The Fourthof Agriculture,
Forestry

Ministryand Fisheries,

Japan(MAFF)

International

Plant

Workshop on Genetic Resources

Genetic

Characterization

Resources:and Evaluation

NewApproaches for Improved Use of Plant Genetic
Resources
National

Instituteof Agrobiological
Tsukuba, Ibaraki, Japan

October

22 - 24, 1996

Sponsored by
Research Council Secretariat
andNational Instituteof Agrobiological

National
National
Japan

International

Resourcesof MAFF
Resources

in cooperation with
Agriculture
Research Center,
Instituteof Fruit Tree Science,andResearch Center for Agricultural

SciencesContents
Page
Welcome Address
Opening

3

M.Nakagahra

5Address

Keynote Addresses
Characterizationand Evaluation
Future Challenges
K. Riley,
Conservation
Question

T. Hosoda

and Genetic

of Plant

Genetic

V. Ramanatha

Characterization

Resources- Present

StatusRao, M.D. Zhou and P. Quekof Plant

Genetic

Resources
H.Morishima

and answers

Topic 1: New and Improved Approaches to Analysis
of Plant Genetic
Resources Diversity
Approaches to Understanding
Genetic Diversity
at the Molecular
Level
S. Kresovich and A. L. Westman
Biosystematics- Implications

for Use of Plant

Conservationof Plant

genetic

D.A.Vaughan,
Evaluationof Interactions

betweenresources:Characterization

N. Tomooka, N. Kobayashi
Plant

Diversity

and A. O. Sari

and answers

47

59

71

and Other Organisms

from Evaluationof Plant

87

Genetic

A.G.Yunus•@and•@M.S.Saad
Question31
43

and evaluation

Y.TosaPlant Breeding Using Improved Information
Resources: Lathyrus as a Model Genus

7

Genetics
Y.Sano•@and•@L-V.Dung

In-situ

and

93
109

Topic2:
Plant Genetic Diversity
Evaluation- GeographicalandEcologicalconsiderations
Geographicaland breeding trends within Eurasian cultivatedbarley germplasm
revealed by molecular markers
P. P. Strelchenko,
N. K. Gubareva, O.N. Kovalyova and A. Graner
Diversity

Analysisand Evaluationof Wheat Genetic

Resources in China

L.H.Li,Y.S.Dong,D.S.Zheng

Crop Genetic

Resources

Diversity

in Indochina

115

and Available

133

Approachesfor

Its Conservation
L.N.TrinhInternational
Collaboration
on Plant Diversity
Analysis
K. Okuno, M. Seki-Katsuta,
H. Nakayama, K. Ebana and S. Fukuoka
In-situ Conservationof Plant
Variation
and Differentiation

Communities: Trendsof Plant Populationsin Studies

157

of Genetic
K.Matsuo

Question

149

and Answers

171
183

Topic 3: Cooperative
Mechanisms to Improve Evaluationof Plant Genetic
Resources
Mechanisms for the Evaluationof Plant Genetic Resources in Japan
H.Seko189
Evaluation
and Characterization
Situation
and Prospects
P.N. Gupta,
Internationalization
Mechanisms

Questionof Eliteto Enhance

and Answersof Plant

Genetic

I. S. Bisht,
Germplasm

Evaluation

Resources

Mathura•@Rai

in India:

Present

and K. P. S. Chandel

199

for Farmers : Collaborativeof Rice

Genetic

Resources
R. C. Chaudhary

221
245

Workshop SummaryGroup Discussion(a)Techniques

K. Riley249

D. A. Vaughan

252

P. Strelchenko

254summaries
Leader : S. Kresovich,Rapporteur:(b)Diversity
Leader : K. Okuno, Rapporteur:(c) Networks
Leader : R. C. Chaudhary,
Closing

Rapporteur:

A.G. Yunus 255

Remarks
H.SekoPictureof Participants

List of Participants

259
261
263

Introduction
Welcome address
Opening address
Keynote addresses

Chairperson
H.SekoWelcome Address
YOSHIHIKO KOTAKA
Research Councilor,
Council's

Secretariat,

Agriculture,

Forestryand Fisheries

Research

Council,Japan

Distinguishedguests, ladies and gentlemen, on behalf of the Agriculture,
Forestry and Fisheries
Research Council, it is my great pleasure to extend sincere
greetings
and best wishes to all participantsin this "MAFF Workshop on Genetic
Resources".
As you are well aware, there is global recognition
that enhanced conservation
and use of genetic resources is crucially
important for present and future generations.
This recognitionis exemplified
by international
trends after the "United Nations
Conference on Environment and Development" in 1992, where "Agenda 21" and the"Convention on Biological
Diversity"
were adopted or signed by many governments.
Since then, the Government of Japan has applied itself to conservationof biologicaldiversityand sustainable
use of its components according to the "National
Strategyof Japan on Biological
Diversity"
adopted in October 1995, which reflects
therequirementsof the Convention.
When we focus on plant genetic resources, these international
efforts bore
fruit at the "FAO 4th International
Technical
Conference", held at Leipzig in June
1996. Japan supported the whole process of the conference, not only financiallybutalso by actively
contributing
to the debate leading to the adoption of the "Leipzig
Declaration"and the "Global Plan of Action". Japan deems it significant
that the"Leipzig Declaration"
expresses each country's commitment to take the necessary
steps towards conservation and use of plant genetic resources in accordance with its
national

capacities.
The Ministry of Agriculture,
Forestry and Fisheries
of Japan, as a ministrysupervising
bio-basedindustry,has been positively
promotinginternational
cooperationon collecting,preserving
and using genetic resources. Holding
thisWorkshop is one example of such efforts. Having considered
recent circumstancesthat "in-situ
conservation"is regarded
as importantin the provisionsof the"Convention
on Biological
Diversity"and new methods of analyzing
biologicaldiversity
at a molecular level are being developed
day by day, we consider it very
important to have a discussion,
among the leading scientistsof relevant fields, on thetheme "NewApproaches to the Characterizationand Evaluation for Improved Use of

Plant Genetic Resources". The discussion will guide us in our future activities
relatedto conserving and using genetic resources.
At this 4th Workshop, reflecting
on our experiences of past Workshops, wehave tried to improve the procedure of holding
it. We have allocateda moreappropriate
meeting room, so that we can have in-depth discussions
in a friendly
atmosphere. We have also provided an excursion to visit a near-by botanicalgardenand, of course, included
a visit to our genebank facilities.
I would like to conclude my address by expressing my sincere desire, that thecollaborative
work over the next 3 days will strengthen our mutual understandinganddevelop warmand lasting friendships,
so that the friendships
among us will finallystrengthen
our cooperation at the level of national governments supervising
genetic
resources related policies.
Thank you very much.

Opening

address

MASAHIRO NAKAGAHRA
Director
Kannondai

General,National

Instituteof Agrobiological

2-1-2, Tsukuba,

Ibaraki

305-8602,

ResourcesJapan

It is a great pleasure for me to welcome you to the National
InstituteofAgrobiological
Resources (NIAR) for this Forth International
Workshop on Genetic
Resources. I very much appreciate
the kindness of participants
who have taken timeout of their busy schedules
to travel here for this workshop. To those who havearrived in Japan for the first time I hope your visit will be memorable and thoroughly
enjoyable.
I would like to thank those organizations
within the Ministryof Agriculture,
Forestry and Fisheries who have supported us in the preparation
for, and holding of,this workshop. Particularly
I would like to thank the Agriculture,
Forestry and
Fisheries
Research Council and our sister institutes
here in Tsukuba for their support.
The topic of this workshop is "New Approaches to the Characterization
andEvaluation for Improved use of Plant Genetic Resources". I would like to make a few
comments related to this theme.
(A) The research environment for the biologicalsciences is currently providing
newopportunities,
almost daily, to better understand life. The biotechnology
revolution
is in progress and this offers many newopportunities
to better understand conserved
genetic resources. NIAR has recently added to its 3 on-going conservation
areas ofplants, microorganisms
and animals, a forth area the conservationof geneticallyuseful biologicalmolecules in the DNA Bank.
(B) A second area which is also in the midst of a technologicalrevolution
is theinformation
sciences. This is having a major impact on disseminationof informationon conserved plant
than ever important

genetic resources. Ease of access to information
makes it more
to ensure that conserved germplasm is well characterised
andevaluated.
In our MAFF Genebank Project we have now linked our system to theInternet so that information
on Plant Genetic Resources in the MAFF Genebank isavailable
to interested
workers world-wide. I should add that there is both an English
and Japanese version.
(C) My last main point is related to a recent visit I made to West Africa. I had thegood fortune

to travel

to rural Benin and was very impressedby the range of cropsand traditionalfarming systems there. I hope that while we keep new technologies
inour mind we also think of germplasm characterizationand evaluation within the wider
context of the environment and farming systems to which improved germplasm isultimately
aimed.
During this workshop I hope that theme of the workshop will help to generate
newideas and cooperative research linkages. In part, because of the participationof
JICA trainees,
we have a greater international
representation
than in previous
workshops, which pleases me very much. Please use the next few days to the
maximum,make many contributions
to the discussions
and ask many question.
Thank you.

Keynote address I
Characterization-Present Status

and Evaluation of Plant Genetic
and Future Challenges

Resources

K. W.RILEY*, V. RAMANATHA RAO**, Z. MING-DE*** and P. QUEK***** Regional
Director,**Senior Scientist,**** Documentation/
information
Specialist,
Office, Asia, the Pacific
Malaysia,

and Oceania

and **Coordinator,

IPGRI

Region(APO),

P.O.Box236,43400

East Asia Office,Serdang,

IPGRI
Selangor

Darul

Regional
Ehsan,

Beijing

Introduction
The role of plant genetic resources (PGR) in the improvement of cultivatedplants has been well recognized.
PGR are conserved so that they can be used. Use
of PGR is a major element in the FAO Commission on Genetic Resources report on
the State of World's Plant Genetic Resources and is emphasized in FAO Global Planof Action (GPA) for the Conservationand Sustainable
Utilizationof PGR for Foodand Agriculture.
Expanding characterization,
evaluationand the number of corecollections
to facilitate
use was listed as one of the 20 priority
activities
in the GPA.
High priority has been given to the development of crop specific characterization
andevaluation programmes to identify useful accessions and for detecting valuable genes.
Such activities
are also consistent with the Convention on Biological
Diversity
underwhich countries agree to conserve, sustainably
use and share the benefits from PGR.
Information about a germplasm accession is essential
if collections
are to beeffectively
conserved, catalogued,
and retrieved
from genebanks. Therefore,characterizationand evaluationof germplasm accessions are essential
both toconservation and use of PGR (Riley et al., 1995).
The International
Plant Genetic
Resources Institute (IPGRI),
formerly International
Board for Plant Genetic Resources(IBPGR),has placed high priority
on the characterizationand evaluationof bothexisting
and new germplasm collections(van Sloten,1987).
Descriptor
lists,
germplasm directories,
core collection concepts, as well as occasional direct support
to countries to assist in characterizationand evaluation has been provided by IPGRI.
Although the proportionof germplasm collections
that have been characterized
andevaluated in the past 15 years, has increased, the report of the State of the World's
PGR for Food and Agriculture(FAO, 1996) reveals that well below half thecollections

Types

in most countriesof Descriptors

have been characterized

to Manageand evaluated.

and Use Germplasm

Collections

Various types of descriptors
are now recognized as necessary to facilitate
themanagement and use of the millions of germplasm samples now held in genebanks
around the world. All new descriptor lists (for example, Descriptors
for Capsicum,1995) now include five types of descriptors.
These are:a Passport descriptors:
These provide the basic information
used for the generalmanagement of the accession (including
registration
at the genebank and otheridentification
information)and describe the parameters that should be observed
when the accession is originallycollected(47 Descriptors).
b Management descriptors:
Provide the basis for the management of accessions in
the genebanks
and assist with their multiplication
and regeneration(31
Descriptors).
c Environmental
and site descriptors:
These describe
the environmental
andsite-specificparameters that are important when characterizationand evaluation
trials are held. They can be important for the interpretationof the results of thosetrials.
Germplasm collectingsite descriptors
are also included
here (48
Descriptors).
d Characterization
descriptors:
These enable a quick and easy discrimination
between phenotypes.
They are generally highly heritable,
can be seen easily by theeye and are equally expressed in all environments. In addition,these may include
a limited number of additional
traits thought desirable
by a consensus of users of
the particular
crop (59 Descriptors).
e Evaluation descriptors:
Many of the descriptors
in this category are susceptible
to environmental
difference
but are generally
useful to crop improvement andothers may involve complex biochemical
or molecular characterization.
They
include yield, agronomic performance, stress susceptibilities
and biochemical
andcytologicaltraits (127 Descriptors).
Each of these 5 sets of descriptors
is important
for the management andrecording of the sample during regeneration,
multiplicationand storage, and finallyfor use, either by breeders and other scientists,
or directly by farmers.

Characterization
Descriptors
Traits required for characterization
are generally highly heritable ones which
are expressed, within acceptable
limits of deviation,over a range of agro-climaticconditions.
This is essential because these traits are expected to help us identify
anaccession and may be used to monitor the identity of an accession over a number ofregenerations.
These generally
include a number of morphological,
botanical
features, with little ambiguity and which can be observed easily.
Characters such asleaf shape, flower colour, seed coat (testa) colour fall into this group. Despite theease with which these could be recorded, there is a need to define the exact (growthstage) time to make the observation and method of recording so that it can be easily
understood by the user community and other evaluators.
Thus, characterizationisprimarily the responsibilityof the genebank curator (van Sloten, 1987) and helps todescribe
the diversity
in collections
and assists the curator to manage thesecollections
effectively.
Evaluation

Descriptors
The second group of characters,
generallyreferred to as the evaluation
descriptors(including
the preliminary
evaluation
descriptors),
have agronomic/economic significanceand are specific to the plant and environment. For a given
species evaluation descriptors
vary in time and space because the needs of crop
improvement scientists
change over time and over geographicallocation.
In general,these are difficult
charactersto deal with mainly because the majorityof theevaluation descriptors
may be controlled
by polygenes (quantitative
characters)
andare greatly influenced
by the environment. There may be the need to test in several
environments or to use statisticalparameters to measure these descriptors.
In the caseof characters dealing with reaction to biotic stresses, factors such as races/biotypes
and host/pest
interactions
would also complicate
recordingof these characters,needing a great deal of sophisticationin techniques used for screening or evaluating.
All this underlines
the fact that the majorityof evaluation
data are more or lesslocation-specificand full evaluation of agronomic performance over many sites canenormously increase the data needed to fully describe an accession. Evaluationisnormally carried out jointly
by breeders and curators with the involvement of plant
protection specialists
or physiologists
in measuring specific traits.

10

Linkages
utmostamong Descriptor
Types
While each of the 5 groups of descriptors
has a distinct
purpose, it is ofimportance that shared databases
be developed
so that all 5 types of

information on an accession can be assessed. For example, the elevation
where a barley accession in Nepal was collected is recorded in passportand district
descriptors.

As location and elevation have strong effects on the different types of barley in Nepal(Rileyand Singh, 1990), both passport and characterization
data are important indescribing
and understanding
barley diversity.
Similarly,
cross-referencing
betweenpassport and evaluation data is needed in order to evaluate for a complex trait suchas cold resistance in barley; a subset of high altitude
barleys would be expected toincrease the likelihoodof identifying
the desirable
trait and thus reduce the cost ofevaluation and selection.

Roleof IPGRIin Supporting

Characterizationand EvaluationofGermplasm
Since its inception,
IPGRI (formerly
IBPGR) has been concerned withenhancing the informationthat accompanies germplasm accessions.
This hasincluded
the productionof crop descriptors,
directories
of germplasm, and direct
supportfor characterizationand evaluationof collections
and support
for
documentation
systems to manage and exchange this information.
Crop Descriptor
Lists
Crop descriptors
have been a central element in IPGRI characterization
andevaluation
activities.
Over 70 descriptor
lists have now been produced (Table 1).
Demand for these descriptor
lists is high and 40 new crop descriptor
lists have beenrequested. A recent survey of users of descriptor lists resulted in a very high response
rate. Information
gatheredfrom Country Reports and other sources, indicatedthat
among the countries that carry out characterizationand evaluation of their germplasm,
92% use the IPGRI descriptors(Thomas Hazekamp, 1996, personal communication).
Descriptor
lists are developedby scientists,
curators and experts who arepresently
working on a given crop who meet to decide on which descriptors
anddescriptor
states to include
for a given crops species.
As far as possible,
thedescriptor
list agrees and complements previous descriptions
that may be already in
use at various institutions.
For example, scientists
from 19 institutions
were involved

Table1. Descriptor

lists

publishedby IPGRI

Anacardium occidentale(1986)
Ananas comosus (1991)
Arachis hypogea (1992)
Arracacia xanthorhiza
Avena sativa (1985)
Beta(1991)
Brassica and Raphanus (1990)
Brassica campestris (1987)
Cajanus cajan (1993)
Capsicum (1995)
Carica papaya (1988)
Carthamus tinctorius(1983)
Chenopodium quinoa (1981)
Cicer arietinum (1993)
Citrus (1988)
Colocasia (1 980)
Dioscorea (1980)
Echinochloa
millet (1983)
Elaeis guineensis(1989)
Elettaria
cardamomum (1995)
Eleusine coracana (1985)
Forage grasses (1985)
Forage legumes (1984)
Fragaria vesca (1986)
Glycine max (1984)
Gossypium (revised
1985)
Helianthus(cultivatedand wild) (1985)
Hordeum vulgare
Ipomoea batatas
Lens culinaris(1985)
Lupinus (1981)
Malus (apple)(1982)
Mango mangifera (1989)
Medicago (annual)(1991)
Musa(1984)
Oryza (1980)
Oxalis tuberosa (1982)
Panicum miliaceum and P.sumatrense
Paspalum scrobiculatum(Kodo millet)
Pennisetum glaucum (1981)(1985)(1983)

andTable 2. (Continued)
Phaseolus acutifolius
Phaseolus coccineus
Phaseolus
Phaseolus(1985)(1983)

lunatus (1982)
vulgaris(1982)

Piper nigrum (1995)
Prunus (cherry)(1985)
Prunus armeniaca (apricot)(1984)
Prunus domestica (plum)(1985)
Prunus dulcis (almond)(1985)
Prunus persica (peach)(1985)
Psophocarpus tetragonolobus(revised,
1982)
Pyrus communis (pear) (1983)
Secale cereale and Triticale(1985)
Sesamum indicum (1981)
Setaria italica
and S.pumila (1985)
Solanum melongena, S. aethiopicum,
S. macrocarpon (and others)
Solanum tuberosum (cultivated)(1977)(1990)

Sorghum bicolor (1993)
Triticum
Aegilops (1989)
Tropicalfruits (1980)
Vicia faba (1985)
Vigna aconitifolia
and V.trilobata(1985)
Vigna mungo and V.radiata (revised,
1985)
Vigna radiata (mung bean) (1990)
Vigna subterranea (Bambara groundnut)(1987)
Vigna unguiculata(1983)
Vitis vinifera (1983)
Xanthosoma (1989)
Zea mays (1991)
In preparation:

Dioscorea,

Fagopyrum esculentum,in agreeing

on a common set of descriptors

Hordeum, Jugulans,Persea americana,

Psidium.

for Capsicum (IPGRI/AVRDC/CATIE,

1995).
Over the past 20 years, the number of types of descriptors
includedin thedescriptor
list has increased from 3 to 5 (management descriptors
and site descriptors
were recently
added).
In early descriptor
lists, the number of descriptors
wereminimized to reduce the burden of characterizationand evaluation.
Gibbons (1987)
reported
just 31 descriptors
in the original
Descriptors
for Groundnuts.
Recentdescriptor
lists now include a more comprehensive set of descriptors
with the 1995
Descriptors
for Capsicum containing
a total of 312 descriptors.
Users of thesedescriptors
are advised to select a limited number of key descriptors
which are mostuseful. Minimum highly discriminating
descriptors
are marked with an asterisk.
Althoughthese descriptor
lists help to standardize
descriptor
information,much of the earlier
characterizationand evaluation
data, recorded prior to thedevelopment of descriptor lists, used different descriptors
and descriptor
states in the
different
genebanks and research stations. Thus there is the need to either transform,
or interpret these data in order to share them among genebanks and users.
DirectSupport for Characterizationand Evaluation
Characterization
activities
must result in detailed
informationon thevariation in the collection
and provide an accurate assessment of the genetic variationthat the collection
represents.
IPGRI has supported characterization
trials to assist
curators for collections
to identify
accessions to help germplasm users select material
with relevant characteristics.
Over the years IPGRI has supported many trials on
different
crops. For example, IPGRI supported the National Hill Crops Improvement
Programme of Nepal to conduct characterization
and rejuvenationof Nepalese Hill
Crops collections:finger millet, barley, buckwheat, amaranths, Panicum miliaceumand Setaria italica (Baniya et al., 1991)
IPGRI also supported
characterizationofAllium fistulosum and many crops collectedin Colombia in 1989. Characterizationof a world collectionof Capsicum in CATIE, Costa Rica was also supported
by
IBPGR in 1989.
During 1990 and 1991, emphasis shifted
towards analysing
thesuccess of the trials and using the data that have been generated.
This would provide
criteria that can be used to direct future support and provide guidelines
for utilizationof data. Since 1991 direct support for characterization
has been substantiallyreduced,but some work has continued. Characterization
trials of maize, okra and sesamecollections
held in NBPGR, India was supported
in 1991 and 1992.
In addition,
IPGRI supported the Chinese Academy of Sciences to evaluate the world safflower
germplasm collection
from 1989-91.
About 1,545 accessions of safflower from 49
countries and 465 accessions from China were characterized
for 50 characters.
IPGRI
has supported the academy to develop and publish a book on the characterization
andutilizationof safflower germplasm (Li et al, 1993).
A project on multiplicationandcharacterizationof buckwheat germplasm resources was implemented by the Instituteof Crop Germplasm Resources of Chinese Academy of Agricultural
Sciences with thesupport of IBPGR in 1990.
Statusof Characterizationand Evaluationin Germplasm

Collections

In spite of the importance of descriptors
for the management and use of
germplasm accessions, surveys of germplasm collections,surveys have revealed that
only a small portion has been properly characterized
and evaluated.
Global estimates(Peeters
and Williams,
1984) are shown in Table 1.
The recent report on the State of the World's Plant Genetic Resources(FAO,1996a)
using data from 153, country reports, reported that much of the world's
ex situ PGR remain poorly documented with only passport data reported for only 37%
of collectionsin national programmes. The extent of characterizationof collections
was found to vary widely. The extent of characterizationin selected
countries thatprovided informationis shown in Fig. 1. The Country Reports used in compilingmuch of this information
cited lack of characterization(and evaluation)
as a major
constraint to use of PGR in breeding programmes (FAO, 1996b).
Reasons for the poor state of characterizationand evaluation of germplasm
collections
may include:-lack of resources or trained staff;-lack of interest from breeders to evaluate germplasm as 1) breeders may possess
their own working collections;2) unwillingnessof breeders to incorporate
genesfrom landraces into elite lines; and 3) lack of information
on genebank material,with existing evaluation data considered to be inadequate
or irrelevantto the plant
breeder (van Sloten, 1987).
- Breeders who do evaluate germplasm, often do not return the data to the genebank
curators, resulting
in lack of accessibilityof the characterizationand evaluation
data that does exist (FAO, 1996a).
The Global Plan of Action suggests a number of measures to improve thecharacterizationand evaluation of germplasm collections
that include closer linkages
with breeders, farmers and private organizations
in carrying out evaluation;
research
into and adoptionof new technologies,
includingmolecular
markers. Othersuggestions
include: improved
characterizationand evaluation
techniques,Figure1 The extent

of characterizationof ex situ collections:

Selected

countries

Table1. Global

estimateson the extent

of documentation

Sampleswith no passport data

Sampleswith no characterization

Sampleswith no evaluationof samples

in genebanks
65%

data

80%

data

Samples with extensive evaluation
After Peeters and Williams
,1984

95%
data

1%

developmentof on-farm evaluation
programmes, trainingof national
staff inevaluation and characterization,
and a step by step programme at the national level tocharacterize
and evaluate germplasm of the important crop species related to theneeds of the different
users of these crops. Finally,
development of core collections
is advocated.
The remainder of the paper will focus on a number of key issues that can
improve the characterization,

Key Issuesof Plantevaluation

for the Improvement
Genetic

and ultimately

the use of PGR.

of Characterization,

Evaluationand UseResources

Issue 1 - Key Descriptors
for Characterization
As earlier pointed out, characterization
is primarily
the responsibilityofgenebank curators using easily observable, highly heritable
traits that are stable acrossenvironments. Characterization
should be therefore be carried out based on the needsof the curators and other users to identify and manage the diversity in the collections.
Taxonomic systems have been developed for this purpose. In many crops, simple anduseful systems of classification
have been developed that rely on only a few simply
inheritedand easily observed traits.
For example, in sorghum, Harlan and de Wet(1972)
developed
a classification
system based on seed, spikelet
and head shapecharacters.
This classification
system can identify the 5 basic races as well as various
intermediate
subraces. This system is widely used by curators, breeders and othersorghum workers. A similarhead and finger characteristics
continued work on systematics

system has been developed for finger millet based on(Prasada
Rao et al., 1992).
There is need forand taxonomy in other crops to develop and promote

simple classification
systems based on a few key descriptors.
This
curators to focus on characterizing
a greater portion of their collections
traits.

would allow
for these key

Issue 2 - Recording Distinctiveness,
Uniformity
and Stabilitya)Heterogeneousaccessions:
Most accessions in any collection
are geneticallyheterogeneous.
This is inevitably
so in out-breeders,
but this can also be true in
in-breeding
landraces
and wild populations.
The variation
presented
by anaccession may be for a few or many characters.
It is necessarythat thisintra-accession
variation
be recorded and noted, but presently
there are nosatisfactory
ways to record such variation.
Most of the current descriptors
do noteven recognize the existence of such variation.
There are various ways in which
the problem is dealt with: (1) to divide the accession into uniform subsamples andidentify
each subsample by a separate accession number, (2) to record the mean on
the most commonstate, ignoring the rarer (that may be of interest)
states, (3) torecord mean and variance for quantitative
traits or the frequency of all qualitative
states, and (4) to record variable traits as variable without any particularscore.
Recent papers by Sapra and Bhag Singh (1992)
and van Hintum (1993)
suggestedmethods for curators to record within-accession
variability.
These are also referred
to by recent descriptor
lists.
From the above discussion,
it is clear that there is no rapid, inexpensiveor
precise method to describe heterogeneousaccessions. There is an urgent need tothink more on this aspect before some of the rarer genotypes or the informationonthem are lost. Current systems of documentationof collections
may even havecontributed
to reducing diversity
among accessions in genebanks since curators
tend to 'purify'
accessions to facilitatecharacterizationand evaluation.
b) Linkages
with UPOV descriptors:
In order to registernew crop varieties,
many
countries
have adopted the guidelinesof UPOV for identifying
a new variety.
Many of the germplasm characterization
descriptors
for a given crop are similar
to those of UPOV. Discussions are now underway to closely link the developmentof IPGRI and UPOV descriptors
to achieve optimalcompatibility,
betweencommondescriptors.

Issue 3 - Evaluate for key traits from the users perspectivea) Key traits for breeders: Evaluation descriptors
are determined by the needs of crop
improvement scientists
interactingwith any genebank. Their needs would
obviously
depend on the breeding
objectives
for any crop species in a given
location. These could be:
1. Improvement of agronomic performance-yield and yield related characters-response to fertilizers-resistance to lodging, shatteringetc.
2.Tolerances/resistances
to biotic stresses such as-disease resistance (fungal,
viral, microbial,
nematode)-insect resistance
3.Tolerancesor resistances
to abiotic stresses such as-drought/heatresistance-photoperiod
sensitivity/insensitivity-resistance to water logging-resistance to adverse soil conditions
4.Quality
characters-improved nutrition-improved cooking quality-improved flavour
Most of these traits are the concern of specialized
disciplines
such as plant
breeding, physiology,
pathology,
microbiology,
biochemistry
and input from all thesescientists
would be necessary to systematicallyevaluate the germplasm. Specialized,
and in some cases sophisticated,screening techniques
have to be developed and used.
The fact that a large percentage
of germplasm collected
has yet to be evaluated canpartlybe explainedby the procedural
difficulties
in effectively
screening
large
collections
for a number of characters.
The efficacy of screening depends on the
optimum and uniform prevalence of a stress factor in the area of evaluationor on theefficiency
with which such epiphytotic
conditions
could be created artificially.
Effective
screening techniques
would be imperative for evaluatinglarge collections
and the objectivesof breeding
in a region would dictate the emphasis placed fordeveloping
such techniques.

In order to maximize progress from breeding, breeders necessarilychooseonly a few key traits on which selection is carried out. As pointed out in the Global
Plan of Action (FAO, 1996b), goal setting is an important part of the breeder'swork,which may involve farmers and other users. The complexityand expense inevaluating collections
can be greatly reduced if curators, breeders and other users set
commongoals. Key traits for evaluation
are often highly location
specific.
For
example, the race of a disease will vary from region to region. Stress factors, such as
time and nature of water stress also change across region. Therefore, different
traitsmay be evaluated using different
methods in different
genebanks.
b. Key traits for farmers:
Direct use of germplasm by farmers is recognized
by
IPGRI and many other genetic resource workers as a valid and potentially
importantmechanism for use. New methods for participatory
selectionandbreeding that include farmers in choosing what traits and germplasm they need is
rapidly
gaining acceptance (Hardon et al., 1995) and have been endorsed in the
Global Plan of Action. The close involvement of farmers and scientists
can resultin effective
evaluation of germplasm using key descriptors
and descriptor
statesthat reflect farmers priorities.
Again such traits may be location specific and in thecase of taste preferences for example, may be conditioned
by many genes.
Issue 4 - Farmers classificationand traditionaldescription
In addition
to the participationof farmers in identifying
key traits forevaluation and selection, it is now realized that in some areas, farmers have developed
distinct
systems of classificationand description.
In the case of classificationof
cassava by the Aguaruna people (Boster, 1985), distinctions
could be made among
landraces
using easily recognizable
traits which were not connected with use.
However, the majority of studies of farmers' classificationand description
have founda utilitarian-basedtaxonomy using traditional
knowledge (Berge et al., 1991).
For
example, farmers in villages
in the midhillsof Nepal will maintain an average of
20-30 distinct landraces of finger millet, and classify them by both easily recognizable
traits such as head type and seed colour, but also by maturity, straw and grain quality.
Similarly,sorghum landraces
in Ethiopia
highlands
are describedby readily
observable traits (Table 3), for example, "moon-like seeds" or "short sorghum witha compact panicle",
as well as for complex inherited
qualitative
traits such as "asTable 3. Selected

Ethiopianvernacularnames of sorghums and their

meanings.

ET S N o.

