

statedplantsof fruit

collected.

cultivatedtrees

predominate

above, a correct comparison can be done only after updatinga general

cataloguethat is planned

for the Future. A Catalogueof Italian

to appear within

Whereas in South Italy

in Souththe checklists

Italy.

But, asand creating

the next two years under the title

Crop Germplasm".

and"Seeds114

Table 3. Summary of plant uses: number of taxa for which a particularKnupffer and Hammer1999)
U se
M edicinal (M .)F ruit (Fr.)
F orage (F o.)
V egetable (V .)S pice,arom atic (Sp .)F ibre (Fi.)
Industri al (I.)
S tarch (S t.)
O il (O i.)
C ereals (C .)T otaluse is reported(afterN um b er of species
S Italy and S icily (1992)
C an d N Italy (1999)
12 1
10 8
90
110
60
8
50
7
14
23
540

140
95
97
124
71
7
51
9
15
22
568

Case studies
As early

as in 1984,

a proposal

was made by us (Perrinoand Hammer 1984)

conserve

the traditional

cerealsemmer and einkorn,which

cultivationin parts of South Italy.

This was not a proposal

for ex situ conservation,in situ approach

under on-farm conditions.

the EC made a positive
In stressing
dealingselection

At that time, no one accepted

power of South Italy,

of introgression

several(see Heiser

introgression

from wild and cultivatedwild and cultivatedspp. (Perrinoand Hammer 1985)of these studies

regions

progressed

genetic

is preferred)

was coined,

generally
Hulledunder

but for an

this idea, and neither

presenting

new evidencerye (Hammer et al. 1985),

and other crops. For a summary

see Hammer and Perrino (1995).

However, the change in conventionaland subtropicalbe foundpapers have been published1973),

from the area, e.g., reporting
Brassica

stillof this new idea.

the evolutionary

with the old problem

could

to

(e.g.,

erosion.

was particularly

Esquivelthinking

came from another side,and Hammer 1988, Altieri

The term "on-farm conservation"and in situ conservation

1989,(todayi.e., tropicalBrush 1989)

with less"on-farm management"(even the term "management" is applied)

promoted by the Convention

on Biological

Diversity(CBD,1992).

wheatsUsing our checklist

method, even further

indicatorsfor rare crops in northern parts of Italyof hulled

wheats in Italy(Perrino

traditional

and threatenedmaterial.

et al. 1981),

observations

were made on hulled(Hammer et al. 1991).

After the re-detection

a programme was establishedThe most effective

wheat asto conserve thismethod was to re-establish

the tradi-tionalforms of use and to develop

1996).

In 1995

a workshop

was carriedout (Padulosi

ment industrial

and scientificprogressin Italy,

Europe started

to work with the hulled

progresssee the paragraphs

in Italy,

new products(Perrino

et al. 1996),

D'Antuonowhich

and Bravi

did not only docu-

but also showed that many other countries

wheatson Triticum

et al. 1993,

again.

As an example for checklists

inand

monococcumand T. dicoccon in Table 4 forcomparison.

Table 4. Two examples for hulled wheat from the Catalogue
and from the Catalogue of Central and North Italy (right)of South Italy

and Sicily(left)

116
A fter H am m er et al. 1992

A fter H am m er et al. 19 99

T riti cu m dicoccon S ch ran k , B aier. F lora 1 T riticum dicoccon S ch ran k, B aier. Flora 1(1789) 389. ‑ T riticum d icoccum S chuebler, (17 89) 389. ‑ T riticum dico ccum Schuebler,
C har. E t descr. cereal, in H ort. T uebing. C har, et descr. cereal, in H ort. T ueb ing .
(1818) 29 (G ram ineae).
(1818) 29 (G ram ineae).
A vena am ericana (B a.), aven a forestiera A dor, adoreum , alica, far (E m .), far, farre(B a.), biada francese (B a.), farra (A p.), farro, (U m .), farro ,sem enspelta am ericana (B a.), speuta (C am .), sp euta D istr.: C e. (T o., E m ., A b., U rn ., M a.), N o.
fran cese (C am .),vena am ericana (C am .)(P i.)
D istr.:A p .,B a., C am .
c.
cD om esticated in Southw est A sia. In the area
D om esticated in S outhw est A sia. In the area know n from 4000‑ 3000 B .C . (Z oh ary andkn ow n from 4000‑3000 B .C . (Z ohary and H opf 1988), frequently in the 4th‑ 3rd century
H opf 1988), frequ ently in the 4 ‑3IU century B .C . (C o stantini 1983). In R om an tim es
B .C . (C o stan tini 1983). Form erly an im por‑ kn ow n as far, ador, sem en , adoreum an d alica
tan t cereal covering large parts of th e area in‑ (Pap a 1996). Form erly an im portant cereal
eluding , e.g ., Sicily. T oday only a relic crop . covering large parts of the area. T oday only a
A certain v ariation can be observ ed still to ‑ relic crop (H am m er et al. 1990a). A certaind ay. T he land‑races are the result of local ad‑ v ari ation can be ob served still today. T heaptation and possibly also introgression s from landraces are th e result of local adaptation andother Triticum spp. (H am m er an d P errino possibly also introgressions from oth er Triti‑
19 84). M ore frequ ently grow n th an T. m ono‑ cum spp . (H am m er and Perrino 1984). M ore
coccum L . but w ith sim ilar distribution frequently grow n th an T. m onococcum L . but(Perrino et al. 19 81,H am m er et al. 19 85).
w ith sim ilar distri bution (P errino et al. 1981,
1988, 1993, H am m er et al. 1985 , 1990a,
L agh etti et al. 1994 , D 'A ntuono and B ravi
1996, Szabo and H am m er 1996).

Abbreviations:Ap. = Apulia, Ba. = Basilicata,Cam. = Campania, Si. = Sicily; Ce. = Central
Italy, No. North Italy; Ab. = Abruzzo, To. = Toscana, Em. =Emilia and Romagna, Um. =Umbria, Ma. =Marche, Fr. Friuli and Carnia, Pi. = Piemonte; C. = cereal

LinosaA checklist
et al. (1997).
important

for the crop plants

Thus, for the firstislandtime,of Linosa island

1) has been publishedan account of the crop plants

has been made. On the other hand, the wild flora

mented. Endemics

are Catapodium

neuradifolium

Delilevar. linosae(Sommier)ssp. algusae(Brullo)

Greuter(Plumbaginaceae),(Brullo(Fig.

and Piccione1980).

Many of the wild species

Severalzwierleinii
Brulloother species

have to be considered(Lojacono)

Brulloby Hammerof this agriculturally
of Linosa

very

was well docu-(Gramineae),

Erodium(Geraniaceae),

Limonium oleifolium

and Valantia

calva

Brulloare rare and/or of phytogeographic
as genetic

resources,

Pign.

(Rubiaceae)
interest.

e.g., Lotus peregrinus

L.

(Leguminosae),

Ononis serrata

Forssk.

(Leguminosae),

Castellia

tuberculosa

be found, excluding

all plants(Moris)Bor(Gramineae).
From the crop plants,
for ornamental purposes

60 species

could

only. A proposal

farm conservation(Hammer et al. 1997).

the flora, vegetationand landscape,approachof nature protection

was made to develop
In addition,a naturalconsideringand crop management in situ. As alreadymentioned,

plants.

integrityof relatedApartsolving

thefound for developingofThus, an integrated

an integratedof nature protection

are important

for the fur-due to the same reason, crop plants

wild species.

practical

management and in situ conservationinterests

introgressions

For nature protection,

endanger the genetic
frominterest

and on-farm management would be an ideal conclusion.
because of the differentof cultivatedinto a centre for on-

the remarkable

reserve could be suggested.

Such an approach is not easy to follow

ther evolution

the islandthat are grownandscientificproblems

on Linosa (see Hammer et al. 1997)

approachwith nature protection.

regardingon-farm

a way has to be

118

UsticaAnother islandof intensive

comprisingmore than 110

ornamentals(Hammer et al. 1999b).

studiedby other researchers,

earliera summary of families

speciesstudieswas Usticaof agricultural
Although

2). A checklist

and horticultural

the cultivated

58 crop plant species

and genera, see Table 5.

(Fig.

plantswas establishedplantsexceptpureof this island

had been

have been newly reported

by us. For

119

Table 5. Index of families and genera of crop plants for Ustica. The plant families are given
in alphabeticalorder. For each family, the genera represented are also listed alphabetically.
The number of species per family or genus is given in brackets, except for the case of one
species per family or genus. Infraspecifictaxa are not considered in the counts (after Hammer
et al. 1999b).
A iz o a c e a e
A n a c ar d ia c e a e
B o r a g in a c e a e
C a c ta c e a e
C a p p a rid a c e a e
C h e n o p o d ia c e a e (3 )C o m p o s ita e (5 )C r u c ife r a e (3 )C u c u rb ita c e a e (5 )E b en a cea e
E u p h o rb ia c e a e
F a g ace ae
G ra m in e a e (6 )J u g la n d a c e a e
L a b ia ta e ( 5 )L au race ae
L e g u m in o sa e ( 14 )C a rp o b ro tu s
R hus
B o ra g o
O p u n tia (2 )C a p p a r is
A tri p lex , B e ta , C h e n o p o d iu m
C y n a r a (2 ), C ic h o r iu m (2 ), H e lia n th u s , L a c tu c a , M a tr ic a r ia
B r a s s ic a , E r u c a , R a p h a n u s
C itr u llu s , C u c u m is (2 ), C u c u rb ita (2 ), L a g e n a r ia , L uff a
D io sp y ro s
R ic in u s
C a s ta n e a
A ru n d o , A v e n a , H o r d e u m , S o r g h u m , T r itic u m (2 ) , Z e a
J u g la n s
M e n th a (3 ), O c im u m , O r ig a n u m , R o s m a r in u s, S a lv ia
L a u rasA c a c ia , A r a c h is, C e r a to n ia , C ic e r , H e d y s a r u m , L a th y m s , L e n s , L u ‑L ilia c e a e (2 )L in a c e a e
M a lv a c e a e
M e li a c e a e
M o ra c e a e (2 )M u sac eae
M y op orace ae
M y r ta c e a e
O le a c e a e (3 )P a p a v e ra c e a e
P itto s p o ra c e a e
P o ly g o n a c e a e
R o s a c e a e (7 )R u ta c e a e (2 )S a lic a c e a e
S o la n a c e a e ( 3 )U m b e llife r a e (4 )V e rb e n a c e a e (2 )V ita c e a ep in u s , M e d ic a g o , P h a s e o lu s , P is u m , R o b in ia , V ic ia , V ig n a
A lliu m (4 ), A s p a ra g u s
L in u m
G o s s y p iu m
M e lia
F ic u s (2 ), M o m s (2 )M u s a (2 )M y op o rum
E u c a ly p tu s (2 )F ra x in u s , J a s m in u m , O le a
P a p a v er
P itto sp o r u m
R um ex
C r a ta e g u s , C y d o n ia , E r io b o try a , F r a g a r ia , M a lu s , P r u n u s ( 7 ) , R u b u s
C itr u s (5 1 R u ta
S a lix
C a p s ic u m , L y c o p e r s ic o n , S o la riu m (3 )A p iu m , D a u c u s, F o e n ic u lu m , P e tr o s e lin u m
L ip p ia , V e r b e n a
V iti s120

Introgression
domestication.
al. 1999b).

has been found to be a spectacular

indicator

A number of such cases could be postulated
Examplesfor ongoing

evolutionandor observed on Ustica (Hammer et

are given in Table 6.

Table 6. Genetic interaction
between wild/weedy
Hammeret al. 1999b);
crops in alphabeticalorder

and cultivatedplantson Ustica(afterF o r m e r in t e r a c tio n
W ild /w e e d y r e la tiv e s
R em a r k s
w e e d y B o r a g o o fi c in a lis P r e s e n tly n o c u ltiv a tio n
B o ra g o off ic in a lis
o n ly n e w v a r ie tie s u n d e r c u ltiv a tio n , s e e d
D a u c u s c a r o ta s s p . w ild D . c a ro ta
S a tiv u s
o n ly b o u g h t fr o m th e sh o p
L . s e r r io la , L . s a lig n a , L . to d a y , th e w ild re la tiv e s a r e a b s e n t o r v e ry
L a c tu c a s a tiv a
V ir o s ara r e , n e w v a rie tie s u n d e r c u ltiv a tio n
L in u m u s ita tis s i‑
L . u s ita tis s im u m s sp . a n ‑d e c lin e o f c u ltiv a tio nm um
L u p in u s a lb u s

g u s tif o liu m , w e e d y L .
u s ita tiss im u mw e e d y L . a lb u s

C rop sA p iu m g r a v e o le n s

P r e s e n t in te r a c t io n
W ild /w e e d y r e la tiv e s
R em a r k s
w e e d y A . g r a v e o le n s
c o m m o n c ro p w ith lo c a l s e e d p r o d u c tio n ,A v e n a s a tiv a

A . b a r b a ta

B e ta v u lg a r is v a r .
C ic law e e d y B . v u lg a r is , B .

B r a s s ic a o le ra c e a

B . f r u tic u lo s av u lg a r is s s p . M a r itim a

C a p p a r is sp in o s a

w e e d y C . sp in o s a , C . r u ‑C ic h o r iu m e n d iv ia

p e s tn s
C . p u m ilu m

P re s e n tly n o c u ltiv a tio n

w e e d y r a c e fr e q u e n tly o c c u rrin g c lo s e to th eg ar d e n s
w ild r e la tiv e re p o rte d b y S a lv a to r ( 1 8 9 8 ) ,th e re a re s till so m e v a r ia b le p o p u la tio n s o f
A . s a tiv a w h ic h m a y b e th e re su lt o f f o r m e r
in tro g re s sio n , in th is w a y e v o lu tio n c o u ld g oon .
w e e d y ty p e s g r o w e v e n in th e g a r d e n s a n d
a re u s e d in th e sa m e w a y a s th e c u ltiv a te do n e s , fre q u e n t in tr o g r e ss io n e v id e n t fro m th ew ild /w e e d y ra c e s to th e c u ltiv a te d o n e s a n dv ic e v e rs ain tro g r e s s io n is p o stu la te d b y th e fa r m e rs
c o n s id e rin g w ild p la n t c h a r a c te r s in th e ir
c u ltiv a te d r a c e s (d o u b tfu l !) , s e lf ‑p o llin a to r ,c h a n c e f o r in tro g re s sio n lo w , m o re o v e r ,c ro p a n d w ild r e la tiv e a re r ar es e v e r a l s p in y p la n ts w ith in th e c u ltiv a te d
ra c es e lf‑p o llin a to r , c h a n c e f o r in tro g r e s s io n lo w ,m o r e o v e r , c ro p a n d w ild r e la tiv e ar e ra r e

121
C icho rium in tybus

C . intybus var. glabratumcross‑pollinator, chan ce for introgression
highF oeniculum vul‑
F . vulga re ssp. P ip eritum
w eedy races are used and protected, theand w eedy ssp. V ulga reprobab ility of in trogressions in both direc‑
ga re ssp. vulgare
tion s is highO lea europaea var. w eedy var. europ aea, var. geneflow from the cultivars to theeu rop aea

sy lv estrisO riganum vulgare

O .vulga re ssp. V irid essp. Viridulum
P isum sa tivum

v/ild/w eedy races is lik ely,but th e cultiv ated
trees are rare tod ay
introgressions easily possible

R osm a rinus offi ci‑

w eedy P . sativum convar. P . sativum ssp .sp eciosum (Stev en)Speciosum
S chm alh. rep orted by C alcara (1842) as P .
b iflo rum
P . sp inosa (P .am ygdali‑ both species grow clo sely together in som e
fo rm is)places, introgression from the cultivated toth e w ild sp ecies is likely
w ild and w eedy R . offt ci‑ exch ange of genetic inform ation in both di‑na lisR uta chalep ensis

na lisrection s
w ild and w eedy R . chale‑ only m inor differences betw een w ild andP yrus com m unisp ensisBecauseof itsconsiderations

cultivated racesrichspeciesand infraspecific

Usticaas an area for on-farm management. The presence

promotes the ongoing

evolution.

Usticais alreadyProtected

NaturalMarinalReserve (Laghetti

integrityof plants

relatedto crop species

easier to unite both protectionin Ustica

diversity,
protected

et al. 1998).
as usually

becauseThis

is worthfurtherof crop plants

strongly

it has the status

does not claim the genetic

under nature protection.

aims, so that there are betterof aprospects

It seems to be

for on-farm management

than in Linosa.

Conclusions
The southern
Mediterraneanpart of Italy(Vavilov1926).

belongs

to the classical

It is characterisedof checklists

this could be confirmed.

al. 1992).

From this number, 202 species
and cultivatedby a richness

More than 500 species

stocksweedy or ruderal
is possible

gene centre of theof crop species.

With the help

have been reported(Hammer etor 36.8 % belong to the local

sense that they can be found there as wild,
tween this material

Vavilovian

plants.

and underlines

flora of the area in the
Genetic

interaction

the evolutionary

beimpor-tance of the area.
Checklists

allow an "anatomical

ies are time and labour
world. The work initially

Harlan (1951).

Such stud-consuming and they do not fit well into our economicallydominatedstartedstudy" of an area following

as a co-operation

project

between gene banks. This is the122part usually

done by persons

and characterisingplant

situ collections

and trying

this

plant

themselvesto collecting,resources,

lookingfor gaps within

genetic

genetic

maintaining,
diversitypresented

in domestication

the typicalof gene banks. They have been engaged

resourcesand on-farm management tools,

tasks

and developing
applicableideas

processes

describing

to find paradigms

work goes far beyond

monitoring

committing

in ex(Hammer 1994).

here inand methods for establishingfor many comparable aspects

But

in situof protection

diver-

sity of life on earth, e.g. Hammer(1996).
References
Altieri,

M.A., 1989.
Conservation

Brullo,Rethinking
Biology

crop genetic

conservation:a view from the South.

vegetazione

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Programma finalizzato

3, 77-79.

S., V. Piccione,

1980.

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Rethinking

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Descrizione

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Calcara,P., 1842.

Castagna,

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Resistanceto rusts

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Evolution

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Das Domestikationssyndrom.

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Hammer, K., 1996.

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38, 191-310.

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1990b.

S. Bullitta

Seedsfrom the Past. A Catalogue

1999a.

Seeds

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A Cata-Bari, 253 pp.

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A checklist

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Hammer, K., G. Laghetti

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32, 1 1-34.

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Hammer, K., G. Laghetti,
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Hammer, K., H. Knupffer

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Hammer, K., H. Knupffer,on the infraspecific

Crop Evol. 44, 285-288.

Hammer, K., 1990.
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Hammer, K., Chr. O. Lehmann and P. Perrino,
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33, 237-267.
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1983.

Kulturpflanzeof the germplasm

collected

Further

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Schrank)

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Kulturpflanze

32, 143-152.

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Hammer, K. and P. Perrino,
Kulturpflanze

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1981,

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1982

and

36, 475-527.

Hammer, K. and P. Perrino,
T. dicoccon

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33, 269-286.

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oases.

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Hammer, K. and P. Perrino,
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Plant

Genetic

Resourcesin situ and on-farm conservation.

Harlan, J.R. 1951.

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Heiser,Introgression

C.B. 1973.

plants

(5).

Italy

Genet. Resources

and Sicily

Newsl.

- Studies103: 19-23

Amer. Nat. 85:97-103

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Hoang, H.-Dz., H. Knupffer
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and K. Hammer. 1997. Additionalnote to the checklistof Korean

Consolidated

Genet. Resources

Crop Evol.

summary and indexes.

44:349-391.
Knupffer,

H. and K. Hammer, 1999.

cultivatedplant

species.

of cultivatedGardens,

In: S. Andrews,

plants:Third

Gardens,

sul farro in Italia.

International

Agricoltura(Italy):collecting

Ricercaand P. Perrino,Plant,,m situ" und in der Genbank.

S., K. Hammer and J. Heller

Papa, C, 1996.

The 'farreRoyal

pp. 215-224.

pp. 154-171.

P., 1982.

Nomenclatura

di T. dicoccon
P., P. Basso,in ambienti(eds.),

1996.

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Schrank)A. di Marzio,
marginali

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ancora coltivati

Botanic

Roy. Bot.

Appeninodelle

resources

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Newsl. 1 16, 12-17.
Ressour-

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Kulturpflanze

35, 109-134.

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Rome, 262 pp.

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In: S. Padulosi

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N. Volpe e G. Laghetti,dell'

Risultati

Crop genetic

crops 4. IPGRI,

landrace

relativa

1994.

Genet. Resources

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de Montelione':

1996,

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of

Taxonomy

Sammlung pflanzengenetischer

sal der Landsorten(sin.

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- ein Reisebericht

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156, 1 19-124.

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Perrino,Third

R., K. Hammer und Chr. O. Lehmann, 1987.

Padulosi,

and C. Alexander

Symposium,

L.F. D'Antuono

G., K. Hammer, G. Olita

Maly,

A databaseKew.

condotte

islandA.C.Leslie

International

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biodiversity:

Kew.

New Taxonomy of CultivatedLaghetti,

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et al.

Schiibler

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1993.

e Selezione

Valutazione

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Infor.

diAgr. XLIX (43),41-44.
Perrino,P. and K. Hammer, 1982.
cultivatedin Italy.

Triticummonococcum L. and T. dicoccon- Presence, collecting

and actions.

Perrino,P. and K. Hammer, 1983.

Sicilian

Perrino,P. and K. Hammer, 1984.

The farro:servation.

Genet. Agraria

38, 303-31

wheat

1.

varieties.

further

Schrank

Genet. Agraria

36, 343-352.

Kulturpflanze

31, 227-279.

information

on its cultivation

are stilland con-125
Perrino,P. and K. Hammer, 1985. Collection
1984.

Perrino,Kulturpflanze

Perrino,of indigenousP., G. Laghetti
1987.

Salvator,material

1981.

of cultivated

Ustica.

Wild

relatives

toringand collecting

Vavilov,

Bull.

plants.

to South

KulturpflanzeCollectionof plant

Italy

1980

for the29, 433-442.

genetic

resourcesin Italy,

In: S. Padulosi

et al. (eds.),

of crops conservation
mission
1998.

1996,

monococcum, T.

pp. 183-192.

K. Hammer, R.Q. Xu and Y. Tsubokura,in Japan with a focus on Vigna species.

to Tochigi,

Ibaraki,Shiga

Annual Report on Exploration

and Tottori

1999.

2. Moni-prefectures.

4th andof Plant

Genetic

and Introduction

15, 63-66.

N.I., 1926.

plants.

of travels

on the taxonomy of faro: Triticum

Tomooka, N., D.A. Vaughan, N. Nakagahra,

Resourcesin South Italy,

Prag, Verlag Heinr. Mercy Sohn.

and T. spelta.

6th - 8th Octoberplants36, 377-390.

Szabo, A.T. and K. Hammer, 1996. Notes
dicoccon

Reportand K. Hammer, 1988.

KulturpflanzeL., 1898.

of cultivated33, 225-236.

P., K. Hammer and P. Hanelt,collectionof land-races

Geographicalappl. bot,

Zohary, D. and M. Hopf, 1988.

regularities

in the distribution

genet, i sel. 17, 3 (Russ.),
Domesticationof plants

411-428of the genes(Engl.

summary).

in the Old World. Oxford.

of cultivatedCases Studies

129

Research on Vigna subgenus

Ceratotropis

genetic

that provide

insights

into in situ conservation

Duncan Vaughan,

Norihiko

Tomooka,

Akito

Dynamics

Laboratory,

Genetic

Kaga,

Kooji

resources

strategies
Doi,and Mun Sup

Yoon1
Crop

Evolutionary

Agrobiological
1 Current

2, National

InstituteofInstituteofResources.
address:

Agricultural
Republic

Resources

Genetic

ScienceResources

Division,and Technology,

Bioresources

Department,

National

Rural Development

Administration,

Suwon 441-707,

and forage

legumes of particularof Korea.

Abstract
Vigna subgenus

Ceratotropis

importance to Asian agriculture.
of island populations,are presented.

food

are discussed.

and species level studies

To illustrateof these diversity

studiesof taxa withinvarious aspects of in situ conservation,

the azuki bean crop complex and a core species collection

The implications

the species discussed

severalThe relevance of population

the subgenus to in situ conservation,
studies

includesfor in situ conservationand have a general relevance to plant genetic conservationof the subgenus

are both specificforresearch. Among theconclusions are:
•E Related

specieshaving the same distribution

based on their evolutionary

may have distinctly

history

or diversity

introduced

level research to understand patternsof diversity

is a prerequisite

different

genetic

variation

into the same area. Thus, population
for sound in situ conservationof wild species.
•E Crop complex populations

where gene flow may occur cannot be easily classified

morpho-agronomic and ecologicalsuch populations.
humanactivity,

Interacting
pollinators,

data needs to be accumulated to understandpests and diseases,

such as,require frequent monitoring to build a picture of
dynamics of target taxa.

may reflect a dynamic evolutionary

situation.

taxonomic groups that appear to be under going rapid evolutionary
In situ conservation

the dynamics ofcomponents within the environment of these populations,the impact of these components on population•E Taxonomic complexity

and abundantIn situ conservation

forchange should be a priority.

sites that have many taxa requiring in situ conservationor contain more130

contain more than one target taxa of a group that may hybridize
strong justification•E Integrating

for establishingin-depthlaboratory

in situ conservation
analysisshould be sought to provide

reserves.

using different

techniqueswith detailedof the taxa and their environment based on field and herbarium studies
developing

sound in situ conservation

knowledge

are the foundationforstrategies.

Introduction
Recent research
that

can be useful

research

on Vigna species

in designingon species

in the subgenus

Ceratotropis

for in situ

conservation.

strategies

providesinformationThis

highlights

paperthat occur on islands

in southern

Japan, the azuki bean complex genepool

and studies

across the whole subgenus.

Laboratory

analyses

molecular

techniquesof researchextensivefield

and herbarium

observations.

and/or locations

to be identified

that would be appropriate

has been the focus

using various

biochemicalandand this has been supplemented

withThese various data enable particular

populations

for long term scientificmonitoringand in situ conservation.
Basedobservations

on surveys of Vigna subgenus
9 species

Ceratotropis

theirhabitats

are protectedfrom destructionThisis undoubtedly

incomplete

and specificvaluable

additional

information

identifiable

habitats,

Buddhist

temples

relatively

safe from destruction.

island,Okinawain Thailand

prefecture,areas andby development

activities(TableCeratotropis,

and V. reflexo-pilosa

protected

areas.

V. exilis represents

one species

thatthe locationofhabitats,

populations

At a site seeing

locationmaintained

local

cutting

officials

to habitat

annuallyof the grass

helps

of Vigna

destruction.

have large areas that are being converted

wild

relativesangularis)

Ceratotropis

in Japan and rice bean (V. umbellata)by their very nature are vulnerableareas a park on Ishigaki

cut the vegetation

to maintainprevent

that

succession

subgenusthat grow on the islandsof subgenus

specieswould

to development

Ceratotropis

grow inAmong these are some populations

since these islands
and weedyof thisthat grows in the grass there.

many populations

areas that are vulnerable

withinIn theseJapan,

hand,

Ceratotropis

1).

areas would provide

1).

(Fig.

of V. riukiuensis

the populationof protectedthat are frequently

eliminate

V. riukiuensis

protected

outcrops

annuallowlandParks or local

on limestone

views of the sea. This

On the otherand field

surveyson taxa in subgenus

Among poorly known taxa in the subgenus
grows in clearlyspecimens

were or are growing in National

therefore
list

herbariumof southern
to grazing

cultigens,in Thailand,
projects.

such

of

Okinawa, Japan,

land. In addition,as azukigrow in disturbed

thebean (V.

habitats

that

131

Table 1. National Parks or other conservation
species have been reported to grow.

areas where wild Vigna subgenus Ceratotropis

S p e c ie s

L o c a tio n

R e f. 1

V . e x ilis
V . e x ilis
V . e x ilis

W a t T u rn M a D e a r, A . S a i Y o k , K a n c h a n a b u ri, T h a ila n d
W a t T h u m K a o C h a a n k , N o n g P a i, K a n c h a n b u ri, T h a ila n d
D e p t. o f F o r e s tr y , T h u m K a o B in , R a tc h a b u r i, T h a ila n d
W a t K o h T h u m T a lu , D o n S a i, P ak T h u a , R a tc h a b u ri , T h a ila n d
W a t K h a o B a n D a i, P e tc h b u ri , T h a ila n d

A 16
A 16

V . e x ilis
V . e x ilis
V . e x ilis
V . e x ilis
V . e x ilis
V . e x ilis
V . e x ilis
V . h ir te lla
V . m in im a
V . m in im a
V . m in im a
V . m in im a
V . m in im aV . r a d ia ta
V . r e fle x o ‑p ilo s a
V . r iu k iu e n s is
V . r iu k iu e n s is
V . r iu k iu e n s is
V . r iu k iu e n s isV . 57?.(n e w )V . sp .(n e w )V . s tip u la c e a
V . s tip u la c e a
V . s tip u la c e a e
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta
V . tr ilo b a ta

W a t K h a o Y o i, K h o o y o i, P e tc h a b u ri, T h a ila n d
W a t B an p o ta v a s , T a y a n g , P e tc h a b u r i, T h a ila n d
T e m p le T h u m P r a N o n , K h a o Y a i C h a a m , P e tc h a b u r i, T h a ila n d
W a t K a o w o n , T . K a o w o n , P r a p u ta ta , S a r a b u ri, T h a ila n d
P u t T a n im e t T e m p le , M ittap a p , M u a k le k , S a ra b u r i, T h a ilan d
N a tio n a l P a rk P a k c h o n g , N a k o rn R a tc h a sim a , T h a ilan d
P h u P h a n N a tio n a l P a r k , S a k h o n N a k h o n , T h a ila n d
N a m N a o N a tio n a l P ar k , L o e i, T h a ila n d
K h o n K a e n , P h u K h ie o G a m e R e s e rv e , K h o n K a e n , T h a ila n d
T e m p le , K h a o K h a t N o n g N a k , N o n g K a e , S a ra b u r i, T h a ila n d
M a e Y o m N a t.P a rk , D o w B o o n , W .Y o m R iv e r a t B a r ite m in e , S o n g
D is t. P ra e , T h a ila n d
K h a w K h ie o N a tio n a l P ar k , S i R a c h a , C h o n b u r i, T h a ila n d
B a n n a K o e n (P a rk ) , I sh ig a k i, O k in a w a , J a p a n
A g a rih e n n a z a k i , M iy a k o is lan d , J a p a n
U g a n iz a k i , Is h ig a k i, O k in a w a , J a p a n
H ira k u b o s a k i, Is h ig ak i , O k in a w a , J a p an
B a n n a K o e n (P a rk ), Ish ig a k i , O k in a w a , J a p a n
G o n g a la w e w a , Y a la N a t . P a rk , H a m b a n to t a , S r i L a n k a
P a r a k r a m a b a h a n , P o lo n n a ru w a , S ri L a n k a
P a ta n a n g a la , Y a la N a t. P a rk ., H a m b a n to ta , S ri L an k a
K o m a w a W e w a , Y a la N a t . P a rk , H a m b a n to ta , S r i L a n k a
R ah u n a N a t. P a r k , K o m a w a W e w a in p lo t R 1 7 , H a m b an to ta ,S o u th e rn D istr ic t, S ri L a n k a
K a ri V illu , P lo t W .9 , W ilp a ttu N a t. P a r k , P u tta la m D ist , S r i L a n k a
R a h u n a Y a la N a t. P a rk , b e a c h E . B u ta w a M o d e ra , S ri L an k a
Y a la N a t. P a rk , n e x t to U ra n iy a L a g o o n , H a m b a n to ta , S r i L a n k a
W ilp a ttu N a tio n a l P a rk a t K a il V illu p lo t W 9 , S ri L a n k a
P a llu g a tu ra i, W e s t o f W ila p a ttu , W e st S a n c tu a ry , S r i L a n k a
P a ta n an g a la , Y a la N a tio n a l P a rk , H a m b a n to ta , S r i L a n k a
J a m b u r a g a l, Y a la N a tio n a l P a rk , H a m b a n to ta , S r i L a n k a
B u tta w a m o d e rn a , Y a la N a t. P a rk , H a m b a n to ta , S r i L a n k a

H refers to data from herbarium specimens,

A+number refers to appendix

references

A 16
A 16
A 16
A 16
A 16
A 16
A 16
A 16
A 16
H
H
H
H
H
H
A 15
A 4
A 4
A 4
A 15
A 17
A 17
A 17
A 17
H
H
H
H
H
H
A 17
A 17
A 17

Fig. 1. Habitatof Vigna exilis at Wat Koh Thum Talu, Ratchaburi,

Thailand.

Inset close up of flower

Fig. 2. V. riukiuensis

growing on cliff tops at the east side of Yonaguni island,

southern Okinawa, Japan. Inset close up of flower.

Fig. 3. Islands

1. Islandof southern

Okinawa, Japan.

populations.

Vigna riukiuensis

and V. reflexo-pilosa

in southern Okinawa

The biology

of populations

that occur on islands

in part,islands

interest,(Vitousek

becauseet al, 1995;

Baldwinnaturalselectionin island1996).

Recentresearch

can enable processes
et al, 1998).

populations
results

have measured the genetic

on different

islands

the sea (Fig.

is a perennial2). Vigna riukiuensis

Taiwan. It is closely

relatednakashimae of northeast

populations

differentiation

species

(Cody

from

and Overton,

riukiuensis

and V.

of these species

3).
in grass, on cliffs

distribution

overlooking

in southern

Okinawa andto V. minima that is most common in Southeast

Asia and V.

these three species

are considereda species

variation

among populations

The genetic

Okinawa has been studied

using

isozymes,

bandingand DNA markers. Two isozymes SDH and Lap were analysed

4 islands,

Ishigaki,variation

while Lap showed 3 phenotypes

Iriomote,

measuredresultingbetween populations

has a rather restricted

Asia. Together

in southernevolutionof Vigna

often growing

the V. minima complex (Tomooka et al., 2000a).
V. riukiuensis

rapid

a lot of research

to be readilyof Lactuca has been reportedof southern Okinawa, Japan (Fig.

Vigna riukiuensis

of evolution

For example,

from studying

reflexopilosa

has generated

Hateruma and Yonaguni (Fig.

populations

from Yonaguni.

riukiuensis

was not found in the populationsin populations

The Lap phenotype

found

total

complex,of

seedproteinfor populations

from3). The SDH isozyme showed nofrom Iriomoteand 2 phenotypes

inin Taiwanese

populationsof V.

from southern Okinawa (Table2).

Table2. Variation in Lap isozyme phenotypes in populations
V.reflexo-pilosa
from southern Okinawa (Modified
S p e c ie s
Is la n daN c oe .. o f

L ap
ADNA analysis,

V . riu k iu en sis
Ish ig ak i
Irio m o te
H a te r u m a
Y o n ag u n i

2
23
1
24

1
6
1
14

V . refle x o ‑p ilo sa
Irio m o te
Y o n ag u n i

5
11

8

revealed

clear

islandto islandarchipelago(Irabu,archipelago(Hateruma,

diversity(5) the genetic

Miyako0.0039and principal

resultssuggestdistancecomponent analysis

that V. riukiuensis

northern

limit.

distribution

as V. riukiuensis

on RAPD and AFLP analysis
Ishigaki,variation(Fig.

suggestwas 0.006

Irabu,

that in southern

4).

1of V. riukiuensis

Populations

from a wider area ofarchipelagoes(Fig.

from the Miyako

were distinct

3)

islandfrom those of the Yaeyama
islands).

Basedon nucleotide

from the two island

distinguishedthe populationsgroups(Fig.

was5). Thebetween the Miyako and Yaeyama archipelagos

perennial,
throughoutwith V. riukiuensis.

often found growing in shaded
the islandsof southernor semiOkinawa

The full range of its distributionis asand east as far as Myanmar. Okinawa, Japan, represents

In southern

Okinawa V. reflexo-pilosa

and showed no variationof 5 populations
Iriomote,4). The intra-specificcomparedclearly

It is foundit grows sympatrically

Okinawa,5
2

is an ancient element of the flora of these islands.

6).

Australia

6

and Yonaguni

barrierover bushes

far south as northern11

between the populations

is a tetraploid,

phenotypes(Fig.

and Tarama islands)

Vigna reflexo-pilosa

occasionally(Ha)variation

Momote, Ishigaki

(Fig.

C

of the Yaeyama and Miyako islandthat there is a floristic

and it is possible

shade,

islands

B

10

RAPD and AFLP, of populations

southern Okinawa includingof V. riukiuensis
andfrom Tomooka et al., 2000).

to 0.039

Tarama and Yonaguni
genetic

diversity(Fig.

seed proteins.

Basedfrom 5 islandsof southern

3) revealed

very little

measured by Shannons

for V. riukiuensis

Okinawa V. reflexo-pilosa

had the same three Lap

for SDH or totalof V. reflexo-pilosa

its(Yoon et al., 2000).

may have been a relatively

diversity

index

These resultsrecent•¡•¥

Fig. 5. Part of a Neighbor-Joining
Vigna riukiuensis

from southern Okinawa prefecture

indicates

the probability

diversity.

(Adapted

Fig. 4. Association

tree showing

supportingthat branch.

genetic

relationshipbetween populations

based on AFLP variation.
Branch length

Number at branchcorresponds

to nucleotide

from Yoon et al. 2000).

among populations

populations,

Miyako populations,

38% of the total

variationof V. riukiuensis.

(•œYonaguni-Ishigaki-Irimote

Hateruma populations).

and PC2 9.9% (modifiedof

PCI accounts

from Yoon et al., 2000)

forFig. 5. Vigna reflexo-pilosa

growing

on a bank between a road and rice
paddy on Yonaguni, Okinawa,
Japan,

introductionin the area and populations

since there is little

are derived

intra-specificvariation.

of analysingof these island

needed for in situ conservationof V. reflexo-pilosa

few populations

the majority

The results

may capture

a. flower and young pods.

from a limited

populations

suggestand V. riukiuensis.

of the genetic

variationsouthern

Okinawa. On the other hand,sincespecific

populations

and Yaeyama archipelagos

conserve the genetic

in both the Mikako

number of founder plants

V. riukiuensis

different

strategies

are

For V. reflexo-pilosaof thisshows genetic

speciesa

acrossdifferentiation

would be necessary

totheir importance,

have

resources of this species.

2. Crop complex populations.
The azuki bean (Vigna angularis)

crop complex.

The crops in the genus Vigna subgenus
been little

studiedcompared to other major leguminous

the taxa presumed

to be the wild

mungo, determined(ChandelCeratotropis

Ceratotropis,

for which detailed

ancestors

et al., 1984).

despitecrops. Only relativelyof mungbean,

V. radiata,

recently

and blackweregram, V.

To date the only crop complex in the subgenus

interand intra-populationis the azuki bean complex, V. angularis(Xu et al., 2000alevel

studieshave been conducted

and b). To discuss

aspectsof in situ

conservationof a crop complex the azuki bean (V. angularis)

crop complex in Japan will be

discussed.
Taxonomicallytwo varieties

angularis,

the cultigen,varieties,

field

populationthat is a reflection

have been recognizedand var. nipponensis,

observations

and laboratory

its wild

analysesin the azuki
relative.

bean complex var.

In addition,

to thesetwohave shown that there are other types ofof the dynamic nature of the azuki bean genepool

in Japan (Xuet al., 2000 a and b).
Wildhabitats

azuki,

Vigna angularisand Solidago

altmisium

var. nipponensis,(goldenis common in disturbed

herbaceousrod) is a common companion annual herb in Japan.

The usual habitatof wild azuki is along river embankments, roadside

fields.

This species

can be classified

succession.

The most northernlatitudeof wild azuki beans is 39•‹N on Honshu, Japan. During the growing

season the habitatof wild

azuki bean may be cut by rotary

Wild plants

are distinguished

as a pioneer

More slender always indeterminate,2.

Seed weight

black;

Smallerand leaves.

flowers

Weeds have various

firstdescribed

phenotypicand the cultigen

twiningdefinitions(Harlanof azuki bean are readily

by Yamaguchi
diversityand wild azuki can ratoon from cut stems.

(1992).

than weedy azuki and seeds are

and de Wit, 1965)

recognized

but weedy plants

andby a number of morphologicaltraits

The weedy type of azuki bean has a greater

range ofthan wild azuki but at the molecularwild azuki. The main charactersby:stems;

is about 30% less and pods 15% shorter

always mottled

weedy populations

cutters

from weedy plants

1.

3.

of secondary

verges and abandoned

level exhibits

less diversity

by which weedy azuki bean can be distinguishedthan thefrom wild

azuki beans are:1.

Stems are stout and may be determinate

2. Seeds

and pods vary in color

or indeterminate;

and seeds

may be mottled

black,

greenish/yelloworand pods 13% longer

thanplum red and pods black or tan;3.

Flowers

are larger

and on average seeds are 43% heavier

for the wild type plant (Tomooka et al. 1999);
4. Leaves are variableof the leaves
Based on field
The first

in size but tend to be heavily

indentedand may approach

the sizeof the cultigen.

observations

and laboratory

may be from hybridization

results

weedy azuki probably

between the cultigen(var.

angularis)

arose in two ways.
and wild plants

(var. nipponensis).
specific

The second is that weedy azuki evolved

habitats

where its larger

Populations

seeds have a selective

directlyadvantage(Vaughanof wild and weedy azuki bean may be either

type or a mixture of both wildcomplex populations

and weedy plant(Xu et al., 2000a,and seem to represent

particularly

types.

b). Complex

dynamic

Thisfrom wild azuki beans toof only wild or weedy plant
later

populations

populations

et al., 2000).

situation

we have called

tend to cover a large area

where gene flow among different

types of azuki occurs.
The implications
that particularof the results

attention

analysis

populationin Tottori

prefecture

to azukibean and plants

geneticallydiverse.

have revealed

in different

interaction

between1990).