V e rn a c u la r N a m e

M e a n in g o f v e rn a c u la r n a m e

E T S 13 47

F e n d is h a

S o rg h u m th a t p o p s

E T S 2 2 83

B is in g a W o r a b e is ah'Hy ey ne an ?a? so rg h u m " ‑ g lu m e s p ro tru d e lik e h a irs o f a

E T S 23 90

S e n d e L e m in e

'W h y ta k e w h e a t" ‑ a s g o o d a s w h e a t fo r m a k in g b re a d

E T S 26 1 1

H af u k a g n eh'Se ah da ma lwe oa ny sm e if I d o n o t h e a d " ‑ e v e ry p la n t p ro d u c e s

E T S 26 24

W o te t B e g u n c h e" M ilk in m y m o u th " ‑ so rg h u m th at is a s g o o d a s m ilk

E T S 28 34

G e b a b ie M u y r a" S h o rt M u y r a" ‑ sh o rt s o rg h u m w ith c o m p ac t p a n ic le

E T S 28 6 1

T in k ish" S w e e t ste m " ‑ so rg h u m sta lk s u se d fo r c h e w in g

E T S 29 70

M archu ke" G a ivros tee ds h o n e y lik e s w e e tn e s s" ‑ sw e e t se e d s c o n su m e d

E T S 3 133

G an S eber" b ermthferepaerokn sctsthe s eos oscla
ftrofey nprmg olyte nuthtaseatio
dt itfon bin
r rem loaakkcs ina thlgbe eloge car anml (cab kelaine yr"g c,‑oitdn utarin
in eg r)E T S 3 14 7

C h e re k it

" M o o n lik e " ‑ s e e d s a re b rig h t a n d w h ite lik e th e m o o n

E T S 3 14 9

D ir b K e te to" T w in se e d e d s o rg h u m "E T S 32 52

W of A y b e la sh" B ird p ro o f

E T S 37 80

A le q u a yw" He igo rse
h in bg e 7a 0n glik. e s e e d s " ‑ v e ry la rg e se e d s w ith 1 0 0 0 s e e d s

E T S 47 62

K itg n A yf e rie

w" Uh nicahfrais idlo oc fa sllyy p reh ilis"fe rre ‑d ntoo t aasffek itg
c ten d (sb yy p Sh trig
ilis ao (kf soitgrgn )h u m )Source:Gebrekidan,

1982

good as wheat for making bread" or "not affected"by (resistantto the parasitic

weed)

striga(Gebrekidan,
1982).
Although such taxonomies have been recognized for many years, renewed
attempts
are now being made to incorporatesuch indigenous
knowledge with
scientificknowledge.
IPGRI is presently
includingindigenous
knowledge
into
standard collecting
descriptors.
Indigenous knowledge about the location and extent
of crop diversitythat farmers maintain in a given area may prove to be the mosteffective way to locate and monitor this diversity.
A new IPGRI project "Establishing
the scientificbasis for in situ conservationof agrobiodiversity"
aims to assess theeffectivenessof using farmers' knowledge to assess and locate such diversity.
InAsia, this project is now under development at sites in Nepal and Vietnam. An IPGRI
project using taro as a model is now underway in Kunming and Beijing in China, tocompare genetic diversity
using farmers description
and using molecular methods.
A key question in such studies is to understand the relationshipbetween farmers or
folk taxonomies, and formal
genetic diversity
analysis.

classifications

includingbotanists'taxonomies

andIssue 5 - Molecular techniques
for characterizationand evaluation
Until recently, most of the characterizationand evaluation of PGR has been
based on recording of either qualitative
and/or quantitative
characters.
Since 10-15
years, more emphasis is being placed on biochemical
characterizationand more
recently on the use of molecular techniques.
The use of morphologicalphenotype forgenotype characterization
has advantages and disadvantages(Ramanatha Rao andRiley, 1994). The multilocus nature of most of these characters provides informationthat is highly useful to breeders. However, the complex inheritanceand interactions
with the environment makes breeding difficult.
The use of gene products (proteins,
peptides)or metabolites(terpenes,
flavonoidsetc.) partly
solved this problem.
Mendelian inheritanceof isozymes makes genetic analysis
still easier.
However,variationin isozymes is often low. Molecular genetic characterization
has several
advantages:
1. no environmental
influence,
2. any plant part from any growth stage
can be used, 3. there is no limit on numbers for analysis,
4. requires only small
amounts of material and 5. DNA is highly stable, even dry samples can be used. The
major practical
disadvantage
is that it is not very suitable for large scale screening.
Experimental
data on nucleotide
sequence variation usually characterize
only small
parts of whole genome, often not related to economicallyinteresting
traits.
Four areas of PGR characterizationin which biotechnology
can be used are:a) identificationof genotypes,
including
duplicateaccessions;b) "fingerprinting"ofgenotypes;c) analyzing
genetic diversity
in collections
or in natural stands and d)assembling a core collection(Dodds and Watanabe, 1990). Many genebanks receive
significant
number of accessions without any relevant passport data. Hence mostgenebanks carry an overload of duplicate
accessions resulting
in increased costs ofmanagement of collections.
DNA fingerprinting
with molecular markers can be very
useful in this case (Watanabe et al., 1995). However, identificationof accessions,especially
identificationcommercial cultivars, though possible,
is yet to be used on a large scale forof duplicatesin collections.
The value of fingerprinting
is more in thearea of varietal identification.
The determinationof the extent of genetic diversity
andits maintenance in collections
can be assisted by analysis of isozyme variationandmolecular genetic variation(Hubby, 1966; Simpson and Withers, 1986; Miller andTanksley,
1990; Clegg, 1990).
Identificationof genotypes, fingerprinting
and study of genetic diversity havebeen carried out using isozyme markers (Jarret and Litz, 1986; Glaszmann, 1988;
Nevo, 1990; Bhat et al., 1992; Lebot et al., 1993) However, in most cases relatively
few loci and alleles have been used in the analysis. Since any method would look ata small part of the genome, there is a need to use a variety of methods (Anderson and
Fairbanks,
1990) and some of the drawbacks with isozyme analysis may be overcome
with the use of molecular techniques.
To get really a complete picture, there is needto combine morphologicaland agronomic evaluation of germplasm with biochemical
and molecular analysis since these studies provide complementary information.
For
detailedreviews see the related references (Peacock, 1989; Anderson and Fairbanks,
1990; Kennard et al., 1994; Ramanatha Rao and Riley, 1994; Clegg, 1993; Watanabe
et al., 1995; Virk et al., 1996).
In evaluating
germplasm, multivariate
analysisof isozyme data can be an
additional
set of criteria to identify
a broad range of diversitythat is needed forscreening for resistances
to stress factors or yield. If one needs to work on a narrow
range of diversity
then isozyme data and RFLPs can help identify
similar or related
germplasm collections.
This is specially
useful when the passport data on area ofcollection

is not available.
At present, the cost of descriptionof a germplasm sample using a molecularmethod is 100 to 1000 times more than for conventional
phenotypic
description(FAO, 1996a).
While molecular methods may prove to be powerful tools forevaluating
germplasm and locating useful genes, such methods are unlikely
economic or practical
for routine characterizationof germplasm.
Issue 6 - Core collections
for improved evaluation
and useThe principal
idea behind the concept of the 'core collections'by Frankel

and Brown (1984).

A core collection

is a limited

to prove

was described

set of accessionsof acrop species and its wild relatives
which would represent,with a minimum ofrepetitiveness,the genetic diversityof a crop species and its wild relatives.
This
subset of the whole collection
would provide potential
users with a large amount of
the available
genetic variationof the crop genepool in a workable number ofaccessions. The main purpose of the core section is to provide efficient
access to thewhole collection
which should be representativeof the diversity
at hand. It would
therefore be useful to plant breeders seeking new characters which require screening
techniques not possible with a large collection.
In the late 1980s, IBPGR had workedon the development of a position paper on core collections,based on literature
thenavailable.
A workshop on 'Core Collections:Improving the management and Use of
Plant Germplasm Collections'
was held in Brasilia
in August 1992 (Hodgkin et al.,1995).
It was clear from that meeting that the core collections
are not forconservation but for accessing and using large collections.
IPGRI has been developing
methodology
for core collection
establishment
in collaboration
with national
programmes. Core collections
may also have a role to play in genebank managementfrom the point of view of distributionof representative
samples. Several studies on
the relevance as well as methodology
for the development of core collections
using
different
types of information,either singly or in combination,
are going on in many
genebanks and universities.
IPGRI is supporting
the Oil Crops Research Instituteof
CAAS in China and National Bureau for Plant Genetic Resources of India to study
on establishmentof sesame core collections.
Core collections
can be developed using different
kinds of informationonthe accessionsof a collection
includingpassport
data, characterizationdata,
evaluation
data, biochemical
and molecular marker data or a combinationof one ormore types of these data. In most cases characterizationand evaluation data (this may
include biochemical
and/or molecular characterization),in combination with passport
data, provided most representative
core subsets (Hodgkin
et al., 1995).
While core collections
may be useful for small breeding programmes, where
fewer accessions and wide diversity
are needed, or where initial exchange between
countries of a representative
sample of diversity
is requested,core collections
cannotreplace evaluation for key traits of the entire collections
as described in issue 3 above.

Issue 7 - New information
tools for better use of characterizationand evaluationdata
It is desirable
to encourage genebanks and users to develop descriptors
andrecord information
on germplasm that suits their own needs as far as possible.
As
pointed out earlier, descriptor
lists help in recording data on a germplasm accession
in a standardized
format for better exchange of this information among genebanks andother users. A number of information
tools are under development that can increase
the exchange and re-use of germplasm data.
a. System Wide Information on Genetic Resources (SINGER). Recently the Genetic
Resources groups in the CGIAR centres scattered around the world, which hold large
collectionsof the major food crops, were brought together under the System Wide
Genetic Resources Programme (SGRP).
A component of this programme, called
SINGER, is linking the informationon the germplasm holdings in these centres, andallowing access to this information
via Internet. The CGIAR has strengthened
its
activities
on genetic resources,
and through SINGER data and informationon allcentres, as well as other CGIAR genetic resources databases,
will become fully
available electronicallyand through other means, to the world community. The datadeliverymechanism preserves the autonomy of existing
Centre databases
andreplicatesthe data at a central node that can be accessed through the Internet. Data
will also be provided on CD-ROM, diskette or as printed output. The Centres havebegun to prepare their databases for linking into SINGER.
b. Data Interchange Protocol (DIP). Within a genebank, germplasm informationona given species is usually recorded in a standardised
format. The Data Interchange
Protocol is an initiative
developed by the IPGRI APO Regional Group. The protocol,
which is under development,seeks to provide a report format that enables a given
genebank to export their descriptor
lists and states in a form that allows recipientstore-use the data with their software. Using this format, germplasm informationhas
been successfullyinterchanged
between the genebanks in Beijing and Tsukuba. The
Regional Information System for Bananas and Plantains,a part of INIBAP, uses DIP
as a tool to facilitate
setting up a network for information exchange among genebanks
in Asia/Pacific.
A recent workshop organized by IPGRI (October
14-16, 1996)
explored how DIP could assist in the sharing and re-use of existing data in genebanks
in information
networking, statisticaland visual analysis,
and electronic
publication.

c. Multimedia for easier access to descriptor information.
The users of genebank
information may increasingly
be farmers who may not readily understand information
recorded in conventional databases. The M S Swaminathan Research Foundation in
Madras, is now compilinggenebank descriptor
information
on seed and plantcharacteristics
using video clips that become part of the descriptor
information stored
in the computer. Video clips of farmers describing landrace characteristics
using theirownterms and language is also included
so that indigenous
knowledge is retained
from the farmer as accurately as possible.
Several other centres are also developing
multimedia systems on computer, to provide precise and easy-to-visualize
information
on germplasm.
d. Geographic
Information
System (GIS).
Another potential
tool for better
visualizationof descriptor
data is GIS, in which different
types of data that have ageographic
reference can be plotted on a map using computers. For example, thegeographic distributionof existing ex situ collections
might be viewed on a map, withthe patterns of diversity
expressed for the various descriptors
for which information
is available.
GIS may also have use in monitoring in situ diversity,using appropriate
descriptor
information
including
ethnobotanicaldata and indigenous
knowledge infarmer-managed systems.
e. Information for the curators. In developing
descriptors,
sufficientemphasis has
been placed on descriptors to assist the curator to conserve the germplasm. However,most documentation systems attempt to provide germplasm users with information toenhance utilizationof the germplasm. There is a tendency to ignore the importanceof the use of descriptors
for accession-identification
purposes. If we ignore thecurators' needs for management of information
to maintain a viable accession, we
may have information but no accession. Similarly
lack of information
can also havethe same effect. The long periods of storage for seeds have resulted in the perceptionthat curators can maintain the germplasm with the current levels of informationcollected
in genebanks.
The need to emphasize
the developmentof storage
descriptors,
genetic drift descriptor
etc., is being addressedin part by the Decision
Support System for regenerationin genebanks, which is presently
being developedby IPGRI.

Issue 8 - The use of Descriptor
Lists
Over the last 15-20 years, a large proportionof genetic resources work has
been internationalized.
The exchange of seed and information
have been extensive
along with collaborative
studiesin genetic
resources that cut across nationalboundaries. This resulted in the need for standardizationin the characters recorded,
the way these are scored and documented - all of which resulted to production of over
70 descriptor
lists (DLs). DLs were also meant to assist the curators in recording
informationon accessions maintainedin the genebank which could be used for
diagnostic
purposes. A third purpose served by descriptor
lists is to provide guidance
to curators or other workers that may not have direct experience to record the mostuseful characters for a given crop.
In general,
the response to IPGRI descriptors
from the major/larger
genebanks and other users has been quite positive. An analysis of 152 country reports
indicated
extensive use of IPGRI descriptors
for characterizationand evaluation 91%. Some studies have also suggested improvements to descriptor lists (Cross, 1992;
Cross et al., 1992). However, it is important to make it clear, especiallyas the recent
DLs are getting more and more comprehensive and complex, that the descriptors
developed for any crop are for guidance and not obligatory,
and a subset of the total
number of descriptor needs to be chosen to suit a given situation.
Additionally,
it mustbe noted that the descriptors
are in a continuous process of refinement.
DLs set a standard so that the data collected
on a crop can easily beexchanged in the future. We need to consider the effect of standardized
DLs onexisting data. There may be many similarities
between DLs and the existing databasesin terms of descriptor
names, however the data or the descriptor
states may bedifferent.
In such a case it may not be cost-effective
to carry out the characterizationand evaluation again. We may have to think in terms of some sort of transformation
or a system like data interchange
protocol (DIP). The DIP format is being developed
precisely
to serve this purpose, placing importance on the information
provider. By
using DIP, a genebank can exchange information with any other genebank without theneed to compile standardized
DLs. Information exchange will encourage developmentof standardized
descriptors
besides providing
users with the information.
Allowing
researchers to develop descriptors can encourage creativity
and breathe new life into
descriptors
states using diverse media, including
video clips and sound recording.

Issue9 - Expanding use of descriptors
through Collaboration
and Networks
Networks for plant genetic resources for food and agriculture
are one of theapproaches for using and conserving these resources. Increased collaboration
among
countries through networking can help ensure more effective management and use of
PGR. No country can rely solely on the genetic resources that are stored or grownwithin its borders and improved use of PGR for the benefit of humankind is necessary
to ensure their continued conservation. Therefore, increasingcollaboration
on PGRis important.
A number of regional and crop networks have been developed aroundthe world that are aimed at improved use and conservationof PGR(Riley,1993).
Increased sharing of germplasm information is a key component of any successful
PGR network. Of equal concern in many networks is to complete the characterizationof ex situ collections
and to carry out evaluation for key traits using commonly agreed
descriptors.
New information
tools can allow these networks to compile andexchange germplasm information more easily.
Conclusions
The importance of adequate characterizationand evaluation data for both theeffective
management and use of PGR is clear. As far as possible,
priorities
need
to be establishedat the genebank level, with decisions made by curators and otherusers on the key descriptors
that can be recorded on the accessions taking existing
resources and needs into account. The descriptor lists developed by IPGRI, can serveas useful guides in standardizing
the way in which the informationis collectedandrecorded.
New concepts and technologies
offer exciting
possibilities
to improved
access and use of germplasm information.
Computers are becoming ever morecommonand able to handle multimedia
data including
indigenous
knowledge about
germplasm accessions
and landraces,
both in situ and ex situ.
Participatory
approaches involving breeders, curators, farmers and other users can help to insurethat the most useful descriptors
and descriptor
states are used in recording
this
information.
Database information can be more easily exchanged, and networks holdthe potential
for insuring the benefits from PGR are realized and equally shared.

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Biotechnology
R.G. Landes,

Keynote address
Conservation
Plant Genetic

II

and Genetic Characterization
Resourcesof

HIROKO MORISHIMA
National

Instituteof Genetics,

Mishima,

411, Japan

Genetic diversity
is defined as genetic variation
within species.
It is ourprecious heritage and essential
for the survival of all organisms on earth. Genetic
diversity
in crop plants is mainly preserved in land races and wild relatives,
and they
are called plant genetic resources (PGR).
Field collection
and preservationin genebanks of PGR has been extensively
conducted at the international,
as well as, thenationallevel.
Genetic characterizationand evaluationof collectedmaterials
areconducted for improved use by breeders and researchers.
In this paper, I will present two PGR issues, mainly based on my experience
with rice species:
1) Loss of genetic diversity(genetic
erosion) occurring ex-situ, as well as, in-situ;
2) The implicationsof phenotypicvariation and molecular variation for evaluation.
These two aspects should be bought together
to make action plans forminimizing
genetic erosion and to enhance use of PGR.
The target taxa dealt with in this paper are Asian cultivatedrice Oryza sativa
L. and its wild progenitor,
O. rufipogon Griffith.
Though they have distinct
speciesnames, they share the common primary gene pool, and form a single biological
species

togetherwith intermediate

or weedy types.

I. Genetic Erosion of Plant Genetic Resources
Ia. Loss of Genetic Diversity
in Gene Banks
Genetic diversity
in crop species is the result of differentiation
during thedomestication
process. A number of mutant genes which are poorly adapted andeliminatedin natural environments have been accumulated under cultivation.
Further,the diversityof crop species might have been enriched by man's intentional
activities
such as breeding efforts. Crop improvement in recent years, however, invariably
has
led to a decrease in genetic diversity
for many species due to the spread of a few highyieldingmodern varieties.
A diversity
crisis was recognized and field collection
conducted and conservation programs have been establishedsince the early 1970's.
It has been claimed that about 2.5 million accessions of PGR are now assembled andpreserved

in national and international
germplasm centers.
I would like to raise the alarm for loss of genetic diversity
occurring in genebanks before reaching the hands of breeders and researchers.
Genetic diversity
isalways threatenedin ex-situ conservation,
not just due to budgetary considerations.
While genetic diversity
can be preserved as "sleeping"accessions in cold rooms forvarying lengthsof time, ex situ methods, such as preservationof plants, seedmultiplication,cultivationfor evaluation and tissue culture result in the loss of genetic
diversity.
During these processes,
in addition
to genetic
and non-genetic
contamination,
genetic diversity
is always subjected
to natural selection.
For
instance, in 1983, we made a trip to Thailand for observationand collectionof wild
rice O. rufipogon.
93 accessions from this trip were registered.
At present, thenumber of accessions for which enough seeds are available for distribution
is only 65.
The reasons for this reduction in number includes nongerminabiltyof the original
seeds, inviable or weak seedlings,
non-flowering under ex-situ conditions,sterility
due to genetic and physiological(late flowering)causes, and low seed productivity.
Even in preserved accessions,selection
for genotypes
adapted to cultivation
andagainst adaptive genes for wild habitats may have resulted in the loss of truly "wild"
genotypes.
This is because landraces and wild species are usually heterogeneous
within accessions. Fig 1 clearly demonstrates
how "cultivation"itself (seedingandharvesting
without any artificial
selection)
worked as a strong selection
pressure(cultivationpressure)
on a wild rice population
shifting
its population
genotype tocultivatedtype over 5 generations(Oka and Morishima, 1971).
Ib. Loss of Genetic Diversity
in Natural Populations
During the last two decades, indigenous varieties or landraces of major foodcrops have been rapidly replaced by modern improved cultivars as large areas shiftedto monoculture.
The proportionof the land planted to local rice varieties
in theMekong Delta between 1976 and 1990 is shown (Fig. 2). Rice cultivation
area in thisDelta increased in the 1980's due to the establishmentof irrigation
systems.
Thischanged deepwater areas to irrigatedrice fields which can be planted to modern highFig 1 Distributionof discriminant scores for distinguishingsild from cultivatedtypes in populations
Oryza rufipogon grown in an experimental
field. (Oka and Morishima,
1971)

Fig. 2 Total rice area and proportion of local variety area(dotted(Source:Agricultural
Office of Hau Giang Province)line ) in Mekong Delta.

ofyieldingvarieties.
The proportion of the rice area planted to local varieties decreased
from 59% to 35% between 1976-1990.
Extinctionof landracesfrom farmers' fields results in the loss of large
amounts of variability
preserved among and within landrace populations.
Fig. 3shows intra-population
diversity
found in two population
samples taken from a
Chinese farmer's seed stock. Seed samples from an upland field showed particularlyhigh levels of diversity,
ranging from Indica to Japonica types, and also from uplandto lowland types (Morishima,1989).
The only way to conserve such diversity
is by
on-farm conservation.
Genetic erosion in wild relativesof crop species is also occurring rapidly innatural habitats
owing to economic development. Asian commonwild rice is widelydistributed
in monsoon Asia. In almost all areas where this species is found thenatural habitatsof this wild rice are threatened
by development
projects.
Many
populations
have been destroyed during the last decade. Further, a large proportionof extant populationsof this wild taxon are not truly wild. They have, more or less,absorbed genes from neighboring
cultivatedrice and become adapted to disturbed
habitats.
In Taiwan it is inferredfrom herbarium specimens that O. rufipogon wasabundant in the 1920s but then declined. The last wild rice populationin Taoyuan,
which was known as the most north easterly site of this species, became extinct in thelate 1970s.
The factors which caused this extinction
are considered
to behybridization
with cultivars, change in water management and water pollution
due tofertilizer
application(Kiang et al., 1979). In Thailand,
we have continued a long-term
observationsof wild rice populations
since 1983 at several permanent study-sites
inthe suburbs of Bangkok (Morishima
et al., 1996).
Fig. 4 shows population
flux as apercentage
of cover observed at our seven study-sites.
Asian commonwild rice isdifferentiatedinto annual and perennial
ecotypes. All four annual populations
wewere monitoring almost completely
disappeared
before 1990. On the other hand,
three perennial
populations
seemed to be relatively
stable and persisted
until 1990.
However, two of these populations
have been destroyed since 1990 by road expansion
and constructionof a petrol station, respectively.
The remaining one still exists but
seems to be in decline probably
due to water pollution.
As a complementaryand supplementary
approach to ex situ conservation,

•œ)Fig. 3 Lowland(Ch54,

classifying

Fig. 4 Populationin the suburb•›) and upland(Ch55,

populations

Indica-Japonica

types and lowland-uplandflux of annual

and perennialof Bangkok.

(Morishima

scatteredtypes.

by the discriminant(Morishima,

1989)

types of wild rice shown by percentage
et al., unpublished)scorescover observedthe significanceof in situ conservation
is well understood (Vaughan and Chang,1992).
The main issues to be consideredin making action plans for in situ
conservation are (1) how to select the site to be conserved, (2) how many and size of
populations
to be conserved and (3) how to manage the population.
Our observations
and results from permanent study sites in Thailand suggest that different
strategies
areneeded for in-situ conservation of plant populations
having different
propagating
systems. Wild relativesof crops usually grow in the habitats
influenced
by humanactivity
to varying degrees. Conservationof genetic diversity
preserved in such
ecosystems may be more difficult
to conserve than natural ecosystems or "nature
reserves". In situ conservation of landraces (on-farm conservation)
seems much moredifficult,because there are various socioeconomic problems to be solved.
II.

Genetic Diversity
Found at the Phenotypicand Molecular Levels
Isozyme polymorphism,and more recently RFLP and other molecularmarkers have been introduced into diversity
studies of PGR. These techniques
haveenabled high resolutionof genetic diversity
in many species. Variation surveys in agiven taxa using molecular markers sometimes yields the same variation pattern asthat obtained from phenotypiccharacters. However, this is not always true. In thefollowing discussion,
I would like to present some examples obtained from our ricestudies, and try to discuss what phenotypicvariation and molecular variation imply,
respectively.
IIa.

Variation
Pattern in Asian Cultivated
Rice and Its Wild Progenitor
It is well known that Asian cultivatedrice can be classified
into two major
varietal groups, Indica and Japonica types. These two groups were clearly recognized
by a particularassociationof several characters, though there are some intermediate
or unclassified
varieties(Oka, 1958).
Since various molecular techniques
werewidely used in PGR studies, many researchers carried out variation
studies in O.
sativa using these new technologies
These analyses based on isozymes (Glasszmann,
1987), nuclear RFLP (Kawase et al., 1991; Wang and Tanksley, 1987), rDNA (Sanoand Sano, 1990), mtDNA (Ishii et al., 1996),
cpDNA (Dally and Second, 1990)
reached essentiallythe same conclusion, that the major variation
found in O. sativa
is represented
by differentiation
into Indica and Japonica varietalgroups.

This

On the other hand, the situationdiffers in its wild
wild taxon contains a large amount of variability

progenitor,
O. rufipogon.
within the species andphenotypicallyperennial and annual ecotypes are recognized (Oka, 1988; Morishima
et al., 1992).
These two types are characterized
by a particularassociationof several
life history traits and are adapted to different
habitat conditions.
Multivariate
analysis
based on phenotypiccharactersconsistently
showed this tendencyof ecotypic
differentiation
towards perennial
and annual types though variationis continuous(Fig. 5a). Analysis of variation
at the isozyme and molecular levels of this species
exclusively
revealed only variation
related to geography,
not perennial vs. annual
variation (Fig. 5b).
The strains from the northern fringe of its distribution(China)and most westerly region (West coast of India) seem to represent two extremes in thisgeographicaldifferentiationof the species.
In general, phenotypicvariationis subjected
to selection,
while molecularvariationis largely neutral to selection,
as argued by Kimura (1983).
Therefore,
phenotypicand molecular variation are considered to reflect the results of selectionaland non-selectionalor neutral processes of evolution,
respectively.
In the case ofcultivatedrice, phenotypicand molecular variation
were largely
non-randomly
associatedwith each other. Both selection
and non- selective
processes must be
involved in indica vs. japonica differentiation.
On the other hand, it is consideredthatin O. rufipogon, perennial vs. annual variation is entirely
adaptive differentiationin
response to habitat conditions,while geographicalvariation
largely reflects isolation
by distance which graduallyproceeded along with dispersalof this taxon in Asia.
We know little
about the molecularbasis underlying
perennial
vs. annual
differentiation.
Even direct sequencing of particulargenes (Barbier et al. 1991; Ooiet al., submitted)
did not reveal differences
between these two ecotype.
Weshould be aware that phenotypicand molecular variation have different
significance,
respectively,in PGR studies.
Conservationists,
breeders
andevolutionists
should understand the inferences revealed at different
levels of variation
and use these information
depending on their purposes.
They may want to elucidate
general variation patterns based on adapted phenotypes,
or distributionof a particulartarget character,or phylogenetic
relationships
which can be effectively
estimatedfrom molecular variation. Studies on the molecular basis of adaptive variation,
which
could give a break through in the use of PGR, are still in their infancy. Since adaptive

Fig. 5 Scatter

diagrams

charactersof Asian wild rice strains(A) and 29 isozymes(B).

plottedby the scores of factor analysis(Cai and Morishima,

unpublished)based on sevencharacters are generally,
geneticallyquantitatively
controlled
QTL analysis assisted
by molecular markers could shed some light on the genetic mechanism of theirvariation and co-variation. A unified approach to quantitativeand molecular genetics
will give us a clearer perspective
for improved use of PGR.
IIb.

Genetic structure of natural populations
Intra-population
genetic diversity,
distribution
pattern of genetic diversity
among and within populationand heterozygosityof individuals
are central problems,
not only for population
geneticists
but also for PGR scientists.
In the case of wild
rice, genetic structure of natural populations
previously
inferred from quantitative
characters was confirmed by isozyme or molecular studies.
To compare the resolving
power among markers at different
levels,parameters for populationdifferentiation(FsT) were computed using phenotypiccharacters, isozymes and RFLPs in seven natural populationsof wild rice. Isozymes
showed a similar level of resolution
for describing
populationdifferentiationas
RFLPs which aremuch more costly and time consuming than isozymes (Fig.6).
Phenotypiccharacters
seemed to have lower resolving power, at least in this case.
Wehave demonstrated that perennial
types are generally
more polymorphic
within populations
than annual ones in quantitative
characters as well as isozymes(Morishima
et al.,1992).
Table 1 shows an example of disease
resistancepolymorphism which is contradictory
to this general trend. Annual populations
weremore polymorphic
in reaction pattern to four races of bacterial
blight disease thanperennial populations(Morishima
and Miyabayashi,1994).
This does not seem an
exception found only in our materials. It was reported that among IRRI accessions
examined, the perennial
group (O. rufipogon) was monomorphic while the annual
group (O. nivara) was polymorphic
in reaction to six Philippines
races of bacterial
blight(Ikeda and Busto, 1990).
Resistancegenes in hosts and virulence genes inparasites
have coevolved interactingwith each other.
Distribution
patternofresistance genes in the natural ecosystem seems to be affected by a complex of biotic
and abiotic environmental
factors.
Various selection
pressures such as frequency
dependent selection and resistance cost could be involved. Thus, distributionof tolerance or resistance genes which are most important for future breeding

patternrequireFig. 6 Comparisonof populationdifferentiation(I) and RFLPs(R) in seven natural(C),

isozymes(Cai

and Morishima,Table

1. Comparison

parameters(FST) estimatedpopulations

by quantitativeof Asian wild

characters

rice.

unpublished)of intra-population

variability

between perennial

and annualtypes of wild riceA v e r a g e g e n e d i v e r s i ty 1 )C o e f f ic i e n t o f v a r ia t io n 2 )N E 8 80 .3 5 0

0 .3 0 8

0 .3 6

C P 2 00 .3 2 7

0 .2 6 3

0 .3 9N E 3

0 .2 0 8

0 .2 1 1

1 .4 3N E 4

0 .1 4 7

0 .2 1 4

1 .3 4

P o p u l a ti o n c o d e

BD Biv er erss itisyta inn cd ee 3x) f o r

P e r e n n ia l

A n nu al

H: computedfrom 9 isozymeloci,

H=l/9‡”

Average of CV for 6 morphological
H1: computedfrom R/S reaction

H' = -‡”plnp, p: frequency(Morishimapi: ith allele

characters.
patternof reaction

and Miyabayashi(1-‡”pi2),

1993)

to 4 pathogen
types.

races.

frequency.

furtherinvestigation.

Conclusions
1. Genetic
genetic

diversity
erosion

2. Genetic
variationis urgently

diversity

found(mostly

3. Diversity
makingof PGR is threatened

neutral)studies
actionex situ

and in situ.

Actionto minimize

needed.
in phenotypiccharactershave

implications

differentof PGR by various

plans

both

to minimize

techniques

genetic(mostlyadaptive)for evaluating
and its synthesis

erosionas welland molecule
PGR.
are importantas to enhanceforuse bybreeders.
References
Barbier, P., Morishima H. and Ishihama, A. 1991. Phylogenetic
relationshipsof annual and perennial
wild rice: Probing by direct DNA sequence. Theor Appl Genet 81: 693-702
Dally, AM. and Second, G. 1990. Chloroplast
DNA diversity in wild and cultivatedspecies of rice(genus Oryza section Oryza ), cladistic-mutationand genetic-distanceanalysis. Theor Appl
Genet :209-222
Glaszmann, J.C. 1987. Isozymes and classificationof Asian rice varieties.
Theor Appl Genet 74:21-30
Ikeda, R. and Busto, G. A. 1990. Resistance of wild rices to bacterial
blight. IRRN 15:3
Ishii, T., Nakano, T., Maeda, H., Kamijima, O. and Khush, G. S. 1996. Phylogenetic
relationships
between cultivatedand wild species of rice as revealed by DNA polymorphisms.
In Rice
Genetics III, IRRI p. 367-372
Kawase, M., Kishimoto, N., Tanaka, T., Yoshimura, A., Yoshimura, S., Saito, K., Saito, A., Yano,
M., Takeda, N., Nagamine, T. and Nakagahra, M. 1991. Intraspecificvariationand genetic
differentiation
based on restriction
fragment polymorphismin Asian cultivatedrice, Oryza
sativa L. In Rice Genetics II, IRRI p. 467-473.
Kiang, Y.T., Antonovics, J. and Wu, L. 1979. The extinctionof wild rice (Oryza perennisformosana)in Taiwan. J. Asian Ecol. 1: 1-9
Kimura, M. 1983. The neutral theory of molecular evolution.
Cambridge Univ. Press, U.K.
Morishima, H. 1989. Intra-populational
genetic diversity
in landrace of rice. Proc. 6th Intl. Congressof SABRAO p159-162
Morishima, H., Sano, Y. and Oka, H.I. 1992. Evolutionary
studies in cultivatedrice and its wildMorishima,

relatives.
Oxford Surveys in Evolutionary
H. and Miyabayashi,
T. 1993. Distribution
populationsof Thailand.

Biology 8:135-184.
of bacterial
blightRice Genet. Newslet.

10:70-72.

resistancegenes in wild -riceMorishima,

H., Shimamoto,Y., Sato, Y.I., Chitrakon,

H. 1996.

Monitoring

wild

Genetics

III, IRRI

p. 377-380.

Oka, H.I. 1958.

Intervarietal

S., Sano, Y., Barbier,P., Sato,

populations

in permanent

study

sites

in Thailand.

In Riceand classificationof cultivatedrice.

Ind. J. Genet. Plant

Breed.

rice

variation

T. and Yamagishi,

18:79-89.
Oka, H.I. 1988.

Originof cultivatedOka, H.I. and Morishima,

H. 1971.

Oryza perennis
Ooi,

K., Yahara,regionT., Murakami,of the Adhl

Vaughan,Sci. Soc. Press/Elsevier,of plant

with

O. sativa.

N. andMorishima,

gene of rice (Oryza
Variation

rice species.

D.A. and Chang,Japan

The dynamics

and its hybrid

Sano,Y. and Sano, R. 1990.
and wild

rice.

Tokyo/Amsterdam

domestication:
Evolution

H. Nucleotide

spp.)(submitted)of the intergenic

spacerCultivation

experimentswith

25: 356-364

regionpolymorphismof ribosomal

in 5'-upstream
DNA in cultivatedGenome 33:209-218
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Botany 40 (4) :368-383
Wang, Z.Y. and Tanksley,
S. D. 1989.
Genome 32: 1113-1118

In situconservation

Restriction

fragmentof rice genetic
lengthpolymorphism

resources.

Economicin Oryza sativa

L.

Questions

and Answers in Keynote addresses

Questions to Dr. Riley
Q: Could you provide a little
directionsof this system?

more information(Vaughan)on the current statusof SINGER

andA. The informationon the accessions in the CGIAR genebanks will soon be madeavailable
via Internet. The SINGER is an activityof the CGIAR system wide
Genetic Resources Program (SGRP)
which coordinates
the Center's genetic
resources activities.
(Riley)C. There is an overlapping
understandingof the words characterization
andevaluation. In an older sense particularly
for breeders, characterizationmeans toidentify
traits useful for agriculture.
However, characterization,
as used by thisworkshop, refers solely to identityof genetic compositionleaving an area toconnect genetic markers to agronomic traits. This area of effort will still require
long hard work, particularly
for the benefit of breeders. (Hayashi)
Q. Out of the two major methods of germplasm characterization,
morpho-agronomic
and molecular the first is cheaper than the second. However, in the first,
environment plays a major role to interact
with the genotype.
Thus morpho-agronomic evaluation is environment specific,
where as molecular markers are
independent,
thus the data can be used globally.
Do you have any comments?
(Chaudhary)
A. It is importantto evaluate germplasm in the environment where it is adapted,inorder to avoid unwanted genotype X environmental
effects. For complex traits,
such as drought resistance,
different
types of resistance will be needed in different
locations,
therefore location specific evaluationis needed. (Riley)
Q. What is your personal opinion
about purificationof germplasm mixtures?
(Ekanayake)
A. As far as possible,
curators should try and maintain the genetic integrityof thelandrace collection
as it comes from the farmers field and as it enters into thegenebank. There are several ways that curators might do this. However, we accept
that there are inevitable
genetic changes in germplasm while it is in the genebank.
(Riley)Question to Dr. Morishima.
Q. Where did you collect the annual

and perennial

rice populations?

What kind ofraces of Xanthmonas campestris pv. oryzae are prevailingthere? (Tosa)
A. Two perennial and two annual populations
were all collectedin the northern
suburb of Bangkok. The results presented were based on reaction to 4 Japaneseraces of X. campestris pv. oryzae. Later we found that these wild rice population
showed very similar reaction patterns to two major races of Thailand.
But I haveno information on the races prevailingin the area where our wild rice plants werecollected.
(Morishima)

Topic 1.
Newand Improved Approaches to Analysis
Plant Genetic Resources Diversity

Chairpersons
K.Riley
F. KikuchiofApproaches

to Understanding

Genetic Diversity

at the Molecular

Level

STEPHEN KRESOVICH and ANNE L. WESTMAN
USDA-ARS, Plant Genetic

Resources

1109

Experiment

StreetGriffin,Georgia

30223-1797,

E-mail:skresov@gaes.griffin.peachnet.edu

Conservation

Unit

USA

Abstract
Effective
conservation and use of crop genetic resources involve asking manyquestionsabout the extent, distribution,and quality(agriculturally
useful phenotypes,
genotypes, and genes) of genetic variation. Only when the appropriate technologies
andmarkers for describing
this variation
are accessible
can such questions be adequately
addressed. Progress will require the integration of technologies
and protocols that provide forthe acquisitionof large quantities
of genetic information
for improved genotype and gene
identification.
Technologies
that provide for high genetic resolution
and throughput
atreasonable costs will find numerous applications
for curators, breeders, geneticists
and alliedscientists
interested
in characterizationof ex situ and in situ diversity,
gene discovery andtransfer, cultivar development, and ultimately
protection of intellectual
property rights.
Introduction
The wise use of plant genetic resources provides
the foundation
for themaintenance and improvement of crop agriculture.
Throughout
the course of history,
plant genetic resources have been acquired, selected, used, and preserved. As the 21st
century approaches,
segments of our society have become keenly aware of the ' value'of ready access to genetic resources. Ex situ conservationof plant genetic resourcesin repositories
has evolved to serve its user community of fundamental
and appliedscientists.
In complement with in situ management of plants within their nativeenvironments, ex situ maintenance will be expected to play a greater role in the future
for conservationof agricultural
biodiversity.
The primary goals of curation include:(1) acquisition,(2) maintenance,(3)
characterizationand evaluation,and (4) utilization(National
Academy of Sciences,
1991).
As will be highlighted
subsequently,
judicious
collection
and analysis
ofmolecular
data
example:-acquisition:

can impact

positively

Data on the diversity

allof theseof existing

criticallyimportanttasks.

For

collections

can be used to planacquisition
strategies.
In particular,
calculationsof genetic distances can be used to
identify
particularly
unique subpopulations
that is underrepresented
in current
holdings.
- maintenance:
Molecular analysis can be used to eliminate
duplicateaccessions in order to better utilize limited funding for conservation. Information may
be applied to monitor management practices. In addition, molecular data may provide
essential
information
for the developmentof core collections
that accuratelyreflect
variation of the entire collection.
-characterizationand evaluation:
The genetic variation within collections(including
phenotypes, genotypes and genes) must be establishedin relation to the total available
genetic diversity
for each species (Schoen and Brown, 1993; Bataillon
et al., 1996).
When available, existing passport data documents the geographic
location where eachaccession was acquired. However, many records are missing or incorrect. Molecular
data may allow for characterization
based on genetic information,
which ultimately
may be more accurate and useful than classicaldocumentation.
utilization:
Users of collections
benefit from genetic informationthat allows themto quickly identify
valuable types and traits. On a more fundamental level, molecular
information
may lead to the identificationof useful genes contained in collections.
As noted previously,
the goals of effective
ex situ curation can be quite
challenging
based on the need to simultaneously
resolve numerous operational,
logistical,
and biologicalquestions.
For curators to make progress, the following
recurring questions must be addressed:
identity:
how to determine that an accession or cultivar is catalogued
correctly, istrue to type, and maintained
properly;
relationship:
how to establish
the degree of relatedness
among individuals
in anaccession or accessions within a collection;-structure:
how to determine the partitioningof variation
among individuals,
accessions, populations,and species; and-location:
how to establish
the presence of a desired gene or gene complex in a
specific
accession,as well as the mapped site of a desired DNA sequence on aparticularchromosome in an individual
or a cloned DNA segment (Kresovich
andMcFerson, 1992).
It is our belief that molecular information
will be of great value to assist
curators in achieving
their collective
goals and solving day-to-day questions.

Markers and Technology
When considering the applicationof molecular markers and technologies
toresolve questions of conservation and improvement, both technicaland operational
issues must be considered.
For example, technicalissues relevant to marker
characteristics
include discriminatory
ability,sensitivity,
reproducibility,
and theability
to be used for further genetic analysis or in diagnostics.
Operational
issuesinclude protocol characteristics,
time, and cost. The ideal molecular marker must be
easy to employ, timely, cost effective, highly informativeand reliable (accurate withthe desired level of precision).
Sample preparationmust be simple and the assay(including
data generation,collection,organization
and analysis)should be suitablefor increased throughput
and automation. A high information
content necessitatesamarker assay that detects high heterozygosityand provides discriminatory
abilityamong closely related individuals,
as well as the generation of data from multiple
genomic sites, using a single assay. Reliability
implies reproducibilityof results fromassay to assay both within and across laboratories,
as well as unambiguous data
analysis.
To date, various constraints have precluded the broad adoption of DNA-based
markers for use in crop conservation
and breeding. However, molecular markers
based on the polymerase chain reaction (PCR) are receiving much attention because
they ultimately
have the potential
for widespread,
low-cost, large-scale
application
suitablefor the multiple
needs of genetic resources conservation
and use. A
PCR-based assay requires only small amounts of crude genomic DNA preparations
from each sample, is a procedure that is not technicallychallenging
or expensive, andprovides accurate results in a single day. In addition,
the assay may readily be scaled
up to handle large numbers through automation.
The subsequent summary of markers and assays for use has been preparedpreviously (Westman and Kresovich, in press). It concisely discriminatesmarker andassay, and how these two particulars
may be integrated
to answer curatorial questions
regarding

how much variation is present and how it is partitioned.
The appropriate markers for a study can discriminate
between entries in anarray, but are not so polymorphic
that important variation is masked by random noise(Brower and DeSalle,1994). Molecular markers range from highly conserved to
hypervariable,

and can be either

proteins

or nucleic

acids.

The nucleic

acids used asmarkers include entire genomes, single chromosomes, fragments of DNA or RNA,and single nucleotides.
A wide variety of nucleic acid fragments are used as markers. While someoccur once in a genome, others are repeated. Many repeated sequences used as
markers are noncoding;others are elements of multigene families.
Some repeatedsequences are interspersed
throughout
the genome, either distributed
randomly or inclusters.
These interspersed
repeats are common in plant and animal nuclear
genomes, and are found in plant (but not animal) mitochondrial
genomes (Palmer,
1992). The chloroplast
genome contains a large inverted repeat (IR); most angiosperm
chloroplasts
have two copies, separated by a short single-copy
region. Repeat lengthand (rarely)
loss of one copy can vary between taxa (Downie and Palmer, 1992).
Much research at present is focused on repeated sequences that occur intandem. The classes of tandem repeats are distinguishedby the length of the corerepeat unit, the number of repeat units per locus, and the abundance and distributionof loci (Table 1). The names for these classes are themselves varied and have been
inconsistently
used, but Tautz (1993) has clarified
the nomenclature.
Tandem repeats were first reported in the literature
as 'satellites'of DNA,detected in CsCl density gradients as fractions with different GC content than the restof the genome (Britten
and Kohne, 1968). These satellites
have repeat units that areusually several hundred nucleotides
long, with thousands of copies at each of several
loci in the nuclear genome. These loci are usually in heterochromatin,
often near
centromeres. Satellite
DNA is present in numerous species. For many satellites,thenumber of loci and number of repeat units per locus vary between species and higher
taxa (Ingles et al., 1973).
Minisatellites(often called variable number of tandem repeat loci, or VNTRloci) are widely used as markers, especiallyin forensics. The repeat units are usuallyless than 100 nucleotides
long, with tens to hundreds of copies per locus. Thousandsof loci in a genome may have similar core repeat units. The number of repeat units at
a minisatellite
locus can vary greatly between individuals
and populations.
First
described
in humans (Jeffreys
et al., 1985), minisatellites
are found in numerous
animal species, often near telomeres.
They are also common in plants (Rogstad,1993), and are often associatedwith satellites
and centromeres. The number of repeat
units

per locus is lessvariablein plants

than in animals,

but is still

high;

plant

Table1. Classesof nucleic

acid

sequencesused as fragment

markers (from Westman and Kresovich,in press).
Marker assay ccS lae qssu e n c e

G e n o m e a # L o c i/ g e n o m e

C o d in g

R e p e a t u n it

re g io n b

le n g th (b p )# T an dem

C sC lD N A ‑D N A

I n situR e stric ‑PC R

u n its/lo c u s

d e n sityh y b rid ‑h y b rid ‑tio n s ite

a m p lifi‑

g ra d ie n t

iz a tio n

iz a tio n

an a ly sis

c a tio n

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

S in g le c o p y

n ,cp , m t

o ne

+

I en ptee rsp
a t e rse d r

n ,m tv a ria b le+v a ria b le

cp

o n e o r tw o+

2300 ,0,0 00 00 ‑no n e to se v e ra l+

9 0 0 0 ‑ 1 1 ,0 0 0

1 0 2‑1 0 4

1 0 0 ‑5 0 0< 20

Iren pv e artet d

T an dem

re p e a t s :n u c le a r r R N A+

g e n e c l u s te rn u c le a r r R N AnIG S s u b re p e a t

1 0 2‑1 0 4 p e r
rD N Alo c u s

S a te ll iteno n e to se v e ra l

2 ‑1 0 0 0

10 3‑1 0 7

M in i s a t e ll it en103

< 10 01 0 ‑1 0 0

+

+

+

M ic ro s a te l li ten

1 0 3‑1 0 5

1 ‑6

5‑100

+

+

+

+

an=nuclear, cp=chloroplast,
mt=mitochondrial.
bmarker sequence present in coding regions (+), noncoding
Appropriate(+) or inappropriate(-) marker assay.

regions(-), either

coding+or noncoding+

regions(+).

minisatellites

are usefulmarkers for variation

between and within

species(Rogstad,1993).
As suggested by their name, microsatellites- also called simple sequence
repeats (SSRs),or simple sequence length polymorphisms(SSLPs)- have very shortrepeat units, no more than six nucleotides
long. SSRs are more abundant thanminisatellites
in noncoding regions of the nuclear genome, and are present in somenuclear genes and organelle genomes (Tautz et al., 1986; Wang et al., 1994). The
number of repeat units per locus is lower for SSRs than for minisatellites,
but canapproach 100 in animals and 50 in plants (Tautz, 1993; Saghai Maroof et al., 1994).
The abundance and polymorphismof SSRs make them particularly
valuable fordescribing
variation between populations
and individuals(Brown et al., 1996).
Like minisatellites,
SSRs were documented first in humans (Tautz et al.,
1986; Litt and Luty, 1989; Weber and May, 1989) and later in plants (Condit
andHubbell, 1991). Plants and animals differ in the abundance of specific SSR motifs in
the genome. In both plant and animal genomes, chromosomal distributionof SSRs
is variable.
Some animal SSRs are found near heterochromatin
or interspersed
repeats, but most are randomly dispersed (Tautz et al., 1986). However, some studies
have locatedplant SSRs near genes, highly
methylatedDNA, satellites,orcentromeres (Bennetzen et al., 1994).
Tandemly repeated genes are also utilized
as markers. Perhaps the mostwidely used are the nuclear genes that encode ribosomal RNA (rRNA) (Hamby andZimmer, 1992). The three rRNA genes are separated
by two internal transcribedspacer regions, generally
a unit that is tandemlyreferred to as ITS1 and ITS2. These genes and spacers form
repeated hundreds of times, at one to several loci in the

genome. At each of these loci, the individual
repeat units are separatedby
nontranscribedintergenic
spacer (IGS) regions.
In the middle region of each IGS are
tandem copies of a short subrepeat sequence. Variation in rRNA gene clusters can bemeasured at several levels, each evolving at a different
rate: (1) the number andlocation of rRNA loci, which is highly conserved;(2) the (more variable)
number oftandem gene clusters per locus; (3) the conserved sequences of the three genes; (4)the variablesequences of the ITS regions;and (5) the highlyvariablenumber of
subrepeats in the IGS region. These features make rRNA gene clusters versatileandinformative
markers for mapping and phylogenetic
analysis(Maluszynska
andHeslop-Harrison,
1993).
Molecular marker assays (Tables

2 and 3) are generallyclassified

by whether

the molecules evaluatedare proteins or nucleic acids, and whether the character
analyzed in a nucleic acid marker assay is the entire genome, a chromosome, afragment, or a nucleotide.
Alternatively,
marker assays can be categorized by the typeof character measured (Avise, 1994). Some methods measure quantitative
differences
between entries in an array. Others measure qualitative
characters,
each with two or
more possible states. Marker assays also differ in the number of loci evaluated per
analysis, whether multiple loci are evaluated simultaneously
or sequentiallyand thetype and amount of informationneeded about the marker loci before conducting theassay.
Choosing
appropriate
marker assays can be challenging,but several
considerations
can make the task easier. Important issues are: (1) what question is
being asked? (2) what level of resolution
is required?
(3) how can the results be
related to characteristicsof the taxa being studied?
and (4) are sufficient
resources
availablein terms of personnel,
equipment,
funding and time? (Kresovich
andMcFerson, 1992).
Summary
The goals and expectations
for analyzing plant genetic variation parallel thoseestablishedacross many other fields of biologicalresearch, from agriculture,
ecology,and evolution to the medical sciences.
In all of these fields, future genetic marker
assays must incorporate
methods to detect, describe,interpret,and store DNA
sequence information. Molecular tools of the future are expected to be user friendly,
accurate, precise, high throughput,
low cost and potentiallyautomated.
DNA sequence informationis the foundation
for developing
and applying
genetic markers to questionsof biologicalvariation,
whether in situ or ex situ.
Researchers who develop and use sequence-based marker assays for quantifying
andpartitioning
genetic variation
will continue to benefit greatly from information
andtechnologies
generated by the international
Human Genome Project (HGP).
In the HGP, technologicalimprovements unanticipatedin 1990 have alreadychanged the scope of the research and allowed for more ambitious approaches andgoals (Collins
and Galas, 1993).
In the plant kingdom as well, progressive
visions ofTable2. Summary of molecularmarker assays used to measure plant

M a rk e r a s sa y

mT oy lep ec uo lef

genetic

variation

G en om es

C h a r a c te ra ss a y e d 3

a n a ly s e d(from Westman and Kresovich,in press).

# C h a r a c te r # L o c i p e r a s s a y c M u lt il o c u s
a n a l y s is ds ta t e s b

I na hn ec re ‑eit

M ic r o c o m p le m e n t f ix a ti o n

p r o t e into ta lr e a c t iv ity

qu anto n e to m a n ys im

M o n o c lo n a l a n ti b o d y a s s a y

p r o t e into ta l+ re a c t iv it y2o n e o r s e v e ra ls im

P r o t e i n e le c t ro p h o r e s is

p r o t e in

n ,cp

e le c t r o m o r p h< 10o n e to s e v e r a l

se q

C s C l d e n s it y g r a d ie n t

D N A

to ta lb u o y a n t d e n s i ty

q uant

m any

s im

D N A ‑ D N A h y b r id iz a t io n

D N A

to ta l△T mq uant

m any

s im

F lo w c y to m e t r y

D N A

n

D N A c o n te n t,
# c h ro m o so m e s

q uant

m any

s im

C h ro m o s o m e b a n d in g

D N A

n

+ ban d

2

m any

se q

co do mD N A ,R N A

n ,cp ,m t

+ r e s tr ic t io n s i te2o n e to se v e ral

se q

co d om

D N A

n ,cp ,m t

+ frag m e n t

2

m an ysim

do mm an ysim

do m

co do mF r a g m e n ts e l e c t ro p h o r e s e d , t h e n d e te c t e d :S i n g le ‑ c o p y o r
lo w ‑ c o p y R F L P a s s a y
M u lt il o c u s r e s t r ic t io n f r a g m e n t a s s a y
A r b i tr a r y P C R

D N A ,R N A

n ,c p,m t+ fra g m e n t

2

D e sig n e d ‑p rim e r P C R

D N A ,R N A

n ,c p, m t

f r a g m e n t le n g t h>2o n e to m an yseq

c od om

D N A ,R N A

n, cp, m t+ fra g m e n t
s ig n a l i n te n s it y2
qu antm an ym an y

sim
s im

d om

D N A

n

+ fra g m e n t

2

o n e to m a n y

se q

d om

D N A ,R N A

n, cp ,m t

n u c le o t id e

4

on e

F r a g m e n t s d e t e c te d d ir e c t ly , w it h o u t e le c tr o p h o re s is :D o t o r s lo t b l o t h y b r id iz a t io n
I n s it u h y b r i d iz a t io n
N u c l e ic a c id s e q u e n c in g

an=nuclear, cp=chloroplast,
mt=mitochondrial.
bquant=quantitative.
cin the nuclear genome. The chloroplastand mitochondrial
genomes are each considereddloci analysed
simultaneously(sim) or sequentially(seq).
cfor loci in diploid(nuclear)genomes, dominant (dom) or codominant(codom) inheritanceas one locus.
of alleles.

d om orco do m

Table 3. Summary of fragment
M a rk e r a ss a y 3

marker assays (from Westman and Kresovich,in press).

T a rg e t

F ra g m e n t# P rim e rs o r p ro b e s

sequ ence b

p ro d u c tio n c

S in g le co p y s e q u e n c e

U orK

R D

v a ria b le<20

R e p e titiv e se q u e n c e

K

R D

1

<20

D A F

U

P CR

1

<1

R A P D

U

P CR

1

0 .5 ‑3

M in ih a irp in D A F

U

P C R

In te r‑re p e a t P C R

U

P C R

A n c h o re d P C R

U

P C R

D es ig n e d ‑p rim e r P C R ,s in g le ‑c o p y o r lo w ‑c o p yseque nceK

D es ig n e d ‑p rim e r P C R ,ta n d e m re p e a t lo c u s
N es te d P C R

AF ra g m e n t

D C

s iz e (k b )R es tri c tio n fr a g m e n t a s sa y

A m p lifi e d fr a g m e n t a s sa y

1

<1

>1

v a ria b le1

v a riab le

P CR

2

<5

K

P CR

2

< 0 .5KP C R th e n
PC R

2 p e r re a c tio n

v a ria b le1

R es tri c te d a n d /o r a m p lifi e d fr a g m e n t a s sa y
C le av e d te m p la te

U orK

R D th en P C R

v a ria b lev a ria b le

C le av e d frag m e n t

U orK

P C R th e n R D

v a ria b lev a riab leS in g le ‑stra n d e d fra g m e n t

U orK

R D or P C R
th e nd e n a tu re

v a ria b lev a riab leaDAF=DNA amplification

fingerprint,

RAPD=random

amplifiedpolymorphic

DNA,

PCR=polymerase
chain reaction.
bU=unknown, K=known.
cRD=restriction
dA=arbitrary,endonuclease
digest.
D=designed,
C=combinationof arbitrary

and designed.

DNA analysis will most likely change the ways in which problems related todescribing
genetic variation are perceived and resolved.
In order to better characterizeplant genetic diversity
and address genetic
resources conservation and use, plant scientists
will need molecular marker assays thatcost effectively
detect and describe DNA sequence variation over many areas of thegenome
This

(both

goal

codingclearlyscientists

and noncoding),

is a challenge,

but

are aware of and build

and approaches

will

for many individuals
much progress

on innovations

be available

for thein a population

or taxon.

made.

as plant

is being

in otherfields,As longa streamof new tools

challenge.

Acknowledgments
The authors

greatlyof the Applied

Genetic

In particular,

we explicitly

and practicalinsights

appreciate

Analysisthe continuing

Laboratory

thankof the Plant

S. E. Mitchell,on genetic

contributions

analysis

Genetic

from the team members
Resources

Conservation

R.E. Dean and C.A. Jesterand genetic

for their

Unit.

theoreticalresources.

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44: 388-396.

S. (Inpress)

Use of molecularmarkerfor description

Ford-Lloyd,

B.V. et al., (eds.)

techniques

Biotechnology

and Plantof
Genetic

Resources,

CAB International,

Wallingford,

UK.

Biosystematics- Implications

for Use of Plant Genetic

Resources

YOSHIO SANO and LE-VIET DUNG
Facultyof Agriculture,

Hokkaido

University,

Sapporo,060-32,Japan

Introduction
Transfer of alien genes into cultivatedspecies often results in a breakdownof the harmonious genetic architecture
which has been called M-V (morphology
viability)
linkage(Grant, 1967).
The complex nature of quantitative
trait loci is a
result of the limited number of chromosomes, and tightly or loosely linked genes tend
to form adaptive sets which have been given various names. These phenomena areclosely related to use of alien germplasm and the genetic mechanisms involved informing of crop gene pools. Genetic comparisons of naturally
occurring variants
among taxa need to be studied to understandthe biological
species concept andinformationon this gives basic information
relevant to understanding
genetic
resources.
We present here our rice experiments in which we investigatedthegenetic mechanisms associated with the formation of gene pools among rice taxa.
Genetic dissectionof M-V linkage in rice chromosomal segments is also preliminarily
presented in relation to their genome architectures.
Classificationas basic information
for genetic resources
Recent studies on genealogy revealed that the appearance of new genes israther a rare event in the evolutionary
process. Most newgenes seem to have evolved
from duplication
following
modifications
to gene expression and the opportunity
tobe fixed in a population
depends upon the environment, as well as interacting
genesets. This view is supported by colinearity,
based on molecular markers, among
cereal genomes (Bennetzen and Freeling, 1993). There are various constraints whichhinder adaptive gene sets changing through natural selection.
Naturallyoccurring
genetic variation is a major contributor to adaptationin organisms. The improvementof agronomic traits have primarily
been accomplishedthrough recombinationofnaturally-occurring
genes rather than a few mutational
events. Hence diversified
germplasm is important for breeding programs.
The fact that it is not easy to transfer useful genes into crop species fromalien taxa suggests that harmonious gene sets are actually orpotentiallymaintainedininterbreeding
individuals,
sharing
the same gene pool. The definitionof widehybridization
depends on breeding objectives
or crop species.
Crosses between
subspecies,
species or genera are often referred to as wide hybridizationand new
technologies
are expected to enhance transfer of alien genes into crop species.
Disharmonious
gene interactions
occur, even after successful
hybridization,
if theparents are geneticallydistant.
Thus, understanding
taxonomic relationships
is aprerequisite
for the use of alien germplasm and we need better knowledge concerning
the genetic basis of species boundaries.
At first, we show an example of confusionin the nomenclature of rice species, since it is subject of continuing discussions in rice
(Vaughan, 1989).
There is a controversy as to whether American wild rice with the AA genome
is a distinct
species,O. glumaepatula.
The American AA genome species are
reproductively
isolatedfrom others, but some samples are sexually compatible with
Asian rice. If all the accessions
are O. glumaepatula,
the sexually compatible
accessions could be used for gene transfer. American accessions preserved at National
Institute of Genetics, Mishima, were reexamined. Hybridization
experiments revealed
that accessions could be divided into 2 groups based on fertility
relationships(Fig. 1).
Hybrids were fertile in crosses within each group but infertilein crosses between
groups. The results could be explained by assuming that all the American accessions
are a distinct
species but are differentiatedwith respect to fertility
relationships
asobserved in Asian wild and cultivated rice species (Oka, 1988).
One of the 2 groups,
however, produced fertile progeny when crossed with Asian wild and cultivatedrice,
it was considered
to be O. rufipogon like and the other was assumed to be O.
glumaepatula.
To look into their genetic divergence at the molecular level, intergenic
spacer(IGS) regions of ribosomal DNA (rDNA) were compared. rDNA is a multigene
familyand the length heterogeneity
results
from repetitionof short repeatedsequences in the IGS regions in rice (Sano and Sano, 1990). The pattern of variation
generallyshows a high level of family homogeneity
within species but a high levelof heterogeneity
between species. There is a possibility
that species specific variation
resolves the discrepancy
mentioned above. Table 1 shows that 6 different
IGS lengthvariants were present in the 28 American accessions examined and 4 out of the 6variants were also present in Asian accessions.
Variants with the same length of
repeats sequences does not always correspond to the same origin.

Fig.1.

Fertility

relationships

Table

1. Intergenic

spaceramong American wild rice accessions

lengthvariation

the AA genome. The variantsof rDNA detectedin American

marked with asterisks

IG S v a ria n t
4 .2 0 k b

with the AA genome.

are present

wild rice accessionsin Asianwithrice species.

N o . o f a c c e s sio n s2

4 .2 5 *7

4 .4 0

1 1

4 .8 5 *2

5 .0 0 *3

5 .3 5 *3

T o ta l

28

The molecular variants could be easily re-evaluatedat the sequence level.
The fine structure in the hypervariable
region were compared among the IGS regions
from the 2 groups by means of the method of indirect
end labelling(Fig. 2). The
structure of sub-repeats is resolved by restriction
enzymes of SalI and HinfI. In Asian
wild and cultivatedrice (O. rufipogon - O. sativa complex), the length variationiscaused by addition-deletion
events of the sub-repeats
marked by SalI and the tworegions between SalI and HinfI are conserved based on current information.
The twolength variants(4.85kband 5.35kb BamHI fragments)from O. rufipogon likeaccessions were analysed and their sub-repeat structures were found to be identicalto those from the Asian rice complex if the lengths were the same. On the other hand,Fig.2. Comparison, of the intergenic

spacerof rDNA in Asian and American rice accessions.

the other two length variants (4.20kb and 4.85kb BamHI fragments) were present onlyin O. glumaepatula
accessions
based on fertility
relationsand the sub-repeat
structures were markedly different
from those of Asian accessions. The central region
in the IGS marked by SalI and HinfI were well conserved among O. glumaepatula
accessions,Indicatinga high level of homogeneityof the IGS region within thisspecies. Although a length variant of 4.25kb was detected in both O. glumaepatulaand Asian wild and cultivatedrice, the sub-repeat
structures from O. glumaepatula
had the conserved central region in the IGS marked by SalI and HinfI showing thatcomparisons of the IGS regions are effective in recognizing
reproductively
isolated
rice taxa. The present results support an assumption that there exists O. glumaepatulaand O. rufipogon
like accessions
in America and the latter might have been
introduced
from Asia (Vaughan,1994).
Morphology-viability
Difficulties
interspecificglaberrima

linkage
in gene transfer

across isolating

hybrid between the two cultivated(Sano et al., 1980).
O. glaberrima

barriers

were demonstrated

in an

rice species, O. sativa and O.
is endemic to West Africa and ischaracterizedby short ligules and fewer secondary panicle branches than those of O.
sativa. The interspecifichybrid between them is male-sterile
but female fertile,andthe hybrid can be backcrossed
as the female parent. About 60 recombinant inbredlines (RILs) were establishedfrom BC2F6 and BC1F6 after backcrossing
and selfing
and the likelinessof a plant to the parent was evaluated based on 8 morphologicaltraits including
4 species-discriminating
traits.
The computed value for the sativa parent is 1.0 and that for the glaberrima
parent is -1.0. Absolute values exceeding 1.0 indicate transgressive
segregation.
The
results showed two different
tendencies depending on the trait examined (Fig. 3-A,
B). Regarding
ligule length, lines similar to the parents were frequent and had ahigher seed production than lines with ligule length intermediate
between the parents.
Primary branch number did not correlate with parental phenotypes.
Transgressive
segregants were observed for primary branch number and they tended to have a low
seed production.
Other species discriminating
traits had a similar tendency as thatfound for ligule length, suggestingthat parent-likephenotypes
are rapidly recoveredin hybrid populations.
The tendency was more clearly observed when the 4 speciesdiscriminating
traits were combined (Fig. 3-C).
The rapid return to the parental phenotype after hybridization
is an exampleof the so-called M-Vlinkage. This trend appears more clearly between species thanbetween varietal groups within species, indicatingthat the genetic factors for themechanism were accumulated as genetic distance increases. Since no abnormality
indevelopment
was detected except for infertility
during the experiments,
genetic
elimination
caused by hybrid
sterility
might be related
to the phenomenon.
Disharmonious
gene combinations
were eliminatedin the population
as fertility
rapidly increased by selfing.
Gene sets for the parental phenotypes might be changed
together with viability
genes such as hybrid sterility.
Disharmonious
interactions
hadto operate between chromosomes as well as within chromosomes since all the traits
examined seem to be controlled
by polygenes.
This assumption is supportedby theresults that the hybrids
recovered fertility
when crossed with the parent having
corresponding
phenotype but not with the other parent.
It should be noted that while
restricted
recombination
occurs in a hybrid population,
genetic homogenization
also
apparently"Selfish"occurs since the recovered parental type is not identicalto the parent type.
DNA such as the gamete eliminators
have been shown to be involved inFig.3. Morphological-viability
rice

species,morphologicallinkageas revealedin the hybrid

derivatives

between

Similarity

or likelinessindex

O. sativa

and O. glaberrima.

traits,1 showing

similarity

to O. sativa

index

and theirmean seed number per plantlines

with different

interspecifichybrids(Sano,

values1990).