Egawa et al.(1990)

wildof complex populations

5 years of observationsof complex

show evidence

populations
collecting

azuki and the cultigen

missions

are commonly grown throughout

betweennipponensis).

We asked the farmers about the local name of this black mottled

showing"kusa-adzuki"

cultivatedcowpea)bean and wildin Hodono, Kami village,

adzuki)in Kamassawa, Oshika

names depending
adzukiupon the people

bean and wild adzuki

many places.

Shifting

When planted

on mountain slopes

with wild adzuki

adzuki bean from Tenryu village

adzukithem a sample. It was called

between cultivatedcultivationin shiftingor "kuro-adzuki"

placesderivedfrom the hybrid

called"nabe-yogoshi",consideredsuch kinds
kuro-adzuki"

to be very suitablebean (V. angularis"kitsune-adzuki"
village.

Thisof blackadzuki bean byin Ota, Tenryu village,(fox adzuki)

and "yama-

mottledaccessionand area, suggestingthathybridization

bean may have occurred(black

to
var.

black

is

spontaneously

inin this district.

adzuki bean may have hybridizedin the mountains.

adzuki)

districtadzuki(weed adzuki)

cultivation,"kage adzuki"(shadow

bean. In Shimoina

et al.,

that is consideredof azuki and soybean used to be practiced

bean that occurs naturally

and considered(EgawaJapan. We werebe a hybrid

variousofreported:a sample of black mottled

given

they plantin Japan thehas been reported

able to obtainadzuki"(mountainof a complex

the fields

parts" Red-seeded azuki bean varieties"nosasa" (wildthatthat farmers rotate yearly

from one year to the next. During germplasm

possibleof the azuki bean complex areshould be placed on in situ conservation

have been shown to be particularly

segregationof populationBlack

adzuki)mottledvarieties

named

are now cultivatedin someprogeny between adzuki bean and wild adzuki
mottledor "sato-adzuki"

varieties(sugarwere also cultivatedadzuki).

for making anko (sweet bean paste)."These varieties

and
are

Considering

strategies

studying

the azuki

population

structure :for in situ conservation

bean complex three featuresappear to be criticala. Disturbance,

mainly by farmers, and their knowledge

in preventing

succession.

In severalof crop complex genepools

for maintainingof the importanceareas of Japan it has been reported

that in earlier

presumed weedy types of azuki bean were gathered(Yamaguchi,

Indigenous

knowledgeof the value of wild

may be one means of protecting

populationsof these plants.

b. Pollinatorsand their protection.

from year to year?
frequently

visit

flowers of V. angularis

instrumental

in pollentransfer(Fig.

gene flow within

and between sympatric

7). There are still

and activityof insects

of pollinators

Xylocopa

flowers for nectar. These

between different

in close proximity

floral features

the species

times

and eaten

and weedy forms of azuki

How do the numbers and species

Bumble bees, particularly

theof this disturbancewhen food was in short supply
1992).

based onfluctuateappendiculata

Smith,and other bees may bemembers of the azuki bean complex growing

have many unanswered questions

on the topic

wild,

However, theweedy and cultivatedon the flower suggests

azuki.

crosspollinationis possibleeven

if only occasional.
c. To understandtimes during

the changes

occurringin populations

scientific

the year and from year to year is necessary.

monitoring
Monitoring

environment of the populationand its general vicinity

are importantFig. 7. Kumabachi

Xylocopa appendiculata

foraging(bear bee),azuki bean (Vigna

angularisvar. nipponensis).

both at differentof the immediate

components to observe.

for nectar in flowers

ofof wild

3. Complexesof species

and complex species

The genus Vigna subgenus Ceratotropis
New information
geographic

data

subgenus

Ceratotropis.

has clarified

the

level

combinedwithmorphologicaland ecowithin

genetic

diversityand species

relationships

information

can help

in developing

guidelines

for in situ

The taxonomy of the subgenus

Ceratotropis

was studied

in detailed

by Tateishi

who considered

consisted

conservation(1985)

at the molecularThisof Ceratotropis

species.

that the subgenus

of 17 species

Since then there have been many germplasm collection
Asia (Appendix)
techniquesand some of these new materials

includingthe

RAPDs (Kaga et al., 1996;

AFLPs (Yoon et al., 2000;

and 15 infra-specificmissions

for wild

have been studied

Tomooka et al., in process),

Vigna species

in detailTomooka et al., 1996),

taxa.

using various

RFLPs (Kaga,and proteinase

ininhibitors

1996),(Konarev

etal.,2000).
Based on morphologicalgeographic(Tateishi,al., in process)studiesand Aconitifoliae
collection

1996),(Tomooka

the species

resultsof the AFLP analysis

distance
0.043

and 0.034,
(and

only

if V. trinervia

complexity,

sinceexcluded

only in sectional., 2000a).

if intra-specific
Angulares

Recent AFLP analysis

of a coresections

distance

between taxa in this section

A general picture

are clearlyreflect

progenitors

areAngulares
taxonomic

of cultigens

intra-specificbut

The genetic

and Radiatae

The results

havingof thedistinguishedAconitifoliae

ranks for wild

are there species

among andbetween taxa in section

The more complex taxonomy of section Angulares

differentiation

relationships

are very different.

is excluded).

taxonomic(Tomooka etRadiatae

between taxa in sections

whereas the genetic

eco-

Angulares,

7). The three sections
the three

et al., 2000),

and molecular

into three sections

and in process).

respectively,
0.015

1988)

(Tomooka et al., in process).

between taxa withindiversity(Egawa,(Konarev

has measured the genetic

is shown (Fig.

based on nucleotide

is 0.028

genetic

et al., 2000aof the three sections

distances

biochemicalhas been divided

in this subgenus

within1955),

cross compatibility

the subgenusof the species

the genetic(Maekawa,

aretaxa (Tomooka etreflectsof the lower levelthat has been revealedofby AFLP analysis(Fig.8).

The complexity
among several species
the subgenus

of section
withinCeratotropis.

Angulares

the section

suggests

The fact that several

and relatively

littlegenetic

this is a rather recently
speciesin section Angulares

differentiation

evolved branch of
are cross

Fig.

8. Unrooted

inferredneighbor-joining

nucleotide

tree, basedon AFLP variation

diversity

was deduced,

for Vigna subgenus

collection.

The three sectionsof the subgenus

and V. trinervia

compatible,

at leastin one direction(Siriwardhane

gene flow may occur in the wild. This may explain
assigning

speciesnames to newly collected

accessions

sectionAngulares

represents

level

sectionAngulares,

Vigna minima and the closely

at the section

riukiuensis

have been referredmolecular

data and morphologicalcharacters,(V. nepalensis)

variablewould best be considered

Moleculartrinervia(Doianalyses

is a species
et al., 2000

Ceratotropis

species

represents

basedintermediate
and in process;et al., 1991),

suggests

the difficultysome workers have had inthat interspecific(Tomooka et al., 1996).
of species

complexes.

relatedspecies

V. nakashimae

that

V. angularisa species

complexbridging

species.

It may be noteworthy

Withintheand V.

based

on bothand a type of wild

Vigna

and that V. hirtella,sectionsa highly

Angulares

this
that

species
V. trinervia

data suggests

in the subgenus

Tomooka et al., in process).

in the section

thata complex species.

between the threeplacedIt appears

a serieson DNA markers and sequence

generallycore

are indicated.

to as the V. minima complex. It is also possible,

found in Nepal
species,from which a matrix of

Althoughmay be particularlyis the only species

that

V.

Ceratotropis
V. trinervia

isimportant

as athat has been

repeatedly
pilosabeen confirmed

as one of the likely(Egawa et al., 1995;

Konarev et al., 2000).

Severalspeciesin thesubgenus

between diploidtaxa may occur (Table

have a situationthat resembles

genome donors to the tetraploid

Ceratotropis

grow sympatrically3). Thus within

the situation

presented

where gene flow between taxa occurs. Thissituation

gene flow has been demonstrated

Vicia

betweenthe subgenus

V. refexo-

and gene flow

Ceratotropis

we mayabove for the azuki bean crop complex
is reported
sativaVicia

where interspecificand V. segetalis(Potokina

et al.,

2000).

Table 3. Wild Vigna subgenus Ceratotropis species
sympatrically
based on direct observations

that have been found growingof the authors.

S p e c ie s

L o c a tio n

R ef.1

V . r e fle x o ‑p ilo sa a n d V . r iu k iu e n s is

K o m i, Iri o m o te , O k in a w a , J a p a n

A 4

V . r iu k iu e n s is a n d V . r ef le x o ‑p ilo s a

BJ a ap na nn a K o e n (p a rk ) , I sh ig a k i , O k in a w a ,A 15

V . a n g u la r is a n d V . n a k a s h im a e

Y a n g p e o n g , K y u n g g id o , R e p u b lic o f K o r e a

V . a n g u la r is a n d V . n a k a s h im a e

N a m y a n g j u , K y u n g g id o , R e p u b lic o f K o r e a

V . a n g u la r is a n d V . n a k a s h im a e

H w a c h e o n , K a n g w o n d o , R e p u b lic o f K o re a(a
V .llr ae sd ciaa pta e ,d V) . m u n g o , V . u m b e lla ta

PR ho ic
u te
h it1,T1 7h ,a ila
5 8 nk dm N . o f N a k h o n S a w a n ,A 8(w
V .mildu )n g o (e sc a p e d ) , V . u m b e lla ta

TR ho au ila
te n1 d2 , 2 2 k m W . o f P h its a n u lo k ,A 8

V . e x ilis a n d V . u m b e lla ta

TK ha ao ila
N ng du , T . P u b P r a , A . R a tc h a b u ri,

A 16

V . e x ilis a n d V . m in im a

WS a ra at bK u ari,o TW h oa nila
, Tn .d K a o w o n , P ra p u te b a ,A 16

V . h ir te lla a n d V . m in im a

B o k L u a , N a n , T h a ila n d

A 16

V . m in im a a n d V . u m b e lla ta

B o k L u a , N a n , T h a ila n d

A 16

V . tr in e r v ia a n d V . r ef le x o ‑p ilo s a

B e n to n g , P a h a n g , M a la y s ia

A 7

V . tr ilo b a ta a n d V ig n a s p .(u n n a m e d )BL au ntta
k aw a m o d e ra , Y a la N a tio n a l P a rk , S r i

A 17

V . s tip u la c e a e a n d V . tr ilo b a ta

L1 ka nm k fa ro m P a ta n a n g a la B u n g a lo w , S r i

A 17

1 References refer to Appendix reference numbers. Information
Dr. Mun Sup Yoon based on his observationsin autumn 2000.

from Korea was providedbyThe implicationsof research

for in situ conservationare:

1. Since this subgenus,

particularly

conservation

would be particularly

into the genetic

diversity

sectionappears to be rapidly

Angulares,

valuableof the subgenus

in areas where different

Ceratotropis

evolving,

in situ,

Ceratotropis

species

grow sympatrically.
2. Focus areas for in situ conservationa. Thailand

and northernshould be:

South Asia for populationsof species

in the section

Angulares;

b. Southern Asia, both Sri Lanka and the southern part of the western Ghats, based on recent
field

surveys(Tomooka

et al., 2000c)

and herbarium

data for sections

RadiataeandAconitifoliae.
3. Since,

V. trinervia

different

sections

conservation

maybe valuableof the subgenus

as a bridging

Ceratotropis,

this

speciesin crosses

betweentaxa in

speciesshouldbe the focus

of bothand research.

Conclusions
Plant genetic
conservation

diversity

in situ.

sources. Conclusions

research

can play a role in developing

However, such researchneeds to integrate

drawn from phylogenetic

analysismarker can be statisticallybiased.

is not correctlyor has undergone repeated

identified

that it is no longer geneticallyStudies

representativeof the genetic

into in situ conservation.
may have distinctly

Similarly,

diversity

Islanddifferent

results

genetic

a prerequisite

for sound in situ conservation

abundantthedynamics

environment of these populations
pests and diseases,

speciesanalysis

Ceratotropis
havingto understandsoaccession.
provideinsights

the same distribution
history
patterns

or diversityof diversity

isof wild species.

where gene flow may occur cannot be easily classified

morpho-agronomic and ecologicalto understandcollected

analysedin the genebank

based on their evolutionary

into the same area. Thus, populationfrom many

if germplasm

cycles of regenerationof related

introduced

Crop complex populations

may be unreliableof taxa of the subgenus

variation

information

tobased on just one type of molecularof the originally

populations

sound approaches

data needs to be accumulatedof such populations.
need to be studied,both within and between years.

Interacting

andover several years

components

such as, human activity,

withinpollinators,theAt theevolutionarypriority

levelof thesubgenus

situation.

In situconservation

particularly
There

at sites where different

is still

Ceratotropis.

taxonomic

much thatV. dalzelliana

have not been studied.
dominant

allelessubgenus

Ceratotropis

taxa grow sympatricallystudiesclarifygroupsdiversity

are not in germplasm

using molecularshouldbe ain the subgenus
collections

markers that readilygene flow among taxa. The different

have not been identified

a dynamic

and gene flow may occur.

is not known about genetic

Population

example, the genera Glycine,

may reflect

for complex taxonomic

and V. khandelensis

would help

complexity

and they
identifyco-genomes in theas has been done for most speciesin, forOryza and Triticum.

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Vol. 16 (in press)in Tottori

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on ExplorationandIn-situ

Conservation

Researchon

Buckwheat(Fagopymm spp.) in Nepal
H. P. Bimb
Biotechnology
Unit,
Council,
Khumaltar,
iscc_nepal2@wlink.com.np

Agriculture
Botany Division,
Nepal Agricultural
Lalitpur
Nepal. Po. Box. 1135. Fax 977 1 521197.

Research
E-mail:Abstract
Buckwheat (Fagopyrum spp.) is a major summercrop of higher hills(above 2000 m) in

Nepal. Nepal is rich in buckwheat genetic resources, however, manyland races are under threat
as the area of production is declining and buckwheat is being replaced with other crops. Two
siteswere selected(2000-3500m)

for in-situ

districts

conservation

researchat Mustang (2800-4000m) and Dolpa

where wild and cultivatedspecies of buckwheat can be found. A recent

survey on these areas revealed that four species
esculentum, F. tataricum, F. cymosum(tetraploid)of Fagopyrum are prevalent in Dolpa, e.g. F.
and F. megacarpum.Only F. esculentum andF. tataricum have been seen in Mustang. Diversity at the sub-species level requires further
investigation.
Analysis of genetic diversityof Nepalese Fagopyrum using appropriate
molecular and/or biochemical

techniques

will be carried out during the project period.

Introduction
Buckwheatmajor portion(Fagopyrum

spp.)of the food requirement

is grown in a barley-buckwheat
altitudes

is a major(abovein highrotationsummer crop in Nepal

hills(above(1000-2000

season as a catch crop in marginalland.

of fodderfor livestock

farmyard

that in return produces

crop followingof hillrice.

commonin Jhapa

farming.

is sporadic

terai)and Chitwan(east

The straw of buckwheat

across(central

manure that playstheterai,

inner terai).

source of rutin that is a medicinal

agent for the treatment

rutin

has been found to be higher

contentof tartary(Kitabayashi

et al., 1995),

hills

consumption.

for local

buckwheat

however, the cultivation

1). The cropand as a mono crop at higher

Below 1000 m, buckwheat

Cultivationam) it is grown both in the autumn

and springrole in the sustainability

2000 m) (Tableat lower altitudes

3000 m). In the mid-hills

and supplies

is a sourcea significant

is grown as a cash

however,

it is more

Buckwheatis a richof vascularin linesof this crop is confineddisorders.

The

from Nepal
to the higher

For many species,
most appropriate

in situ or on-sitemethod of conserving

evolutionary

processes,very diverse

in its topography,

genetic

resources.

phapar)climatebuckwheat(tite phapar).

with altitude

ranging

while

and agro-ecology.

tartary

withand terai.

growthhabit

species.

In contrast,

flowers.

It is mostly(F. esculentum)
buckwheat(Baniyagrown as a summer crop in the high hills

is referred(F. tataricum)

to as sweetis calledin the highet al., 1995).

Common buckwheat isand mountains and as an autumn and winterstem. It has pink

buckwheatcultivatedis mainly

in highflowers

self-pollinatedhills

are not grown in the terai. Yield

1. District

bitterand mid-hills

and is an out-crossing
with greenish

wise area, production

to whiteas a summer crop. Bitterof bitter buckwheat

is higher

and productivity

types

of buckwheat1989).
(Fig. 1).

D is t r ic t

A r e a (h a .)P r o d u c t iv ity (M t/h a )T o ta l p r o d u c tio n (M t)

M u sta n g

845

1 .9 91 6 8 6 .6

M an an g

240

1 .0 12 4 2 .4

S o lu k h u m b u

250

2 .4 01 2 2 .7

J u m la1 10

1 .3 01 4 3 .0

H u m la500

1 .0 05 0 0 .0

D o lp a

2 10

1 .4 33 0 0 .0

Source:

ADO, Annual Report,

1998.

Since the area of production
many landracesof buckwheat

is declining

and new crops are replacing

may disappear.

buckwheat

genetic

maintainedby farming communities

input requirementsofthan common

buckwheat. Both species have been grown in Nepal for a long time (Ohnishi,
Tablepreserves

The most commonautumn type has indeterminate

red pigmented
bitteris theIt is also rich in buckwheat

Commonbuckwheat is importantfrom 300-4200m

crop in the mid-hills

habitatto adapt to environment changes. Nepal is

The commonbuckwheat(mithein the naturala crop gene pool. In situ conservation

which allows plants

buckwheat

buckwheatconservation

Thus, collection

resources is an urgent priority.

buckwheatand conservationof

To date the crop is cultivatedandin the mountains where the crop is adapted

whereare low. Though the significancediversity

has been discussed

for a long period,

initiated

in Nepal. Research to improve understandingof in situ conservationof genetic

research on this topic is only now beingofFig. 1. Map of Nepal showing districts

in situ conservation
research

is required.

where buckwheat

Thus, a projectprogram on in situ conservation

be undertakenby the Nepal Agricultural

Agriculture,

Forestryand Fisheries

budgetis covered

throughthe International

objectives

are:project

has been formulatedon "Collaborative

resourcesin Nepal" jointly

toResearch Council(NARC)and the Ministryof(MAFF)of the Government of Japan. The project

Genetic

fund from the Government of Japan

Resource

Institute(IPGRI).

•E To enhance and support a framework of knowledge on agricultural
buckwheatand its wild relatives•E To develop scientificmonitoring

diversity

is focused.

of plant genetic

for three years by the special
Plant

in-situThe project

biodiversityofin Nepal.

basis for their in situ conservationof plant populations

by analyzing

with appropriate

andmolecular/biochemical

techniques.
•E To broaden the use of buckwheat

diversity.

Activities
A planningmeeting

Following

this,

Division,

Khumaltar,

was held on 4 September

1998 to finalizea review meeting was held in October,
Nepal in which two sites (Mustang

the project

proposal.

1998 at the Agriculture

Botanyand Dolpa)

were selectedforin situ

conservation

researchendorsed by Dr. S. Miyazaki
Instituteof Agrobiology

November, 1998)(Table2). The letter(on 14 October,

1998)

Resources (NIAR),

for and on behalfof agreement(LoA)

was duly

for and on behalfof the National

Japan and by Dr. Upendra Mishra (on 12

of NARC, Nepal. A review of the current status of

buckwheat research in Nepal was preparedcollection,

preservationand evaluation.

activities.

Accordingly,

participatory

in Dolpa(2000-3500

which covered the past works on germplasm

This was used to determine
rural appraisal(PRA)m) and Mustang (2800-4000

the sites for projectsurvey for villagem) including

selection

Upper Mustang wasundertaken.
Table 2. Ecologicalparametersof buckwheat

growing areas in Nepal, 1997.

D istric t
A ltitu d e (m )M u stan g

13 7 2 ‑8 1 6 7

T em p era tu re ( ℃ )M in im u m
M a x im u m
17 .7
5 .5

D o lp a

1 2 2 5 ‑7 6 2 5

1 6 .6(Source:

Nepal District

Profile,Materials

and methods
Two districts

reviews on bitter

6 .9

1998,

research(Bimb,

production

and diversity.

botanist,plant breeders,technical

constituted

to review the literatureliteraturesfrom secondary

agriculture(VDC)

4 9 9 .4

L an d ty p ereg
W aterim e s

S ilty lo a m

Irrig
R a inafed
te d

RS itrev eram

S ilty lo a m

R a in f e d

ADO Annual Report, 1998)

areas of buckwheat

format to obtain

13 2 1 .7

S o il ty p enamely Mustang and Dolpa were selectedbuckwheat

geneticist,

R (ma inmfall)

specific

information.
officers

and local leader

farmers.

reported

that these districts

A survey team included
officers

weretaxonomist,

and outreach research officer

wasand conduct PRA. The team reviewed the available

information.

development

1999)

for PRA study. Past

The team also developed
In the field,(ADOs),

the team obtained

chief of the village

different

kinds

information

development

of
from

committees

Results
Summaryof the buckwheat cultivation

practices

is given (Table

3).