"Selfish"between species without changing theirsignificanceremained to be elucidated.

and -1 to O. glaberrima.

elements

taxonomic

statusthe two cultivated
was computed
Frequency

fromof B2F6

are shown.

could enhance introgression
althoughits full biologicalComplexity
of QTLs
The time to flowering
is a major adaptive factor which enables rice plants tocomplete their life cycle appropriately
in relation
to the latitudeat which the rice
evolved (Oka 1988, Chang et al., 1969).
The inheritanceof heading date is of apolygenic
nature and as a result hybrids show continuous variation (Akemine andKikuchi, 1958).
Recent interest has focused on dissectionof quantitative
traits into
major loci and it has been pointed out that there are often regions of the genome thatcan account for large portions of phenotypicvariation.
One of the major QTLs forheading date in rice seems to be present on chromosome 6 (Yokoo et al., 1982, Yanoet al., 1996) although it is not easy to address whether the regions with major effects
are due to the action of single genes (orthologous)or clusters of genes. We compared
the genetic complexityof heading date in relation to chromosome 6.
T65 (Wx-pat) has an alien segment of chromosome 6 introduced from the Indica
variety Patpaku by backcrossing.
Based on the segregation
pattern in the F2 of[T65wx] x [T65 (Wx-pat)], the introduced segment seemed to carry Se-1 judging fromthe linkage intensities(Fig. 4-A).
Further, 5 different
recombinant inbred lines(RILs)
for heading date were detectedin the later generationsof the hybrid,suggestingthat at least 3 genes might be located on the introduced
segment. One of
the RILs showed transgressive
segregation,
since it headed much later than theparents. Genetic experiments revealed that another RIL (Type II) carries a recessive
gene, se-pat (tentatively
designated)present on the segment (Fig. 4-B). All the
involved genes seemed to be responsible
for photosensitivity
since the number of days
to heading of all the RILs was reduced by short day treatment, showing a cluster ofrelated genes on the segment. The gene se-pat maybe widely distributedin Oryzaspecies since a similar recessive gene for photosensitivity
was detected in backcrossed
populations
including
Indica type of O. sativa, O. rufipogon and O. longistaminata.
Interestingly,
a short day treatment at the early stage of development delayed heading
in se-pat homozygotes indicating
an age-dependent
expression for photosensitivity.
The results confirm that a gene complex on chromosome 6 results in a range ofvariationin heading date by recombining
genes on the segment after hybridization.
Polygenic traits are controlled by the interactionof numerous genes whose effects are
essentiallyinterchangeable
and small relative to environmental
sources of variation.
The present results is an example of loosely linked genes on a chromosome segmentthat has the potential
to adjust heading
date of a hybrid population
to different
environments,

throughthe reconstructionof the genie content.

Genome architecture
Accumulated evidence at the molecular level confirms that the genomes of
cereals have similar gene composition
and map colinearity.
In addition,
genomes arecomposed of a mixtureof conserved

and variable

parts,as shown in the multigene

Fig.4.

Segregation
and Japonica

patterns for heading
types

of O. sativa.

time showing

the complexity

T65 (Wx-pat)is a near-isogenic

segment of chromosome 6 from Indica
from T65 wx x T65 (Wx-pat)indicatingtype (Patpaku).
a recessive

on chromosome 6 between
line

of Japonica

Type II is a recombinantgene, se-pat(t),Indicatype withinbredline derived

for photosensitivity

different

sequencesin higherafrom Se-1 on chromosome 6.

familyof rDNA. Single

genes

are surroundedby repetitive

organisms and they interact with other genes in a coordinated
way resultingindevelopmental
processes. Since the number of chromosomes are limited, single genes
are linked with many other genes. These considerations
lead us to examine whatkinds of genie differentiation
are involved in parallel
fine-structure
mapping withoutchanges in genie order. We attempted
to compare naturally
occurring
variants
between different
taxa in order to examine the biologicalsignificanceof adaptive
gene complexes on a chromosome. Differences in allele frequencies
on chromosome
6 have been repeatedly
reported with respect to varietal differentiationin rice. The
loci are wx, C, alk, isozymes and so on as shown (Fig. 5). We took advantage of thisFig. 5. Genetic

differentiation

observed

upper position

show differencesin the allelicand the genes in the lower position

on chromosome 6 among related
frequency

are responsible

between Indica

for reproductive

taxa of rice. The genes in theand Japonica

barriers

operatingtypes of O. sativa
between the related

taxa.

information to dissect M-V linkage in rice. The allelic differences
detected between
different taxa might suggest the presence of the mechanisms which act to conserve theco-adapted genes from destruction through recombination
after hybridization,
if any.
As mentioned, clustered genes for photosensitivity
are located on chromosome 6 andthe differencein flowering time partly acts as a premating reproductive
barrier.
Internal
barriers
play a significant
role in the genetic
changes of hybrid
populations.
Genes controllingthese internalbarriers
were also detectedonchromosome 6 of rice (Morishima et al., 1992). In addition
to genetic differentiations
for hybrid sterility
and heading date, we recently found that a cluster of genes
responsible
for cross-incompatibility
in rice are located on chromosome 6 (Sano,
1992).
Reduced seed setting was first found when a segment of chromosome 6 wasintroduced from O. rufipogon (W593 from Malaysia)
into O. sativa (T65wx). Whenthe plant carrying the introduced
segment was pollinatedby T65wx, it frequently
produced aborted and inviable
seeds while the reciprocal
cross between the same
parents
showed normal seed setting.
Genetic
experiments
showed that theincompatibility
system is controlled
by three genes, Cinf, Su-Cinf and cinm. Cinf andcinm specify cross-incompatibility
in the female and male reactions, respectively,
andSu-Cinf suppresses the action of Cinf. Therefore, unidirectionalcross-incompatibility
occurs when megaspores expressing Cinf are fertilizedwith pollen grains from plants
homozygous for cinm. It seems that Cinf is rare in rice accessions
but Su-Cinffrequent in Indica varieties of O. sativa. This causes a difference
plants between Indica and Japonica types.

in response

to Cinf

The genie effects in the heterozygotes
revealed that
all the three genes
controllingthe cross-incompatibility
system act sporophytically,suggesting their gene
expression occurs before meiosis. Cyto-histologicalobservations showed that aborted
seeds in the cross incompatible
system are caused by retardationof the endosperm 4-5
days after fertilization.
The question arose as to how these genes cause defective
seeds after fertilizationeven though they are expressed before meiosis and they giveno adverse effect on selfing. This implies that the female and male gametes are notgeneticallyequivalent
and an interaction
among the genes enables plants to recognize
their mates through abortion of seeds.
A summaryof gene divergence responsible
for reproductive
barriers detected on
chromosome 6 of rice is shown (Fig. 5, after Sano, 1993).
It is not clear if this region
has a disproportionallylarge role in isolating
rice taxa. We think that various
chromosomes play a role and each of them has the potential
to produce a high levelof variation by recombination.
The segment of chromosome 6 we examined confirmsthat genie differentiation
is distinct
in rice and the segment is able to respond indifferent
ways in hybrids
dependingon their genetic content.
Further genetic
comparisons using molecular markers are expected to throw light on the genetic
systems involved in the formation of crop gene pools.

Acknowledgments
The seniorauthoris indebted

to the late H. I. Oka for his invaluableadviceandencouragement.
References
Akemine, H. and Kikuchi,

Genetic

variability

among hybrid

under various

environments.

In Studies

on the Bulk

R. Takahashi

and K. Kumagai,

Yokendo,Tokyo, p.89-105.

Bennetzen,

J.L. and Freeling,

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Chang,

F. 1958.

M. 1993.

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in rice by the basic

as a single

system:Methodof rice plants
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grown

Eds. Sakai,

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vegetative

phase

Component

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from seeding
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18:79-91

toGrant, V. 1967.
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betweenmorphology

H., Sano, Y. and Oka, H.I.

relatives.

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in Evol. Biolin plant

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studiesAm. Nat. 101:125-139

on cultivatedOrigin of cultivatedrice. Jpn. Sci. Soc. Press, Tokyo, JapanThe genie nature of gamete eliminatorin rice. Genetics
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Sano, Y. 1992.

Genetic

42:561-572
Sano, Y. 1993.
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Constraints
In

Internat.

in usingof chromosome 6 between

wild and cultivatedwildlack of basic

and wild rice species.

relativesin breeding:Crop Sci. I, Ed. D.R. Buxton,

U.S.A. p.437-443
Sano, Y. and Sano, R. 1990. Variationof the intergenic

variations

in backcrosscharacters.

Jpn. J. Genet. 55:19-39

knowledge

spacerDNA in cultivatedderivatives

Genetic
betweenstudiesof speciationin cultivatedrice. 2. Character

Oryza sativa and O. glaberrima: M-V linkageof heading-conferringof natural

Philippines

F. 1982.

day length.

The genus Oryza L.: current

IRRI, Manila, Philippines
Vaughan, D.A. 1994. The wild
Manila,Wisconsin,of ribosomal

III. IRRI, Manila, p.650-656
Yokoo, M., Toriyama, K., and Kikuchi,changeson crop generegionKurata,heading

D.A. 1989.

Jpn.J.Breed.

Crop Sci. Soc. Amer., Madison,

Yano, M., Yoshiaki,H.,
Kuboki, Y., Lin, S.Y., Nagamura,Y.,
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to seasonal

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Genome 33: 209-218.

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Vaughan,and its wild

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Oka, H.I. 1988.
Sano, Y. 1990.

comparisonsrice

relativesof rice:

Responses

N., Sasaki T.,and Minobe Y.
time in rice. In:Rice
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A genetic

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IRRI Res. Paper

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No 38.

In-situ conservation of plant genetic
Characterizationand evaluation

resources:

DUNCAN A. VAUGHAN1, NORIHIKO TOMOOKA1, NOBUYA KOBAYASHI2
and ALI OSMAN SARI3
1 National
2 Experimental

Instituteof Agrobiological

Resources,

Farm, Kobe University,

Japan

3 The Aegean Agricultural

Research

Institute,

Tsukuba,Izmir,

Ibaraki

305, Japan

TurkeyAbstract
Long term support for conservation rests largely on the perceived benefits that accrue
from that support. One way in which ex-situ conservation has provided a return on investment
is as a result of characterizationand evaluation. Populations conserved in-situ also provide aliving laboratory which can provide insights into population
structures, evolutionary
andecological dynamics. This paper focusses on issues related to the conservation and evaluationof germplasm conserved in-situ. Among the important issues which germplasm conserved
in-situ can give insights
into are related to sustainability
and resolving,scientifically,
biodiversity
paradigms.
Introduction
Emerging
models for the conservationof plant genetic
resources are
increasingly
comprehensive.
At the molecular level, advances in biotechnology
areenabling
germplasm banks to isolate
and conserve molecules
such as, DNA
sequences. On a global scale remote sensing technology enables identificationof rareor threatened
habitats
and thus specificareas for environmental
protection.
Ex-situ models for conserving plant genetic resources based on genebanks
are nowincorporating
various types of in-situ conservation.
The harmonizingof theFAO International
Undertaking
on Plant Genetic Resources for Food and Agriculture
and the Convention on Biological
Diversity
is one example of how conservation ofplantsof agricultural
importanceare viewed within
the concept of overall
conservationof biodiversity.
Advances in biotechnology
have made it possibletotransfer
genes between almost any organism. Consequently
useful genes foundanywhere in the ecosystem may be used in agriculture.
This requires that conservation
be comprehensive
can be conserved

and that the continuum of ways in which plant genetic resourcesfrom ex-situ to in-situ
be encompassed in practicalconservation

program.
Another trend in conservationof plant genetic resources can be called thecrop improvement/crop
conservation
loops. Examples are abundant of genebanks
repatriating
germplasm to areas where indigenous
germplasm has been lost, forexample, in Cambodia after the period of war there (IRRI, 1995).
The opposite
process is also occurring where, for example, community germplasm projects, suchas those in North Cotobato,
the Philippines,
find that local germplasm is not
represented
in the genebank. This germplasm maintained
in-situ but not in thegenebank can be sent to genebanks to improve the representationof the ex-situ
genebank collections(Salazar,
1995).
Country reports to the FAO Technical Conference held in Leipzig,
in June1996, had many examples of the synergistic
relationshipbetween crop improvement
and germplasm conservation both ex-situ and in-situ. The relationshipbetween thegenebank and local communities in some countries, for example, Sierra Leone andEthiopia,
is very much related to crop "improvement" (Country reports of Ethiopia
and Sierra Leone to the FAO Technical Conference, available
on the Internet).
In
some countries, particularly
those whose agriculture
has been affected by war, ex-situ
genebank collections
are an integral part of agricultural
restoration.
Genebanks canplay a role in finding varieties lost in one part of a country but present in another andhelp reintroducethat variety where it is lost. Plant breeders and scientificplant
breeding
may or may not be involved in the process. Emerging models of plant
genetic resources conservation link in-situ and ex-situ conservationand in-situ andex-situ breeding.
An example of the linkage between in-situ/ex-situ
conservationon the one
hand and breeding on the other is provided by the narrowly endemic giant sequoia,
Sesquoiadendron giganteum (Lindl.)Buchh.
of California,
U.S.A. Based on isozyme
variability
populations
this species appeared to be inbred. It was recommended thatgeneticallydistinctsequoia populations
be maintained in ex-situ nurseries where they
could inter-breed,
and more vigorous outcrossed,
hybridseedlings,
rather thanseedlings
collected
in nature, would then be planted in reforestation
programs
the geneticallyinvigorated
species can be maintained
in-situ (Fins and Libby,
The objectiveof this paper is to highlight
where characterization
evaluation
within the context of germplasm conserved in-situ can contributewhere1982).
andto theoveralluse of agricultural
plant genetic resources in a sustainable
way.
Populations
and communities conserved in-situ can furnish material or act asa laboratory/monitoringsite which may be characterised
and evaluated to answer
questions which cannot be answered with germplasm conserved ex-situ. Central to thevalue of in-situ conservationsites is that biodiversity
conserved in-situ can help
unravel evolutionary
and ecologicalprocesses fundamental
to global sustainability.
In this paper we will highlight
three issues.
1. Enhanced information on spacial patterns of genetic diversity;
2. Temporal changes in genetic diversityof populations/communities;
3. The relationshipbetween genetic diversityof plants in relation
to other organisms
and ecologicalfactors.

1. Spacial

Patterns of Diversity
Plants conserved in-situ permit details of spacial patterns of diversity
to beanalysed, both at the populationand community level, in a way which is not possible
to do for material
conserved ex-situ. Population
genetic diversity
is criticaltounderstanding
evolution since populations
are the basic unit of evolution(Harper,1977). Spacial diversity
is relevant to sustainableagricultural
production systems andhas received increasing
attentionin the on-going debate regarding
the value ofgeneticallyheterogeneous
populations(e.g. Trenbath, 1974; Tilman et al.,1996).
At the individual
populationlevel species differ in genetic diversity
as a resultof many factors such as breeding system, population
size and age (Matsuo, thisvolume; Loveless and Hamrick, 1985). Among the perennial
wild relativesof rice
genetic polymorphism
based on RAPD banding of outcrossing
Oryza rufipogon (AAgenome) is very high at the individual
populationlevel, particularly
in areas where
this species grows sympatricallywith rice (Fig. 1a). Inbreeding
Oryza officinalis (CCgenome) is also a perennial, diploid species but has very little genetic polymorphism,
as revealed by RAPD banding, at the individual
populationlevel and also between
populations
over a wide area of west Malaysia (Fig. 1b). Clear polymorphic
bandingdifferences
begin to emerge for this species in populations
from geographicallyisolatedareas in east and west Malaysia (Fig. 1b) (cf. discussion
by Okuno in thisvolume on Aegilops in Central Asia and Caucasia).
Studies of spacial genetic variation
provide information
relevant to:Fig.1

Variationin polymorphism

Data analysed

using

Both dendrograms

at the DNA level (RAPD)NTSYS software and the DICE coefficientwere create using(A) O. rufipogon. Nine plants
on bands polymorphic(B)

O.officinalis.

banding

patterns

for two Malaysian

revealed

Oryza species.

was used to prepare

the matrix.

the UPGMA method.

from one Malaysian

population

could be uniquely

identified

by 15 primers

only

based

by 7 primers.

Based on 28 polymorphic

bands

were found among the 48 plants

revealed

analysedfrom 8 populations.

15 different

-samplingwhen collecting(e.g.

Brown and Munday, 1982);-development of core collections(both ex-situ and in-situ);-comparative and detailed
studies give insights
into the genetic characteristics
ofspecies which may be valuable for crop improvement. In the case of Oryza, forexample why has the CC genome of Oryza repeatedly
undergone allopolyploidy
events leading to stable new species but not the AA genome of Oryza? Why do Vigna
species in the tropics show a great deal of genetic variation
within and between
populations.
Whereas, temperate species such as Vigna angularis var. nipponensis
show relatively
littlepopulation
variation
despitegiving rise to a cultigen
andbelongingto a crop-weedy wild species complex (Tomooka et al., 1998);-detailedstudies at the population/site and regional level can reveal associationsofcharacters with ecologicalconditions(Annikster
et al., 1991);-studies of many traits may reveal useful spacial differencesamong agronomicallyuseful traits (Brown et al., 1978). The reasons why there are spacial differencesamong traits can lead to a better understandingof genetic diversity
in relation
toenvironmental factors. Such knowledge may also lead to an understandingof how to
deploy genetic diversity
from breeding programs.
Recently we found intra-population
variation during a collecting
mission for
Vigna genetic resources in Japan. In Mie prefecture,
Japan, a small populationofweedy Vigna angularis
had two plants with unusuallylarge and plentiful
root
nodules(Vaughan
et al., in preparation).
However, other plants in close proximity
did
not have any noticeableroot nodules (Fig. 2). The reasons for this intra-population
diversity
is now under investigation.
2. Temporal

Changesin Genetic

Diversity,

a. Bottlenecks
A recent paper has challenged
conventional
thinking
regardingthe genetic
consequences of a population
bottle neck by suggesting
that, in some circumstances,a genetic bottleneck
can lead to increased
genetic diversity(Carson,1990).
Crop
domestication,
represents
a genetic bottleneck(Tanksley
and McCouch, 1997), andcan lead eventually
to genetic diversity
between wild, weedy and cultivated(Pickersgill
and Heiser, 1976; Beebe

not found in the wild, particularly
if geneflow
relativesis possible
after domestication
et al., 1997).
Ex-situ conservation
involves

applyinga severe bottleneckthe domestication
process.

to populations
An accession

but lacks the dynamics which are seen inconserved ex-situ represents a population

which has undergone a severe bottleneck.
The consequences of this can lead to
immediate or almost immediate extinctionof geneotypes
due to non-compatibility
with the environment to which the genetic resources are taken (Morishima,
thisvolume). Evaluation of a range of genetic parameters can enable us to determine thegenetic consequences of the ex-situ conservation process (Breese, 1989). In-situ sites
enable the consequences of natural genetic bottlenecks,
such as colonization,
to befollowed. Early stages of colonization,
may give information
which is pertinent
tosuccessful long term ex-situ conservation and help in understandingof quantitative
genetic characterswhich are usually been neglectedduring characterization
andevaluation of plant genetic resources conserved ex-situ.
Polans and Allard (1989) have furnished empirical data on what happened aftera genetic bottleneck.
They restricted
the population
size of Lolium multiflorum for 4
generations.
The genetic consequenceswere generally
what might have been
expected, for example, a loss of allozyme alleles.
However, in some of theirexperimental
populations
there was an increase in genetic variance of quantitative
traits. One explanation for this counter-intuitive
result is that "the increase may result
from conversion of balanced epistatic
variance to additive
variance that becomes
immediately
available
to selection".
Such information
is useful because of the rapid
advances in both understanding
and using various types of genetic traits both incultivatedand wild relativesof crops (Tanksley
and McCouch, 1997). Plant genetic
resources conserved in-situ can provide the materials which can enable more complex
genetic
characterizationand evaluation,
not possible
with germplasm conserved
ex-situ.
b. Rapid evolution
Weeds, though not exotic, are very useful for studying
evolutionarychange.
While plant breeding
representsmans process for speeding up evolution,in natural
conditions
plants can evolve very quickly.
Rapid evolutionof plants in natural
conditions
may provide useful information
for adaptive plant breeding.
Cody and Overton (1996) have demonstrated very rapid evolution as a result of
naturalselectionin Lactuca.

Islandpopulationsof Lactuca murialis

evolve distinctlylarger achenes and a smaller pappus than mainland populations.
which differenceswere clearly detected was only 5 generations.

The time scale overOur recent studies on the evolution and diversity of weedy rice in Malaysia haveshown that this weed has emerged rapidly from cultivatedrice over about a 5 year
period. The rapid emergence of weedy rice is likely to be the consequence of strong
artificial
selection
for shattering
due to the practice of volunteer seeding (allowing
shattered seeds to contribute to the subsequent crop). This practice was most prevalent
in the mid 1980's and weedy rice was recognised by 1990, three years after thispractice peaked in MUDA the main rice growing area of Malaysia (Abdullah
et al.,
1996; Watanabe et al., 1996). Weedy rice is highly heterogeneous (Fig. 3) and showsgroups which may indicate the weed has arisen several times or that it is in an earlystage of differentiation(Vaughan et al., 1995). Similarly,
Oka and Morishima (1971)
reported
that indica-japonica
differentiationin rice could occur after only 7
generations.
Evaluators of germplasm and plant breeders screen their germplasm accessions
or segregating
populationsin adverse conditions
to enable selection
for complex
traits. However, if germplasm or segregating
populations
are removed from adverse
conditions
complex traits may be quickly lost (see Morishima this volume). In-situ
monitoring studies can furnish material which can provide answers to questions on
how rapidly or slowly populations
adapt/evolvein response to particularfactors.
Comparison of the genetics
of complex traits as they occur naturally
and whenremoved from the stresses to which they are adapted may be useful informationforboth conserving genes ex-situ and plant breeders.
3. Interactions
a. Allopatricresistance
In-situ conservation
provide the opportunity
to evaluate interactions.
It hasalmost become a principal of PGR work that one looks for resistance genes in centersof diversity
or where pest/pathogens
and crops occur sympatrically.
However,
resistance which is derived from co-evolution is essentiallyof the gene-for gene type(see Tosa this volume) and therefore
likelyto be readily
overcome by thepest/pathogen
in an agricultural
setting. Agriculture
is replete with examples of single
gene resistancebreaking down (Bonman et al., 1992). Correcting
this can be veryFig. 2. Two plants from one small populationcollected

Fig. 3. Fieldin Mie Prefecture,of rice in the MUDA irrigation

heterogeneous

weedy rice.

of a weedy form of Vigna angularis

Japan, showing

variationin nodules

area of Peninsular

Malaysia

on the root system.

heavily

infested

withTable 1. Examples(adaptedof host plantfrom Harris,

resistancewhich apparently

evolvedin the absenceof the pest or virus

1975)

H o stP e st o r v ir u s

R e sista n c e r e fe r e n c e

M a lu s sy lv e str is M ill.

E m p o a sc a fa b a e

S c h o e n e a n d U n d e rh ill (1 9 3 7 )Z ea m ay s

O s trin ia n u b ila lis

P a in te r ( 1 9 5 1 )G ly c in e m a x

E p ila c h n a v a r iv e s tr is

K o g a n (1 9 7 2 )O ry z a s a tiva

h oj a b la n ca (v iru s )L in g (1 9 7 2 )R u b u s sp .

A m p h o ro p h o ra ru b i

K n ig h t e t a l.. ( 1 9 6 0 )Fig.4.

Distributionof the green leafhopper

found in O. sativa accessions
spp. basedon Nasu, 1969)(Nephotettix

originating(fromspp.)In

Asia and regionalfrom Asia conserved

Vaughan,variation

at IRRI (distribution

in resistanceof Nephotettix

1991)

expensive. Finding durable resistance,
by definition,
takes a long time.
Allopatricresistanceis fortuitously
derived from pleiotropic
effects of genes
maintained due to natural selection
pressures unrelated
to the pest/pathogen(Harris,1975). Such resistance may therefore be difficult
for the pest/pathogen
to overcome.
Harris (1975)
has given many examples of successful allopatricresistancefor insects(Table 1). In rice, resistanceto the green leaf hopper is found in varieties
where thepest is not found (Fig. 4). In addition,
resistanceto rice hoja blanca disease, of theAmericas, was found in japonica
cultivars
which evolved in Asia where the virus is
not present(Vaughan,

1991,

Vaughanet al., 1997).

Seeking resistance genes in centers of diversity
may be counter productive. By
studying the processes of evolution in a comparative way new concepts may emerge
to add to Vavilovian Centers of Diversity and the Gene-for-Gene hypothesisof Flor.
Populations
in-situ at centers of diversity,
centers of cultivation
or edges of diversity(distribution)/cultivation
can provide material which will enable such new concepts
to emerge. A knowledge of distributionof allopatricand sympatric resistance can help
determine what populations
to conserve.
b. The costs of resistance
IR8, which had very few genes for resistance to pests and diseases, was quickly
followed by a series of IR varieties which had an increased numbers of genes for pest
and disease resistance.
However, for a long time IR8 remained the highest yieldingof the IR varieties- in the absence of pests (Chandler,1979).
Estimates of the average
selective
penaltiesof resistanceto 3 races of Rynchosporium secalis in a barleycomposite cross were 12, 24 and 9% per generation (Webster et al., 1986). If thefitness costs reported by Webster et al. are typical something must be occurring which
reduces this cost. High costs of resistance have implications
related to the distribution,
search for, use and deploymentof resistance genes.
To unravel the intra and inter-population
distribution,seasonal and long term
fluctuation of resistance genes in relation to pest/pathogen
dynamics in nature, longterm monitoring experiments
will be required (Burdon and Jarosz, 1986). In-situ
conservation sites can be used as experimentallaboratories
for such studies and may
furnish the type of informationnecessary for more sustainableagricultural
systems.
c. How is diversity
arranged
Ecologists
are beginning

to promote stable communities?
to obtain much data related to species

richnessandconsistentecological
function and productivity(Hanski,1997). Recent experimentresults concluded
that species richness and diversity
enables ecosystems to function
more consistently(e.g. Naeem and Li, 1997). In depth crop experiments have shownthat species richness leads to increased ecosystem productivity(Tilman et al., 1996).
Such results provide scientificbacking for the value of biodiversity.
Results of studying
ecosystems suggest that habitat
contributor
to the generationof species. The organisation

diversity
is an importantof different
habitatsmayalso be important. One example is the study of different
types of rain forest habitat in
the Cameroons (Smith et al., 1997) This study showed that geneflow from relatively
species poor ecotone habitats were one factor in generating
rain forest biodiversity.
Such experiments
to unravel in an holistic
and scientific
way issues related tobiodiversity
conservation
are criticalin helping formulate policy and strategies
forglobalconservation.

Conclusions
In this paper we have touched on a number of issues related
to thecharacterisationand evaluationof genetic resources from an in-situ conservation
perspective.
Enhanced gene-ecologicalunderstandingof PGR is fundamentallythe"in-situ perspective".
To paraphrase Harris (1975)
characterisationand evaluation of
PGR should involve " the minimum expenditure of money, time effort and materials".
What trends of the future will enhance characterisationand evaluationof PGR
conserved

in-situ(and ex-situ).

1. Techniques
In the future rapid, cheaper and safer methods which supply more information
can be expected which will enable greater through put and allow populations
in-situ
to be more easily studied (see Kresovichs this volume; Zheng et al., 1996; Ishii et al.,1990).Theability to take laboratory
methods to the field, particularly
DNA extraction,
will enable a wealth of new gene-ecological
data to be accumulated.
A majorconstraint
at present is the cost of some chemicals involved in new technologies,
such
as DNA amplification

enzymes.

2. Statistics.
Statisticalmethods which permit analysis
and synthesis
need to reach a newlevel of sophisticationin order understandecosystems, which are among the mostcomplex systems known (Maurer, 1998).
To study genotype
and environment
interactions
large number of replicatesare necessary. A paper dealing with diversity
and sustainability
in the North American prairie
ecosystems required
147 plots
involving
21 replicates(Tilmanet al., 1996).
Statisticalmethodologies
permit complex relationships
to be reliably
analysed relatively
cheaplywhich canwill also beneeded.
3. Shifts

in focus, from major genes to quantitative

trait loci (QTL),

from cultigens

towild species.
Tanksley and McCouch (1997) have presented several strong arguments why
QTLs and wild species will be important in the next century. Not only are QTLs now
readily
analysed,
their complexity
and unequal importance is being revealed.
In
addition,
it is now clear that wild species have "hidden" superior alleles which canbe introduced into elite breeding lines. Genebank curators struggle to conserve wild
genetic resources ex-situ because many wild species produce very few or no seeds
in ex-situ conditions.
In-situ conservation
is a essential
component of wild PGR
conservation (Brown et al., 1997;Morishima
this volume; Vaughan, 1994)
4. Research beyond Centers of Diversity.
While centers of diversity
are a logical laboratory
for PGR workers it isnecessary to consider other areas where plant genetic resources may be equallyimportant. In rice, New Guinea is not generally considered particularly
important andvery few collecting
missions for Oryza germplasm have occurred there. However,
New Guinea is the region with greatest Oryza genome diversity(Vaughan, 1991).
Similarly
Madagascar is not well known for Vigna genetic resources but is the sourceof one of the most important sources of resistanceto seed pests (Tomooka et al.,1992). In the future in-situ research at edges of genetic diversity
as well as centers ofdiversity

will

increase.

5. In-situ and ex-situ conservation.
While conservation is simplified
into in-situ and ex-situ conservationin reality
there are many types of conservation
which incorporate
aspects of both in-situ andex-situ conservationsuch as botanic gardens. In the drive to find long term safe
conservation
at reasonable cost a range of different
approaches
to conserve genetic
resources are being explored. It is now clear that characterisationand evaluationofgenetic

resources

conserved

in-situ,by for example medical

to pay for this conservation.
In the 1980's an eminent plant breeder

and scientist

companies,

is helping

was asked why at one of themost prestigious

university

on agricultural

ecology.

grown". Perhaps
now trying

agriculture
The reply

in the 1990's

to incorporate

holistic

conservation

evaluate

PGR

offacultieswas " Well

greaterin thein thisecological

USA had no academicStateawarenessonly

coursecorn and soybeans

exists.

PGR

thein-situ

ecologicaldimension

PGR.

In-situconservation

research

to characterize

as wellas geneticallywill

a trendecologicallyintoscientists

bemodels

in

are
are

for

the
and

the

new

millennium.

Editors
publications

note:

This

paper

was updated

that

appeared

afterduringthe paper

the editorial

process

was originally

written.

to takeaccountof relevant

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DNA isolationforEvaluationof Interactions

Y.TOSA
Laboratory of Plant Pathology,

between Diverse Plants

Faculty of Agriculture,

Kobe University,

and Other Organisms

Kobe

Abstract
Resistanceto the wheatgrass powdery mildew fungus, Blumeria graminis f.sp.
agropyri has been found in wheat. Four genes in wheat that control this resistance have been
designatedPm10, Pm11, Pm14 and Pm15. Subsequently,
four avirulence genes were foundin f.sp. agropyri that correspond to these resistance genes. These results suggest that this typeof resistance is controlled by the gene-for-gene relationship.
The geographicaldistributionofthese resistance genes showed a pattern that corresponds to Vavilov's gene center theory. The
possible
relationshipbetween the diversityof plant genotypes and the establishmentof
specialized
forms of the parasite is discussed.
Introduction
Plantsare surroundedby many microorganisms,
which are air-borne orsoil-borne,
pathogenic
or non-pathogenic.
For plants
the plant
pathogenic
microorganism is one of the selection
pressures that have affected
their evolutionary
processes. Plants have modified themselves to resist such microorganisms
or adapted
themselvesto coexist with them. Conversely,plant pathogens
have survived under
severe selection
pressure from plants. Plant pathogens
have continuously
modifiedthemselves to overcome plant resistance,
which has resulted in the establishmentofhost specific forms of the pathogen. In this paper an example of such interactions
will
be described,
those between Blumeria graminis and graminaceous plants.
Blumeria graminis(=Erysiphe
graminis),the causal agent of powdery
mildew,Isolates

is found on such graminaceous plantsfrom these hosts are morphologicallyas wheat, barley, rye, oat, wheatgrass.
the same, but distinct
in their hostranges; isolates
from wheat are parasitic
on species of the genus Triticum only, while
those from barley are parasitic
on species of the genus Hordeum only. Suchhost-specificforms on wheat, barley, rye, oat and wheatgrass are designatedas forma
specialis(f.sp.)
tritici,
hordei,secalis,avenae and agropyri,
respectively.
This
relationship,
forma specialis- genus specificity,
is very strict, at least in Japan. How
has this

strictrelationshipevolved?

Identificationof Genes Controlling

Powdery Mildew Fungus
For analyses
f.sp.

triticithe Resistanceof Wheat to the Wheatgrass(wheatfungus)mildew

and f.sp.

agropyri(wheatgrass
mildew fungus) were chosen since they are inter fertile (Hiura, 1978).
First, genes that controlled
the resistanceof wheat to the wheatgrass mildew fungus
were determined(Tosa et al., 1987, 1988; Tosa and Sakai, 1990).
Generally,asusceptible
wheat cultivar
is necessary for such an analysis,
but common wheat
cultivars tested were all resistantto the wheatgrass mildew fungus. So, we crossed thewheatgrass mildew fungus with the wheat mildew fungus, produced their F1 hybrids(Tosa, 1989a) and used some of them for analysis.
The common wheat cultivars
tested were Triticum aestivum 'Norin 4','Chinese Spring1, 'Norin 10' and 'Red Egyptian'(Table 1). These varieties
were allsusceptible
to the wheat mildew fungus, isolate Tk-1, and resistant to the wheatgrass
mildew fungus, isolate
Ak-1. When inoculatedwith a hybrid culture,
Gw-34,
however, Norin 4 was resistant
but Chinese Spring was susceptible;
this hybrid
culture revealed differences
detectable
at the phenotypiclevel between these two
cultivars and made genetic analysis possible. When an F2 population
derived from thecross, Norin 4 x Chinese Spring, was inoculated with Gw-34, resistant and susceptible
seedlings
segregatedin a 3:1 ratio, suggestingthat a major gene is involved in theresistanceof Norin 4 to Gw-34. This gene was found to be locatedon thechromosome ID and was designated Pm10. Pm11 in Chinese Spring, Pm14 in Norin
10 and Pm15 in all three cultivars
were identified
in a similar way.
What kind of resistancegenes are Pm10, Pm11, Pm14 and Pm15? Since
Norin 4, Chinese Spring and Norin 10 are susceptible
to the wheat mildew fungus
these genes must be resistancegenes to the wheatgrass
mildew fungus, Blumeria
graminis f.sp. agropyri.
Evidencefor Gene-for-gene Relationshipin forma specialis- Genus Specificity
Flor(1956)
proposed
an hypothesisthat "for each gene that conditions
reactionin the host there is a corresponding
gene in the parasitethat conditions
pathogenicity".
This hypothesis(gene-for-gene
hypothesis)
is now widely accepted
as a basic concept that explainsrace - cultivar specificity.
Does the forma specialis
genus specificity
also follow the gene-for-gene relationship?

TABLE1

A methodfor detecting

wheat

genes

for resistanceto f.sp.

agropyriWeexamined segregationof virulence on wheat cultivars in the F1 population
derived from the cross, the wheatgrass mildew fungus x the wheat mildew fungus(comprising
240 cultures),and in an F2 population
derived from a cross between two
representative
F1 cultures (Tosa 1989a, 1989b). All results obtained in these analyses
supported
the hypothesisthat forma specialis- genus specificity
follows thegene-for-gene relationship.
Avirulence genes corresponding
to Pm10, Pm11, Pm14
and Pm15 were detected,
and designatedPpm10, Ppm11, Ppm14 and Ppm15,
respectively(Tosa 1989a, 1989b; Tosa and Sakai, 1990).
The genetic mechanisms of the forma specialis- genus specificity

aresummarized as follows. The wheatgrass mildew fungus carries the avirulence genes
Ppm10, Ppm11, Ppm14 and Ppm15. When this forma specialis
is placed on wheat,these avirulence
genes induce the expressionof Pm10, Pm11, Pm14 and Pm15,
respectively,
resultingin resistantreactions. The rye mildew fungus (f.sp. secalis)does not carry Ppm10, Ppm11 or Ppm14. However, this forma specialis
carries
Ppm15, which induces the expression of Pm15, resulting
in resistantreactions inwheat (Tosa, 1994). On the other hand, the wheat mildew fungus carries none of theseavirulence genes and, therefore, can parasitize
wheat.
Relationshipbetweenthe Diversityof Plant

Genotypes

and the SpecificityofParasitism
As mentioned above, the parasitic specificity
of each forma specialis
is very
strict in Japan. However, Eshed and Wahl (1970) and Wahl et al. (1978) reported thatthe formae speciales
possessed wider host ranges in Israel than elsewhere. The
differencein the degree of specificity
between Japan and Israel may be attributable
to the diversityof plant genotypes.
To test this assumption,
we examined thegeographicaldistributionof Pm10, Pm11, Pm14 and Pm15 using 360 landrace wheat
cultivars
collected
from various areas of the world.
To determine the genetic constitutionof 360 cultivars is a very laborious task
if conductedby the traditional
method (i.e., crossing plants).
Thus, we applied thegene-for-gene relationshipto the identificationof resistance genes. The outline of thismethod is illustrated
in Fig.1. In the gene-for-gene
system there is one-to-one
correspondence
between resistancegenes and avirulence genes. Therefore, if a test
cultivar carries a resistancegene corresponding
to Ppm10, we can conclude that theFig.1

A method for identification
relationship.

of resistancegenes using

the gene-forgene

resistancegene is Pm10 (Tosa and Sakai, 1991). There is no need to cross plants.
Instead, you produce an hybrid population
among which avirulence genes segregate
and inoculate test cultivars
with the population.
Various genotypes occurred around Israel, or near the center of diversityofcommonwheat (Transcaucasia)(Tosa et al,. 1995).With
increasing distance from thisarea, however, the diversity
decreased. In the east, for example, Pm10 and Pm15
prevailedwidely, and genotype [Pm10 + Pm15] was predominant while Pm11 wasrarely found. This was a typical pattern that follows Vavilov's gene center theory.
Rye, wheatgrass and other species may also show similar patterns of distributionofgenotypes.
The differencein the diversityof host genotypes between the primary centerof diversity
and Far Fast may be closely related to the degree of parasitic
specificity
between Israel and Japan. We also suggest that the spreading of host plants from theirprimary center of diversity
played a role in the establishmentof the strictly
specificforms of the parasite. Probably, primitive formae speciales of B. graminis developedaround the Middle East, but each of them comprised diverse genotypes since hostgenotypes were diverse there. However, as hosts spread from their center of diversity
genotypic diversity declined, which in turn decreased the parasite diversity,
resultingin the establishmentof the strict forma specialis- genus specificity.
Acknowledgements
I would like
professor

to thankof Okayama

Specialthanksthroughout

Dr. S. Mayama, Professor

University,and Dr.H.

of Kobe University,

Heta, Okayama

are due to Dr. H. Ogura, Emeritus Professor

University,of KochiDr.U.

Hiura,

for valuable

University,

Emeritussuggestions.

for continuous

supportthe course of this study.

References
Eshed, N. and Wahl, I. 1970. Host ranges and interrelationsof Erysiphe graminis hordei, E. graminis
tritici and E. graminis avenae. Phytopathology
60:628-634.
Flor, H.H. 1956. The complementary
genie systems in flax and flax rust. Adv.Genet. 8:29-54.
Hiura, U. 1978. Genetic basis of formae speciales
in Erysiphe graminis D.C. In The Powdery
Mildews. (Spencer,
D.M., ed.), Academic Press, London, pp. 101-128.
Tosa, Y. 1989a. Genetic analysisof the avirulenceof wheatgrass
powdery mildew fungus on common
wheat. Genome 32:913-917.
Tosa, Y. 1989b. Evidence on wheat for gene-for-gene

relationshipbetweenformae speciales

ofErysiphe graminis and genera of gramineous plants. Genome 32:918-924.
Tosa, Y. 1994. Gene-for-gene interactions
between the rye mildew fungus and wheat cultivars.
Genome 37:758-762.
Tosa, Y. and Sakai, K. 1990. The genetics
of resistanceof hexaploid
wheat to the wheatgrass
powdery
mildew fungus, Genome 33:225-230.
Tosa, Y. and Sakai, K. 1991. Analysisof the resistanceof Aegilops squarrosa to the wheatgrass
mildew
fungus by using the gene-for-gene
relationship.
Theor. Appl. Genet. 81:735-739.
Tosa, Y., Tsujimoto,H. and Ogura, H. 1987. A gene involvedin the resistanceof wheat to wheatgrass
powdery mildew fungus. Genome 29:850-852.
Tosa, Y.,Tokunaga, H. and Ogura, H. 1988. Identificationof a gene for resistanceto wheatgrass
powdery
mildew fungus in the common wheat cultivar
ChineseSpring. Genome 30:612-614.
Tosa, Y., Kusaba, M., Fujiwara,N., Nakamura, T., Kiba, A., Noda, T., Furutsu, Y, Noguchi, H. andKato, K. 1995. Geographicaldistributionof genes for resistanceto formae specialesof ErysipheWahl,graminis in common wheat. Theor.Appl.
Genet. 91:82-88.
I., Eshed, N., Segal, A., and Sobel. A. 1978. Significanceof wild relatives
other wild grasses in cereal powdery mildews. In The Powdery Mildews.
AcademicPress,

London, pp. 83-100.

of small grains and(Spencer,
D.M., ed.),Plant Breeding Using Improved Informationfrom Evaluation
Genetic Resources: Lathyrus as a Model Genus

of Plant

A.G.YUNUSand M.S.SAAD
Center for Tropical Crop Germplasm, Department of Agronomy and Horticulture,
Universiti Pertanian Malaysia, 43400 UPM Serdang, Selangor, Malaysia
Abstract
Information
on genetic diversity
and biologicalrelationships
are important for
efficientuse of germplasm by plant breeders.
Multivariate
analysisof morphologicalcharacters will group related species together, the scanning electron microscopy (SEM) can
reveal further variation from pollen morphology and the seedcoats. Isozyme electrophoresis
can detect polymorphism.
Interspecifichybridizationand karyotype analysis can reveal
biologicalrelatedness.
From crossability
studies, gene pools of the cultivatedspecies can beobtainedand the genetic resources of the crop identified.
The implicationsof these
information is discussed in terms of germplasm use.
Introduction
The breeder will have a wider range of choice in selecting
the appropriate
kinds of diversity
for his breeding
programmes if more genetic diversity
is available(Hawkes, 1983).
It was noted by Allard (1970)
that genetic
variability
can beobtained from both natural and domesticatedspecies, within populations
and between
different
geographicalareas. Diversity
in crop plants is artificial
selection
by farmers
to different
culturaland ethnic preferences(Hawkes,1983),
as well as natural
selection
in response to geographical,
climatic
and edaphic
features.
Frankel andSoule (1981)
explainedthat diversificationof crop gene pools was mainly due todispersal
and the introgression
from wild and weedy relatives
and this has enrichedand broadened the scope for selectionof adaptations.
They however warned that due
to intensive
agriculture
and scientificbreeding,
the infraspecificdiversityof cropplants has decreased. Harlan and de Wet (1971)
grouped crop germplasm resources
into primary, secondary and tertiary
gene pools. The primary gene pool comprises
the biological
speciesthat includes
the cultivatedand the spontaneousraces. The
secondary gene pool includes those species that can be crossed with the primary genepool with at least some fertility
in the Fi. The tertiary
gene pool is more remotewhere gene transfer is not possible
unless special techniques(e.g. embryo rescue,chromosome doubling,
the use of bridging
species etc.) are used.
In this paper we shall attempt to illustrate
the importance of information
genetic
sativusdiversityand biologicalas an example.

relationshipto the breederonby using the crop Lathyrus

Evaluationof Genetic Resources of Lathyrus sativus
Lathyrus sativus L. is in the genus Lathyrus which contains 160 species and
45 subspecies(Allkinet al., 1986), divided
into 13 sections (Kupicha,
1983).
L.
sativus is placed in Section Lathyrus along with 33 other species.
Its widelydistributed
due to its use as a forage legume. Its use as a pulse is mainly confined to
India where it occupies an area of more than 1 million hectares (Lai et al., 1986).
In
India it is one of the most reliable grain crops and may be the only food available
insome areas when famines occur. This can result in excessive consumption and may
provoke the neurologicaldisorderknown as lathyrism(Ganapathy
and Dwivedi,1961).
There is a good scope for selection
and development of varietiesof low
toxicity(Kaul et al., 1986).
L. sativus is undoubtedly
a grain legume withconsiderable
potential
for improvement and one of the steps in this process is theevaluationof the germplasm resources of the crop.

1. Multivariate
Analysis of Morphological
Characters
Multivariate
analysis
was carried out on 271 herbarium
specimens
representing
29 species out of 34 species in the genus Lathyrus Section Lathyrus(Yunus, 1990).
Fourteencharacters(Table1) were analysed
to determine
morphologicalvariationwith the aim of determining
the species that may be moreclosely related to L. sativus. The data were analysed using the techniquesof Cluster
Analysis,(Euclidean
Distance plus Wards Method)and Principal
Components
Analysis, using the Clustan IC Computer package (Wishart,1978).
The phenogram
formed after Cluster Analysis is shown in Fig. 1. At a dissimilarity
coefficientof 36.5
eight clusters were formed (Table 2).
based

The analysis
identified
ten species as having a close affinity
to L. sativus
on morphologicalcharacters.
These species were L. amphicarpos, L.

Table1. Charactersused in Multivariate

Analysisof Lathyrus

SectionN o.

C h a r a c te r s

1

L e a f le n g th (s u b te n d in g flo w e r)2

L e a fle t le n g th3

L e a fle t b re a d th4

P e tio le le n g th5

L e a fl e t n u m b e r (p a ir)

6

L e a fle t s h a p e

7

L e a f v e n a tio n

8

S tip u le le n g th

Fig. 1. Phenogram

9

S tip u le b re a d th10

F lo w e r n u m b e r/p e d u n c le

11

P e d u n c le le n g th

12

F lo w e r le n g th13

C a ly x le n g th

14

C a ly x te e th len g thformed after Cluster

OTUs with 14 characters

Analysisof morphological(Euclidean

distance

data of the Cluster

Lathyrus

plus(Herbarium

survey).

Ward's Method)of all 271

Analysis(Yunus,1990).

Table2. Species

composition

formed by cluster

all 271 OTUs with 14 characters
S p e cie sanalysisof morphological(Euclidean

distanceplus Ward's method)data at a dissimilarity

A c c e s s io n scoefficient1

L .a m p h i c a r p o s

55

L .a n n u u s

31

5

L .b a s a l ti c u s

2

2

L .b le p h a r i c a r p u s

99

L .c a s s iu s

65

L .c h lo r a n t h u s

42

L .c h ry s a n th u s

3

L .c i c e r a

26

L .c i r r h o s u s

5

L .g o r g a n i

13

L .g r a n d if lo r u s

7

L .h e te r o p h y l lu s

5

L . h i e r o s o ly m i ta n u

4

L . h i r s u tu s

18

4

L . h i r tic a r p u s

22

L . la t ifo l iu s

8

1

3

L . ly c i c u s

3

1

2

L . m a r m o r a tu s

8

L . m u lk a k

1

L . o d o r a tu s

6

L . p s e u d o ‑ c ic e r a

6

L . r o tt u n d if o l iu s

6

L .s a ti v u s

47

21

L .s te n o p h y ll u s

55

L .s y l v e s tr is13

L . tin g it a n u s

10

L .t r a c h y c a r p u s

2

L .t u b e r o s u s

13
3

T o ta l

27 1

of 36.468

C lu s t e r s f o r m e d

e x a m in e d

L .u n d u la tu s

of

2

3

4

25

1

6

8

2

7

2

1
1

7

7

1

1
18

5

4

6
7
4

1

4
12

2

1

3

8
1
6
5

1
5
25

1

5

6

2
1

1

1

1

8
1
13

99

4 1

52

13

1

2

28

26

6

6

marmoratus, L. pseudo-cicera
demonstrated that species
the same cluster.

and L. stenophyllus.

Principal

which were presumed to be closelycomponents
relatedanalysis

were broadlyin

2. Scanning Electron Microscopy(SEM)
Besides the charactersused in the multivariate
analysis,
variability
can beshown in the species of Lathyrus Section Lathyrus using seedcoats and pollen as
observed under SEM (Yunus, 1996).
Seedcoatsand pollen morphology
were
observed using the 'Hitachi
2300' SEM. Untreated specimens were mounted onaluminium stubs using double tape and "sputter" coated with gold. Twenty-one
species of Lathyrus Section Lathyrus were studied for seedcoat charactersandseventeen species were analysed for pollen morphology.
Seedcoats
The seedcoat characteristicsof the 21 species are summarised (Table 3).
Papillose
testa ornamentation,
the standard pattern occurred in 16 species (76 %) of
Lathyrus Section Lathyrus observed. In five other species, L. annuus, L. cassius, L.
chrysanthus, L. hierosolymitanus
and L. hirsutus, secondary features were observed
and the papillae
became distorted.
This ornamentation
could be observed at a lower
magnificationand different
pattern types could be seen in these five annual species.
In 8 species with papillose
testa ornamentation
formed low mounds and differedslightly
from L. sativus.
From the seedcoat characteristics
under SEM the species which can beconsidered closely related to L. sativus are L. amphicarpos, L. basalticus,
L. cicera,
L. marmoratus, L. pseudo-cicera, L. chloranthus
and L. tingitanus.
The first five of
these species were also closely relatedcharacters from the herbarium survey.

to L. sativus

basedon other morphologicalPollenfeatures

Variationin size, shape and exine ornamentation
are summarised (Table 4). The size of the pollen

variableand the shape ranged
The mesocopial
ornamentation

was observed and thein the section was quite

from elliptic
to rectangular-elliptic
was either reticulate
or rugulate.

and rectangular.

Table 3. Characteristics
electron microscopy.

of seedcoatsin 21 speciesof Lathyrus

SectionLathyrus

observed under scanningS p e c ie s

H a b itT e s ta o r n a m e n ta tio n

L . a m p h ic a rp o s

A nn u al

P ap illo s e

L . b a s a ltic u sA

P ap illo s e

L . c h lo ra n th u sA

P ap illo s e

L . c ic e raA

P ap illo s e

L . m a r m o ra tu sA

P ap illo s e

L . p se u d o ‑c ic e r aA

P ap illo s e

L . s a tiv u sA

P ap illo s e

L . tin g ita n u sA

P ap illo s e

L . g orgo n iA

P ap illo s e , lo w m o u n d s

L . o d o ra tu sA

P a p illo s e , lo w m o u n d s

L . c a ss iu sA

M o u n d s, rid g e s

L . c h ry s a n th u sA

M o u n d s, rid g e s

L . h ir su tu sAM o u n d s, rid g e s

L . a nn uu sAA re o late m o u n d sA

P itte d m o u n d s

L . h ie r o s o ly m ita n u s
L . c irrh o s u s

P e re n n ia l

P a p illo se , lo w m o u n d s

L . h e te ro p h y llu sp

P a p illo se , lo w m o u n d s

L . la tifo liu sp

P a p illo se , lo w m o u n d s

L . ro tu n d ifo liu sp

P a p illo se , lo w m o u n d sp

P a p illo se , lo w m o u n d s

L . sy lv e strisp

P a p illo se , lo w m o u n d s

L . tu b er o s u sp

P a p illo se , lo w m o u n d s, b e a d e dsu b sp . m in ia tu ss u b s p . ro tu n d if o liu s

In reticulate
in rugulate
pollenirregularlypollen the sculpturing
elements formed a netlike
the elements that formed the pattern were thickpattern whereas
and distributed(Faegriand Iversen, 1975).
From the analysis of pollen characters,
seven species were found to be closely
related to L. sativus, namely L. amphicarpos, L. basalticus,
L. cicera, L. marmoratus,
L.hirsutus,
L. odoratus and L. tingitanus.
The survey also showed three species (L.

Table 4. Characteristics

of pollenin 17 speciesof Lathyrus

SectionLathyrus

under scanningelectronmicroscope.
S p e c ie s

P o la r a x is x

P /E

E q u a to r ia lr a tioS h a p e (E q u a to ri a l v ie w )M e so c o lp ic
o r n a m e n t a tio n

a x is (m m )L . a m p h ic a rp o s

37 x 24

1 .5

R e c ta n g u la r‑e llip tic

R u g u lateL . b a sa ltic u s

43 x 25

1 .7

R e c ta n g u la r‑e llip tic

R u g u lateL . c ic e r a

43 x 24

1 .8

R e c ta n g u la r‑e llip tic

R u g u la te

L . h irs u tu s

42 x 23

1 .8

R e c ta n g u la r‑e llip tic

R u g u la teR u g u la te

L . m a rm o r a tu s

45 x 29

1 .6

R e c ta n g u la r‑e llip tic

L . o d o ra tu s

45 x 29

1 .6

R e c ta n g u la r‑e llip tic

R u g u la te

L . p s e u d o ‑c ic e r a

53 x 3 1

1 .7

R e c ta n g u la r‑e llip tic

R u g u la te

L . tin g ita n u s

43 x 32

1 .3

R e c ta n g u la r‑e llip tic

R u g u la teace. no .4 04

43 x 29

1 .5

R e c ta n g u la r‑e llip tic

R u g u lateace.n o .4 29

46 x 24

1 .9

R e c ta n g u la r‑e llip tic

R u g u la teR u g u lateL . sa tiv u s

ace.n o .4 30

43 x 26

1 .7

R e c ta n g u la r‑e llip tic

ace.n o .4 34

43 x 24

1 .8

R e c ta n g u la r‑e llip tic

R u g u la teace.n o .4 35

43 x 27

1 .6

R e c ta n g u la r‑e llip tic

R u g u la teace.n o .4 68

43 x 25

1 .7

R e c ta n g u la r‑e llip tic

R u g u la teace.n o .50 742 x 24

1 .8

R e c ta n g u la r‑e llip tic

R u g u la teace.n o .55 8

39 x 26

1 .5

R e c ta n g u la r‑e llip tic

R u g u la tea ce . n o . 5 8 040 x 25

1 .6

R e c ta n g u la r‑e llip tic

R u g u latea ce . n o . 5 8 840 x 25

1 .6

R e c ta n g u la r‑e llip tic

R u g u lateL . c a ss iu s

44 x 25

1 .8

R e c ta n g u la r‑e llip tic

R e tic u la te

L . ch lo r a n th u s

50 x 26

1 .9

R e c ta n g u la r‑e llip tic

R e tic u la te

L . ch ry s a n th u s

42 x 27

1 .6

R e c ta n g u la r‑e llip tic

R e tic u late

L . g o rg o n i

55 x 252 .2

R e c ta n g u la r‑e llip tic

R e tic u late

L .a nn u us

35 x 27

1 .3

R e c ta n g u la r

R e tic u lateL .

37 x 25

1 .5

R e c ta n g u la r

R e tic u lateL . c irr h o su s

32 x 25

1 .3

E llip tic

R u g u lateL . la tifo liu s

37 x 30

1 .2

E llip tic

R u g u la teh ie ro so ly m ita n u s

pseudo-cicera, L. cassius and L. gorgoni) which were similar in other morphologicalcharacters to L. sativus but differedin the pollen characteristics.
Analysisof seedcoats and pollen under SEM included
four species (L.
chloranthus,
L. tingitanus,
L. hirsutus and L. odoratus) which were not analysis formorphologicalcharacters (herbarium
survey). These 4 species appear to be closely
relatedto L. sativus.

3. Isozyme Electrophoresis
Isozyme polymorphisms
were determined within L. sativus (Yunus et al.,1991)
as previous morphologicalstudies (Jackson & Yunus, 1984) had shown that
this species is clearly differentiatedinto several distinct forms. Analysis was carriedout on 52 accessions which represent all the three flower types (blue, blue and whiteand white) and also a single plant with pink flowers. The accessions represent a wide
geographicalrange. Horizontal
starch gel electrophoresis
was carried out and six
enzymes were selected for detailed analysis after a preliminary
survey of 13 enzymes,
since they gave consistent results with this species.
Young leaves were used for extraction and absorbed on wicks before samples
were run on the starch gel. Staining
was carried out to demonstrate the banding
patterns which were assessed and polymorphism
calculated.
The six enzymes assayed were 6-phosphogluconate
dehydrogenase(6-PGD),
malate dehydrogenase(MDH), peroxidase (PRX), isocitrate
dehydrogenase(IDH),
glutamateoxaloacetatetransaminase(GOT) and galactose
dehydrogenase(GD).
Polymorphism
was observed for all six enzymes, and much polymorphism
wasrecorded for PRX and 6-PGD, while there was little polymorphismfor GOT. Thereis no apparent correlation
with morphology.
Furthermore,
the isozyme variation
could not be explainedby geographicaldistribution.
Althoughno formal genetic analyses of the isozyme

banding

patterns

wasmade, analogy with similar systems in closely related genera such as Pisum, Lens or
Vicia and other plant species enabled an estimate of the extent of genetic variation atparticularisozyme loci. The banding patterns which were observed represent allelicvariation

at severalloci.

4. Crossability
Studies
A total of 14 species were available
for crossing experiments to determine
biologicalrelationships.
Interspecifichybridization
was carried out to reveal thebiologicalrelationshipsof L. sativus with wild species in Section Lathyrus in an
attempt to define the gene pools of this under exploitedpulse (Yunus & Jackson,
1991).
Fifteen wild species from Section Lathyrus were used in the interspecificcrosses. The crossing technique used was that described by Cruickshank (1984).
The
sepal covering the keel was folded back, the keel excised and the anthers removed.
A stigma covered with pollen was removed from the male parent and rubbed on the
stigma of the female parent. After pollination
the remaining flower parts were kept
intact and covered with parafilm to avoid dehydration,
as well as contaminationbyforeign pollen.
Developing pods could be seen in successful pollinations
after oneweek, when the parafilm was removed. Successful crosses were indicatedby theformation of pods and seeds. However, in only two combinations,
namely L.
amphicarpos x L. sativus and L. cicera x L. sativus, were verified hybrids obtained(Table 5). Seeds were obtained from the crosses L. sativus x L. gorgoni and L.
latifolius x L. sativus, but following germination, the Fi hybrids were inviable.
In thecross L. chloranthus
x L. sativus, the F1 seed failed to germinate.
In othercombinations no hybrids were formed but producing empty pods or totally shrivelled
seeds or pollinations
failed completely.
Although the gene pool concept of Harlan and de Wet (1971)
for the classificationof cultivatedplants,of genetic
resources,
for purposes

was considered

the system is of equal value in considerationof theirclassification,
evaluation
anddocumentation (Smartt, 1990).
In terms of defining
the gene pools of L. sativus on
the basis of interspecifichybridization
reported in this paper, it was suggested that
(Yunus and Jackson 1991), the gene pool concept of Harlan and de Wet (1971)
wasinadequate
to encompass the range of interspecificrelationships
between Lathyrus
species.
Smartt (1980)
suggestedthat a quaternary
gene pool be introduced
toaccommodate the related species which form effective
genetic barriers but whose
resources may eventually
be exploitedby the techniquesof genetic engineering.
Afurther modification
was also suggested by Smartt (1986)
to provide even greater
distinction
within the tertiary
gene pool, where the order of the gene pool is equatedwith the relative
degree of effectivenessof the interspecificisolatingmechanisms.