Land preparation
One or two ploughingsof land. In high hills
firstploughingby bullocks

is done dependingon the altitude(more than 2600 m to 3800 m), farmers usuallyis done to tillthefallow

land following

and type

plough twice. The

compostingand second

Table 3. Indigenous

technical

knowledgeof buckwheat

cultivation(Farmer's

perception)

C u ltu r a l P r a c tic e s

D is tr ic t
P la ne t.
tim

M(t/h
/fe ar nta u) r e(kS ege /dh a r) a tepP rlaontetc tio n

I r ri g a tio n

H a r v e s t t im e

M u sta n g

J u lyA u g u st75

6 0 .10 0

wAc urripthotw
mrmys,o,rm

Tw o

D o lp a

M na ey ‑Ju

3 0 ‑3 5

8 0 :1 0 0

In both districts
seed source is local, land is ploughedthinning is done once and weeding twice.

H a r v e s t m e th o d

T h r e sh in gmM eillthino dg s

S to r a g e
m e th o d

O c t‑N o v

C u t th e p la n t b y
sic k le fr o m th eb a se o f w h o lep lan tB y b e atin go n th e sto n e
o r flo o r o rb e a tin g
w ith stick s

Ja to (lo c a l st o reg ri n d er ), G h a tta(w ate r
m ill
T u rb in e ), D h ar
K u tn e M ish in(ri c e m ill)

E a rth e n p ot
and
w ood
box

A u g ‑O ct

C u t t h e p la n t b ys ick le fr o m th eb a s e o f w h o lep la n t

B y b ea tin g
w ith stic ko n m at

G h atta
J u to )P il m eth o d
M ud
p la ste re dea rthpot
w o o d en b o x

twice, there is no seed treatment,( lo c a lsowing is by broadcasting,Y ie ld
ran gem t/h a1‑12

1 6 .2 5

com m onu se
R o tim ixedw ith D a l
s a la dv e g eta b lea n d m ix e dw ith m o ist
s au c e
R ot i la g er,
v e g eta b le,sa la dD h e su a n d
m ix e dw ith ot h e r
flo u rploughing

is applied

after broadcasting

the seeds followedby levelingto ensure seeds

are well covered with soil.
Composting
In general,chemicalfertilizer

farmers from Jumla district
fieldsreported

is not appliedthat had applythat urea promoted sterility

in buckwheaturea at 15-20
and bitterness

cultivation.

However,to bitterbuckwheatkg/haof grains.

The majorityof hill

farmers apply manure and compost at the rate of 5-30 Mt/ha. Farmers from Mustang
use manure in small amounts from horses and mules.
Seed rate
Seed rate varies from location
lower

elevations

to higher

buckwheat is 80-100kg/ha
Planting

mountainsand for bitterand ranges from 60 to 100 kg/ha

respectively.

The seeding

rate

at

for sweet

buckwheat 60kg/ha.

time
Buckwheat can be planted

irrigated
bitterto location

conditions
buckwheat,crop generally
secondat various times. In Mustang this crop is grown underwhereas in Dolpa it is grown in non-irrigated,seeds are sown in late July and harvested

fields.

For

at the end of October. The

matures between 90 and 100 days. Commonbuckwheat is sown in theweek of August and harvestedby the third

week of October.

buckwheat is sown during May to June and harvested
Planting

slopingtime is adjusted

In Dolpa

between August and October.

to avoid rain during the flowering

periods,otherwise

sterility

results.
Thinning

and weeding
First weeding is done one month after sowing at the 3-5 leaf stage of the crop.

During this period,

plantsof uprooted plants

are collected

livestock.

are thinnedout to obtain optimum plant population.

for vegetablepurposes

A second weeding follows at flowering

Leavesand stalks are used as fodder for

time.

Plant protection
The crop was observedwithpowdery

mildew

infection.

Similarly,

foliardamage due to cutworm was obvious, however, farmers were not much concerned withthe damage as they perceived
plant

protection

it not to be significant.

measures in buckwheat.

problem and cause significant

There is no evidence of the use of

Farmers reported

that wild bear are a realdamage to the crops in the high hills.

Harvesting
Farmers harvest
plantswith a sickle,

the harvested

plantsin severe shattering

when approximately

70% grains

become brown. They cut

3 to 4 inches above ground level. Most farmers make a bundle ofand keep it uprightin the field

Losses due to shattering

for drying.

Any delay in harvesting

and thus incurring

significant

reduction

may resultin grain yield.

can be between 25 and 50%. To avoid this loss, farmers start

harvesting

the crop before sunrise when there is sufficientThreshing

and winnowing
Dried plants/bundles

moisture.

are spread over a mat or cloth and then they are beatenwith wooden sticks

for threshing.

supo (flatmade of bamboo) to separate grains from inert materials.

bold

structureand heavy grains

following

During windy days farmers winnow with the help ofare separated

duringwinnowing and stored

Healthy,

as seeds for theyear.

Storage
Seeds are thoroughly

dried in the sun before storage. Dried seeds are examinedby womenfarmers by pressing

the grains

between teeth. Local methodsof storage

include(a) Seeds

are stored for one year in earthenpots,

mud plasteredpots,

tin boxes orwooden boxes.
(b) Grains

for food consumption

are stored in a wooden box or in an underground pit.

Such pits are dug close to the house in an upland area where water stagnationdoes

not occur. The size of a pit depends on the quantity of grains to be stored. Usually,
the dimension of a pit is 1x1x2 m3 with a capacity of one ton of grain. The bottom,
side walls and top are covered with Bhojpatra
to protect

grains from moisture. The top is thickly

and a flat stone is placed
reported(Himalayanover it to protectthat they can safely

silverplasteredgrainsbirch, Betula utilis)(up to 30 cm) with mudfrom rain and pests. Farmers

keep grains in such pits for 8-10 years without

insectinfestation.
Flour milling
Traditionally,
circulargrainsare milled

in a water millflat stones, the lower is stationary

and in between these, grains are put through(Ghatta)and the upper is manuallya hole for milling).

or in a Janto (tworotatedclockwise*=LowUsea. Humanconsumption
Buckwheatgroups.

is usedBitterin various

buckwheatways in different

communities

that4). It is importantfood preparation

buckwheatkinds

of localDhindo(flouris mixed with water and well cooked),ryale

roti (pancake)are available

and beer. In Mustang,to prepare

thickporridgeto make roti that is a very popular

bitterand pancakes.

diversity
includeDhenshu (very(Table5).

roti (chapati),
thickchapati),is mixed with

Sugar is addedin bitter

food preparations

calledbuckwheatsweet

buckwheatleaves to preparesoup

as Dhapra. Tender leaves andusing buckwheat

Roti and Dhindo, Salad
Thick porridgeand pancake, Soup of dried leaf powder
Flour mixed with meat soup
Roti, Vegetable,
Salad, Dry Vegetable,
Dheshu,Bhate

Dolpa

these

the food culture

baby food in the Jumla area. The Tibeto-Burmese

with beans and yak meat. This is locally

Mustangand cultural

buckwheatcommunity at Mustang use dried powder from bitter
Table 4. Comparative

to understandgrow and maintain

Different

buckwheat

ethnicity

is used for animal feed at Mustang whereas it is the main

source of human diet at Dolpa (Tableof farming

ecosystems,

and mixed with bean or dal,Beer, Fresh

Table 5. Comparative value of buckwheat in terms of social,and economic diversity in high mountains.

flour

with

sait,

food, genetic

D is t r ic t

Sd oivceiar ls ity

F o o d c u lt u r e

dV iva re ier stity y

E c o n o m i c im p o r ta n c e
a t lo c a l m a r k e t

N o . o f la n d r a c e s g r o w n inc o m m u n it y o r v ill a g e

M u s ta n g

G u run g

F o o d c o n s u m p tio n

M i th e *Sw eet‑ H igh

2

T h ak u ri

Tu srea tio n a l/R e lig io u s

T ite * *B it te r ‑ L o w

N ag ark ot,C h u san g ,K agb en i

B i s taM e d ic in a l u s e

W ild * *K am iC o m m e r c ia l u s e

T a r e li

F o o d c o n su m p tio n

M ag ar

Tu rsea tio n a l/R e lig i o u s

K am iM e d ic in a l u s e

D o lp adiversity,

LR oamn ipa natuhwa na ,gJh o n g ,S w e e t ‑L o w

3

T ite ***

B h a t e ‑H ig h

B h a te * *(R i c e b u c k w h e a t )B it te r ‑H ig h ( e s p e c i al ly
in T ib e t w ith s a lt o r
g o a ts o r s h e e p )S a h a r ta r a , R a s i, J u p h a l,
P h ok sun do ,R um ag aon ,L a u n a o n , K n ig a o n , S a ld a n g ,C h a rk a , D u n a i, M aj h p h a l**=Medium diversity,***=High

diversity

shoots are used as a green vegetable

and salad and consumed with yoghurt.

driedleaves are also used as vegetable

Tareliand Magar use bitter

also tastes

bitter.

after cooking.

Ethnicbuckwheat to prepare alcoholInterestingly,

bitterbuckwheatflourgroups such as Thakuri,

and chhang (local(half-cooked

feed bees in the rainy and winter season when there is a scarcity
the fields.

Farmers of Mustang take out the blood
types of buckwheat based on bitterness
Bitterness
Varietyslurry)

beer)

that

is used toof blossom flowers infrom yak and mix with bitter

buckwheat flour to make a slurry for the preparationdifferent

At Dolpa,

of localsausages.

Farmers rank

as follows:High ========================->Low

name Chuchche > Syangre>Kalo khishe>Seto

Chuchche is considered

to be the bitterest

khise>Mithe

followedby syangre, kalo khise, seto khise

is used as a fodder especiallyfor goats and sheep in Mustang

and for cattle in Dolpa;

and used as a beddingin the livestockbelievefodder is nutritionaland mithe (common buckwheat).
b. Fodder and manure uses
Bitterbuckwheatthat buckwheat

Khise phapar is reported
phapar.

The flour

slurry (Kundo)
increaseto be highly

to livestock

buckwheatthat is given to the lactatingproduction

shed. Farmers

and produces heat in the body of livestock.

palatable

and husk of bittermaterial

followedby mithe and tite

are mixed with water to prepareanimals,akids and calves in morning hour toand fat content of milk. In Dolpa, the straw of bitter

buckwheatisused as a green manure.
c. Medicinalvalue

Farmers in the high hills

and mountains believe

headachesand hemorrhage

problems.

jaundice.

In Mustang, farmers reported

agent to help heal bone fractures.
intestinal

disordersthese ailments.
d. Religious

It is alsothat bitter

buckwheat alleviates

used to control

that buckwheatflour

gastric

patients

with coughs,

and fever are administered

bitterbuckwheatin keeping

colds,

pneumonia,

to help recovery frombody warm.

value

Dough of buckwheat flour is used to make pseudo monks (idols)
Dhami/Jhankri
duringandis used as a plastering

In Dolpa,

It helps

acidity(priest)the Purnima (fullin a ritualof spiritual

moon day) festival,

treatment

againstby Lama anddiseases.

women farmers take bitter

In Dolpa,

buckwheatasfalahar(similarto other fruits).

Poppedgrains are offered to God in the Chaite

Dashainfestival.
Buckwheat diversity
A multidisciplinary
officers recently

surveyed (in September

Four buckwheat
esculentum,

species

F. tataricum,

Fagopyrum species
trekkingteam of taxonomist,
1999)

were observed

geneticist,duringthesurveyperiodof Juphal,

Majhphal,Rasi, Dunai and Sahartara

Vyasgaad(a stream).

These

species

Agricultural

BotanyDivision,

Khumaltar,

was observed in plants
identification

will

be carried(Prof.

Ohmi Ohnishi

are wellpreserved

Nepal.

and grains of bitterof species(Courtesy

and sub-species

Hari Bimb, Mr. M.L. Vaidya

Fukuoka)along theVDCs (Fig.

afterof Dr. H. Nakayama)in the herbarium

Agronomic

and morphological(Tableof wild and cultivatedbuckwheatof scientists

5, 6,

from Japan

and Nepal (Mr. Bimal Baniya

and Mr. D.R.Sharma).

at theand sweet buckwheat

out in November 1999 by a team consistingand Dr. ShuichiF.

two wild

F. cymosum is abundantFig. 2. Fagopyrum cymosumat Dunai, Dolpa, Nepal

7). Further

namely,F. cymosum and F. megacarpum. The latter

2) whereas F. megacarpum was only observed on the route to Sahartara

diversityand technical

Mustang and Dolpa district.

were found in Dolpa district.

routes and slopes

plant breeders

The team will also collectand Dr.
seedsamples for genetic diversity

analysis

to be conducted by Dr. Bimb in December, 1999

in NIAR, Japan using isozyme and /or molecular(RAPD)

techniques

F lo u r

Table 6. Types of buckwheat
D is tr ic t

V ap re .
tyla
L on cd a rl a c e

cG o rloa inursG h ra ap ine

T a s teM u stan g

S w eetG h y ab re

B lack /b ro w n

T rian gu lar

S w eetB itter

D hop

G re y/b la ck

S len d e r

B itterc o lo u r
P in k/R ed /W h iteG reen /R e d

W ild

B an b h ad e

S w eetM ith eB lack /b ro w n

S w eetB itte r

GDS yh anoe resugre,,

B h a teSKT etoashu
alo kknhhgiseise,

B lackG rey
B ro w n
B lackW h iteD o lp a

B o ldR ou n d
S len d er
B old /e lon
gatedcS otelom u r

Mfo rillin
fl o gu r

R ed

E a syW h ite/y ellow

L e ss easy

W h ite

R ed

E asy

B itter
B itterle ss

Y e llo w
W h iteY ello w ish

L e ss e a sy

T astyW h iteR ed

hN uoollincok ing
gm illing
rice
fo r

W ild

Conclusions
Nepal is rich in buckwheat

diversity.

known from Nepal, F. esculenturn,

F. tataricum,

Small

scale

buckwheatfarmers living
resources

for their

managing and exploiting
enhanced. Extension

in remoteanalysis
Coordinatingof theirand economicallyto encompass value addedof buckwheat

are

hills

are dependent

knowledge

about

agro-ecosystemonconserving,needs to be

to enable farmers to become expert at growing a healthy

buckwheat crop in an ecologicallyThus, the efforts

Farmers

germplasmof buckwheat

F. cymosum and F. megacarpum.

areas of the highlivelihood.

the local

efforts

To date, four species

populations

and developing

links

into

sustainable

activities

in-situas wellconservation

with other in situ projects

exchange experiences

and technicalknowledge

long term conservationof agro-biodiversity

manner are needed.
as genetic
researchdiversity

are needed.

and NGOs to share andfor mutual benefit and for broader andare hoped for in the future.

Acknowledgements
The author is gratefulworkshop. He is thankful
acknowledges
Botany, Nepal

to MAFF, Japan for the financial

to Dr S. Miyazaki

for the sincere

supportfor his constantsupport to attend thisencouragement. He alsoof Mr. B. K. Baniya, Divisionof Agricultural

Table 7. Distributionof diversity

eaAcol zgloorogn ic‑e

M u sta n g>MH 2igo6uh0n0hta‑3
illin5 a0s n0dm

D o lp a

H> 2ig2 h0 0h ills
m .a an.s.l.
d M o u n ta in s

Source:Nepal District

Profile,1998

L o c a tio n
A ltitu d e (m .)L o c a tio n (vNillao .)g e & W a rd

V a r ie ty

K e y in fo r m a n t

Jh ark o t
N a g a rk o t
R an ip u w a
L o m an th an g
S a h artara ‑8 , D u n a i‑5 ,T rip u ra ‑6 , & Ju p h al‑1 ,P h o k su n d o ‑8 , M ajh p h a l

M ith eT ite

2 0 0 0 ‑3 8 0 0

D o rje G u ru n g ,L ah u re G u ru n g
N u rb an g S h erp a

T ite ,
M ith e ,B h a te2 0 0 0 ‑2 9 0 0

M an B ah ad u r
T h ak u ri, B a g h
B a h ad u r S h a h i
T ik a B a h a d u r
B o h a ra , H a sta
B ahadurB ud dha,
S a m p a ti
B ud dha,
W an g e l L am a ,C h a n d ra D atta
K h a tri

Im p o rta n t
tr a its
G h y ab re , D h u p ,M ith eG h o re , S y an g re ,D h e sh u ,T a sh in g , M ith e
B h ateU se o f sp ec ific v a riety
M e d ic in a l v alu e+ L o c a l
c o n su m p ti o n u se fo r fo o dv e g etab le an d liv esto ck u se
L o c a l c o n su m p tio n
M ed icin al/R elig io u s, p la n ts
a re u se d for liv e sto ck ,c o o k e d w ith ri c e

References
Annual Report. 1998.

Agriculture

Development

Office,Dolpa, Nepal.

Annual Report, 1998.

Agriculture

Development

Office,Mustang, Nepal.

and NR Dhungel.

1995.

Further(Fagopyrum

spp.)

Baniya,BK, DMS Dongol
evaluationof Nepalese

304 in Current
International

buckwheatcharacterization
landraces.

Pages

295-

Proceed.

6th

Advancesin Buckwheat

Research.

Vol. I-III.

Symposiumon Buckwheatin Shinshu,

24-29

Matano and A. Ujihara,eds.).

Shinshu

University

andAugust1995(T.

Press.

Bimb, HP. 1999.

Current

statusof buckwheat

submitted

to theprojecton "Collaborative

research

of

plant

genetic

in

Nepal",

NIAR,

Japan.

T Hiroseand M Minami. 1995.

Varietaldifferences

andfor rutin content

in common buckwheat,F. esculentum

conservation

research

resourcesin Nepal.

A review reportprogramon in-situ(unpublished).
Kitabayashi,

H, A Ujihara,

heritability

Moench.

Jpn J Breed. 45:75-79.
Nepal District
Ohnishi,

Profile.

O. 1989.
southern

1998.

His Majesty's

Government of Nepal.

Cultivatedbuckwheatand their wild relatives

562-571

in Proceed

China.

Buckwheat.

Pages

Oreal, USSR.

in the Himalayas

4th International

Symposium

andonIn-situ

conservationin Chile:

Collaborative

Takanori

Sato

National

Research

research on wild relatives
research

between Chile and Japan

Instituteof Vegetables,

Ornamental

514-2329,

Japan.

Currently:

Japan International

ResearchCenter for Agriculturalof in-situ conservationof wild crop relatives

1-2, Tsukuba,

Ibarakiof tomatoes

Plantsand Tea (NIVOT),

SciencesAno, Mie,(JIRCAS),

Ohwashi

305-8686

Abstract
The objective
its natural habitat
conservation,

an evolutionary

plant

genetic

dynamic state. Through the combination

diversity

can be effectively

America is a major center of crop origins.
speciesthat are native to southwestern

the northern part of Chile.

America. Lycopersicon chilense(3,600

to 1998.

Field

Japan, and Instituto

observations

prevail,relatedbetween the coastal
research project

Research Institutede Investigaciones

and resultsof nine closely

and L. peruvianum grow inm). An in-situ conservationof tomato in Chile was conducted between Nationalfrom 1996

The Andean region of SouthIn northern Chile extremely dry conditions

Ornamental Plants and Tea (NIVOT),
Chile,

maintained.

inof both in-situ and ex-situ

The genus Lycopersicon consists

plain at sea level and the Andean foothills
wild relatives

is to maintain genetic diversity

on theof Vegetables,

Agropecuariasof this basic

research(INIA),

on wild genetic

resources of the genus Lycopersicon from northern Chile will be discussed.
Introduction
Ex-situconservation

However, recently
international

the importancefora. Genetic

for many traits(Vaughanof crops to be a useful
maintaining
genetic

has contributed

diversity

breeding

resource.

diversity.

diversity

can be effectively

Theregiondomesticated.
L. pimpinellifolium,of

All the wild relatives
L. cheesmanii,

to the development

conservation

has become an important

topic

compared

relativesof cultigens

and Okuno, 1996).

of the genetic

Andeanof in-situ

significantly

is smallNew breeding

In-situ conservation

Throughboth

in-situwith theirtechniques

enableof wild relativesof agriculture.

wild

in

wild relatives
is essentialand ex-situ conservation,

for
plant

maintained.
South

Americaisa regionof tomato, Lycopersicon
L. parviflorum,

where

many crops

wereesculentum

var. cerasiforme

andL. chmielewskii,

L. pennellii,

L. hirsutum,L. peruvianum,and L. chilense,

are native

southern

Colombia

to northern

Galapagos

islands)

to the eastern foothills

in the northern
distribution

part of Chileand characterized

A collaborative

in-situ

betweenobservation

sites

research project

NIVOT, Japan,
in northernwere to measure the genetic
three years. In addition,of the Andes. L. chilenseareas of Chile
diversity

analysisof L. chinensis

diversityand the climate

1). Theplan to 3300m. These two

The objectivesof L. chilense

wasconditionsof the researchand L. peruvianum over

diversity

Finally,of tomato in Chileon ecogeographic

the habitats

and L. peruvianum.
in northernBasedbetween genetic

paper we describe(Photo.

self-incompatibility.

were selected.

of populations

the relationshipwas to be examined. In thisprevailon these wild relatives

and INIA, Chile.

theand L. peruvianum growdry conditions

by gametophytic

They occur from

Ocean coast (including

is from sea level on the coastal

are outcrossing

initiated

America.

and from the Pacificwhere extremelyof these two species

species

Field

Chile

to southwesternand eco-climaticand discuss

factors

resultsof RAPD

betweengenetic

the relationship

Chile is discussed.

observations

1996
Based on the results of a planning trip in 1995 the route of the expedition in 1996 wasdetermined. A field trip from 13th November to 1st December 1996 had four members,
Takanori

Sato(NIVOT),

AngeraE.

Pezoa(QUILAMAPU-INIA),

Pedoro Leon L. (CRI

organized

the observation

to establish

city of Chile

to Copiapo

Duringsites
fields.

initially

were added over the period

roadsides,

coastalareas(80m),

from the fixedfrom a further 40 sites during
17 samplesof otherT.

LA PLATINA-INIA).

The first

missionwas

14 observation

the expedition

sites
These

areas (3,300m),

and 97 fruit

observationfrom Arica the northernmost

1).

mountainous

Solanaceae

Matusof the project.

A total of 255 leaf samples

were collectedIvansites, in an area extending

770 km south (Fig.

the expedition,(INTIHUASI-INIA),

sites.

and two further

sites

dried

includedwastelands,

samplesof L. chilense

A further

165 fruitconsisting

genera (Table

were selected1). Leaf samples

and cultivatedand L. peruvianum

samplesof 148 samples

riversides,

were collectedof Lycopersicon

were pressedandand dried

between paper towels.
The middleof November is the beginningrainfallcan occur at elevations

speciesseems to depend

higherof the summer season in northern Chile andthan 2500m. In this area the growth of Lycopersiconon the amount of rainfall

in the rainyseason(DecembertoFig. 1. Map of South America with inset of expeditionroutes and sample sites in Chile

Table1 Resultsof explorationand collectionof wild tomato in Chile
Samples(1996)for DNA analysisa cc
N ue mssiob e nr so flea
N ufm sab em rp olef

N u itfru
m bsamer op fleL y cop e rsic o n p e ru via n u m

1 18
30

80
17 5

25
72

S o la riu m ly c op ersico id es

7

0

0

S o la rium rick ii
S o la rium sp p .

4
1

0
0

0
0

C a p sicu m p u b es cen s

5

0

0

16 5

2 55

97

S p e c ie s

L y co p ers ico n c h ilen se

T o ta l

Table

2 Number of leaf and fruit
years in Chile (1996-1998)

samplescollected

S ite sN o . o f le a f s a m p le s ( N o . o f c o lo n ie s )(A ltitu d e )2)

1 9 9 6 1)

1 9 9 7 1)

19 9 8 1)AB( 1 ,3 4 5 )(2 8 0 )

2 1
18

6
12

C
D
O
E
F
G
H
I

(7 3 0 )(3 ,0 5 0 )(3 ,2 9 5 )(2 ,4 5 0 )(2 ,3 5 0 )(2 ,4 3 0 )(2 ,5 7 0 )(2 ,7 2 0 )

20
20

22
26
10
14
27
13

8
12
2 3 (< 1 0 0 ) 3 )
2 5 (> 2 0 0 )
2 2 (< 10 0 )
12
2 5 (< 1 0 0 )926

2 5 (> 1 0 0 )

J
K
L
P
M
N

(2 ,8 7 0 )(3 ,0 6 0 )(3 ,1 8 0 )( 3 ,3 3 0 )(2 ,8 7 0 )(8 0 )

24
18
4

2 5 (> 1 0 0 )
2 5 (< 5 0 )

21
2 1

26
22
1
16
25
21

1 9 (< 4 0 )
2 5 (< 1 0 0 )
2 3 (< 3 0 )

2 55

2 67

278

T o ta l

24
24
7
13
20

from each site for three
N o . o f f ru it s a m p le s
1 9 9 6 1)

1 9 9 7 1)

19 9 8 1)

3
3

1
1

0
0

20
18

1
3
10
0
6
0

19
16
1
0
15
6

2

2
10
9

7
0

13
8
0
4
10
1

97

60

96

11
19
7
1
0
0
9
0

: Date of explorationin 1996 were from Nov. 12 to Dec. 1.
Date of explorationin 1997 were from Nov. 19 to Dec. 7.
Date of explorationin 1996 were from Oct. 12 to Nov. 19.
: meter above sea level.
: Number of plants in fixed sites.

0612

Photo. 1. Habitatsof wild relativesof tomato in Chile,bloom near Arica. 1b. Solanum lycopersicoides

by the roadsidechilense

growing in a rock crevice in very dry conditions

3,295masl. 1d. Lycopersicon

Belen, 3,050m

asl.

chilense

1a. Lycopersicum peruvianum in fullat point

near Putre. 1c. Lycopersicon
O near Central

growing as a weed in terraced

fields

Chapiquina,

at point

D nearFebruary).

While autumn (March-May)the lower elevations,fruitsis a better time to collectcan still be found at the beginning

trip followed

route CH11 from Arica to Putre at 3,415m

at 4,000m.

L. chilense

and the often

sympatric

fruitsof Lycopersiconat allof summer. The first part of theand then to the border with Bolivia

species

were1b) However, plants

were notfrequent

along the roadside

fruiting

at this elevation

at this time. This may be because the rainy season had not begun andplantsat this elevation

were not vigorous.

roads are locatedbetween 3,000 and 3,600m (Photo,

Solanum lycopersicoides

During the rainy season, mountain slopes

can hold more water than other mountain slopes.

In northern Chile,

L. peruvianum was found only in restricted(lessthan 1,200m)

and on the coastal

plain

besidesroads,

L. peruvianum

was found in Arica,

Paposo,and Taltal.

valleyswasteland.

peruvianum plants that grow near Arica differof this species

where

distribution

the road crossed

areas of lower parts ofaround and in fields

and

The habitof L.

from those around Paposo, the southern

extentbased on observations

during the mission.

At site N near Paposo,a dried river.

L. peruvianum was found in both the lower reachesof theriver, 100m from the seashore,and in the upper reaches. These plants are presumably

derivedfrom plants

in the upper mountains, however they differ

in many charactersfrom those in thenorthern Andean mountains of Chile.
Populations
populationof wild

were sampled.

Lycopersicon
Sites

were selected

DNA samples were extracted(Santiago)

by Jorge Gabriel

generally

consistto represent

from dry leaf

and Patricioof few plants

so all plants

broad eco-geographic

samplesin adiversity.

at CRI LA PLATINA-INIA

Hinrichsen.

1997
During the second year of the project,same 14 observation

leaf and fruit samples

were collectedsites as 1996 from November to December (Appendix

ecologicaland meteorological

data was gathered

for the fixed

Takanori

Sato(NIVOT),

AngeraE. Pezoa(INTIHUASI-INIA),PLATINA-INIA),

Claudio

Villegas(CRI

LA PLATINA-INIA).

sites.

B). In addition,

MissionLuis

from themembers wereMeneses(CRI

The winter temperaturesLAin

1997 was cooler than average in northern Chile.
Site observations(Site(Appendix

A; Table 2)A) Compared to 1996 fewer and smaller plants

corn field.

In 1996,of L. peruvianum were found around theL. peruvianum was found in corn fields,

only found growing along with a small irrigation

but this year small plants

weresystem.

(SiteB) This site was in the lower reaches of Rio Lluta and L. peruvianum was growing infallow

land (Photo,1a). Althoughmany plants

with flowers

were found, only one fruit wasfound. Farmers in this area have a corn or onion and fallow land rotation.
peruvianum can survive as a weed in cultivated(SiteC) Locatedbesidethe road and abundant garden products

D) L. chilensebean, alfalfa

such as vegetablesfrom which Brassica

However, most plants were floweringand fruits

that L.

of irrigationwater

for long periods.

at the bottom of a deep ravine and there was plentyof L. peruvianum were in a field(Site

fields

It is possible

and fruit trees. Many plants

crops had recently

were too young to sample.

was growing in a stone wall that formed the terrace

and corn at an elevation

been harvested.

of 3050m (Photo.

1d).

Althoughof fieldsof broad

many L. chilense

plantswere growing at this site most were still young.

(SiteO) This site was the dry bank of an unpaved road but there were many small plants

chilenseand 10 fruits

plants(Photo,(SiteE) Only desiccatedF) Large

riverbed.

were collected.

Some ratoon shoots were coming from old L. chilense

plantsbeside1c).

dry ravine. Rainfall
(Site

of L.

the paved road along aat this site does not occur annually.

plantsof L. chilenseThis riverbed

many L. chilense

were found at this site locatedplantswere found growing

pool may retain water throughout

at thissite around in a pool of athe summer season. The leaves ofat this site were small and shrunken. Generally,

L. peruvianum grows

at below 1,200m, but at this site a L. peruvianum like plant

was growing at 2,350m.

This

plant had completely

Stems, leaves and fruits

werehairy and adhesive

different

characteristics

from L. chilense.

and stems were creeping.

These characteristics

peruvianum samples found during the mission in northern
On the way to site G, L. peruvianum, L. chilensefound growing sympatricallywere similar

along a dry riverbank.

are different

from other L.

Chile.
and Solanum lycopersicoides

were

The L. peruvianum plants at this location

to those at site F. This was the first time we had found these three speciesallgrowing together.
(SiteG) The number of L. chilenseriverbed(Site

plantsover the previous

site and most plants had no shoots. We could not find any flowersI) L. chilense

was growing in a dry riverbed

area of 100m by 30m. The wild relatives
Chile.

had increased

However, at this site seedlings

a big taproot.

There were plants

with flowers,

dryor fruits.

at the bottom of a shallow

of tomatoes grow as perennialof L. chilenseyear at thisravine over anplantsin northernwere found with small, young leaves andbut plants were small and only 13 fruit samples

were collected.
(SiteK) This site was besidean unpaved road and slope on the way to the bottom of a large

and deep gorge. We found two typesof L. chilense

plants.

One type had young shoots

regenerated

from the stem of old plants.

The other type appearedthat this area had rain during the rainy season, since a landslide(SiteL) Thisfour leaf samples

A new site (P) near site L with similar

providedinsufficientP) Thissmallgorge has similar

characteristics

were collected.

(Sitesteep gorge without

M): This small,
(Salt

(Site

geographic

features

was found since site L

samples.

samples of L. chilenseRiver).

large plants

was seen close to the site.

and no fruits. In 1997 only 1 leaf sample wasobtained.

SaladaIt appearedsite is a small gorge, about 20m deep and 10m wide and lacks water in the dry

season. In 1996 we obtained(Site

to be seedlings.

to site L. 16 leaf samples

and 4 fruit

water in the dry season is a tributaryof the Rio

We found small L. chilense

plantson the steep sidesof the gorge andin a remote part of the gorge.

N) Thisdry riverbed at 80m had L. peruvianum in its upper and lower reaches.

were many L. peruvianum plants

at N point, but most plants

and plants with fruits were being attacked
Taltal,locatedby insects(Tableabout 90 km south of site

There

were too young to have fruits

2).

N, is the southernmost

site where L.

peruvianum was found growing in Chile.
267 leaf samples
extracted

and 60 fruit samples

from dry leaf samples

were collected

in 1997(Tableat CRI-LA PLATINA INIA (Santiago)

2). DNA wasby Maria Herminia,

Humberto Godofredo and Patricio Hinrichsen.
1998
In 1998,
October(NIVOT),

we planned to collectleaf and fruit samples from the same 14 sites, from 12th

to 19th November in 1998 (Appendix
HiroshiMatsunaga (NIVOT),

Pezoa (INTfflUASI-INIA),

C). The mission

members were Takanori Sato

Horacio Lopez (CRI LA PLATINA INIA),

Luis Meneses (CRI LA PLATINA INIA).

The 1998

Angeraexpedition

was carried out about one month earlier

than in previous

in the number of leaf and fruit samples

collected

during the three years. It appeared

was no major variation in the climate

of northern

Chile over the three years. However, basedon informationfrom an Ecuadorian professor

years. There were no major changes

in 1997 the climate

affectedby El Nino. Northern Chile was unaffectedprevailed(Tablethat thereof Ecuador and Peru wasby El Nino and stable climate

conditions

2).

(Site

A) L. peruvianum was growing by a simple irrigation(SiteB) L. peruvianum

was growing besideuncultivated(SiteC) An uncultivatedfieldwith L. peruvianum over an area of 100 X 30m. 19

fruit samples were collected.

was dottedsystem and plants were small.
fieldsas a weed in crops (Photo. 1a).

*(p)

(Site

D) L. chilense

1d).

The first

was growing in a terraced fieldsampling

locationfound growing in terraced
had no irrigation.
(Site

was at 3,135m

of corn and broad bean as weeds (Photo.
and the lowest

at 3,085m.

L chilense

field with crops but was not found in near by abandoned

wasfieldsthat

16 fruit samples were collected.

O) This site was the bank of an unpaved mountainsidewere small compared with the plants

at other sites(Photo,road. L chilense

plants at this site1c). L. chilense

plantswere along

an unpaved road.
(SiteE) Thishabitatof the plants

was a rocky bank besideof L chilensea paved road. The site was very dry and somewere dead presumably

because of hightemperatureand lack of

rain.
(Site

F) Plants

chilense

were growing around a puddle

plantshad grown well during

in a dry riverbed

by a road crossing.

the rainy season since most plants

The L.

were large. 15 fruit

samples were collected.
(SiteG): L. chilense

plantswere scattered

(Site

I):

plantswere scattered

L. chilensealong a dry riverbed.
along a dry riverbed

and plants

were relatively

small.
Table

3 Average plant size of samples (LycopersiconchilenseandL. peruvianum) collectedfrom each site in Chile (1998)
S it e ssN a om .op fle(wP climad nt)ht

hP ilga hn t

sD tei am m ( aem ttmbe ar) so ef(c m )
A(p )*8

6 6 .4 +‑ 7 0 . 0

2 2 .5 +‑7 .5

8 .6 +‑

6 3

B

(p )

12

7 9 . 6 +‑1 8 .13 4 . 1 +‑5 .2

7 .7 +‑

1 .2

C

(p )

2 3

2 5 0 .3 + ‑ 4 4 . 1

5 0 . 5 +‑2 1 .6

1 9 .4 +‑

1 .0

D

(c )

2 5

5 8 .0 + ‑7 .1

1 9 .5 +‑

5 .0

E

(c )

12

8 2 .5 +‑ 2 6 .3

4 3 .4 +‑1 2 .91 7 .1 +‑

5 .4

F

(c )

2 5

2 1 4 .8 +‑ 4 1 .7

6 9 .8 +‑7 .8

3 1 .2 +‑

4 .S

G

(c )

9

1 1 7 .8 + ‑ 8 7 .0

5 3 .3 +‑1 7 .3

1 5 .9 +‑

6 .7

H

(c)

I

(c)

2 5

6 2 . 4 +‑1 4 .73 9 .0 + ‑4 .2

5 .3 +‑

1 .2

J

(c)

2 5

5 6 . 0 +‑1 0 ,5

3 4 .6 + ‑5 .7

1 1 .7 +‑

2 .3

K

(c)

2 5

7 9 . 0 +‑1 8 .03 1 .4 +‑3 .5

1 0 .6 +‑

1 .6

L

(c )
1 .3

8 8 .2 +‑1 4 .9

M

(c )

2 5

6 1 .4 +‑1 2 .44 7 .4 + ‑6 .9

8 .0 +‑

N

(p )

2 3

1 1 7 .8 +‑ 2 3 .5

4 4 .3 +‑6 .2

1 0 .8 +‑

1 .8

O

(c )

2 2

4 0 .7 +‑8 .0

2 6 .0 +‑3 .9

8 .2 +‑

2 .3

P

(c )

19

1 0 5 .0 +‑ 2 8 .7

5 4 .7 +‑1 0 .3

6 .5 + ‑ 0 .9

: Lycopersicon

peruvianum(c) : Lycopersiconchilense(SiteJ):This

outsidesite

was a gently,

Coscaya village.

(SiteK): ThisP):site was a steep slope besideThischilense

rocky wasteland.

L. chilense

an unpaved road at the edge of a deep gorge. 9

were collected.

site was a small, dry gorge 20m deep and 10m wide. 12 leaf samples

were collectedM): This(Salt

River).

site was a small, steep, dry gorge that formed a tributary(SiteN) At this site L. peruvianum plants grew along the dry riverbed

25 leaf samples and 6 fruit samples of L. chilenseto near the shoreline.

12 fruit samples were collected

L. peruvianum plants

have a creeping

250cm (Tableof L.

but no fruit samples.

(Site3). However, L. chilenseto the amount of available
DNA was extracted

were collected.
from the upper reaches

in 1998.

plantsat F point were the largestof 31mm (Table

plantswas up tobeing 215cm

3). A plant size appears towater.

from dry leaf samples

Claudio

Narvaez, Genevieve Merabachvili

Genetic

diversityof L. chilenseto the Rio Salada

habit and at site C the canopy width of plants

across and 70cm tall with a basal stem diameterbe related

was growing in and

10 fruit samples were collected.

fruit samples of L. chilense(Site

sloping

at CRI LA PLATINA-INIAand Patricio(Santiago)

byHinrichsen.

and L. peruvianum in northernChilebased on RAPD

analysis.
Leaf samples were collectedof wild relatives
humidity.

from 14 fix pointsof tomato are soft and easily

At the start of thiscollaborative

witherbetween 1996 and 1998. Leaf samples
and decay in warm conditions

project,

we testedand high

two methods for keeping

samples for DNA extraction.

The first method was to maintainhumidity

The second method was to keep leaf samples at low humidity

and low temperature.

and room temperaturearound 10-30•Ž.

samples should be pressed
DNA was extracted
method.

A greater

extracted

DNA as templatea final concentration

Based on the results

it was concluded

and kept dry because of the lower humidity
by a modified

DNA oligomerthat leaf
bromide)than normal was used. In order to use

DNA for PCR (Polymeraseof 5 pmol/ml.

of highin northern Chile.

CTAB (Cetyltrimethyl-ammonium

amount of 2-mercaptoethanol

primers for PCR. Reaction

leaves in conditions

leaf

chain reaction),

DNA was adjusted(12)Gene) were used as

mixtures for PCR were 1.25ƒÊlset (Nippon
X 10 reaction

buffer,1.0ƒÊl

to
2.5

ƒÊl

mMdNTP, 0.125

ƒÊl
ƒÊl.

and 6

template

94•Ž-30s,

45 cyclesof denaturation

elongation

at 72•Žfor 3 min followed

by 72•Žseparatedalongsidea molecular

1.5% agarose

Tth DNA polymerase(TOYOBO),

DNA, total mixture was 10

gel containing

primer,

3.125

PCR amplification

at 94•Žfor 30s.,

distilledwater

was performed, at firstannealingat 40•Žfor 2 min. andproducts

weremarker (ƒÉEcoRI & HindIII)by electrophoresisonbromide.

out on a Takara PCR

weightethidium1

for 7 min. Amplification
PCR was carried

Thermal Cycler MP with 96 well sample blocks.
Randomly amplified
binarypolymorphic

matrix (1 for the presence

Pairwisedistance(similarity)

package(Rohlf1998).

and 0 for absence

matrices

Dendrograms

methods with arithmetic

DNA (RAPD)

averaging)bands were scored and recordedof a band at a particularwere computed using NTSYS-pc
were createdclusterposition).

ver. 2.0 softwarewith UPGMA (unweighted

analysis(Sneathand Sokal

as

1973,

pair

group

Swofford

andOlsen 1990).
L. peruvianum and L. chilense
intra population

genetic

samplesin 1997

collected

diversity

have strict

gametophytic

was found. Results

are shown (Fig.

2). Results

self-incompatibilityof cluster

analysisusingof 1997

showed that L. peruvianum

plantsaround Arica (Sites

A, B, and C) are in the same group. This result

genetic

diversity

to geographic

to be different
Samplesis related

between L. peruvianum

location.

Also morphologicalsome of thesuggests

charactersthatwere seenin the Arica area and those from the Paposo area.

from site D are L. chilense.

Sites

these were in the same group, but L. chilense
peruvianum at site N. Genetic diversity

and thus

A, B, and C were of L. peruvianum andat site D belonged

to the same clusterof L. peruvianum within

as L.

site N was low, but one

sample from site N was an unusual sample.
Morphologicaldiversityof wild relatives

Ninetyaccessions

collected

cultured1999).

by the INIA/JICA

in a glass house. MorphologicalIn the case of wild relatives

descriptors

and units for evaluating

results

for four descriptors,

leafletnumbers, are shown (Fig.

chilense

is generally

we could

greater

not distinguishlengthof tomato.
collaborative

survey was based on standardof tomato, additional
the wild relatives
3a to d). Diversity

project

were

descriptors(MAFF,

are necessary.