Table 5. Interspecifichybridization

C rossbetween Lathyrus
N o. ofL . s a t iv u s x L . g o r g o n i

P od ssativus and 15 wild
P od sp o l l in

w ith

a t io n sseed s3 3

3

1

speciesin Section

Lathyrus

R em a rk s

S e e d g e r m in a t e d , b u t s e e d l i n g
in v i a b l e

L . s a t iv u s x L . a m p h i c a rp o s

12

1

0

x L . h a s a lt ic u s

10

3

0

x L . c ic e r a

3 1

1

0

x L . h i e r o s o ly m it a n u s

2 7

1

0

x L . h irs u tu s

13

1

0

x L . a nn u us

6

0

0

x L . c a s siu s

7

0

0

x L . c h l o r a n th u s

13

0

0

x L . c h r y s a n th u s

10

0

0

x L . l a tif o li u s

8

0

0

x L . m a rm o r a tu s

1

0

0

x L . o d o r a tu s

2 5

0

0

x L .p s e u d o ‑c ic e ra

11

0

0

x L . t in g it a n u s

16

0

0

L . a m p h i c a r p o s x L . s a t iv u s

17

11

1 1

L . c ic e r a x L . s a ti v u s

15

15

6

L . la t if o l iu s x L . s a t iv u s

10

1

1

L . c h lo r a n t h u s x L . s a t iv u s

11

1

1

L . a n n u s x L . s a ti v u s

3

2

0

L . m a r m o r a tu s x L . s a t iv u s

18

15

0

L . p s e u d o ‑ c ic e r a x L . s a t iv u s

14

6

0

L . ti n g i ta n u s x L . s a t iv u s

19

1

0

L . c a s s i u s x L . s a ti v u s

5

0

0

L . c h r y s a n th u s x L . s a t i v u s

8

0

0

L . h i e r ox sLo ly. sma it ta
iv un su s

1 6

0

0

L . h i r s u tu s x L . s a ti v u s

8

0

0

L .o d o r a t u s x L . s a ti v u s

12

0

0

L . b a s a lt i c u s x L . s a tiv u s

7

1

1

L . g o rg o n i x L . s a tivu s

20

9

1

E m p ty p o d s o r s h r iv e l le d s e e d s

F e r ti li z a t io n f a i le d c o m p le t e ly

V e r i f ie d h y b r id s o b t a in e d

S e e d g e r m in a te d , b u t s e e d l in g
in v i a b l e
S e e d o b t a i n e d , b u t d id n o t
g e r m in a te
E m p t y p o d s o r s h r i v e l le d s e e d s

F e rt il iz a t io n f a il e d

F 1 s u s p e c t e d s e lf

Based on the study by Yunus and Jackson (1991) the different
assigned to the three gene pools, as shown in Table 6.
5. Karyotype Analysis
Chromosome morphology

Lathyrusof L. sativus and other species

speciesin Section

were

Lathyrus

was studied
to determine
whether chromosomal differences
are relatedtohybridisation(Yunus, 1990).
Fifteenspecies from Section Lathyrus were used in karyotype
analysis.
Seeds were germinated in an incubator at 20•Ž and sown in vermiculite.
Roots werepretreatedin water at 0•Ž for 24 hrs and fixed in a fresh solution of absolute ethanol,
chloroform and glacial acetic acid in the ratio of 3:1:1 respectively
for 24 hrs. The
roots could be stored in 70% ethanol after washing in tap water. Further treatments
were hydrolysis
in IN HCI for 8 minutes at 60•Ž in a waterbath and after washingwith tap water the roots were stained with Schiff's
reagent for at least 15 minutes or
longer. Root tip squashes were made after macerating in a drop of 45% acetic acid.
All the species of Lathyrus analysed had 14 chromosomes and they were either
metacentric or submetacentric(Table 7). Secondary constriction
and satellites
wereobserved in some species. The majority of the species were asymmetrical. The studyof chromosome morphology has shown that L. sativus was similar to L. amphicarpos,
L. basalticus, L. cicera, L. gorgoni and L. marmoratus but differed from other species
because of the presence of secondary constrictions
or great differencein chromosome
length.
There is a correlation
between karyotypes
and the relationshipbetweenspecies in Section Lathyrus. In particular,
the karyotypesof L. sativus was similar to
L. amphicarpos and L. cicera, with which F1 hybrids were formed, as well as L.
basalticus,
L. gorgoni and L. marmoratus but which showed a lesser degree ofrelationshipwith L. sativus.

GeneralDiscussion and Conclusions.
Studies of morphologicalvariation
through multivariate
analysis(Cluster
Analysisand Principal
Components Analysis)of the species of Lathyrus Section
Lathyrus agree broadly with classicalapproach of Kupicha (1983).
Ten species were
found to have close morphologicalaffinity

to L. sativus.

Table 6. The germplasm

resourcesof Lathyrus

sativus,and ordinationsuggested

G en e p o ol

O rd in a tio n

C o n s titu e n ts

S p e c ie sI‑A

1 s t o rd e r

C u ltig e n

L . s a tiv u s

I‑B

2 n d o rd e r

W ild c o u n te rp a rt

U nk n ow n

II3 rd o rd e r

C ro s s c o m p atib le s p e c ie s p ro d u c in g

L . a m p h ic a rp o s

m o re o r le ss fe rtile h y b rid s

L . c ic e ra

C ro s s c o m p atib le s p e c ie s p ro d u c in g

L . g orgo n iv ia b le b u t ste rile h y b rid s

L . la tifo liu s

C ro s s c o m p atib le sp e c ie s p ro d u c in g

L . c h lo ra n th u s

III

4 th o rd e r
5 th o rd e r

by Smarttbased on the gene pool concept

de Wet (1971),

of Harlan and(1986).

in v ia b le h y b rid s
6 th o rd e r

O th e r re la te d s p ec ie s n o t p ro d u c in g

L . an n uu s

a n y h y b rid s

L . b a s a ltic u s
L . c a s s iu s
L . c h ry sa n th u s
L .h ie ro so ly m ita n u s
L . h irs u tu s
L . m a rm o ra tu s
L . o d o ra tu s
L . p s e u d o ‑c ic e ra
L . tin g ita n u s
O th e r S e c tio n L a th y r u s

7 th o rd e r

D is ta n tly re la te d s p e c ie ss p e c ie s( ?)O th e r L a th y ru s se c tio n s

These are L. amphicarpos, L. basalticus,
L. blepharicarpus,
L. cassius, L.
cicera,L. gorgoni,
L. hirticarpus,
L. marmoratus, L.pseudo-cicera
and L.
stenophyllus.
In addition
to these, L. choloranthus,
L. hirsutus, L. odoratus and L.
tingitanus
were identified
as similar
to L. sativus from the characteristicsof theirseedcoats and pollen under SEM and with the ten species already selected formed thebasis for crossability
studies.
L. amphicarpos and L. cicera are closely
relatedbiologicallyto L. sativus and hybrids with some fertility
were obtained.
These two
species were placed with L. sativus in the arrangement of species in Section Lathyrus
by Kupicha (1983).
Close morphologicalaffinities
between L. sativus and L. cicera
were shown by Jackson and Yunus (1984)
and indicatedby Zohary and Hopf (1988).
The resemblance between L. sativus and L. cicera was earlier noted by Davis (1970)

Table 7. Karyotypes

in Lathyrus

S p e c ie s

SectionLathyrus
C h ro m o so m e len g th

C h ro m o so m e ty p e(A rb itra ry u n its)2

M etacS ub ‑

e ntric

m etacen tric

1

6

nd

S atellites

M ea nR an g e(n = 5 )c on striction s

T .F
% *

A n n u a ls (D e lic a te )#L . a m p h ic a rp o s

y es

110

100 ‑13 1

37

7

130

11 8‑136

39

L . c ic e ra

1

6

107

100 ‑123

39

L . g o rg o n i

1

6

135

1 11‑157

40

95

8 8 ‑1 0 9

38

134

13 1‑137

46

L . b a s a ltic u s

L . m a rm o r a tu s

1

6

L . p s e u d o ‑c ic e ra

1

6

y es

yes

L . sa tiv u s
a c c .n o .4 0 4

7105

9 0 ‑1 2 039a c c .n o .4 2 9

7

y es

125

1 15‑13537

a c c .n o .4 3 0

7

y es

11 1

10 3‑13637

a c c .n o .4 3 4

7

y es

12 1

1 17‑12 838a c c .n o .4 3 57129

10 9‑13936

a c c .n o .4 6 8

7

y es

129

1 17‑140

38

y es

124

1 12‑14038

a c c .n o .5 0 77a c c .n o .5 5 87117

1 12‑12238

a c c .n o .5 8 0

7123

1 16‑13637

a c c .n o .5 8 87123

10 7‑13 9

37

39

A n n u a ls (S tu rd v) #L . an nu us

25

L . c es s iu s160

15 2‑17 17146

12 2‑16 139147

13 0‑166

40

L . ch lo r a n th u s

25

L . h ie ro so ly m ita n u s

1

6

yes

yes

13 1

124 ‑15343

14 1‑163

36

L . h irs u tu s

1

6

154

L . o d o r a tu s

1

6

159

13 9‑166

37

7

yes

158

13 9‑174

34

6

yes

2 06

1 9 7 ‑2 2 137

sum of short arm lengths

to the total

L . tin g ita n u s
P e re n n ia l (S tu rd v )L . la tifo liu s

1* Total form (T.F.)

is the ratio

chromosome length# Classificationin floral(Huziwara,

by Kupichacharacteristics

The wild origin

in percentageof the total

1962)(1983)

but in fruit L. sativusof L. sativus

is still

was more similar

unknown but the Balkan

to L. amphicarpos.

peninsula

was indicatedas a centre of domesticationof L. sativus by Kislev (1989)
who suggested
that asearch for the living wild progenitorof L. sativus should be made in this area, whichis also one of the places where L. amphicarpos and L. cicera are also native (Allkin
et al., 1985).
Pollen and seedcoats of L. sativus, L. amphicarpos and L. cicera as seen
under SEM cannot be differentiatedand the karyotypesof these three species are
strikingly
similar.
The resultsof crossability
studies
showed that the combinationL.
amphicarpos x L. sativus was more successful than L. cicera x L. sativus based on thepercentage
of hybrid seeds formed. However, the F1 of the latter combination
had
better pairingof chromosomes (Yunus and Jackson, 1991).
The most likely wildprogenitor
of the cultivatedgrass pea cannot be fully verified
on the basis of present
information.
The variation
in karyotypesof species in Section Lathyrus is generally
correlatedwith morphology
and crossability
and was consistent with the work ofDavies (1958)
and Yamamoto et al. (1984).
The results of intraspecifichybridization(Yunus and Jackson, 1991, however,revealed the presence of a barrier to gene flow among L. sativus accessions fromdiverse geographicalareas, but this could not be related to the different
forms ofspecies based on flower colour (Jackson and Yunus, 1984). The different
forms ofL. sativus were not correlated with phenotypic
isozyme polymorphism which was also
not related to geographicalorigin of the accessions,as opposed to that reported inLens (Skibinski
et al., 1984)
and Vicia (Amet, 1986).
L. sativus was highly
polymorphic
for two enzymes, namely Px and 6-PGD but lower for IDH, GD, MDH
and GOT.
The germplasm profile of L. sativus demonstrated
that the most economical
ways of using the genetic resources for its improvement is to exploit the first order
gene pool where the cultigen
itself is highly variable.
The second order gene pool,
the wild counterpartis still unknown. The third order gene pool consistsof twospecies with only some fertility,
indicatingthat gene transfer will be difficult.
Varieties
of L. sativus with low toxicity are available(Kaul et al., 1986) and perhaps
other agriculturally
useful characters can be found through evaluation of the cultigen
beforeusing other distantly

relatedspeciesof whichthere are very many in Lathyrus.

Acknowledgement
Wewould like to thank Universityof Agriculture
Malaysia) for the permission to present this paper.

Malaysia(Universiti

In Genetic

resources

Pertanian

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Questions

and Answers

in Session

1

Questions to Dr. Kresovich
Q. Can molecular markers be used to establish
definitetaxonomic relationships
whichwill not require subsequent revisions?
(Yunus)A. I don't expect any analyticalmethod to 'finalize' systematic
relationships.
However,
because evolution is dynamic, I expect biochemical
and molecular methods to be
useful in dissecting
ecologicalprocesses affecting
populationdifferentiation
andsubsequent speciation
events. (Kresovich)
Q. What is your personal opinion concerning purificationof germplasm mixtures?
(Ekanayake)
A. Genetic structure of accessions should be maintained
as close to the original
sample as possible,
recognizing
our limited
genetic
understanding
and theassociated costs of regeneration. However, we must acknowledge the limitationsof ex-situ maintenance. (Kresovich)
Q. Referring to my earlier question to Dr. Riley. Morpho-agronomic characterizationis sensitive
to environmental
factors while molecularcharacterizationisexpensive. Can you comment?
A. I think understanding
the genetic basis of characters and traits is critical
for thefuture. Any assay contributing
to this understanding
will have value. As I noted
asking the right questions
will lead to effective
use of the right analyticaltechnique. (Kresovich)
Q. What is a "core collection"?
Could you please explain the possibility
or utilityofmolecular analyses for determining
core collections.(Kikuchi)
A. The core collection concept was proposed by Brown and Frankel in the 1980's. To
foster improved use of large collections,
Brown and Frankel suggested
that asubset of accessions be establishedthat represent the collections
diversity.
Basedon calculations,
it was hypothesizedthat a core collectionof 10% of theaccessions
could represent
as much as 70% of the allelicvariationof thecollection.
Neutral biochemical
and molecular markers are effective for quantifying
andpartitioning
genetic variation
and would be useful when deciding which entries
warrant inclusion in the core. High throughput,high resolution
typing of genetic
resources aids curators to understandgenetic relationships
among numerous
populations
and individuals.
(Kresovich)Q. I would like to ask you about molecular techniques to characterize
and evaluate
diversityof plant genetic resources in the future. (Okuno)
A. Newer molecular techniques
will continue to be developed which are quicker,
more accurate and precise, more reproducible
and cheaper. Continuingresearch
will be fruitful.
More importantly,
the technique of choice will be dependent on
what the biologicalquestion is and what level of genetic resolution
is needed tosolve the question. The future is bright because of technologicaladvances. I
expect improved genotype and gene discovery
to occur. The techniqueandtechnology
are important, but the ultimate generationof genetic informationismost critical.
(Kresovich)
Questions to Dr. Vaughan
Q. Are there intermediatetypes between wild and cultivatedtypes or weedy types of
Glycine? (Morishima)
A. There is a type of Glycine which has been considered to be a weedy type in China
called "G. gracilis".
This appears to have an intermediate
plant type, but seems
to be restricted
to China. In addition,soybeans in the US were first used
extensively
for fodder and the plant type is more similar to G. soja than cultivars
used for its seeds. (Vaughan)
Q. In soybeans have any alleles been found which are specificto wild or cultivatedsoybean? (Morishima)
A. Yes (Shimamoto)
Q. Distinct
merits of in-situ conservation
still suggest a need for a close linkage
between phenotype, genotype and molecular level of evaluation. In addition,
close
complementarity
between in-situ and ex-situ conservation.
Could you comment.
(Hayashi)A. Studying genetic resources in-situ provides research opportunities
quite distinct
from germplasm held in ex-situ collections.
I make a clear distinction
betweengenetic
resources in-situ and genetic
resources conserved in-situ.
Genetic
resources conserved in-situ are not usually within a strict conservation
program.
The types of in-situ
Certainly
genetic"conservation"
are more variable than ex-situ conservation.
resources in-situ can furnish excellent
material for gaining

betterof the genetic

understanding

relationships.

We need to balancescientificendeavours to ensure that attributes,

such as what governs phenotypic

plasticity,

are not ignored as our studies at the molecular level attain greater precision.
While ex-situ and in-situ conservation,
and all the types of conservation
inbetween, complement each other, I think we are perhaps not yet fully aware how
germplasm conserved in-situ can be more than potentially
useful material forplant breeders.
In addition,
genetic
resources in situ can be of use inunderstanding
critical issues related to such topics as sustainableagriculture
andenvironmentallysafe agriculture(e.g. issues related to the release of transgenic
germplasm). (Vaughan)
C. In situ conservation
is important to know the crop we want to conserve, be it
cross- pollinatedor propagatedasexually
like sweet potatoes,
then in-situ
conservation can be effective. In addition one should document the culture of thecommunity in which the crop grows, the farming system, their conservation
andmarket demand so that these will be factors for sustainablein-situ conservation.
In-situ conservation,
however, should be backed up with ex-situ conservation.
(Mariscal)
C. As you rightly say in-situ conservation of germplasm in farming communities (socalled on-farm conservation,
which includes not just cultivatedplants but also
wild and weedy relatives
in and around the field)
requiresmultidisciplinary
collaboration
for fully appreciating
the in-situ conservation system(plantscientists-geneticists,taxonomists, social workers and anthropologists).
Sweet potatoes is
one crop in which such an inter-disciplinary
approach has been very successful
in enhancing our knowledge of the whole system. (Vaughan)
Q. Highest yields for rice are outside the area of origin. Is there a case for in-situ
conservation outside the area of origin. (Chaudhary)
A. Yes. Papua New Guinea is an area diverse in sweet potatoes but not where the crop
originated.(Riley)
C. Regarding in-situ conservation,in the Chiloe area of Chile, I noticed that farmers
plant imported
breeds of white potato. The breeds, such as "Desiree"forcommercial production,
while they purposefully
grow native primitive
varieties
like "Papa Cacho" (meaning horn shaped potato)
for their home consumption
because of its good taste and suitability
for traditional
recipes.
To support this on-farm conservation UACH (Austral Universityof Chile) anda nongovernmentalorganization
CET (Center of Technology
and Research - a
farmers cooperative)
have a special program. They are collecting
diversityofpotatoes grown in farmers backyards,
classifying
them in field experiments
andselecting them for redistribution
to farmers the improved lines that are more stableproducers and can resist climatic disasters or newly occurring biohazards.(Suzuki)
Questions to Dr. Tosa.
Q. What do you mean, genetically,
by wild plants are less specialized
hosts?
(Vaughan)A. For example, some individualsof Aegilops spp. are susceptible
to two or moreformae speciales,
so they are less specialized
hosts as individuals.
Further,
individualsof Aegilops spp. show various patterns of reaction to formae speciales;
one shows the wheat type pattern and another shows the rye type pattern etc. So
the genus Aegilops is less specialized
host as a population.
(Tosa)Q. Could you explain the merits of in-situ conservation from the stand point of the"gene for gene theory". (Kikuchi)
A. Around the center of origin of host plants, there are diverse genotypes of theirpathogens (diverse avirulence genes), which produce diverse selection pressure,and diverse "niches". Therefore, in-situ conservation
at the center of origin may
be an easy method to maintain the diverse corresponding
resistance genes. (Tosa)

Topic2:

Plant Genetic Diversity
Evaluation
GeographicalandEcologicalconsiderations

Chairpersons
H. Shimamoto
P.N.Gupta-

Geographicaland Breeding Trends within EurasianGermplasm Revealed by Molecular Markers
P. P. STRELCHENKO,

CultivatedBarleyN. K. GUBAREVA, O. N. KOVALYOVA and A. GRANER*

Instituteof Plant

Industry*FederalCentre for BreedingD-85461

Grunbach,(VIR),

44 Bolshaya

Research

Morskayaon CultivatedPlants,Street,Institute

190000St.Petersburg,for ResistanceRussiaGenetics,

Germany.

Abstract
Knowledge of genetic variability
within a crop species is invaluable
for its
improvement. Restriction fragment length polymorphisms(RFLPs) and hordeins have beenused to characterize genetic diversityof 93 barley cultivars and landraces originatingfromdifferent regions of Russia and neighbouring countries. The RFLP banding patterns from 70
clone-enzyme combinations(41 map-based DNA clones, restriction
enzymes Eco RI and
Hind III) yielded in total 335 polymorphic
fragments. These were used to generate a genetic
distance matrix, which was used in both cluster and principal
coordinate
analyses. Bothanalyses
clearly separated
all accessions
into two major genetic groups, which are
geographicallylinked with oriental and occidental
regions of Eurasia. This confirms theexistence of two principal
paths in the evolution of cultivatedbarley. The occidental-typegroup consisted of more accessions and were clearly divided into two-rowed and six-rowed
forms on the basis of spike morphology. Among major genetic groups, further sub-groups
were apparent. These were cultivars with a similar pedigree background which clustered
together. The use of RFLP and hordeins analyses for determining
barley genetic variability
are discussed.
Introduction
Genetic improvement of crops by man can be regarded as directed
evolutionacting upon the existing
genetic variability
in the germplasm. In order to optimize andaccelerate
breeding,
it is essential
to screen, evaluate
and classify
the genetic
variability
availablein the germplasm. This is especially
importantfor collecting,
maintaining
ex-situ and studying plant genetic resources in national and international
germplasm programs.
Assessment of genetic variability
between individuals
and populations
hasbeen based on the analysis of pedigree records, morphologicaltraits and more recently
on molecular markers. However, pedigree data of lines are not always available.
For
example, landraces represent a large part of germplasm collectionsof many crops andmay be a rich source of genetic variation
for cultivar
development.
Moreover,pedigree data do not account for the effect of selection, mutation and random genetic
drift. Use of morphologicaltraits for plant diversity
analysishas been criticizedbecause genetic control is largely unknown and expression depends on environmental
factors. Among biochemicalmarkers, polymorphic
proteins such as isozymes andstorage seed proteins have been successfully
used in different
crops to characterize
genetic variation in numerous taxonomic and population genetic studies (see Konarev
et al., 1996, for review). However, proteins often failed in the classificationof crops
because of the small number of available
marker loci, which provided only poor
genomic coverage. Recently
DNA-markers such as restriction
fragment length
polymorphisms(RFLPs) and random amplified polymorphic
DNA (RAPD) are being
successfully
used for assessment of genetic diversity
in cultivatedplant species. Suchmarkers have the advantage of being generally
independentof phenotype
and, if
representativeof the entire genome, can provide a comprehensive
survey of thegenetic variation present in a sample of cultivars.
In barley (Hordeum vulgare L.) high-density
genetic marker maps are being
constructed using both RFLP and RAPD markers (Graner et al., 1991; Heun et al.,
1991; Tragoonrung et al., 1992; Graner et al., 1993; Kleinhofs et al., 1993). Recently,
several studies have examined the genetic variation in cultivatedand wild barley with
RFLP (Graner et al., 1990; Liao and Niks, 1991; Pecchioni et al., 1993; Zhang et al.,
1993; Melchinger
et al., 1994) and RAPDs (Dweikat et al., 1993; Tinker et al., 1993;
Gonzales and Ferrer, 1993; Song and Henry, 1995). However these studies werelargely restricted
to the analysis of elite barley germplasm adapted to Western Europe
or North America. But, cultivatedbarley is one of the oldest, most widely grown andpolymorphic
crop species and was domesticatedin Asia and principal
centers of its
genetic diversity
are situatedthere. N.Vavilov was the first to begin
world-widecollecting
and studying of genetic diversityof many crops including
barley. On thebasis of his work principal
world centers of barley diversity(gene-centers)
weredetermined by him (Vavilov,
1926). Afterwards,
Vavilovs ideas were developed bymany researchers at VIR (the Vavilov Instituteof Plant Industry).
Lukyanova et al.
(1990)
proposedan eco-geographicalclassificationof barley. According to thisclassification

the present
Russia occupies

centers of barleya considerable

diversity
are shown (Fig.1).
part of Eurasia with many different

Fig.1.

Globalcenters

3 -West Asian;of barley

diversity(Lukyanova

4 -Central

Asian;

5 -East Asian;

et al., 1990):

1 -Abyssinian;

6 -Europe-Siberian;

2 -Mediterranean;

7 -New World.

agro-ecologicalregions. Russia borders on the primary centers of barley diversity
andRussia has a long history of barley cultivation.
Consequently
a high level of barleygenetic diversity
is expected in Russia. The most representative
germplasm collectionof Russian barley, which includes several thousand accessions collectedduring thiscentury in different regions of Russia and neighbouring
countries, is being preserved
at VIR.
In the present study, we assayed 93 cultivatedbarley cultivars and landraces
originatedfrom different
regions of Eurasia. Our primary objectives
were to (i)estimate the genetic relationshipbetween barley accessions based on RFLP patterns,and (ii) compare the possibilities
of RFLP and hordeins analyses for determining
barleygenetic

variability.

Materials
and Methods
Plant Material
In total 93 barley accessions

including

54 cultivars(Table1) and 39 landraces(Table 2) were used in this study. The 82 cultivars and landraces were selected fromthe VIR germplasm collection
to represent
wide geographic
diversitypresent in
Russia and other countries of the former USSR. There were 39 two-rowed and 43

Table.1.
N oBarleycultivars

C u lt i v a r *used in thisstudy.

V IR g e n e b a n k

B o tan ic alc a t a lo g n u m b e r

v a r ie t ie sn u ta n s

P e d i g r e e /B a c k g r o u n doR reigg iino n o f

T w o ‑r o w e d
1

V ik in g

2 47 00

D o m e n x I n g r id

V y a tk a

2

V y a t ic h

2 68 23

B r ig i tt a x L u c h

3

R isk

2 93 52

C o m p l e x h ib r i d ( K m 1 1 9 2 , T e m p ,4

A u k sin y a i 3

2 8 117

C a rin a x T ap p a 2 6

L i th u a n ia

5

Z h o d i n s k ii 5

2 73 72

M a s u rk a x K m 1 1 9 2

B y e lo r u s s ia

6

T alo v s k ii

2 62 6 1

U nkn o w n

V o ro n e z h7

L y u b im e t s 1 0 8

2 73 73

U nk no w n

L u ts k

8

K h a rk o v sk ii 8 22 73 78

U n i o n x C h e r n o m o r e ts

K h a rk o v

9

D o n e ts k i i 6 5 0

1 83 3 1

S p a rt a n x M e d ic u m 5 1 3

D o n e tsk

10

O d e s s k ii 3 6

1 99 34

D o n e ts k i i 6 5 0 x S te p o v y i

O d e ssa

11

O d e s s k ii 1 0 0

26 8 64

H2( (4Mm 4 lexd3 ic6M 4ue6md2 i/6c1 u34m4) x1 3H 4 ip) ) rxo l(yS) lax v (uN t icu tha nx s

12T em p

22 05 5C h e m o m u ta n t o f K ra s n o d a rs k ii 3 5

K ra sn o d a r

13

P r ic u m s k i i 2 226 18 0m e d ic u m

L i n e ‑ 1 4 0 9 4 x L i n e ‑9 9 4 3

S ta v ro p o l

14

N u ta n s 1 1 5

19 35 5n u ta n s

S e le c t io n f ro m l a n d r a c e ( A r m e n ia )A rm e n ia

15

K v ant

27 55 8U n kn ow n

E k a t e r in b u r g

16

I lm e n

26 96 8P e r o g a x K r a s n o u f i m s k i i9 5

C h e ly a b in s k

17

O m sk ii 8 0

2 6 17 9m e d ic u m

P a li s s e r x O m s k ii 1 3 7 0 9

O m sk18

K r a s n o y a r s k ii 8 0

2 7 10 2

n u ta n s

S ‑8 0 x U n a

K ra sn o y a rs k

M osko w

H ip r o l y , M o s k o v s k i i 1 2 1 )m e d ic u m

19

E r o fe i

2 92 2 1m e d ic u m

K e y s to n e x L u c h

K h a b a ro v s k

20

P r im o r s k ii 8 92 70 55

V I R k ‑ 1 9 6 6 0 x U s s u r i is k i i 8

V la d iv o s t o k

2 1

G ra n a l

2 93 4 2

n u ta n s
u sb i n e r m e

O l im p x ( V I R k ‑ 2 1 6 8 3 x k ‑ 1 9 9 9 1 )K az a h sta n

22

T s e l in n y i 2 1 3

2 8 0 15n u ta n s

S e l e c ti o n f r o m T s e li n n y i 5

23

M e d ic u m

173 86m e d ic u m

S e l e c ti o n f r o m T u r k i s h l a n d r a c e

24

A l e x is

(2 9 5 7 8 )25

A r a m ir

26

U rse l

27
28
29
30

89 55

(V IR K ‑6 8 5 7 )
1 6 2 2 d 5 x T ru m p f

F ra n c e(2 1 8 7 5 )V o l la x E m ir

G e rm a n y(2 9 5 5 8 )A ra m ir x T r u m p f

A re n a(2 8 9 4 7 )A u fh a m m e r 3 9 /6 8 x H 4 6 4I s a r ia

(1 8 3 0 7 )D a n u b i a x B a v a r ia

A u s t r ia

s w U n u m l i‑ A rp a

19 177S e le c t io n f r o m M o r o c c a n l a n d r a c e

U z b e k is ta n

sw N u ta n s 2 7

16 33 5S e le c t io n f r o m l a n d r a c e ( U z b e k is t a n )n u ta n s

*w

Table.1.

Barleycultivars

used in thisstudy.

(Continued)No

C u ltiv a r*

V IR g e n e b a n k

B o ta n ic a

P e d ig re e /B a c k g ro u n d

3 1

w Ig ric a ta lo g n u m b e r v a rie tie s
M a lta x ((A u re a x C a rste n s 2 z lg .) x
(2 4 9 9 5 )e re c tu m

32

w T rix i

ab se n t

33

w M a lta

(2 1 8 2 7 )R e g io n o fo rig inH o lla n d

In g rid )((M a lta x V o lla ) x (T ria x E m ir)
n u ta n s

G e rm an y((C a rste n s 2 z lg . x A u re a ) x D e a ) x
H e rfo rd ia

S ix ‑r o w e d
34

P o la rn y i 1 4

15 6 19p a llid u m

S e le c tio n fro m la n d ra c e (K a re lia )M u rm a n s k

35

B e lo g o rs k ii

22 08 9p a llid u m

C h e rv o n e ts x K e y s to n e

L e n in g ra d

36

P a llid u m 4 5

1 185 6+ rik o te n se
p a llid u m
S e le c tio n fro m

37

G e lio s

28 93 6rik o te n se

(N u ta n s 3 2 x P a llid u m 1 2 5 ) x A th o s

O d e s sa

38

A g ul 2

27 64 9n c o te n s e

A g u l x K e y s to n e

K ra sn o y a rsk

39

V IR ‑6 52 183 3S e le c tio n fro m B ee c h e r (I sra e l)

U zb e k is ta n

40

sw G ia g in sk ii 3 9 5

18 12 2S e le c tio n fro m C h e n a d 3 9 5

K ra sn o d a r

p a llid u mla n ra c e (S a ra to v )S a ra to v

(R u m a n ia )4 1

sw K ru g lic 2 1

13 03 1S e le c tio n fro m la n d ra c e (K ra s n o d a r)

42

w R o sa v a

27 40 4O d e ss k ii 8 6 x O k sa m y t

43w P a llid u m 4

13 03 6S e le c tio n fro m la n d ra c e (K ra s n o d a r)

44

w K le p e n in s k ii

25 30 245

w S ilu e t

27 70 446

w V a v ilo n

29 36 1p a p a lle lu m

O d e s sa

V in e s c o x A lm a z

K rim e a

R o s to v s k ii 1 5 x Z im ra n

R o sto v( M e te o r x K N IIH 8 4 /II) x (A g e r 3 1 x K ra sn o d a r
M 13 )

47

w S k o ro h o d

29 40 448

w K ra sn o d a rsk ii

16 94 8p a llid u mSrege leioc ntio) n fro m la n d ra c e (C a u c a su s

M e te o r 5 7 x M 1 3 (m u ta n t o f R e g ia )49

w P rik u m sk ii 4 327 55 3p a ra lle lu m

F ‑2 1 7 9 x F ‑ 1 1 4 0 9

S ta v ro p o l

50

w A r a rati 7

25 99 4p a llid u m

M u ta n t o f K a le r (A rm e n ia )A rm e n ia

29 29

5 1

w N a h ic h e v an d a n i

13 24 8S e le c tio n fro m la n d ra c e (A z e rb a ij an ) A z e rb a ija n

52w V o g e ls an g e r(1 9 9 2 7 )(4 4 ‑0 1 3 x P e ra g is X II) x H a u te rs

53

w B ru n h ild

a b se n t

B a rb o x B a n te n g

54

w M am m ut

(2 7 0 9 9 )G e rm a n y

G o ldp a llid u m

V o g e lsa n g e r G o ld x (M a d ru x
W ss h .3 8 2 /4 9 1-winter;sw - semiwinter.

G e rm a n y

Table2. Barley

N o .

landraces

used in thisV IR g e n e b a n k

study.