The newof tomato is shown (Appendix

D). Thedescriptorsof a rachis truss, sepal length,

fruit weight,of morphologicalthan for L. peruvianum in northern
L. chilense

researchfrom L. peruvianum

Chile.
based

and compoundcharacteristicsof L.

For some accessions,on morphologicalFig. 2. Dendrogram of 92 Lycopersicon accessions based on genetic diversity
based onRAPD analysis(1997).
Leaf samples were collected from A, B, C, D and N fixed sites in
1997. A, B, C and N are L. chilense and D is L. peruvianum.

Fig. 3. a. Length of truss rachis of Lycopersicon peruvianum and L. chilense collectedfrom Chile (1995).
b. Length of sepal in L. peruvianum and L. chilense collectedfrom Chile (1995).
c. Fruit weight of L. peruvianum and L. chilense collectedfrom Chile.
d. Number of leaflets in a compound leaf in L. peruvianum and L. chilense collectedfrom Chile.

characteristics.

Changes in wild relatives
Exploration
1996

and 1997.

of tomato populationsin 1998

based on the field

was carried out about one month earlier

The time was aimed to coincide

The difference

these three years did not markedly

the total

affectexcept for the fruit samples in 1997 (Table

highlandAn agro-climatic

map of northernof northernis highland

Chile

Andes above 3,800m

temperaturesin the oceanic tropicalinin time of the field

number of leaf samples

surveysduringand fruit

samples2).
Chiledesertis shown (Fig.

(2,000m)

at night.

and oceanic subtropical4). The middle

range (plain

and an area of limited

are too cold for wild

can drop to below freezing

than explorations

with the start of the rainy season, but rainfall

only occurred once during the trip in 1997.

color)

survey.

relativesTheof tomato to grow, because

However, there is no rainfall
desertrainfall.

below 2,000m

area. In areas below 3600m L. chilenseand L. peruvianum can grow.
The annual mean minimum and maximumair temperaturesnear the collecting

sites

are shown (Fig.

maximum air temperaturein Putre5). This(3,530m)

figureshows that the mean, minimum andis rathermaximum,minimum and average air temperature

over four years 1995-98
year round. The mean
respectively.

The

Azapa area (250m)

near to Arica is warmer from November to March (summer season)

than

May to September(wintercollecting
Putre

sites

is shown (Fig.

(3,530m),

but theresummer season (January
The humidity

in the highlands
is related

and humidity

is no rain in the Azapa area (250m).
to March)these climate
for L. chilense
above freezing.

some sites seedlings

is high,but otherwise

conditions,temperatures

humidity

In Putre humidity
is between

water is sufficient.

above freezing

L. chilensein theto the sea.

and summer rainsystems in the lowlands

are the minimumof tomatosources of water such as irrigationand L. peruvianum are generally

and low temperatures.

in35 and 40%.

and L. peruvianum. Plant size of wild relatives

were found, these plants

conditions

near the6). Between January and March (summer) there is rainfall

to the amount of summer rain, alternative

and temperature

available

The annual mean precipitation

and water from rivers or irrigation

requirements

severe drought0•Ž and 10•Ž,in the Azapa area is high between 60 and 70%, due to its proximity

Considering
habitatseason).

is 18•Ž,

uniform

perennial.

water
At

had large roots and this may be to withstandThus, propagation

by seeds may occur if

Fig. 4. Map of agroclimatic

divisionsin northernChile(adaptedfrom Novoa et al., 1989)

Fig.

5. Annual maximum, mean and minimum temperaturenear fixed points(PutreAzapa).

Fig. 6. Annual mean precipitationand humidity

near fixed sites (Azapa

and Putre)andConclusions
After three years of collaborative
1) The weather conditions
2) Rainfall

is limited

3) L. chilense

researchin northern

we can summarized the results

Chile are very stable;

and uniform;and L. peruvianum are mainly perennial

4) L. peruvianum

as follows.

generally

plants in northern Chile;

grows at less than 1,200m

besidefields,roadside,and seasonalrivers;5) L. chilense
seasonal

grows at more than 1,200m

and less than 3,600m

besidefields,road andrivers ;6) There is a lot of water in the lower reaches of a river, but in some case, the salt content
very high and wild relativesof tomato cannot use such water. In fertileof tomato can grow as weeds besideor in fields,isregions

wild relatives

however, if farmer discontinue

cultivation

other weeds become dominant.
Northern Chile is thought

to be good region for in-situ conservation

L. peruvianum. The weather conditions
disruption

by natural

or artificial

The degree of genetic

forces is relatively
diversityeach site and between different
between genetic
it willbe possible

speciesin Chile.

diversity(RFLP)and ecologicalself-incompatibility

for the S gene by PCR-RFLP
character
This

joint

years is insufficient
factors.

It willconclusions.

factors

program of in-situ

analysis.

Genetic

has been carriedfragment

plantsdiversity

to continue

thisand thatof wild

lengthcharacterized

tomatopolymorphismby gametophytic

are highly

polymorphic

genetic

diversityof S-RNase is an interestingandmarkers (RAPDs).

over three years (1996-1998).

time to analyze the relationshipbe necessaryconservationin PCR we can expect to detectcompared to random polymorphism
project

atthat the relationshipwill be clarified,by the S gene. S gene products
primers

a population

diversity.

and L. peruvianum are outcrossing

S-Rnase. Using S-RNase specificstableIt is anticipatedand environmental

to measure genetic

that is regulatedof environmentalof the tomato within

be monitored.

a practical

and

low

such as RAPD and PCR-restriction

are useful techniques
L. chilense

and the possibilityof wild relatives

sites will

to develop

DNA analysis

are very stableof L. chilense

between genetic

work for several

diversity

However, threeand ecologicalmore years to obtainfirm

Acknowledgments.
We are gratefulproject

to Dr. Alberto

for in -situ conservation

Dr. Shigeru

Suzuki(Leader

am grateful

Mr. Tatemi

Cubillosfor organizingof wild relativesof tomato in Chile.

of the former JICA/INIA
Yoshidaour collaborative

project)

for supervisingtheresearch

We are also gratefulfor his helpfullaboratory

tosuggestions.

in NIVOT

absence. We thank all the members of NIVOT, INLA, and NIAR for supportingI

duringmythis project

References
FAO. 1996.

The state of in situ management (Chapter

genetic

resources

for food and agriculture.

FAO, Leipzig,

MAFF (Ministryof Agriculture,

Forestry

Resources:

Characterizationand evaluation

plant genetic

resources.

MAFF, Japan. 256 pages.

Novoa, R. et al. 1989.

Rohlf,

Mapa Agroclimatico

Instituto

de Investigaciones

Santiago,

Chile.

J. K. 1998.

Numerical

two). The state of the world's

and Fisheries,

Germany, 17-23,
Japan).

1996.

- new approaches

de Chile.

EditedAgropecuarias,

taxonomy and multivariate

plant

1996.

Plant

Genetic

for improved

use ofby Novoa, R. and Villaseca,S.

INIA

Ministeriode Agricultura.

analysis

system. Exeter, Publishing,Ltd., New York.
Sneath,P. H. A. and R. R. Sokal.

Swoffordand Olsen. 1990.
M.Hillis

Vaughan,1973. Numerical

Phylogenic

and C. Moritz.

conservation

reconstruction.

Sinauer Associates.

D. A. and K. Okuno. 1996.

taxonomy. Freeman, San Francisco.
In Molecular
Sunderland.

systematic.

Editedby D.

pp.411-50.

Plant

genetic

resourcesin situ. Res. J. Food Agric.

19:16-22.

(in Japanese)

for agriculture

and their

*:AppendixA

Collecting

S it e s A lt i t u d e(m )sites for leaf and fruit

L oa tnitguidtue d e/P Sr otav tien c esamplesof LycopersicontNo ew a nr e /s tv i l la g e

in Chile(1996-1998)

L o c atio n

SC po ellc ei ce ts e d

C o rn fie ld

L . p e r u v ia n u m

A

1 ,3 4 5

S : 1 8 ‑ 1 9 '3 7 .5 "W : 6 9 ‑ 4 9 '5 7 .9 "T a ra p a c a

B

2 80

S : 1 8 ‑ 2 4 '3 7 .9 "T a ra p a c a

A r ic a

S i d e o f f ie l d

L . p e r u v ia n u m

T a ra p a c a

1 8 d e S e p t ie m b r e

U n c u l t iv a te d la n d

L . p e r u v ia n u m

S : 1 8 ‑2 7 '5 4 .2 "W : 6 9 ‑3 1 ' 1 6 .5 "T a ra p a c a

B e le n

T e r r a c e f ie ld

L . c h ile n s e

S : 1 8 ‑2 8 ' 1 3 . 1 "T a ra p a c a

C e n t r a l c h a p iq u i n a

R o a d s id e

L . c h ile n s e

R o a d s id e

L . c h ile n s e

W : 7 0 ‑ 0 9 '5 8 .0 "
c

7 3 0

S : 1 8 ‑3 4 '5 4 .2 "WD

3 ,0 5 0

O *3 ,2 9 5: 7 0 ‑ 0 0 '2 7 .0 "W : 6 9 ‑ 3 1 '4 0 .0 "E

2 ,4 5 0F

2 ,3 5 0

S : 1 8 ‑2 6 '4 1 .5 "W : 6 9 ‑4 5 '3 6 .7 "T a ra p a c a

S : 1 8 ‑4 6 ' 0 1 .6 "T a ra p a c a

T im a r

B a n k o f d rie d r iv e r

L . c h i le n s e

T a ra p a c a

E s q u in a

B a n k o f d rie d r iv e r

L . c h ile n s e

L . c h i le n s e

W : 6 9 ‑4 1 '4 6 .2 "2 ,4 3 0

G

S : 1 8 ‑5 4 ' 0 6 .4 "W : 6 9 ‑3 7 ' 2 6 .6 "H *2 ,5 7 0

S : 2 0 ‑0 0 '3 6 .2 "W : 6 9 ‑ 1 2 ' 1 2 .7 "T a ra p a c a

P a rc a

R i v e r s id e &ro a d sid e

I

2 ,7 2 0

S : 2 0 ‑0 3 ' 4 0 .9 "W : 6 9 ‑ 1 3 ' 1 8 .7 "T arap aca

M a m in a

D rie d riv e r b e d

L . c h i le n s e

J

2 ,8 7 0

S : 1 9 ‑ 5 1 ' 5 3 .5 "T arap aca

C o sc a y a

S lo p e o f w a ste la n d

L . c h i le n s e

T arap aca

U sm a g a m a

R o a d s id e

L . c h i le n s e

A n to f a g a s t a

S a n P e d r o d e A ta c a m a

S m a l l r a v in e

L . c h i le n s e

A n to f a g a s t a

S a n P e d r o d e A ta c a m a

S m a l l r a v in e

L . c h i le n s e

A n to f a g a s t a

A i q u in a

R o a d sid e

L . c h i le n s e

A n to fa g a sta

P ap oso

D rie d riv e r b e d

L . p e r u v ia n u m

W : 6 9 ‑0 8 ' 1 9 .7 "K

3 ,0 6 0

S : 1 9 ‑4 3 ' 4 4 .2 "W : 6 9 ‑ 1 2 ' 3 9 .2 "I *3 ,1 8 0

S : 2 2 ‑ 5 4 ' 3 3 .4 "W : 6 7 ‑ 5 8 ' 1 8 .5 "P *3 ,3 3 0

S : 1 9 ‑ 4 3 ' 6 6 .6 "W : 6 9 ‑ 1 2 ' 8 4 .5 "M

2 ,8 7 0

S : 2 2 ‑ 1 7 '2 6 .7 "W : 6 8 ‑ 2 0 '4 5 .0 "N

80

FinallyS : 2 4 ‑ 5 2 '5 7 .4 "W : 7 0 ‑ 3 1 '0 7 . 1 "O and P sites were added in the collection

sites

insteadof H and L sites.

Appendix

B Itinerary

and resultsof mission

to collectD a teR o u te1

N o v .1 9 (W e d .)S a n t ia g o ‑ A r ic a2

N o v .2 0 ( T h u .)A r ic a ‑ M o l in o s ‑ R io L l u t a ‑ A r ic a

trip of wild Lycopersiconin Chile
S i te s(1997)
slN ae mao f. polef s

sfN ra muo i.ptol ef s

A

6

1

R

12

1
1

3

N o v .2 1 ( F r i .)A r ic a ‑ Q u e b .A z a p a ‑ A r i c a ‑ P u t r e

C

22

4

N o v .2 2 ( S a t.)

P u tr e ‑ C e n t r a l C h a p iq u i n a ‑ B e le n ‑ P u t r e

D

26

3

O

10

10

5

N o v .2 3 ( S u n .)P u tre ‑ A ric a

E

14

0

6

N o v .2 4 ( M o n .)A ric a ‑ T im a r ‑ C o d p a

F

27

6

7

N o v .2 5 (T h u .)C o d p a ‑ E s c u in a ‑ C o d p a

G

13

0

8

N o v .2 6 ( W e d .)C o d p a ‑ I q u iq u e

9N o v .2 7 (T h u .)I q u i q u e ‑ P o z a A l m o n t e ‑ M a m in a ‑ I q u iq u e

T

26

2

I q u iq u e ‑ H u a r a ‑ P a c h ic a ‑ C o s c a y a ‑ I q u i q u e

J

26

13

1 1 N o v .2 9 ( S a t .)I q u iq u e ‑ U s m a g a m a ‑ I q u iq u e

K

22

8

12

N o v .3 0 ( S u n .)I q u iq u e ‑ C h u q u ic a m a t a ‑ C a l a m a

13

D e c . 1 ( M o n .)C a la m a ‑ S a n P e d r o d e A t a c a m a ‑ Q u e b .E l Z a r z o ‑ C a l a m a

10

N o v .2 8 ( F r i .)14

D e c .2 ( T h u .)C a la m a ‑ A iq u in a ‑ C a la m a ‑ A n to f a g a s t a15

D e c . 3 (W e d .)A n to f a g a s ta

16

D e c . 4 ( T h u .)A n to f a g a s t a ‑ P a p o s o ‑ T a lt a l

17

D e c . 5 ( F r i .)T a lta l

18

D e c . 6 ( S a t .)T a lt a l ‑ C o p i a p o ‑ L a S e r e n a

19

D e c . 7 ( S u n .)L a S e r e n a ‑ S a n t ia g o

T o tal

H

L

1

0

P

16

4

M

25

10

N

2 1

1

15

2 67

6 0

Appendix

C Itinerary

D a teand resultsof mission

to collect

Lycopersiconin ChileR o u te(1998)
S i te slN e ao f. o f

sfN ar omu i.tpolsf

sam p le s1

O c t .2 7 (T h u .)S a n tia g o ‑ A r ic a2

O c t .2 8 ( F ri .)A r ic a ‑ M o l in o s ‑ R i o L lu ta ‑ A ri c a

A

8

0

B

12

0

3

O c t .2 9 (S a t .)A r ic a ‑ Q u e b .A z a p a ‑ A r ic a ‑ P u tr e

C

23

19

4

O c t .3 0 (S u n .)P u tr e ‑ C e n t r a l C h a p iq u in a ‑ B e le n ‑ P u tr e

D

25

16

O

22

1

P u tr e ‑ A ri c a

E

12

0

5

O c t .3 1 (Mo n .)

6

N o v . 1 ( T h u .)A rica ‑ T im a r ‑ C o d p a

F

25

15

7

N o v .2 (WC o d p a ‑ E s c u in a ‑ C o d p a

G

9

6

8

N o v .3 (T h u .)C o d p a ‑ I q u iq u e

9N o v .4 (F r i .)I q u i q u e ‑ P o z a A lm o n te ‑ M

I

25

2

e d .)

a m in a ‑ I q u iq u eH10

N o v .5 ( S a t .)I q u i q u e ‑ H u a r a ‑ P a c h ic a ‑ C o s c a y a ‑ I q u iq u e

J

25

10

11

N o v .6 ( S u n .)I q u i q u e ‑ U sm a g a m a ‑ I q u i q u e

K

25

9

12

N o v .7 ( M o n .)I q u i q u e ‑ C h u q u i c a m a ta ‑ C a la m a13

N o v . 8 (T h u .)C a l a m a ‑ S a n P e d r o d e A t a c a m a ‑ Q u e b .E I Z a r z o ‑ C a l a m a

P

19

0

M

25

6

N

23

12

14

27

96

14

N o v .9 ( We d .)

C a la m a ‑ A i q u in a ‑ C a l a m a ‑ A n to f a g a s t a15

N o v . 1 0 (T h u .) A n to f a g a s t a ‑ P a p o s o ‑ T a l ta l

16

N o v . 1 1 ( F r i .)T a l ta l

17

N o v . 1 2 ( S a t .)T a l ta l ‑ C o p i a p o ‑ L a S e re n a18

N o v . 1 3 ( S u n .)L a S e r e n a ‑ S a n ti a g o

T o ta lLAppendix

D. Descriptors

for wild tomato

1)
2)
3)
4)
5)
6)
7)
8)

Length of truss rachis (cm)Angle between a truss rachis and stem (•‹)
Numbers of flowers on a truss
Length of a truss (cm)Length of peduncle (cm)Numbers of bracts on a truss
Length of the major axis of the biggest bracts
Length of the minor axis of the biggest bracts

9)
10)
11)
12)
13)
14)
15)
16)
17)

Sepal length (cm)Exserted length of style from anther (mm)Presence/absenceof curve in a tip of sterile
Fruit length (mm)Fruit width (mm)Fruit weight (g)Intensityof anthocyanin in fruit
Presence/absenceof stripe in fruit
Fruit hairiness(Absent,
Slight,Present)

18)
19)
20)
21)
22)
23)

Number of leaflets in a compound leaf
Number of the smallest leaflet between main leaflet
Leaflet length (cm)
Leaflet width (cm)Depth of lobation in leaflet (mm)Presence/absenceof a leaflet leafstalk

1)
2)
3)
4)

Length between stem and the first ramification
Angles at a fruit mature stage;
Number of ramificationsof a truss;
Length between the first ramificationof trussof tip;5) Pedicel length from rachis;1) •`8) : Average of 2nd and 3rd truss;18) •` 23) : The biggest
leaf;

in a truss (cm)in a truss (cm)tipof truss or the firstpeduncle;or the firstand onepeduncleSpatial

andtemporallandracesof aromatic rice in the Red River Delta, Vietnam

revealed

by RAPD markers

ShuichiFukuoka1),

Kazutoshi

Okuno1)

1) National

Instituteaspectof genetic

variation

Tran Danh Suu2), Luu Ngoc Trinh2),

of Agrobiological

Resources,

ScienceVietnam2) Vietnam Agricultural

Institute,

TsukasainNagamine1),

Japan

Abstract
The genetic
River Delta,

structureof landraces

Vietnam, has been analyzed.

morphologicallyvery similar.

The analysis

of grainamong aromatic
polymorphism

Plantsin the Red

observations

revealed

that the aromatic landracesof the same landraces

in a field

were morphologicallyuniform.

arecharacters

did not show any distinctgroupslandraces.

landrace

were collected

to analyzeDNA

and 1999.

RAPD

Leaveson a single plant basis
Polymorphic

was determinedDNA) analysis

to be 10. Baseddifferencesin genetic

annually.

Based on these results,of eachfrom several farms for each landraceof aromatic rice. The best number of single
field

Field

locatedshape and some morphologicalrice(Random Amplified

of aromatic rice in Nam Dinh province,

relationships

revealedplantheterogeneity

among plants
the strategy

withinand among landraces

heterogeneityof a landraceperdata the heterogeneity

among fields

andthat the geneticof fieldschangesof lowland

rice issamples to estimate

on two years'
suggests

in 1998

to conserve landrace

structure

populations

discussed.

Introduction
Genetic diversity

in crop plants

and future crop improvement
landraces

needs. Genetic

and their wild relatives.

heterogeneous

landraces

ecosystems. Conservation
Genetic diversity
Ex situ conservationis necessary

Biodiversity

by geneticallyto furnish

diversity

in crop plantsof crops is declining

uniform improvedof genetic diversity

new genes to meet the current

cultivars

is one of the crucial

resources maintains

populations

found indue to replacementofand the destructionofissues facing

can be conserved ex situ or in situ (Astley,of plant genetic

is mainly

agriculture.

1992, Frankel et al. 1995).
in habitats

other than

those in which they evolved

or became adapted.

rice ex situ in last three decades.
genetic

diversitya living

library

Accessions
as genetic

preserving

genetic

continuing

selectionpressures.

approaches

have previously

In situ conservation

drift and contamination.

resources

involves

and genetics(Vaughanand Chang 1992,

consideredin making an action plan for in situ conservationconserved,

how to manage the population

Currently

research

limited.

The objective

variation

in a closelythat addresses

genetic

resources in Vietnam was initiatedexploration

for rice landraces

between the Ministry
Agriculturalcollecting

to in-situ

and spacial

to upland

and most lowland

to be

the sites to be

aspectsof rice (Changof genetic

to be productive.
1930's(Khoi

and isConservationof riceet al, 1990).

and Fisheries(MAFF),

these missions.

Most landraces

in a joint

project

Japan and Vietnamof cultivatedandof rice in Vietnam are adapted

rice areas are dominatedsuggestRecently

by improved

These observations

it exists

areas. MAFF initiateda joint

projectconservation

research in Nam Dinh province

locatedin the Red River Delta. The objective

this project

was to analyse the genetic

diversity

the need to conserve genetic

cultivars(Okuno et al. 1998).
in lowland

is

1976)

Vietnam. More than 1 100 accessions

duringissuesconservation

parts of Vietnam was undertaken

wild rice were collected
rice culture

is likely

Forestry(VASI),

the environmentof rice.

in the early

to variousof Agriculture,

Science Instituterelatedthe area of maximumdiversity

an area where intensivetoand ex situ

One of the scientificis to focus on temporal

therefore

politics,

is how to identify

questionsgroup of landraces

Northern Vietnam is within

is subject

1984).

to society,et al. 1997).

isand how to monitor changes from year to year.

the scientificof this paper
relatedBellonthatroles of in situ

(Ingram and Williams

many factors related

can looseIn situ conservationin an environment

The dual and complementary

been discussed

in conservingof rice conserved ex situ conservation

due to such factors
plant

There has been much progresswithdiversity

where

VASI to conduct

in situof aromatic rice landraces

farmers over several

years. Genetic

variation

withinNamDinh province

was analyzed

using Random Amplified

Based on the results

principals

maintainedby theand among aromatic rice landraces

that may help in conservation

DNAs (RAPDs).

of rice genetic

diversity

in situand Methods

Research sites and plant materials
Landraces of aromatic rice in Nam Dinh provinceinPolymorphic

are discussed.
Materialsofhave been chosen for thisstudy.

NamDinh province is an area of lowland rice withinsoutheast

from Hanoi. The area of rice cultivation

rice cultivars

are improved

the Red River delta,

about 80 kilometersin Nam Dinh province is 82,000

cultivars

while

autumn rice cultivars

are landraces

toleranceof deep water that inundates

1.1,000

ha in the province.

landracesin Nam Dinh are Tam xoan and Tam tieu. Other landraces

ha. Spring
due to their

The two major aromatic riceof aromatic rice are Tamthorn, Tam co ngong, Tam cao cay and Tam nghen and are only cultivatedon a limited

NamDinh. Farmers save their own seeds for planting

farmers exchange

annually

in additionarea ofseeds based on farmer interviews.
Three landraces
seven farms during
tieu,

of aromatic rice, Tam xoan, Tam tieu and Tam thom were surveyed in

1998(Table1,Fig.

1). Four landraces

Tam thom and Tam co ngong and an improved

reference,

were surveyed from 14 farms in 1999

collected

from 20 plants

of aromatic

non-aromatic rice cultiver(Table1,Fig.

1). Leaves

at each farm in 1998 and from 10 plants

farm were used for the pilot

rice, Tam xoan, Tamstudy in the first year to determine

in 1999.
optimalQ5, as awere randomly

Twenty plants

sampling

pernumber andten plants per farm were used in further experiments.

MorphologicalsurveyMorphologicalcharacteristics

awning and apiculus
individuals

withincolor

such as, culm and panicle

were recorded

to observe

length,morphological

grain

size and ofdifferencesamong

DNA extraction.

Total

a field in 1999.

DNAextraction and RAPD analysis
Leaves were dried at 60•Žfor two hours and stored at -70•ŽDNA was extracted

using

the CTAB method describedƒÊl,DNA amplification

was performed in a volume of 10ƒÊM

10mM Tris-HCl
and dTTP),
reaction

landraces

containing

10ng of genomic DNA,of dNTPs (dATP,

and 0.3u of Taq DNA polymerase(Perkinmix was covered with a drop of mineral oil. The Programmable
PTC-100/200)(3min).

was programmed for 45 cycles

Amplified

DNA fragments

agarose gel with TAE buffer.
illumination

by Murray and Thompson (1980).

50mM KC1, 2mM MgCl 2, 0.21ƒÊM of primer(MJ Researchand 72•Ž(pH8.3),until

with UV light

were separated

Each gel was stained

and photographed.

of aromatic rice were used to amplify

at 93.5•Ž

FifteenElmer ABI).

(1min.),

bromide,decamer primers

The

Thermal Controller
36•Ž

by electrophoresis

with ethidium

DNA fragments.

dCTP, dGTP

(2min.),
on 1.4%visualizedpolymorphic

by
among

Fig. 1. Field

survey and sampling

sites

(A-E)

in Nam Dinh province,

Table

1. Aromatic rice landraces

surveyed in Nam Dinh province,

VietnamL o c atio n 1)

V ari ety

D istrictC o m m u n e V illa g e

9 8T X O ‑1

T am x o a n

H a ih au

H a ip h o n g

A n le9 8T X O ‑2

T am x o a n

H a ih au

H a ian

A nd aoB

9 8T X O ‑3

T a m x o an

H a ih au

H a ito a n

X om 7c

C ode N o.

A

9 8T T I‑1

T am tie u

H a ih au

H a ip h o n g

A n leA

9 8T T I‑2

T a m tie u

H aih auH a ito an

X om 3c

9 8T T H ‑1

T a m th o m

M y lo c

M yhung

X om 6D

9 8 T T H ‑2

T a m th o m

M y lo c

M yhung

X om 5D

9 9 T X O ‑1

T a m x o an

H aih auH a ip h o ng

A n leA

9 9 T X O ‑2

T a m x o an

H aih a u

H aiph on g

X om 4A

9 9 T X O ‑3

T a m x o an

H aih a u

H aianB

9 9 T X O ‑4

T a m x o an

H aih a u

H aianA ndaoD oi 19 9 T X O ‑5

T a m x o an

H aih a u

H aito an

X om 7c

9 9 T X O ‑6

T am x o an

H a ih a u

H aito an

L ie n tienc

9 9 T T I‑ 1

T am tieu

H a ih a u

H aip h o n g

X om 3A

9 9 T T I‑2

T am tieu

H a ih a u

H aia nA n d aoB

9 9 T T I‑3

T am tieu

H a ih a u

H aito a n

X om 3c

9 9 T T I‑4

T am tieu

H a ih au

H a ito a n

L ien tienc

9 9 T T H ‑1

T am th om

M y lo c

M yhung

X om 6D

9 9T T H ‑2

T am th o m

M y loc

M yhung

X om 5D

9 9T C O

T am c o n g on g H a ih au

H a ip h u o n g X o m 4

E

9 90 5Q 5 (Im p ro v e d ) H aih au

H a ian

B1)Corresponding

to the site in Fig. 1

D oi 13

VietnamB

Data analysis
Polymorphic
for missing

DNA bands on the gels were scored 1 for presence,

data. The dissimilarity

Dissimilarity

index was calculated

0 for absence and 3

as follows:index =1- 2 Nij / (Ni + Nj)

where i and j are samples, Nij is the number of share bands and Ni, Nj are total number ofbandsof samples

Heterogeneityi and j. Thisof landraceindex

was calculated

for all

RAPD bands in a farm was estimated

pair

wise

comparison.

by average dissimilarity

among the samples in a given farm as follows:‡”

Heterogeneity= DSij/n

where i and j are samples, DSij is the dissimilarity
number of samples in the farm. Dissimilarity
calculated
Clusterby average dissimilarity
analysisindividual

index of between sample i and j. n is totalof landrace RAPD bands between two farms wasof pair wise comparison between the samples in two farms.

was conducted to create a dendrogram showing genetic

relationships

plants or farms using computer software NTSys ver 2.02h (Applied

the unweighted

among

Biosystems)byand grain size characterizedinpair group method (UPGMA).

Results
Morphologicalvariation

Morphologicalwithina farmvariationin culm length,each farm is shown (Table 2). Landraces
the improved cultivar
to discriminate
landraces

Heterogeneity

optimalin landraces

98TXO3plantsduplication

permittingfromto each other and were difficult

deviations

for grain characteristics

However, culm lengthand panicle

inlengthcompared to the improved cultivar.

plant number for heterogeneity

and 98TTH2).

markers. The average estimatesdistinctusing different number of plants

based on RAPD markers consisting

farms (98TXO2,

Standard

Q5 were similar.

in a field estimatedTo determine
subsets

were quite similar

by plant and seed characters.

had a larger standard deviation

lengthof aromatic rice were morphologically

Q5, but the landraces

and improved cultivar

panicleestimationof 5, 10, 15 and 20 plants were created for three
Each data set was created by random selection

from the raw data with 20 individuals
for each plant sampling

shown (Fig. 2). The average heterogeneity

in a farm, ten data

analyzedusing 8 RAPD

numbers and their standard

among the three farms was consistentoferrors areand standard

Fig. 2. Heterogeneityof aromatic rice landraces

samplesizes.

Average of ten data subsets

98TXO3and 98TTH2.

Table 2. MorphologicalC od e N o .

charasteristics

Bars indicate

of aromatic

estimatedusing RAPD markers based on four

are indicatedstandard

for threefarms;

98TXO2,error.

rice in Namdinh province,

VietnamV a ri e ty

C u lm

P a n ic a l le n g th (c m )G r a inG ra in w id th ( m m )L /W

99 T X O ‑ 1

T am

x oa n

105

+‑

8 .7

ab c

2 7 .4 +‑1 .5 8abc d

7 .5 + 0 .1 0

c de f2 .5+‑ 0 .0 7

a2 .9 8ab c

1 9 .1ab

99 T X O ‑2

T am

x oa n

106

+‑

4 .6

ab c

2 8 .8 +‑2 .8 1ab c

7 .4 +‑ 0 .0 7

abc d

2 .5

+‑ 0 .0 6

a2 .9 4ab cd

1 8 .7a99 T X O ‑3

T am

x oa n

114

+‑

6 .3

a

2 9 .9 +‑1 .4 8ab cd

7 .5 + 0 .2 0

b cd e2 .6+‑ 0 .3 2

ab2 .8 7

ab cd e1 9 .8ab

99 T X O ‑4

T am

x oa n7 .5 +‑ 0 .2 2

b cd ef

2 .7+‑ 0 .0 6

ab2 .8 3b cd e2 0 .0a b

99 T X O ‑5

T am

x oan

99

+‑

5 .3

c

2 6 .7 +‑1 .5 3a7 .4 + 0 .1 7

b cd

2 .6

+‑ 0 .0 9

ab

2 .8 9ab cd e1 9 .2

ab

+‑

4 .9

b c

2 9 .1 +‑1 .2 5abc

7 .4 +‑ 0 .1 3

b cd

2 .7

+‑ 0 .0 5

ab

2 .8 0b cd e1 9 .7a b

7 .3 +‑ 0 .1 1

ab c

2 .7

+‑ 0 .0 7

ab2 .7 3cd e

1 9 .5a b

7 .7 +‑ 0 .1 6

d ef

2 .5

+‑ 0 .1 8

ab

3 .1 0a

1 9 .2a b

7 .2 +‑ 0 . 1 1

a

2 .6

+‑ 0 .0 5

ab2 .7 1de

1 9 .0a b

le n g th ( c m )

N A

N A

99 T X O ‑6

T am

x oa n

10 1

99 T T I‑1

T am

tie u

109

+‑

5 .2

ab

2 9 .3 +‑1 .3 0

a b

109

+‑

6 .9

ab

2 5 .9 +‑2 .4 3c d

le n g th (m m )

2

L *W

3

9 9 T T I ‑2

T am

tie u

9 9 T T I ‑3

T am

tie u

9 9 T T I ‑4

T am

tie u

99

+‑

6 .6

c

2 8 .7 +‑2 .8 4ab c

7 .6 +‑ 0 .1 7

cd ef

2 .6

+‑ 0 .0 4

ab2 .9 5ab cd

1 9 .3ab

9 9 T T I ‑5

T am

th o m

10 1

+‑

8 .7

b c

2 6 .4 +‑3 .4 2b cd

7 .3 +‑ 0 .3 2

ab c

2 .8

+‑ 0 .0 8

bc

2 .6 5e

2 0 .4b

9 9 T T I ‑6

T am

th o m

10 5

+‑

4 .4

ab c

2 5 .1 +‑2 .6 8d

7 .2 +‑ 0 . 1 1

ab

2 .7

+‑ 0 .0 7

ab2 .7 0de

1 9 .5ab

2 7 .8 +‑2 .2 4ab cd

7 .8 +‑ 0 . 1 2

ef

2 .6

+‑ 0 .0 4

ab2 .9 9ab

2 0 .2

ab

f

2 .9

+‑ 0 .1 1

99 T C O

T am

co n g o ng

N A

N A

99

+‑

4 .7

c

73

+‑

2 .7

d

99 Q 5Q 5

2 1 .5 +‑0 .4 5e

7 .8 +‑ 0 . 1 8

c

2 .6 7e

2 2 .7c

9 9 T X O ‑1 6

T am

x oan

10 5

+‑

7 .9

a

2 8 .4+‑2 .1 1a

7 .5+‑ 0 . 1 6

a2 .6 +‑ 0 .1 5

abc

2 .8 9ab c

1 9 .4a

9 9 T T I‑1 4

T am

tie u

10 6

+‑

7 .5

a

2 7 .9+‑2 .5 1a

7 .5+‑ 0 .2 7

a2 .6 +‑ 0 .1 8

ab

2 .8 9ab

1 9 .3a

9 9 T T H 1, 2T am

th o m

10 3

+‑

7 .0

a

2 5 .7+‑

3 .0 7b

7 .3+‑ 0 .2 4

a2 .7 +‑ 0 .0 9

ac

2 .6 7ac

1 9 .9a

9 9 T C O

T am

co n g on g

99

+‑

4 .7

a

2 7 .8+‑2 .2 4a

7 .8+‑ 0 . 1 2

b2 .6 +‑ 0 .0 4

a2 .9 9a

2 0 .2a

9 9 Q 5

Q 5

7 3

+‑

2 .7

b

2 1 .5+‑

0 .4 5c

7 .8+‑ 0 . 1 8

b2 .9 +‑ 0 .1 1c

2 .6 7c

2 2 .7b1) The data followed by same letters are not significantly
2) The ratio from grain length to grain width.
3) The product of grain length and widthdifferent

accordingto the Turkey (HSD)

test

Heterogeneity

in afield

Heterogeneity(Tableand its annual change
in a field

was estimatedusing ten plants

based on 32 RAPD markers

in heterogeneity

among farms both in 1998

3). There was a five to six folds difference

and in 1999 although

the range of heterogeneity

among farms was different

Among landracesof aromatic rice, heterogeneity

in a field was significantly

differentamongcultivars

4).

were found between locations(Table5). The(TableSignificant

differences

samples with codes 98TXO2 and 99TXO3, 98TXO3 and 99TXO5,

98TTH2 and 99TTH1 werecollected

from the same farm in 1998 and 1999. Heterogeneity

similarwhile in the other two pairs, three to seven folds difference

samples collected

in 1998 and 1999.

Genetic relationship(Tablebetween two years.

of 98TXO2

and 99TXO3 waswas observed between the

3)among landraces of aromatic rice

210 individualsof aromatic rice landraces

clusteranalysisto reveal genetic(Fig.3).

The samples collected

relationshipand improved rice cultivar

among the samples collected

in 1998 and 1999 were analyzed

from the same farm but sampled in different
togetherand only distantly

relateddistinctgroups based on DNA variation

years. Landraces

to the improve cultivar

dissimilarity

withina landrace

dissimilarity

withinand among landracesin this study.

or a sampling locationand locations

togetherwere subjectedfrom different

including

to

farms

the samples

of aromatic rice were clustered

Q5.

Landraces

did not show any

Significant

differencein the degree of

was observed (Table4, 5). The degree ofwere of a similar

range.

Discussion
The present study analyzed
aromatic landraces
conservation

genetic

variation

withinusing RAPD markers to enhance understanding

research. Landraces

variation

between morphologicalmaintained

recommendations
Morphologicaldistinctof genetic

of aromatic rice in Nam Dinh province

River Delta was chosen for present study. Some morphologicalrelationshipand among farms of lowland ricein aromaticand DNA variation
rice landraces

aspect of in situ

locatedon the Red

data was used to determine

and to determine

in the researchthethe nature of genetic

area and to provide

somefor in situ conservation.

characteristics

of landraces

The analysis

of grain shape and some morphologicaldifferencesamong landraces

characters

did not show anyof aromatic rice. However, the aromatic landraces

wereTable 3. Heterogenity
Vietnamin a farm and dissimilarity

among farms of aromatic rice in Nam Dinh province,
D i s s im i la ir ni ty
c lut od ith
n ge Qo th
5 e( rx lf aO r‑2m ) 2s

fDa ir sm s is m e xilca eri pt yt Qto5 t h(xe 1 0o t‑2h )e2r)

H e te r o g e n e i ty
C o de N o.

V a ri e t y( x 1 0 ‑2 ) 1 )
M ean

M

ax

M

in

M ean

M ax

M in
2 .2

9 8T X O ‑1

T am

x oan

1 .6

abcd

1 1 .85 7 .62 .2

9 .4

1 3 .5

9 8 T X O ‑2

T am

x oan

1 .3

ab c d8 .1

6 7 .71 .7

5 .0

1 1 .91 .7

9 8 T X O ‑3

T am

x oan

3 .8

d e fg

1 0 .06 2 .84 .4

7 .2

1 0 .64 .4

9 8 T T I‑ 1

T amti e u2 .2

ab cde1 1 .35 7 .92 .2

8 .8

1 2 .22 .2

9 8 T T I‑2

T amti e u2 .0

ab cde8 .7

6 7 .71 .9

5 .6

1 2 .3

1 .9

98 T T H ‑1

T am

th o m

0 .7

a

8 .2

6 8 .70 .7

5 .0

1 0 .00 .7

9 8 T T H ‑2

T am

th o m

0 .7

a

8 .3

7 0 .1

0 .7

5 .0

9 .9

0 .7

9 9 T X O ‑1

T am

x o an6 .0

g

1 0 .06 7 .35 .2

7 .0

1 0 .65 .2

9 9 T X O ‑2

T am

x o an

5 .0

fg

9 .6

6 9 .14 .2

6 .4

1 1 .5

4 .2

ab cd

99T X O ‑3

T am

x oan

1 .8

8 .2

6 5 .91 .7

5 .1

1 2 .1

1 .7

9 9 T X O ‑4

T am

x oan

2 .0

abcd e8 .3

6 9 .62 .1

5 .1

1 1 .02 .1

9 9 T X O ‑5

T am

x oan

1 1 .4h

1 4 .46 7 .59 .9

1 4 .5

1 4 .59 .9

9 9 T X O ‑6

T am

x oan

1 .1

ab

8 .5

6 6 .71 .7

5 .4

1 2 .6

1 .7

9 9 T T I ‑1

T amt ie u4 .0

d e fg8 .6

6 4 .33 .6

5 .7

1 1 .4

3 .6

9 9 T T I‑ 2

T amt ie u3 .5

c d fg

1 0 .26 3 .54 .4

7 .4

1 0 .74 .4

9 9 T T I‑ 3

T amt ie u1 .9

ab c d e

8 .5

6 5 .92 .0

5 .5

1 2 .42 .0

9 9 T T I‑4

T a mti e u1 .9

ab cde9 .1

6 6 .91 .7

6 .0

1 3 .0

1 .7

99 T T H ‑1

T a m

th o m4 .7

e fg

1 0 .06 8 .73 .8

6 .9

1 2 .33 .8

9 9 T T H ‑2

T a m

th o m

3 .8

e fg

9 .6

6 7 .03 .8

6 .6

1 2 .03 .8

99 T C O

T am

co n go ng

2 .8

bcd ef

1 0 .37 1 .23 .4

7 .1

1 4 .5

3 .4

99Q 5

Q 5

1 .8

a b cd

6 6 .37 1 .25 7 .6

1) Heterogeneity
is based on average dissimirarity
among individuals
in a farm
The data followed by same letters are not significantly
different according to the Turkey (HSD)
2) Dissimilarity
between a given farm to the other farms
Table

4. Dissimilarity

withinand among aromatic rice landraces
No fu fam rm
b er

V a ri e ty

D iss(xim 1 ilarity
0 ‑2)1)

D iss im ila rity to th e o th e r v ar ie tie s (x 1 0 ‑2 )2)
M ean

M ax

M in

x o an

9

6 .8

7 .0

7 .7

6 .5

T a m tie u

6

5 .4

6 .4

6 .7

6 .0

T a m th o m

4

3 .1 b

6 .5

6 .9

6 .0

T am c o n go ng

1

2 .8

7 .1

7 .7

6 .5

20

5 .8

6 .7

7 .7

6 .0

T am

to ta ltestb1 ) Dissimilarity
is based on average dissimilarity
among given landraces
The data followed by same letters are not significantly
different according
2) Dissimilarity
between a given landrace to the other landraces

to the Turkey (HSD)

test

Fig. 3. Genetic

relationships

province,

Vietnam.