B o ta n ic a l v a r ie t ie sc a t a lo g n u m b e r

Y e ar o f

R e g io n

re c e iv in go f o r ig in

R e m a rk s *T w o ‑ ro w e d1

3 2 2 22

1 64 1 1

19 2 1

K a re lia

19 3 8A r k h a n g e ls k3

4 5 4 14

1 64 1 0m e d ic u m

19 2 3V o lo g d an u ta n s

19 3 85
6

5 0 3 4m ed icu m

19 2 3S m o le n s k

2 1 8 2 0

n u ta n s

19 7 2M7

2 9 4 6n u d u m

19 1 4

K rasn o y a rsk

h8

18 0 5 9e r e c tu m

9

5 2 7 9n u d u m

19 2 3K a z a h s ta nh

1 0

18 3 6 2

p e r s ic u m

19 5 4

1 1

1 17 4 9

p e r s ic u m

19 2 9K y rg h y z s ta n

1 2

14 9 2 3

n u d u m

19 3 4T u r k m e n is t a n

1 3

2 9 0 4n u ta n s + p a ll id u m

n u ta n s+ i n t e r m e d iu m

( s ix ‑ ro w e d )( s ix ‑ ro w e d )ak h a c h k ala

19 5 119 14

h , sw
s w

S ix ‑ ro w e d
14

16 8 8 115

9 3 3 81 6

9 5 3 71 7

9 8 2 7

19 4 4M u rm a n sk

19 2 7

K a r e lia

c o e le s te

19 2 7A r k h a n g e ls k

p a llid u m

19 2 7V o lo g d a

6 4 2 019 3 8V y atk a19

9 4 2 319 2 7

K o m i2 0

9 5 1 1

19 2 7

K o s tro m a

2 1

1 1 9 7 0

1 9 4 9K az an

2 2

4 9 7 219 2 2O m sk2 3

1 6 4 7 81 9 3 8

Ir k u ts k

2 4

2 9 1 0 2

1 9 8 62 5

4 8 2 5

1 9 2 3C h it a

2 6

1 0 6 9 3

1 9 2 7

Y a k u tsk

2 7

1 1 0 7 5c o e le s te

1 9 2 7

S a k h a lin

2 8

5 0 9 2p a llid u m

1 9 2 3K a za h stan

2 9

4 8 4 7p a llid u m + n u t a n s (tw o ‑ ro w e d )1 9 2 33 0

1 0 8 7 7

p y ra m i d a t

1 9 2 63 1

1 6 4 6 8n ig r u m

1 9 3 83 2

3 0 3 8re v e l a t u m

1 9 1 73 3

1 7 2 2 7p a l lid u m

1 9 4 718

1

p a ll id u m( p a llid u m )T u r k m e n is ta nhU z b e k is ta n

3 4

1 1 75 5n ig r u m

1 9 4 9K y rg h y z s ta n

35

3 1 1 8c o e le s t e

19 1 7

T a d z h ik is t a n

3 6

10 6 2 8a n c o b e re n s e

19 2 83 7

2 14 7 7p a ll id u m

19 6 53 8

8 12 33 9

6 12 8* w- winter,n ig r ip a llid u m

sw - semiwinter,h - hulless.

+ p a llid u mhhh

19 2 6A z e rb a i ja nw

19 2 4T u r k m e n is ta nw

ƒÉ

six-rowed accessions, among which 49 were landraces or cultivars
derived by
selection from landraces. Also in this study 11 well known West European spring andwinter cultivars from different germplasm groups were included.
group were kindly provided by German breeders.

Seeds of the latter

Hordeins Electrophoresis
Hordeins were extracted from crushed single seeds with 40 ml of 6M urea.
After centrifugation
the supernatantwere used for electrophoresis.
Hordeinelectrophoresis
was carried out in slabs of 6.5% PAGE 0.013M acetic acid pH3.2
during 4-4.5 h (U = 600 V and I = 20-25 mAper slab). After electrophoresis
gels were
stained with 0.075% Coomassie G-250 in 10% trichloroacetic
acid and photographed.
RFLP Analysis
Leaf DNA was extracted from 2-to 3-week-old seedlings(bulks of 20-25
seedlings
per accessions).
Isolationof genomic DNA, digestion
with restriction
enzymes, electrophoresis
in agarose gels, Southern blotting
onto nylon membranes,
hybridization
with 32P-labelled
DNA probes, autoradiography,
and post-hybridization
washes for stripping of probes were performed as described in detail by Graner et al.
(1990).
DNA was separately
digested with restriction
enzymes Eco RI and Hind III.
Electrophoresis
was performed in gels 20 cm long and 15 cm broad with 20 lanes andtwo rows of wells. Digested DNA of all accessions was loaded on six differentgelseach including
two check varieties('Igri'and 'Alexis')
and a lane of phage DNA
digested by Hind III. For detection of restriction
fragments, we used 41 anonymousclones previously mapped, mainly, single-copy DNA clones, from Hordeum vulgare
L.(Graner et al., 1993). The clones were selected to provide a fairly uniform coverageof the barley genome with at least five clones per chromosome (Fig.2).
Thirty-fivewere genomic DNA clones (with MWG, ABG and WG prefixes) and six were CDNA
clones (with CMWGand ABC prefixes).
Data Collectionand Statistical

Analysis

Hordein and RFLP patterns on autoradiographs
for each clone-enzyme
combinations(CEC) were usually scored by assigning a number to each band. For
subsequent numerical analyses, data were coded in binary form, i.e., presence orFig.2. Chromosomal location
top. Clone

designation

Igri/Franka

map.

of DNA clones assayed.
according

Chromosomes are oriented

to Graner et al. (1993).

Distanceswith the short arm onin cM are presented

from

absence of a band in a line was coded by 1 or 0, respectively.
Only polymorphic
bands
were includedin the raw data matrix. This matrix was used to generate a genetic
distance matrix using Nei's (1972)
distance:

where dij is the genetic distance
between accession i and accession j , xki is the iallele frequency
at locus k and n is the total number of loci. Dendrograms were
produced using unweighted
pair-groupmethod, arithmetic
average (UPGMA)
clusteringand scatter diagrams resulted from principal
coordinate analyses (PCA) on
the genetic distance matrix. The normalized Mantel statistic(Z) (Mantel,
1967) wasused to compare the genetic distance matrixes generated from RFLP and hordeinselectrophoresis
data. The program NTSYS-pc version 1.8 (Rohlf,
1993) was used togenerate the distance matrixes for UPGMA clustering,
the PCA analysis,
and thematrix comparison.

Results and Discussion
Variation for RFLPs and Hordeins
Altogether,
we analyzed data from 77 CEC. Seven CEC showed completely
monomorphic RFLP patterns. The DNA clones used in this study detected on average
4.9 (ranging
from 2-13) polymorphic
fragments per CEC for a total of 335
polymorphic
fragments from 70 CEC. Restriction
enzymes EcoRI yielded158
polymorphic
fragments from 35 CEC s and Hind III yielded
177 polymorphic
fragments from 35 CEC s. Typical RFLP patterns obtained are illustrated(Fig.3).
All
93 accessions could be distinguishedwith the set of 335 polymorphic
fragments.

Fig.3.

Restriction
fragment
cultivars
and landraces

length polymorphism
banding
patterns
obtainedon selectedwith Hind III and barley genome DNA clone MWG938.

EurasianFrom hordeins electrophoresis
patterns 42 polymorphic
bands were includedin the raw data matrix. Twenty-seven accessions (29.0%)
were polymorphic
andconsisted of 2-3 biotypes based on hordeins analysis
of 20 seeds of each accession.
For further analysis the main protein phenotypes
from each accession was selected.
Thus, among 93 accession 71 different
proteinphenotypes
were determined.
Thirty-fouraccessions formed 12 groups. Each group contains 2-4 accessions withidentical
pattern.
Clustering of Barley Accessions Based on RFLPs
The relationships
between 93 barley accessions based on RFLP genetic
distance measurement were analyzed by UPGMA clustering.
All accessions, except
for the hulless six-rowed landrace (acc. 10628) from Central Asia (Tadzhikistan),were separated into two major clusters (Fig.4).
Cluster A comprises mainly landracesfrom Central Asia, Siberia and the Caucasus regions. This cluster consists of 19
landracesand 3 cultivars
derived by selection
from landraces.
It includes
both
two-rowed and six-rowed accessions and all of the analyzed hulless forms. Except forthe six-rowed landrace (ace. 6128), from Central Asia (Turkmenistan),
there are two
major sub-clusters:
one is geographicallylinked with Central Asia and another ismore widespread.
Cluster B is larger and consists of 5 sub-clusters(Fig.4).
Most accessions arein sub-clusters
6 and 7 and are from a wider geographic
area and distinguishable
mainly on the basis of spike morphology. Sub-cluster
7 consists of two-rowed West
European spring cultivars('Alexis','Arena', 'Isaria','Aramir' and 'Ursel')andlandraces and cultivars from different
regions of Russia. The Russian cultivars
havepart of their pedigree from Western Europe, Eastern Europe and Canadian cultivars('Trumpf,'Ingrid','Isaria','Emir','Masurka','Chenad','Diamant','Gatway','Keystone' and others). Sub-cluster
6 consists mainly of six-rowed barley accessions
which can be divided into three groups. One group includes spring landraces
relatedto cultivars'Belogorskii','Agul 2' and 'Erofei'.
They have the Canadian cultivar'Keystone'in their pedigree.
The second group includes
winter cultivars
fromWestern Europe ('Vogelsanger
Gold', 'Mammut' and 'Brunhild')and some winter
Russian cultivars
possibly
related to them. The third includes
landraces 4847 and11970
and cultivars'Pallidum
45' and 'Kruglic
21' possibly
relatedto cultivar

Fig.4. Dendrogram constructed from the restriction
Eurasian barley accessions.

lengthpolymorphism

genetic

distancesmatrixof 93

'Giaginskii

395', which was derived from Roumanian cultivar 'Chenad 395'.
Sub-cluster
5 comprises West European two-rowed winter cultivars'Igri','Trixi'and 'Malta', which have different
pedigreefrom the above mentioned West
European six-rowed winter cultivars. This sub-cluster
also includes Russian cultivar'Gelios'
related to cultivar 'Emir'.
Sub-cluster
4 includes cultivars'Nakhichevandani'
and 'Pricumskii
22',which are evidently
relatedto cultivar'VIR-65'
selectedfrom Israeli
cultivar'Beecher'.
Finally,
sub-cluster
3 includes landraces 18362 and 11749 and cultivars'Medicum 8955' and 'Unumli-Arpa'. The latter ones were derived by selection
fromTurkish and Moroccan landraces, respectively.
The principal
coordinate
analysis(PCA) is independent
from UPGMA
clustering,but their results were similar (Fig.5).
Some of the variation(45.5%)
wasaccounted by the first two principal
coordinate(PC) axes. Most of the variation(28.4%)was explained by the first PC, which clearly divided the analyzed accessions
in two groups (A and B, see dotted line). These groups correspond exactly to clusters
A and B on the UPGMA dendrogram. The second PC explained
17.1% of variationand clearly divided two-rowed and six-rowed accessions comprised in the group B
into two sub-groups. This dividingof accessions according to spikelet
rows is moreclearly
shown by a PCA plot, than by a dendrogram. On the PCA plot two-rowed
accessions
from sub-clusters
3 and 4 are locatedin sub-group of two-rowed
accessions, but six-rowed cultivars'Gelios'and 'Polarnyi'
are located in sub-groupof six-rowed accessions. Only two cultivars do not group according to spikeletrowsof the ears: two-rowed 'Erofei'and 'Malta',
they are located in the sub-group of
six-rowed accessions.
The results of RFLP analysis confirm the existence of high genetic diversity
present in Russian barley. This study reveals the existenceof two major genetic
groups in the analyzed material.
Together with the West European cultivars,
themajority of Russian cultivars and landraces form a large and heterogeneousgroup (B).
The second group, which was identified
in this study (A) includes
a group oflandraces predominantly
originatedfrom Central Asia. Vaviliov (1926)
was the firstto point out exotic charactersof barley from Central and East Asia. The reason forthis distinction
is geographicalisolationand evolution
in the agro-ecologicalconditionsof the region(Vavilov,1926).
The hypothesisof independentFig.5. Plot of the principal coordinate scores from the restriction fragment length polymorphism genetic distances matrix of 93 Eurasian barley accessions.
Some of the variation (45.5%) is accounted for by the two axes.

domesticationof barley in oriental and occidental
regions of Eurasia was suggested
by a number of researchers (see Takahashi,
1955, for review). Recently Zhang et al.
(1994)
using isozyme and ribosomal
DNA markers showed both broad genetic
diversityof cultivatedbarley from Tibet and considerable
oriental-occidental
differentiationof barley. In our study both cluster and PCA analyses of RFLP dataclearly
separated
all accessions
into two major genetic
groups, which are
geographicallylinked with oriental and occidental
regions of Eurasia. This confirms
the existence of two principal trends in the evolution of cultivatedbarley. It is likely,that the broad clustering
into oriental and occidental
accessions reflects historicallydifferent sources of germplasm contributing
to the two groups. According to a modern
classificationof global centers of barley diversity(gene-centers)
adopted by VIR wemay connect the above mentioned germplasm groups to Europe-Siberian(cluster B)
and Central Asian (cluster A) centers (Fig.1).
Central Asia might represent a valuable
source of germplasm to increase the variabilityof barley. Group B in our study clearly
dividedinto two sub-groups consisting
predominantlyof two-rowed and six-rowed
accessions. Tinker et al. (1993)
using RAPD markers differentiated27 barleyaccessions into two groups, two-rowed and six-rowed forms. Similar results using
RFLPs were obtained by Melchinger et al. (1994) in the analysis of European barley
germplasm. The only exception was the position of a two-rowed winter forms, whichclustered
together with six-rowed winter cultivars.
In our study this group ('Igri','Trixi' and 'Malta')
formed rather distinct sub-clusterin cluster B. There are several
classification
systems of cultivatedbarley in which on the basis of spike morphology
two principal
sub-species(two-rowed
and six-rowed)
are determined(see
Trofimovskaya,
1972, for review). It should be noted, that accessions of group A wereboth two-and six-rowed forms, but there is no order to their clustering.
Moreover,genetic distinction
between accessions with the same number of rows in the spike, but
belongingto different
groups was shown by both clusteringand PC analyses. We
propose the existenceof two principal
trends in breedingof occidental-typeofcultivatedbarley. However, there maybe some exceptions, for example, the 'Malta'group of cultivars,
which possibly
have hybrid nature and derived from crossing
two-and six-rowed forms. Apart from above mentioned 'Igri','Trixi' and 'Malta'group of cultivars
related to 'Malta' there are several groups of related(Fig.4).
In two-rowed sub-cluster 7 the most interestinggroup includes

accessions
both WestEuropean ('Alexis','Arena', 'Isaria','Aramir' and 'Ursel')and Russian cultivars
related to them ('Lyubimets
108', 'Ilmen', 'Krasnoyarskii
80' and 'Auksinyai3'). In
six-rowed sub-cluster
6 there are two groups of accessions.
One includes both
cultivars with 'Keystone' pedigree background ('Belogorskii','Agul 2' and 'Erofei')and 10 landraces (from 16881 to 4972). All of these accessions originatedfrom thenorthern regions of barley cultivationin Russia (northern
Europe and Siberia).
Another group includes both West European ('Vogelsanger
Gold', 'Mammut' and'Brunhild')and 8 related Russian winter cultivars
were from southern regions. In
cluster A group consistingof 5 closely related hulless landraces from Central Asia
and Siberia can be seen. Another one includes 5 two-rowed accessions (from Nutans
27 to 2904), which on the PCA plot are quite close to the two rowed accessions ofgroup B (Fig.5).
The third group consists of 7 six-rowed landraces and two-rowed
cultivar Nutans 27. All are linked to Central Asian origin.
Comparisons
between Genetic
Distances
Based on RFLP and Hordein
Electrophoresis
In this study we attempted to compare the use of RFLP and hordeins analyses fordetermining barley genetic variability.
For this purpose for 93 analyzed accessions thegenetic distance matrixes obtained separatelyfrom RFLP and hordein electrophoresis
data were compared. The normalized Mantel statistic
obtained from this comparison
through 500 random permutationsof matrices was low (r = Z = 0.18) but highly
significant(p = 0.002). UPGMA clustering
based on hordeins electrophoresis
data
showed a picture of the accessions grouping (Fig.6) principally
different
from the one
received from RFLP data (Fig.4).
But there are several groups of related accessions(markedby grey bands),
which have the same grouping
on the dendrogram
constructed from RFLP data. In our study among 93 accessions 71 different
proteinphenotypes were determined which indicates the high level of hordein polymorphism
and its potential
usefulness for barley cultivar identification.
Taking into account therelative simplicity
of isolationand electrophoresisof hordeins,
this methodologicalapproach is valuable
for solving many practicalproblems in breeding,cultivar
identificationand seed control. But the possibilityof using hordein electrophoresis
data for studying genetic relationshipsof different
barley cultivars are limited due tothe smallnumber of locidetermining
hordeins.
There are only two

Fig.6. Dendrogram constructedbarley accessions.

from the hordeins

polymorphism

genetic

distancematrix of 93 Eurasianhordein-determining
loci in the barley genome, which are localizedon the short armof 5th chromosome and positioned
at a distance of about 15 cM from one another(Graner et al., 1993). Unlike hordeins, RFLP fragments detected by a single clone
represent both different alleles and different loci and the abundance of RFLP-markers
permitsa representative
samplingof the whole genome. For these reasons,
RFLP-based
genetic distances
providea truer estimateof the actual genetic
relationshipbetween barleyaccessions.

Conclusions
In conclusion,
our results of studying
a diverse collectionof barley fromdifferent
regions of Eurasia are in accordance with recent investigations
in barley(Melchinger
et al., 1994) that RFLPs are suitable to (i) define a germplasm groupmore clearly,(ii) assign lines with unknown or incomplete
pedigreerecords toestablishedgroups, and (iii) identify
diverse germplasm sources. RFLP analysis ofbarley cultivars and landraces from different countries of Eurasia made it possible
toconfirm the existence of two principal
trends in the evolution of cultivatedbarley,which are geographicallylinked with oriental and occidental
regions. Also in thisstudy.breeding
trends were observed, such as sub-grouping of oriental forms and theirfurther sub-groupingto groups of cultivars with similar pedigree background.
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Diversity

Analysisand Evaluationof Wheat Genetic Resources in China

L. H. LI, Y. S. DONG and D. S. ZHENG
Instituteof Crop Germplasm

Resources,

ChineseAgricultural

Sciences,

100081,

China

Beijing

Academy of

Abstract
A great number of wheat genetic

resources are found in China and more than 40,000

accessions are conserved in the National Crop Gene Bank. The diversity of these wheat genetic
resources
was evaluatedfor the following
characteristics:
distribution
and growthenvironments,
species of wheat and its relatives,
genetic diversity
in agronomic characters,
grain quality,
resistanceto diseases,
pests and environmental
stresses, and crossability.
According to the results of diversity
evaluation,some suggestions
for future collection,conservation,
study and use are discussed.
Key words: Wheat genetic resources, diversity evaluation and analysis, China.

Introduction
Biodiversity
preservation
is essential
for man's survival. Food supply andpopulation
growth are not in balance thus food crops closely linked with mankind's
survival should be given priority for evaluation and conservation. Diversity evaluation
and analysis of crop genetic resources can address the following
issues: (1) to makea correct strategy for collection,conservation,and use; (2) to find genetic diversity
centers for various crops; (3) to broaden the genetic base of crops and steadily
increase food production.
Wheat is one of the most important food crop worldwide. Many internationaland national research programs are engaged in wheat improvement. High yielding
wheat cultivars
have been released to farmers. However, modern cultivars
havenarrowed the genetic base of wheat. Thus, the diversity
evaluation
and analysis ofwheat, especiallylocal varieties and their wild relatives,
have become an urgent task.
China is one of the secondary centers of wheat diversity
and has abundant
genetic resources, including unique subspecies, local varieties and wild relatives. This
paper gives an overview of the major progress in the diversity
evaluationof wheat
genetic resources in China.

•Ž
•Ž;

Collection

StatusIn the 1950's, the Chinese government organized the collectionof localvarieties around the country. After the founding of the Instituteof Crop Germplasm
Resources, Chinese Academy of Agricultural
Sciences in 1978, further plant
explorationand collection
were carried out in some regions. To date, a total of 42,777
accessionsof wheat germplasm have been collected(Table1). These accessions
included local varieties and improved cultivars of commonwheat (Triticum aestivumL.), 18 other species and subspecies in Triticum, 190 species and subspecies from 14
genera related to wheat, and some special genetic resources such as aneuploids,
malesterile lines, constitutethis collection.
So far, almost all local varietiesof common
wheat have been collected and are conserved, but there is a need for further collectionof wild relatives.
Distribution

and Environment
Wheat is cultivatedthroughout China. The diverse topographicalfeatures andclimate of China influence the diversityof wheat. They are lowland basins such as at
the Tuloufan Basin, at 150 m below sea level, in Xinjiang,
plain, mountains andplateaus, such as the Qinghai-Xizang
Plateau at an altitude of 4000m. The climate is
generally
temperate monsoon but, due to the geographic
situationand the diverse
topography,
there are regions with unique local climatic conditions.
The mean annual
temperatures vary from -5.8
to 26.4
average annual precipitation
ranges from
3.9 mmto 6,558 mm.There are about 40 soil types, mainly black soil, brown soil,
yellow soil and red soil where wheat genetic resources grow.

Species

Diversity
The species(Table2).
Triticumof wheat and its wild relatives

distributedin China

are shownL.
Six speciesof this genus are found in China, including
T. aestivum, T.
turgidum, T. durum, T. compactum, T. orientale
and T. polonium. More than 96% of
local varieties
are commonwheat, T. aestivum; 2% are T. turgidum and T. durum; T.
compactum and T. polonium are less than 1% respectively;
T. orientale
are very few.
Three subspeciesof T. aestivum, indigenous
to China, have been recognized:

Table1. Collectionsof wheat genetic

resources

S p e c ie s

in Chinaa c cN e os .s ioo fn s

I m Op rroigv ine d
L o calF o re ig n

T r i ti c u m a e s t iv u m

3 7 ,3 9 8

1 3 ,9 0 2

O t h e r s p e c ie s in T r i ti c u m

2 ,1 9 1

68 6

1 ,5 0 5

W il d r e la t iv e s

2 ,2 3 7

1 ,7 8 7

4 50

G e n e t ic s t o c k s

9 5 1

74 1

2 10

4 2 ,7 7 7

1 7 ,2 1 6

T o ta l

Table 2. Speciesof wheat and its wild

relatives

distributed

9 ,7 2 1

9 ,7 2 1

1 3 ,7 7 5

1 5 ,9 4 0

in China

G enu sNs p oe c. oief s

G eno m e

P lo id y

L if e c y c l e

M a ti n g

Ma r ea ja o r d i s t r ib u tio n

T r it ic u m

6 *

A B D

4 x ,6 x

A n nu al

S e lfA l l C h in a

A e g i lo p s

1

D

2x

A n nu al

S e lf

X in ji a n g

S e c a le

2

R

2x

A n nu al

C ro s s

X in jia n g

E re m op y ru m

4

A B C #

2 x ,4 x

A n nu al

S e lf

X in ji a n g

H o rd e u m

8H I

2 x ‑6 x

pA en ren nu na liaolr

S e lf

CN ho irntha w e s te rn

A g r op y ro n

5P

2 x ,4 x

P e re n n ial

C ro s s

I n n e r M o n g o li a

R o e g n e r ia

70

SH Y P

4 x ,6 x

P e re n n ial

S e lf

A ll C h i n a

E ly m u s

12SH

4 x ,6 x

P e r e n n ia l

S e lf

S ic h u a n

E ly tr ig ia

1

SSX

6x

P e r e n n ia l

C ro ss

X in j ia n g

L ey m u s

9N X

4 x‑ 1O x

P e r e n n ia l

C ro ss

X in j ia n g

Pa cs ha yt hs y r o s t‑

4

N

2x

P e r e n n ia l

C ro ss

X in j ia n g

H y s tr ix

2

?

?

P e re n n ia l?

?

subspecies

indigenous*, including

three

and uniqueand ssp. tibetanum#, the genomes are different

from those of Triticum

to China,ssp. yunnanense, ssp. petropavlovsksyi,

(1) Yunnan wheat, T. aestivum ssp. yunnanense King, which consists of 16 varieties,
distributed
in shallow
gulliesand open areas in forests on high mountains
between 1,500-2,500
m, in the lower reaches of the Lancang River and the Nu
River in Yunnan province, southwestern
China. Double ditelosomic
analysisindicatedthat 8 chromosomes showed some differencesfrom those of Chinese
Spring (CS) (Chen et al., 1988; Huang et al., 1989).
(2) Xinjiang
wheat, formerly
identified
as a distinct
hexaploid
species,T.
petropavlovsksyi
Udacz. et Migush., now recognized as a subspecies by Y. S.
Dong, and consists of 7 varieties.
This subspecies
is found in the agriculturalareas in the west part of Talimu Basin, Xinjiang.
Cytologicalanalysis of crosses
with common wheat and tetraploid
species indicate that this subspecies
may bederived from natural hybridization
between T. aestivum and T. polonium (Chenet al., 1985).
(3) Tibetan weedy wheat, T. aestivum ssp. tibetanum Shao, consists of 23 varieties.
Distributedin the upper reaches of the Lancang River, the Nu River and theYaluzangbu River valleys, Tibet, between 1,700 - 3,600 m (mostly 2,300 m).
Double ditelosomic
analysis
showed that its chromosome constitution
wasessentiallythe same as CS except that 7BS usually
failed to pair (Chen etal.,1991;
Huang et al.,1981).

Wild

Relatives

China is one of the major distribution
areas of wheat relatives,
including11
genera and about 120 species. Through collecting
expeditions
over the past 15 years,
a number of sizeable
collections
have been established,including
3 perennial
Triticeae nurseries located in Beijing,
Sichuan, and Xinjiang
respectively.
Aegilops L.: Only one species,Ae.
tauschii (2n=2x=14, DD) was proved to be anative Chinese species. It grows in the natural vegetationof the Yili river valley,
which lies between the mountains west of Mount Tianshan in Xinjiang.
When theelevation
rises to 1,420 m, Ae. tauschii and Bromus gedosianus compete well withother tall grasses, and formed a dense steppe community of about 15 hectares (Yenet al., 1984). Moreover,Ae. tauschii was also found as weeds of winter wheat fields
in Henan and Shaanxi provinces. Species of this genus are noted for their resistanceto powdery mildew. Amphiploidsof tetraploid
wheat with 10 species
were

•Ž

synthesizedby chromosome autoduplicationof hybrids (Xu and Dong, 1992). Also,
some newgenes resistant to powdery mildew were found in accessions of Ae. tauschii(Kong, 1996).
Secale L.: 2 annual species occur in China and both have the R genome. S. cereale
was planted in only a few mountain areas. S. sylvestre is widely distributed
as a weedof the winter wheat fields in Xinjiang,
but a natural populationof about 1 hectare wasfound in Habahe county, Xinjiang.
The collectionsof this genus from Xinjiang
showed high tolerance to cold and drought.
Eremopyrum (Ledeb.) Jaub. et Spach.: 4 species occur in China and they havethe A, B, C genomes (Sakamoto, 1967) and two ploidy levels, diploid and tetraploid.
They are annual and only found in Xinjiang
and Inner Mongolia. It is a typical genus
in semi-desert
vegetationand species have a short life cycle in early spring. Somecollections
are highly resistantor immune to powdery mildew. Hybridsof common
wheat with E. orientale and their derivatives
were obtained (Zhang, 1996).
Hordeum L. consists of 3 annual species and 5 perennial
species in China, with
the H and I genomes and three ploidy levels - diploid, tetraploidand hexaploid.
They
are mainly distributedin the northwestern China, and usually grow in saline swamps.
Agropyron Gaertn. consists of 5 perennial
species in China, with the basic P
genome and two ploidy levels, diploidand tetraploid.
The tetraploid
species are themost commonand are distributed
in most areas of China but are most abundant inInner Mongolia and Xinjiang. Species of Agropyron are noted for their high tolerance
to cold and drought, and have moderate tolerance to salinity.
For example, somecollections
growing in Xinjiang
and Inner Mongolia can complete their life cycles
without any rain at all and can survive temperatures as low as -44
due to wind andlack of snow. In order to transfer desirable
traits, such as tolerance to environmental
stresses and resistanceto diseases from this genus into wheat, some intergenerichybrids
and their derivatives
between common wheat and tetraploid
species ofAgropyron have been obtained (Li and Dong, 1990, 1991, 1993; Li et al., 1995).
Roegineria
C. Koch, consists of about 70 species in China, with S, H, Y, P
genomes and two ploidy levels, tetraploidand hexaploid.
This genus is the mostcommon,complex and the largest genus of Triticeae in China, and most of these
species are endemic to China. Some species of this genus showed wide adaptation
anda high level of cold tolerance.

Elymus L. consists of 12 species in China,levels, tetraploidand hexaploid.
These species
particularly
Sichuan province.

with S, H genomes and two ploidy
are mainly found in western China,

Elytrigia
Desv.: Only one species of this genus, E. repens (2n=6x=42, SSX) is
found in several regions of China.
Leymus Hochst.:Ninespecies of this genus occur in China, they have the N, X
genomes and 4 ploidy levels from 4x to 10x. Most species are be found in Xinjiang.
Their perennial
habit, large seeds, tolerance
to salinity,
alkalinity
and cold, andresistance to diseases have made some species of Leymus attractive to wheat breeders.
Hybrids
of common wheat with L. multicaulis
and their derivatives,
show highresistance to BYDV and tolerance to salinity,
have been obtained by our laboratory.
Psathyrostachys
Neveski consists of 4 species in China. They are all diploidandwith a basic genome N. Three species are mainly found in Xinjiang,
but also sparsely
in Gansu and Inner Mongolia. P. huashanica can only be found in the Huashan
Mountain, Shaanxi. The plants of this genus showed characteristics
of cold resistanceand tolerance to poor soil. The hybrids and their derivatives
between commonwheat
and P. juncea have been obtained (Chen et al., 1988).
Hystrix Moench: 2 species, H. duthiei (Stapf)
Bor and H. komarovii (Roshev.)
Ohwi, were described in Flora of China (Guo, 1987), and are distributed
sparsely in
China. However, no accession has been collected
yet.

Diversity

Evaluation
A primary objectiveof germplasm collection
is to ensure the continued
availabilityof germplasm suitable for the development of stable, productive and highqualitycultivars(Damania,
1990). Therefore,
all collectionsof wheat genetic
resources were evaluated for agronomic characters,
grain quality,
resistancetodiseases, pests and environmental stresses, and crossability.
Agronomic Characters
The agronomic characters
evaluated
included
ecotype,headingdate,maturation period, plant height, spike length, awn length, awn color, glume color andgrain color, number of grains per spike, weight per 1,000 grain.
Date of maturity. Early maturity is one of the distinguishingfeatures of the local
Chinese wheat varieties.
250 local winter wheat varieties that have a growth period

of less than 252 days were analyzed in detail after all were evaluated for date of
maturity in Beijing(Song et al., 1995). These 250 varieties
were divided into fourtypes according to the length of various physiologicalperiods (Table 3). The earlier
maturing varieties had a short period from flowering to maturity. For example, thevariety Sanyuehuang needed 241 days from emergence of seedlings to maturity, but
the period from flowering to maturity was only 25 days.
Table 3. Classification
developing
C la s s i f ic a ti o nof 250 winter wheat varieties
periods

with early maturity

D e v e lo p m e n t p e r i o daccordingto days of various

D ay sN o . o f v a r ie t ie sI

PE hm aesreg e n c e o f s e e d l in g s ‑ V e g e ta t iv e

1 8 0 ‑1 8 5

43IIV e g e ta t iv e P h a s e ‑ S p i k e e m e rg e n c e

2 0 ‑2 552

I II

S p ik e e m e rg e n c e ‑A n t h e s i s

3 ‑5

14

IV

A n th e s i s ‑ M a tu r it y

2 7 ‑3 2

41

Semidwarfness.
The commonwheat genetic resources were planted in various
ecologicalregions and about 200 accessions with plant height below 60 cm were
found. Among them, about 62% were insensitive
to GA3. Moreover, by family
analysis,
there were 5 semi-dwarf categories
or dwarfing genes which led to thesuccessful development of semidwarf cultivars in China (Jia et al., 1992). They are:(1) Suman 86, carrying 2 pairs of GA3 insensitive
semidwarf genes, Rht1 and Rht2;(2) St2422/464,
originatingfrom Italy, and bearing 1 pair of semidwarf genes similar
to that of Saitama 27 with weak GA3 insensitivity
designatedas Rht1S; (3) 2 Chinese
varieties,
Huxianhong and Youbao, each carrying Rht2; (4) Funo, Abbondanza andother derivativesof Akagumughi,
each carrying
1 or 2 pairs of GA3 sensitive
dwarfing genes designatedas Rht8 and Rht9; (5) Tom Thumb and Aibian 1, carrying
Rht3 and Rht10 respectively.
In general, they were used in hybrid wheat development
and recurrent selection
for semidwarfness.
Grain weight per spike. Grain weight per spike is mainly determinedby twofactors, number of grains per spike and weight per 1,000 grains. Through selection
over several years, a great number of local varieties with over 60 grains per spike havebeen obtained, but the weight per 1,000 grains was usually less than 35g.