Table 5. Dissimilarity
L o c a tio namong aromatic rice landraces

within and among locations
S ite 1

No uf mfa brme r

D iss im ila rityfrom different

farms in Nam Dinhof aromatic rice
D is sim ilar ity to th e o th e r lo c a tio n s(x 1 0 ‑2)3)(x 10 ‑2)2

M ean

M ax

M in

7 .6

8 .9

6 .6

H a ih a u ,H a i p h o n g

A

5

6 .4a

H a ih a u , H a ian

B

4

3 .7

5 .7

6 .6

5 .2

H a ih a u , H a ito a n

c

6

6 .5a

6 .7

7 .8

5 .7

M y lo c , M y h u n g

D

4

3 .1c

6 .4

7 .0

5 .2

H a ih a u , H a ip h u o n

E

1

2 .8c

7 .0

8 .9

5 .2

to ta l

20

5 .9

6 .7

8 .9

5 .2

b

1) Corresponding
to the mark in Fig.1
2) Dissimilarity
is based on average dissimilarity
among individuals
The data followed by same letters are not significantly
different
3) Dissimilarity
between a given farm to the other farms

in each of locationaccording to the Turkey (HSD)test

quitedistinctfrom the improved

discriminatedcultivar

based on morphological

Q5 (Tablecharacters(Frankel

farmers comments measurements failed

to discriminate

significant

weremorphologicalcharacterizationdifferences

in individual2). Usually

et al. 1995).
clearlyobserved

farms is necessary

landraces

In this study

the landraces

among

to clearly

can readily

understandbedespitestudied.

Sincesome farms,

furthermorphologicalvariationfrom one farm to another.
Farm level variation

for morphologicalcharacters

Heterogeneity

for agriculturally

important

et al. 1995).

In this study, standard(Frankelwere generally

larger in landraces

grain size similarvariation

there are no special criteria

deviations

is not rare phenomenon in landracesof culm and panicle

and improved cultivars.

on the different

for selecting

characteristics.

seeds for plantingthat farmer selection

influences

Mixturesin landraces

consistingwith a different

contributeto stability

Heterogeneityof plants

when populations

plantsthevariation

in morphologicalphysiologicalstressescharacters.

charactersmay(Wolfe1985).

for such traits is a topic for further studies.

at the DNA level and its change from one year to the next

study,

estimates

while those foryear. No evidence wasare exposed to environmental

RAPD markers were used to estimate
initialfarms

Based on farmers interviews,

the followingthat suggested

evaluation

withinThe main reason is probably

obtained

Furtherlengthof aromatic rice than in the improved cultivar,

between landraces

effect of environmental

traitsthe optimum sampling

heterogeneity

withinnumber from a farm was determinedusing ten, 1 5 or 20 plants per farm gave similar results
were used in subsequent

heterogeneity

experiments.

of three farms when eightcompared. This result

suggests

determinationof heterogeneity.

only for relative

comparison

and among farms. As an

Significant

markers (Fig.

2). Since

based on independent

differences

trials,tenwere detectedin the2) and 32 markers (Table3) werethat the number of markers is importantHence, the value obtained(Fig.

in the present

factor in accurate
study should

be usedamong farms.

The present study indicatedsignificant

variationin heterogeneity

among farms (Table

Surprisingly

for some landraces heterogeneity

was lower than in the improved cultivar,

observation

might suggestof aromatic landraces

that heterogeneity

3).

Q5. Thison some farms has been lost.

Since the area of each farm was small and seeds for the next years plant were usually taken froma small number of plants

it is possible

genetic

drift occurs. This strongly

suggeststhat DNAmarkers are usefulfor selecting

and monitoringin situconservationsites.

heterogeneity

in a farm based on polymorphic

morphologicalvariation.

of heterogeneity

based on RAPD makers, 99TXO-5, had a low level of morphologicalThis observation

might suggestthat most of variations

in morphologicalcharacters.

understoodin the context of the evolution
and morphologicalThe biologicalplantings

Consequently,
selection

are generally

annual genetic

and farmers' selection.

source is an important
farms studied

and maintained

Occassionally,factor when monitoringin heterogeneitywith increasing

are needed.
issue. Seeds for the

by farmers

in Nam Dinh.

contributedby both naturalfarm level genetic

in landraces

monitoringof eachfarmers acquire seeds from other farms. Seedchange. Among the three

was observed in two farms. Thechange was observed had used self selected

seven times increaseof the associationis largely

for two years, a large increase in heterogeneity

farms where little

monitoringstudiescharacteristics

change in farm landraces

variation.

of DNA variation is poorly

at the farm level is an important
selectedlevel

at the DNA level is independentsignificanceof crops, further

or physiological

Annual change in heterogeneity
next years

detectedofwith the degree ofFor example, the farm where aromatic landraces had the highest

to variation

marker genotype

DNA markers did not correlate

The level

seeds. The reason for the four tofrom two farms is unclear.

sites is needed to understandSuccessive

short term genetic changes

on a farm.
Genetic relationshipamong farms of landraces

Landraces did not show any distinct
this

study

(Fig.

3). However, significant

landrace or a sampling location

can be largely

selection

due to differences

variation

withindissimilarity

difference

attributedin the degreein the microclimate.

withinand among each of landracesof this researchvariationinwithina4, 5). Since research sites are all within an

and climaticto genetic

of dissimilarity

conditions

are similar

drift or farmers' selection,
The present

over the area, therather than natural

study demonstrated

each landrace was almost the same level as among landraces.

the aromatic landraces
Hypothesis

groups based on DNA or morphological

was observed (Table

area of diameter 30Km and geographicaldifferencesof aromatic riceand locations

were similar.

that genetic
The degree ofThis suggests

thathave a narrow genetic base.

and future work

From the present

research using DNA markers, some knowledge

and its change was obtained.

Based on these results,

recommendations

on genetic

structurefor selectionandmaintenance of the seeds to maintain
Since the present

study revealed

diversity

detected

characters

rice landraces

can be made.

such as grain shape are highly

here may not be correlatedwith traits

concerned within the market.

To determine the factors responsiblelevel, long term research is required.

for change of genetic diversity

To clarify

how DNA level variation

several questions

remain. Research related

Twoapproaches

are a genetic map based approach

map basedof aromaticthat morphologicalhomogeneous, genetic variation
grain quality

genetic

approach

involves

overtime and to determine

to genome analysis

identification

linkage

map and genome sequence information.

screening

differences

stressesand identifying

is related

to adaptation

may help answer these questions.
approach. Genetic

that are heterogeneousor changed

for long term adaptation

using fine genetic

An eco-physiologicalapproach

involves

among individuals

in a population

againstenvironmental

the genes important

for adaptation.

Progressin genome informatics

enhance this kind of study. Throughgenetic diversity

at the DNA

and an eco-physiologicalof genes

the genes significancedetectedsuch studies,

knowledge

and long term adaptation

will be achieved

Preservation

diversityor biologicalon the biologicalfor efficientwill

meaning ofconservationof genetic

diversity.

References
AstleyD. (1992)

of genetic

and accession

integrity.

Field

Crop Res. 29:

205-224
Bellon M.R., J.-L. Pham and M.T. Jackson (1997)
Pages 263-289
Ford-Lloydin Plant Genetic

Chang T.T. (1976)

attacked

Changedby the rust pathogen

The origin,evolution,

and African rices. Euphytica
FrankelConservation:conservation:a role for rice farmers.

The in-situ approach. Ed. N. Maxted, B.V.

and J.G. Hawkes. Chapman & Hall

Burdon, J.J., J.N.L. Thompson, (1995)
marginale

Geneticof resistancein a population

Melampsora lini. J. Ecology

cultivation,

25:425-44

of Linum

83: 199-206

disseminationand diversification

The conservationof biodiversity.

of Asian

1O.H., A.H.D. Brown, JJ. Burdon (1995)
Univ. Press, Melbourne. 299pp

patterns

Cambridge

Ingram C.B., J.T. Williams
in Crop genetic
Williams.

(1984)

resources:

In situ conservationconservationand evaluation,

in Rice germplasm

collecting,Nationalpreservation

Murray MG, Thompson WK (1980)

ed. by J. H. W. Holden and J. T.

Rapid

collecting

activities-Vietnam.

Pages 27-28

and use. IRRI, Los Banos, Philippines.
isolationof highmolecular

weight

plant

DNA.

Acids Res 8: 4321-4325

Okuno K., M. Katsuta, H. Nakama, K. Ebana and S. Fukuoka (1998)
on plant diversity
and evaluation.
Parodaof crops. Pages 162-179

George Allen and Unwin, London

Khoi N.D., B.C. Buu and L.N. Trinh (1991)

Nucleic

of wild relatives

analysis.

Pages

158-169

in Plant

genetic

International
resources:

collaboration
Characterization

NIAR-MAFF Japan.

R.S. and R.K. Arora (1991)
Genetic Resources

Plant

Conservation

Genetic

Resources:

Generalperspective.

In Plant

and Management. Ed. R. S. Paroda and R. K. Arora 1-18

IBPGR ROSSA, New Delhi
Vaughan D.A., T.T. Chang (1992)

In situ conservationof rice genetic

resources.

Eco. Bot. 46:

368-383
Wolfe M.S. (1985)
for disease

The current status and prospects
resistance.

Ann. Rev. Phytopathol.

of multiline
23: 251-273

cultivars

and variety

mixturesClosing

Remarks

In-situ conservation

research:

Concluding

remarks

Hiroko Morishima
Saiwai-cho

15-2, Hiratsuka-shi,

formerly National

In-situresources

Institute

Kanagawa, Japanof Genetics,

conservation

Mishima

has become a significant

as a necessaryconservation

254-0804,

part of conservation

complement to ex-situ conservation.

research" was organized as a satellite

duringin-situ conservation
the workshop.

were presented.

These presentations

examples of conservationof variation

dynamics

necessaryinternational

research projects

I would like to make the following(1)

On-farm conservation

habitats

are quite distinct

of in-situ

interdisciplinary

disciplines.

For in-situ

evolutionary

processes determining

generalizationof methodologies

of wild

population

for genetic

linkingboth biological

species,

understanding

and meta-population

reserve conservation

may be difficult.

and domesticatesapplicable

to bothOptimum strategies

and sometimes weedy relatives,

the ecologicalandspecies-specificfor conservation

andshould begroup may be useful for otherof many crop species often grow in morewhich are affected by human activity.
that has been given relatively

and socio-economic

dynamics may be essential.

are highly

life-history.

relativesin their native

For on-farm conservation of crop diversity,groups with similar

habitats

andbeing conducted.

of wild species

target taxa, though experience in a particularor less disturbed

Many national

are currently

sought for respective(3) Wild progenitors,

anddata on populationconservation.

and conservation

are requiredconservation(2) It seemed that techniques

scientificthough there are some commontechniques

approaches

interestpoints.

cases such as the methods to assess genetic variation.
truly

in-depth

on in-situ conservationof crop diversity

issues,a number of

research is moving from simple survey andtechniques

focussingprovidingthat there is now broad international

stage to getting

for developing

Workshopin the various parts of the world.

Conservationin its infanton "In-situ

A broad range of research areas

were truly international,

It was encouraging for the participants
monitoring

genetic

Keynote addresses and case studies were presented

research conducted

research on in-situ conservation.

This workshop

session of 7th MAFF International

on Genetic Resources the main theme being "Wild legumes".
involvingof plant

littleGene flow between wild or weedy
considerationin relation

to in-situ

conservation

might play an important role in maintaining

such populations

to go through extinction

evaluate and use such populations

genetic diversity.

/ recolonization

It is not unusual for

cycles in vulnerable

in the context of in-situ conservation

habitats.

is a challenging

Howto
problemfor researchers.
4) Several advantages
researchof molecular technique

as in other fields.

population

structure,develop

have led to their widespread

It is a powerfulestimating

in-situ conservation

the patternprograms.

tool for constructing

diversity,

adaptive diversity

which can be assessed by quantitativeof the limitationsof the information

The advances in researchin-situ conservation,

have revealed

but the obtained

results

and theirconservationconservation after collecting

Basic research and conservation projects
be modified

factors togenetic approaches.

Weshould be awarefrom the neutral molecular markers.
many problems involvedin the various facets ofhave not been fully used for designingis urgently

all information

ascertaining

however, does not always reflect

plans. While, genetic erosion in farm-land as well as in native habitats
proceeding,phylogeny,of gene flow which are all important

Molecular

obtained

use in conservationneeded.

conservationof wild relatives

are rapidly

It may be late to start practicalnecessary for designing

an ideal conservation

should go hand in hand and the projectscheme.

scheme should

when necessary.

The workshop provided
knownfor in-situ conservation

an opportunity

to perceiveof plant genetic resources.

what is known and what needs to be
Time allottedto this session was quite

limited,though the theme has covered a broad range of topics. Many researches and conservation

projects

presented

in this workshop and conducted by others are stilla future workshop on in-situ conservation
greater depth.

will be organizedgoing on. It is hoped thatby MAFF to explore the topic inPhotographsof some authors

and participants

Photo.1.
Some of the workshop participants

at Tsukuba mountain Temple during a post-workshop

excursion.
Backrow:
Takanori Sato (pages
Peersak Srinives,
Middle165-186),

Akihiko

H.P. Bimb (pages

Fujii,

151-163),

Nigel Maxted (pages 83-109),
Faithi

Michiko

Yoshino,Hassan, Akito Saito

row

Paul Gepts, Devra Jarvis, Elena Potokina,

Than Htay Oo, Maria del Rosario Josefina

Munoz,Ramanatha Rao, Daniel Debouck (pages

Castro19-39)

Front row
Desiree Hautea, Hakim Kumiawan,Indra Kalyani Wasala, Maria Teresa Federici

Rodoriguez

Photo.2
Karl Hammer(pages 111 -125)examining

wild azuki beans in Tottori prefecture,Japan

Photo.3
Nigel Maxted (pages 83-109)
prefecture,

preparingherbarium specimens of Vicia species at dusk in Nagano

Japan.

Photo.4
Barbara Pickersgill
beans in Tottori(pages 3-18)
prefecture,hunting,

Japan.

with her camera, insects visiting

flowers of wild azuki

Workshop participants
and contributors
to the 7th MAFF
International
Workshop on Genetic Resources"In-situ conservation Research" Proceedings

List of ParticipantsA b e ,Ju n

H o k k a id o U n iv e rs ity , J a p a n

A s a n u m a , S h u ic h i

JIR C A S ,J ap an

B im b , H . P .

N e p a l A g r ic . R e s e a r c h C e n te r , N e p a l

C a s tr o M u n o s , M a r ia d e l R o s a ria

U n iv . N a c . A g a ria , la M o lin a , L im a , P e ru

C h ib a n a , T ak a s h i

N IA R , J a p a n

D e b o u c k , D a n ie l

C IA T , C a li, C o lo m b ia

E b a n a , K a o ru

N IA R , J a p a n

E g a w a , Y o s h in o b u

J IR C A S , J a p a n

F e d e r ic i R o d r ig u e z , M a ria T e r e s a

IN IA , L a s B r u ja s , U r u g u a y

F u j ii, A k ih ik o

H o k k a id o U n iv e r s ity , J a p a n

F u j ii, Y o sh ih a m

N IA E S , J a p a n

F u k u o k a , S h u u ic h i

N IA R , J a p a n

G e p ts , P a u l

U n iv e rs ity o f C a lifo rn i a , D a v is , U S A

H a ra d a , K yu u y a

C h ib a U n iv e r sity , J a p a n

H a u t e a , D e sire e

U n iv e rs ity o f th e P h ilip p in e s , L o s B a n o s , P h ilip p in e s

H is a d a , M ig ih ik o

N IA R , J a p a n

H o s h in o , T su g u o

J IR C A S , J a p a n

I g u c h i, S h u n ta r o

Ib a ra k i U n iv e r sity , J a p a n

I s h im o to , M a sa o

C h u g o k u A g r ic u ltu r a l E x p e rim e n t S ta tio n , J a p an

J a r v is , D e v r a

IP G R I, R o m e , Ita ly

J a v a r d y , F iro u z e h

N a tio n a l P la n t G e n e B a n k o f Ira n , Ir a n

J in n o , H ir o n o b u

H o k k a id o P r e f e c tu ra l P G R C e n te r , J a p a n

K a d o w a k i, K o u ic h i

N IA R , J a p an

K a g a , A k ito

N IA R , J a p a n

K a t o , K u n ih ik o

N IA R , J a p a n

K a t su r a , N a o k i

N IA R , J a p a n

K a t su ta , M a s u m i

N A R C , Jap an

K aw ab e , S u su m u

N IA E S ,Jap an

K ik u chi,F um io

T okyo A gricultural U niversity,JapanK itam ura, K eisuke

N A R C ,JapanK ob ayashi,H irom i

N IA R , JapanK ojim a,T oshio

Ibarak i U niversity,JapanK ok ub un ,M akiei

JIR C A S ,JapanK u m am oto ,M ak oto

A FF R C Secretariat, M A FF ,JapanK u m iaw an ,H akim

R esearch. Institute for Food C rop B iotech ., Indon esia

M atsuo , K azuto

N IA E S ,JapanM axted , N igel

B irm ingham U niversity,U K

M iu ra ,K iyoyuki

N IA R ,Jap an

M iyazak i, Shoji

N IA R , Jap an

M orish im a, H iroko

form erly N IG , Jap an

N agam in e, T sukasa

N IA R , Japan

N akayam a, H iroki

N IA R , Japan

N akayam a, Shigeki

N IA R , Japan

N ish ikaw a, T om otaro

N IA R ,JapanO b ata, T aro

N IA R ,JapanO k ada, K ensuk e

JIR C A S ,JapanO k un o, K azutoshi

N IA R ,Jap an

O o, T han H tay

S eed B ank, M yanm ar

O okaw a, M asao

N IA R , JapanO ono ,K iyoharu

N IA R , Japan

P otok in a, E lena

N .I. V avilov In stitute for P lant Industry, R ussia

R am an ath a R ao, V .

IPG R I‑A PO ,M alaysia

Saito ,A kito

JIC A ,Jap an

Sak am oto ,S hinichi

H okk aido U niversity,japan

Sato ,T ak anori

N IV O T , JapanS hiin a, T su gio

N IA R ,Jap an

S him am oto, Y osh iya

H okkaido U niversity,Jap an

Sh irata ,K azuto

N IA R ,Japan

Srinives,P eersak

K asetsart U niversity, T hailan dT om ooka ,N ori hiko

N IA R , Japan

V au ghan , D uncan

N IA R , Japan

W asala,In dra K alyani

Plant G enetic R esources C enter, Sri L anka

Y am ad a, T oshiaki

N A R C ,JapanY am am oto, A kio

A gricultural Structure Im provem ent B ureau, JapanY osh in o, M ichik o

H okkaido U n iversity ,Jap an
A cron ym s

A FF R C S ecretari at

SA ecretar
griculture,i at (Jap
Forestryan) and Fisheries R esearch C ouncil

C IA T

C entro(Colom Internacibia)
on al de A gricultura T ropical,

IN IA

Insti
(U
rugu
tutoay)N acional de In vestigati on A gropecuaria

IP G R I

Intern ational Plant G en etic R esou rces In stitute(headquarters Italy)

IP G R I‑A P O

PIntern
acificational
O fficePlant(M alaysia)
G en etic resources In stitute ‑ A sia

JIR C A S

SJapanciencesInternation(Japan) al R esearch C enter for A gricultural

M A FFM inistry(Japan) of A gricu lture, Forestry and F ish eries

N A RCN ational A griculture R esearch C enter (Japan)

N IA E S

N ational(Japan) In stitute of A gro‑E nvironm ental S ciences

N IA R

N ational In stitute of A grobiological R esources(Japan)N IG

N ational In stitute of G en etics (Japan)

N IV O T

NO rinmation ental
al R esearch
P lants andInstitTuteea (ofJapan
V egetables,)MAFF International

Workshop on Genetic Resources Series

1. Root and Tuber Crops

2. Soil Microorganisms

3. Animal Genetic

4. Plant

Genetic

Resources:

Resources:

NewApproaches

Efficientconservation

Characterization

for Improved

5. Diversity

and Use of Agricultural

6. Genetic

Diversityand effectiveand Evaluation:

Use of Plant Genetic

Resources

Microorganisms

and Conservationof Animal Genetic

Resources:

1. Improvement and conservationof genetic

diversity

2. Currentof effective

use of sperms.

statusand perspectives

7. Part 1. Wild Legumes*
Part 2. In-situ*Published

Conservation

separatelyuse

Research*

in livestock.

Editors
Editorin Chief:

Kiyoharu

OonoManaging editors : Duncan Vaughan
Norihiko

Tomooka

Akito Kaga
Consulting

Editor:Shoji

MiyazakiPublished December 2000

Research Council Secretariat of MAFF and National Institute ofAgrobiological Resources Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan

ISBN 4-931511-05-8