*:Grain Quality
Variation on content of protein

and lysine.

were analyzed for content of protein
was 15.1% and 0.438%, ranging

and lysine. The mean value of protein and lysine
from 7.5 to 28.9% and from 0.25 to 0.8%,

A total of 20,184

accessionsof wheatrespectively.
Among accessions determined,
protein content of 1,637 accessions
exceeded 18% and lysine content of 1,988 accessions
was above 0.5%. The
correlation
between protein and lysine content and the effects of ecologicalfactors
on protein and lysine content have also been analyzed (Li, 1992).
Variationin grain hardness and sedimentation
value. Grain hardness (groundtime method) of 21,509 accessions and flour sedimentation
value (Zeleny method) of
11,286 accessions of wheat were determined, and accessions of high quality wereidentified(Liet al., 1993). The range variations
in grain hardness and sedimentation
value were 8.5 - 619.4 ml and 4.0 - 62.0 S, respectively(Table 4). The correlationofgrain hardness and sedimentation
value with ecologicalfactors was also analyzed.
Table 4. Variation
C h a rac te r

on hardness

G e kr min dp sla s m

and sedimentation

value of wheat genetic

resourcesa Nc c oe .s so iof n s

M ean

R an gew N ito h. oh fig ahc cq eusasliito yn *s

C o m m on w h eat

2 1 ,2 8 5

2 4 .0 0

8 .5 ‑ 6 1 9 .4

5 ,2 4 5

L ocal8 ,6 3 4

2 2 .9 4

8 .5 ‑ 2 1 4 . 1

2 ,5 4 8

Im p ro v e d

4 ,5 3 2

2 5 .3 8

9 .8 ‑ 4 2 9 . 1

63 9

F o r e ig n

8 ,1 1 9

2 4 .3 6

1 0 .0 ‑ 6 1 9 .4

2 ,0 5 8

2 24

1 3 .8 3

1 0 .3 ‑4 1 .5

18 0

2 1 ,5 0 9

2 3 .8 9

8 .5 ‑ 6 1 9 .4

5 ,4 2 5

1 1 ,2 0 0

2 4 .5 1

4 .6 ‑ 6 2 .0

49 0

L o cal

5 ,2 9 8

2 4 .1 2

5 .0 ‑ 5 4 .072

I m p ro v e d

3 ,1 4 3

2 4 .9 4

4 .0 ‑ 6 2 .0

17 3

F o re ig n

2 ,8 4 5

2 4 .5 3

4 .0 ‑ 6 2 .0

24 5

86

1 6 .9 4

7 .8 ‑ 3 4 .8

0

1 1 ,2 8 6

2 4 .4 5

4 .0 ‑ 6 2 .0

49 0

15 seconds

and sedimentation

H a rd n e s s (S )O t h e r s p e c ie s
T o ta l
S e d i m e n t a ti oC no vm a mlu oe n ( mw hl)e a t

O t h e r s p e c ie s
T o ta l

The criterionof high quality
is hardness
value more than 40 ml, respectively.

less thanBread baking quality.
During the past decade there has been substantial
researchon the role and genetics of high-molecular-weight(HMW) glutenin in bread-making
potential(Payne et al., 1984). Studies of the compositionof HMW glutenin subunitsof the Chinese wheats and its relation to bread baking quality were made by several
institutes.
In a study by Mao (1992),
the HMW glutenin subunits compositionof
5,071 commonwheats, comprising 936 local varieties,
2,307 improved varieties
and
1,828 foreign introductions,were determined by SDS-PAGE. It was observed that the
3 categories of wheats differed
subunits (Table 5).

quite significantlyin the distribution

Table 5. Distribution pattern (%) of high molecular weight glutenin
V a rie tya c cNe so sio. o nf s

G lu ‑A 1

pattern

subunits in 5,071 commonwheats

G lu ‑B 1

N u ll1

2*

7+8

of HMW

G lu ‑D 1
7+ 9

22

2 + 12

5 + 10L o cal

9 36

8 8 .66 .5

5 .2

8 4 .74 .92 .7

9 4 .33 .7

Im p ro v e d

2 ,3 0 7

5 7 .42 7 .6

1 5 .04 2 .04 1 .92 .8

7 3 .7

5 .7

1 ,8 2 8

5 4 .5

2 7 .3

1 8 .32 5 .23 7 .2

1 1 .74 6 .44 5 .9F o re ig n
in tro d u c tio n

Resistance to Diseases
About 23,000 accessions of wheat genetic resources were screened for resistanceto diseases such as rusts, powdery mildew, wheat scab, barley yellow dwarf virus(BYDV), and root rots.
Powdery mildew. Powdery mildew is one of the major wheat diseases in China.
The screening of germplasm for resistance to powdery mildew was done under natural
epiphytotic
conditions
and inoculationof seedlingsand detached leaves using isolatesof known virulence in the various regions. Of the 3,441 accessions of local varieties
screened, 6 accessions with immunity to powdery mildew were obtained and theirresistantgenes identified(Sheng et al., 1992). The results indicatedthat the resistantgenes carried by these 6 accessions were different from those previously
known, anddesignated
XBD. In wild relativesof wheat, about 700 accessions,
includingabout
100 species and 11 genera were screened for resistanceto powdery mildew(Table
6)(Zhou et al., 1993; Wang et al., 1994).

Table 6. Screeningof wild relativesof wheat for resistanceto powdery mildew
N o . ac c e s sio n s sc re e n e d

G enu sN o . s p e c ie s s c re e n e d

T o ta l

Im m u n e%

A e g ilo p s

14

1 18

63

5 3 .4S e c a le

2

38

38

10 0 .0

E re m op y ru m

4

14

10

7 1 .4

H ordeu m

15

33

19

5 7 .6

A gropy ron

4

32

4

1 2 .5R o e g n e ria

21

16 1

47

2 9 .2E ly m u s

20

2 33

50

2 1 .4E ly trig ia

2

8

4

5 0 .0L eym us

7

83

14

1 6 .7

P s a th y ro sta c h y s

2

6

1

1 6 .7T h n o p y ru m

6

6

5

8 3 .3P s e u d o ro e g n e r ia

5

5

4

8 0 .0T o ta l

10 2

7 37

25 9

3 5 .1

Wheat scab. Wheat scab is a major wheat diseases in China. For screening
resistance to wheat scab, a national cooperative
group was establishedand a total of
34,571 accessionsof wheat genetic resources were screened. No wheat variety was
found to be immune to the disease.
However, 1,765 accessions were resistantor
moderately resistant to infection
development
when Sumai 3, a cultivar noted for its
tolerance to wheat scab, was used as control variety (Table 7) (National
cooperative
group for study of wheat scab, 1984). Some accessions of R. kamoji and R. ciliaris
were also reported to be highly resistantto wheat scab and are being used for wheat
improvement (Liu et al., 1990).
Table 7. Screening of wheat germplasm for resistance to scab*
S p e c ie s

N o . o f a c c e s sio n s

T r i ti c u m a e s t iv u m

R e s is t a n t o r m o d e r a te r e s i s t a n t%

L oc al

1 3 ,1 10

4 70

3 .4 4

Im p ro v e d

1 0 ,3 2 4

1 ,1 5 5

1 1 .1 8

F o re ig n

9 ,1 8 4

137

1 .4 9

1 5 ,5 7 0

00

W h e a t r e l a t iv e s #26

2

7 .6 9T r it ic a le

170

1

.5 9

1 ,7 6 5

5 .1 1O t h e r s p e c i e s in T r i ti c u m

T o ta l

3 4 ,5 7 1*, Sumai 3, a notablecommon wheat cultivar#, includingfrom Aegilops,

speciesfor its tolerance

Secale and Dasypyrum

to scab, as control

speciesBYDV. BYDV is recognized
as a major pathologicalconstraint
to cereal
production in northern China. To date, no accession, including
all species of Triticum,
was found to be resistant
to BYDV, and only a few showed moderate tolerance to thedisease.
However, resistanceand immunity to BYDV were found to be widelydistributed
among the indigenous
wild relativesof wheat in northern China (Table 8)(Dong

et al., 1992;

Zhou et al., 1993;

Xu et al., 1994).

Table 8. Screening of perennial Triticeae for resistance to barley yellow dwarf virus strain PAV
N o . a c c e s s io n s s c r e e n e d
G en usN o . s p e c ie s s c r e e n e d

T o ta l

Im m u n e%

H o rd e u m

3

2 4

1 1

4 5 .8A g ro p y r o n

5

3 3

5

1 5 .2R o e g n e r ia

15

10 0

13

1 3 .0E ly m u s

10

17 0

6 1

3 5 .9E ly tr ig ia

2

6

3

5 0 .0L ey m u s

7

7 1

2 9

P s a t h y r o s ta c h y s

2

5

1

T o ta l

4 4

40 9

12 3

4 0 .820 .0
3 0 .1

Resistanceto Pests
About 1,000 accessions of commonwheat were preliminarily
screened forresistanceto three kinds aphids,
Toxoptera graminium Rond., Macrosiphum
granarium Kirby, and Rhopalosiphum padi L. Only a few of accessions were foundto be moderately

resistant

to aphids(Ma, 1986;

Tong et al., 1991).

Resistanceto Environmental
Stresses
Evaluationfor environmental
stresses included
drought, salt, coldness andwater logging. About 16,000, 3,300, 3,000 and 1,500 accessionsof wheat genetic
resources were screened for the above mentioned environmental stresses, respectively.
Some accessions with high resistance to environmental
stresses were identified(Dong
et al., 1992; Xiao et al., 1995).

Crossability
Chineselocal wheat varieties

are noted for theirhighcrossability

with rye.

Among 864 local varieties
tested, 50 had a crossability% significantly
higher thanthat of Chinese Spring (CS). 19 of them showed a crossability
with rye of 90% ormore. Genetic analysis on a selection, J-11, of a local variety from Sichuan province
where CS originated,revealed that it carried a new gene for crossability,
kr4, locatedon chromosome 1A. So those local varieties
with a crossability
percentage
significantly
higher than CS might have carried 4 recessive kr genes (Luo, 1992; Luoet al.,

Diversity

1992,

1993;

Zheng et al., 1992).

Analysis

Before 1995 the major tasks for wheat genetic resources workers werecollection,conservation,and evaluation of wheat genetic resources. Nowdiversity
among collections
can be analyzed using biochemical
and molecular techniques. Most
past studies dealt with the use of biochemical
and molecular markers for identificationof alien

genes or chromosome fragments in wheat background.
To collect and exploit Agropyron and Roegneria genetic resources, isozyme
variation of 7 different enzymes encoded by 28 and 26 presumptive loci, respectively,
were analyzed using leaf extracts and polyacrylamide
gel electrophoresis(Li et al.,
1994, 1995).
Variation
was found among isozyme loci both within and amongaccessions. This suggested that a new approach to collect and use wheat germplasm
should be made based on species with the different
mating systems.
Gliadin variation on 38 accessions of Aegilops tauschii were analyzed by acidpolyacrylamide
gel electrophoresis(Zhang et al., 1995). The results indicatedthat
gliadinpolymorphism
was closely related to collection
sites, i.e. Middle East >Former USSR > Xingjiang > Henan and Shaanxi. The same results were obtained by
RAPD analysis (Kong, 1996).
Using 31 RAPD primers, 4 species of Eremopyrum were analyzed. The results
indicatedthat 85.7% were polymorphic
and some bands were genus-specific
RAPD
markers. Also, the genetic relationships
among species has been determinedusing
clusteranalysis(Zhang,1996).

Future Plans
Exploration
and Collectionin Xinjiang
Xinjiang is the largest province in China with a total area of about 1,600,000

km2.

It has a very cold wintertemperature
coldin China

resistantandand extremely

occur in Xinjiang.

Localdrought

tolerant.

Moreover,that

genetic

resources

indicated

province

might

be a major

explorationand collection

Diversity

mostwheat varieties

diversity

centerthroughout

therefore,

preliminaryof species

are found

for wheat

Xinjiang

will

and the highest
are extremely

evaluation

for wheatin Xinjiang

andgenetic

be conductedthisresources.

Thus,from 1997

to 1999.

Analysis

For the future
one of the major
traitshot summer. Both the lowest

will

studies,tasks

be tagged

diversity

using
with

analysisbiochemicalmolecular

among accessions
and molecularcollected

techniques.

will

becomeSome important

markers.

Use of Diversity
The local

wheat

varieties

evaluatedwithimproved

wheat cultivars.

Transferring

wheat

shouldbe continued,

althoughobtained(Liand Hao, 1992)

and withdesirable

desirable
manycharacters
genesintergenericwill

be crossed

from wild

relatives

into

derivatives

have

been

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Crop Genetic Resources Diversity
Conservation
LUUNGOCTRINH
Vietnam Agricultural Sciences Institute,in Indochina

and Approaches

for Its

Vietnam

Abstract
Indochina is one of the most floristicallydiverse regions of the world. There are
specific factors related to history, geography, ecology and the socioeconomy of the region thathave created the rich crop genetic diversity.
There exist approximately
1000 crop species
belonging to 100 genera in the region. There are important crops, that are endemic and possessa high degree of genetic diversity in Indochina, such as rice, banana, coconut, taro, yam, grape
and lemon. Several crops introduced from the other continents, but adapted to the diverse
ecological conditions of the region, also have a relatively high degree of genetic diversity, they
are sweet potato, corn, cassava, coffee and orange.
Approaches for conservation of crop genetic resources in the region are discussed inthis paper. The main method for annual food crops is ex-situ conservation in the genebank,
but this needs to be complemented with on-farm conservation. For vegetables, perennial fruits
and cash crops, the principal approach is in-situ conservation, in which conservation in homegardens play an important role.
I. The main features of plant genetic resources in the Indochina
region.
The southeast Asia region is considered
to be one of the most diverse in plant
genetic
resources. Indochina,
consisting
today of Cambodia,
Laos and Vietnam,
possess not only Southeast Asian diversity
but also the particularfeatures,includingboth tropicaland temperate plant genetic resources.
The following
historical,
geographical,
ecological,
economic and social
factors account for the diversityof plant genetic resources in Indochina.
-Historicalfactors. In prehistoric
times Indochina was linked with Indonesia
and Malaysia
by land bridges.
This resulted
in an interchangeof plant genetic
resources

over what is now continental
and insular Southeast
Asia.
In the past, the Vietnamese
lived in the southern part of the Yangtse riverdelta. Due to war they moved south to establish
the Red River Delta civilization.
When moving to the south, the Viet dwellers brought along with them various crop
species which originatedfrom northern areas which is now China.

-Geographicaland ecologicalfactors.

Indochinalies

at the end of two

mountain chains which stretch from China and India-Myanmar. Thus, the Indochina
flora are greatly influenced
by those from South Asia as well as East Asia.
The Indochina
sea is the gateway between the Indian and Pacific Ocean,
between Asia and Oceania. Many new crops have been introduced into Indochina bysea.
Indochina is situated in the tropical region and is influenced
by the monsoon.
The north of Vietnam is characterized
by a subtropicalclimate with some featuresof a temperate climate in high mountain areas. The plant genetic resources, therefore,
involve tropical,
subtropicaland temperate species.
-Socioeconomic factors. There are approximately
one hundred minorities
living in Indochina.
Diversity
in minorities
generates crop diversity,
particularly
cultivar diversity
within each crop species. Indochina has a traditional
agricultural
economy. The traditional
agriculture,with a low degree of urbanization,
has resulted
in crop genetic diversity
being maintained
and relatively
little genetic erosion has
occurred for some crops.
Plant genetic resources of Indochina consist of three components:a) indigenous
species;b) introduced
species from South China and South Asia;c) introduced
species from the other continents.
Lecomte, in his voluminous work published between 1907 and 1941, gave an
inventory and describedmost of the plant species existingin Indochina.
Ho (1991)
stated that there are at least 12000 plant species in the Vietnamese flora, among which
the author described,
with illustrations,
10500 species. According to Ho, Indochina
is one of the most floristicallydiverse regions on our planet. Ho (1991)
cites thefollowingcomparisons; Canada, with an area 30 times bigger than Vietnam has only
4500 plant species;
the North America continent has a little more than 14,000 plant
species. In Southeast
Asia, in both Indonesia
and Malaysia, which have an area six
time bigger than Vietnam, there are about 25,000 plant species.
II. Crop Genetic
genetic

Resources

Diverse floristic
genetic
resources in Indochina.

in Indochina.
resources
Vavilov,

are the main factor creating diverse crop
Zukovski,
Zeven and others agree that

Indochina
is where several crops originatedcrop species (Paroda and Arora, 1991).

and is a center of genetic diversity
forAnother factor which accounts for thediversified
crop genetic resources in Indochina is the ancient agricultural
civilizationof the Indochinese
people. Khoi (1995)
reported that in Vietnam 734 crop species
belongingto 79 genera exist. Crop groups of Indochina include:
Crop group
Number of species-Starchy food crops
39-Non-starch food crops
95
Fruit crops
104-Vegetables
55-Spice crops
39
Beverage crops
12-Fiber crops
16-Oil crops
44-Perfume crops
19-Cover crops to rehabilitate
eroded land29
The principal
crop species of Indochina with their degree of genetic diversity
are listed (Tables
1, 2, 3 and 4).
As an example of genetic diversity
analysisof a crop species and its wild
relatives,
rice in Vietnam will be given. Rice is an economicallyimportant crop in
Indochina
and the staple food of the people. In Indochina
there are five wild Oryza
species: O. granulata, O. nivara, O. officinalis, O. rufipogon and O. ridleyi (Vaughan,1994). Among these O.nivara and O. rufipogon is considered to be the direct ancestorof cultivatedrice, O. sativa. Rice shows maximumvarietal diversity
in a broad region
from Nepal to northern Vietnam (Chang, 1976). There is a high degree of genetic
diversityof upland rice in northern Laos and northern Vietnam, as well as, deepwater
rice in southern Cambodia and southern Vietnam. Classification
based on isozyme
patterns has shown that in Vietnam 89% of varieties
are indica,
9.5% are japonica
and 1.5% are as yet unclassified.
Further, the aromatic rices of northern Vietnam arejaponica
rices (Trinh et al., 1994). A new allele of the isozyme locus Enp - 1 wasfound in traditional
rice germplasm of Vietnam from the north to the south ofcountry(Trinh
et al.,1993).

Table 1. Main food crop species
C om m on n a m e

in Indochina
S c i e n t ifi c n a m e

A rro w ro o t

M a ra n ta a r u n d in a c e a

D e g r e e o f d iv e r s it yS e c o n d a ry

B la c k b e a n

V ig n a c y li n d r i c a

S e c o n d a ry

C an naC a n n a e d u l isS e c o n d a ry

C a s sa v a

M a n ih o t e s c u le n ta

S e c o n d a ry

C o rnZ ea m a y s

S e c o n d a ry

G ro u n d n u t

A r a c h is h y p o g a e a

S e c o n d a ry

L e sse r y a m

D io s c o r e a e s c u l e n t a

P r im a r y

M u ng b ean

V ig n a r a d i a t a

S e c o n d a ry

R ic eO r y z a s a ti v a

P r im a r y

S e s am e

S e s a m u m in d i c u m

S e c o n d a ry

S w e e t ‑ p o t a toI p o m o e a b a ta ta s

S e c o n d a ry

T a roC o lo c a s ia s p p

P r im a r y

T a roX a n th o s o m a s p p

S e c o n d a ry

T a roA m o rp h o p h a ll u s s p p

P r im a r y

Y am

D io s c o r e a a la ta

S e c o n d a rv

Table 2. Main vegetable
C om m on nam ecrop species

in Indochina

A m a ra n th

S c ie n t i fi c n a m e
A m a r a n th u s s p p .

D e g r e e o f d iv e r s i t y
S e c o n d a ry

A r o m a t ic g o u r d

L u f f a c y lin d r ic a

S e c o n d a ry

B it te r g o u r d

M o m o r d ic a c h a r a n ti a

P rim a ry

B o t tl e g o u r d

L a g e n a r ia s ic e r a r i a

S e c o n d a ry

C h i li

C ap sic u m a n n u m

S e c o n d a ry

E g g p la n t

S o la n u m u n d a tu m

S e c o n d a ry

E g g p la n t

S o la n u m m e l o n g e n a

S e c o n d a ry

G a r li c

A l li u m s a t iv u m

S e c o n d a ry

T o s s a j u te

C o r c h o r u s o l it o r iu s

P rim a ry

P u m p k inC u c u r b it a m a x im a

S e c o n d a ry

R ig id g o u rd

L u ff a a c u ta n g u a

S e c o n d a ry

S h a ll o t

A l liu m a s c a l o n ic u m

S ec o n d a ry

S p in a c h

S a u ro p u s a n d ro g y n u s

P r im a r y

W a t e r c o n v o lv u lu s

Ip o m o e a a q u a tic a

P r im a r y

W e ls h

A ll iu m fis t u l o s u m

P r im a r y

W h it e g o u r d

B e n i n c a s a h is p i d a

S e c o n d a ry

Table3. Main perennial

fruit crop species

in Indochina

C o m m on na m e

S c ie n tific n a m e

D e g r e e o f d iv e r s ity

B anan a

M u sa s p p .

P rim a ry

C a ra m b o la

A ve r rh o a c a ra m b o la

P rim a ry

S ta r ap p le

C h ry so p h y llu m c a in ito

S e c o n d a ry

D u ria n

D u r io z ib e th in u s

P rim a ry

G ra p e fru it

C itru s p a ra d isi

P rim a ry

G u ava

P s id iu m g u a va

S e c o n d a ry

J a c k F ru it

A r to c a rp u s h e te ro p h y llu s

S e c o n d a ry

L em onC itru s a u ra n tifo lia

S e c o n d a ry

L em onC itru s lim o n ia

P rim a ry

L itc h i

L itch i c h in e n s is

S econ dary

L o n g an

E u p h o ria lo n g a n

S e c o n d a ry

M an d a rin

C itru s re tic u la ta

S econ dary

M an g o

M a n g if e ra

S e co n d a ry

O ra n g e

C itru s s in e n s is

S ec o n d a ry

P e rs im m o n

D io sp y r o s k a k i

S ec o n d a ry

P o m e lo

C itru s g ra n d is

S ec o n d a ry

R a m b u ta n

N e p h e liu m la p p a c e u m

P rim a ry

S o u rs o p

A n n o n a m u r ic a ta

P rim a ry

S u g a r a p p le

A n n o n a sq u a m o sa

S e c o n d a ry

W ate r m e lo n

C itr u llu s la n a tu s

S e c o n d a ry

Table

4. Main perennial

cash crops in Indochina.

C o m m on na m e

S c ie n tific n a m e

D e g r e e o f d iv e r s it y

C oco nu t

C o c o s n u c if e r a

P rim a ry

C o ffe e

C o f f e a r o b u s ta

S e c o n d a ry

C o t to n

G o ssy p iu m s p p

S e c o n d a ry

C y p e ru s G ra ss

C y p e r u s t e g e t if o r m isS e c o n d a ry

J u teC o r c h o r u s c a p s u la r isS e c o n d ar y

M u lb e r r y

M o r n s a u s t r a lis

S e c o n d a ry

S u g a r c an e

S a c ch a ru m sp p .

S e co n d a r y

T e a se e d

C a m e ll ia s a s a n q u a

S e co n d a ry

T ea

C a m e ll ia s i n e n s i sP r im a r y

T un sA le u r ite s s p p .

S e c o n d a ry

III. Approaches
to conserving crop genetic resources in Indochina
Conservation,
includingsustainable
use of plant genetic resources areinternational
issues. Based on the natural and socioeconomic conditions of the region,
we propose the following
approaches to conserve each crop species group:
1. Annual Food Crops.
This is economicallythe most important group of crop species, at the sametime is under the most serious threat of genetic erosion. Seeds of these crops are ofan orthodox nature and are well suited to preservation in cold storage conditions. The
main approach for conserving these crop species is ex-situ in the genebank. To
prevent erosion of genetic diversity
during the process of cold seed storage
preservation,
which is considered "static conservation", ex-situ conservation must becomplemented by on-farm conservation.
Genetic conservation on the farm must beorganicallylinked with rehabilitationand conservation of traditional
cultural practices
and traditionalfarming systems (Tuan and Trinh, 1996).
2. Vegetable Crops.
There are two kinds of vegetables
in Indochina,
the temperate and thetropical vegetables. Varieties of temperate vegetable crops cultivatedin the region are
mainly high yielding new varieties which have seeds which show orthodox behavior
in cold storage conditions. Their method of conservation,
therefore,
can be the same
as for annual food crops.
Tropical vegetable species are more widely distributed.
They are principallyused in rural areas and are the main source of vegetables
for the rural people. Their
genetic erosion is still low. Thus, the most suitable method for their conservation is
in-situ.
In-situ conservationof genetic resources in home gardens is appropriate
provided suitable
monitoring is undertaken.
3. Perennial Fruit Crops.
Due to economic development in the region, the demand of people for fruit
has been increasing.
This situation
creates favorable conditions
for the developmentof fruit production
and consequently
conservationof fruit crop genetic resources isnecessary. Almost all the fruit cultivars
in Indochina
are traditionalvarieties
andprincipally
cultivatedin home gardens. Therefore,
in-situ conservation
in homegardens should be the main approach for fruit crop conservation.
It can becomplemented by in-situ conservation
through establishmentof plantationsof fruitcropsor by fieldin publicparks.

4. Perennial
quicklygenebanks

Cash

Crops.

Like

fruit

crops,

since

the recent

regionhave been introduced

cash

crop

approachcultivarsof theirhome gardens
plantations

throughthe production
economic

are composed
conservation
example

fruit

crops

from diverse

ecologicalof cashcrops

in Indochina

has developed

renovation.

from otherThe

continents

of localfor coffee

exampleareas

majorityof cashcrops

some hundred

years

ago. Thus,and newly

must be the in-situ

for same crops (for(for

planting

for coconut

introduced

method.

the

The

main

It can be performedinand cashew)

ones.

in theor by developing

and rubber).

References
Chang T. T, 1976. The origin, evolution, disseminationand diversificationof Asian and African Rice.
Euphytica 25: 425 - 441
Ho, Pham Hoang 1991. Cayco Vietnam Mekong Printing, Santa Ana, USA (in Vietnamese)
Lecomte M. H., 1907 - 1941. Flora general de l' Indo - China. Masson et Cie Editeur, Paris, France.
Khoi, Nyugen Dang, 1996 - Report of the Project Director.Page
10-26 in Plant genetic resources inVietnam. Proceeding of the National Workshop on Plant Genetic Resources. 28 - 30 March
1995, Hanoi, Vietnam.
Paroda R. S., Arora R. K, 1991. Plant genetic resources: General perspective.
In: Plant genetic
resources, conservationand management. Malhotra Publishing
House, New Delhi, India
Trinh, Luu Ng, B. C. de los Reyes D. S. Brar and G. S. Khush, 1993. A new allele of Enp-1 in rice
germplasm of Vietnam. Rice Genetics Newsletter 10:85-85,
Rice Genetics Cooperative.
Trinh, Luu Ng, Dao The Tuan, D. S. Brar, B. G. de los Reyes and G.S. Khush, 1995. Classificationof
traditional
rice germplasm from Vietnam based on isozyme pattern. Pages 81-83 in Vietnam
and IRRI: A partnershipin rice research, 81 - 83. IRRI, Philippines.
Tuan, Dao The and Luu Ngoc Trinh, 1996. The biodiversityof agro-ecosystem and sustainable
development.
Pages 107-111
in Plant genetic resources in Vietnam. Proceedingsof theNational Workshop on Plant Genetic Resources, 107 - 111. 28 - 30 March 1995, Hanoi,
Vietnam.
Vaughan D. A. 1994. The Wild Relativesof Rice: a genetic resources handbook. IRRI, Manila, The
Philippines

International
KAZUTOSHI

Collaboration

on Plant Diversity

OKUNO, MASUMI KATSUTA, HIROKI

Analysis
NAKAYAMA, KAORU EBANA andSHUICHI FUKUOKA
Laboratory
Nationalof Plant
InstituteKannondai

Genetic

Diversity,of Agrobiological

2-1-2, Tsukuba,

IbarakiDepartment of Genetic

Resources

I,

Resources
305, Japan

Introduction
Research on plant evolution involves determining
where regions of genetic
diversity
are located and how crop landraces and their wild relatives
with distinct
characteristics
are distributed.
This research helps to determine target regions for
ex-situ and in-situ conservationof plant genetic resources.
Centers of genetic
diversity may be a candidate for explorationand collectionof plant genetic resources.
Edges of genetic diversity
are also important due to unique germplasm which may
exist in such locations.
Cryptic characters which are difficult
to evaluate morphologically,
includereproductive
barriers
and polymorphismat the peptide
and DNA levels.
Polymorphism in seed storage proteins, isozymes and DNA furnishes genetic markers
for diversity
analysisand are generally
free from artificial
selection.
Variation inphysiologicaland morphologicalcharacters may be the result of artificial
selectionor natural selection in specific environments. Artificial
selection for physiologicalandmorphologicalcharacters occurs in crops, but not in their wild relatives.
Research on
genetic variationin cryptic,
physiologicaland morphologicalcharacters can help
improved understanding
about genetic diversity
and phylogenetic
relationships.
The
recent advances in plant genome research have given genetic diversity
analysis newtechnologies
for molecular characterizationof plant genetic resources.
For the past two decades Japanese scientistsof the Ministryof Agriculture,
Forestry and Fisheries(MAFF) have collaborated
with more than 40 countries toconserve plant diversity
and exchange information
on germplasm. The collaborative
exploration
missions and collaboratorsthat the Laboratory of Plant Genetic Diversity,
NIAR, has participatedin since 1984 is shown (Appendix).
Since 1989, we have takena part in research collaboration
with scientists
in Pakistan,
Russia, central Asian
republics

and Vietnam to analyze and to conserve diversity
This report deals with the results of internationalof plant genetic resources.
collaboration
on diversity

analysisof plant

genetic

resources.

Plant

Genetic Diversity
in Pakistan
Pakistan shares commonborders with Afghanistan,Iran, Tajikistan,
China
and India, and is partly included in the Central Asian center of crop genetic diversity.
Ancient trading routes, such as the silk road, have contributed
to the introductionofvarious kinds of crops to Pakistan from the East and West. Crops are adapted to thevariable geography and climate of Pakistan.
Explorationin Pakistan was undertaken to investigate
the current situationof plant genetic resources over a wide area in 1989 and 1991. It was recognized
during the explorationthat Pakistan, in particular
the mountainous valleys in northern
Pakistan, still has a broad diversity
for cereals and food legumes. In total, crops from
15 families,
42 genera and 57 species were collected(Okuno et al., 1995). Major
samples collected were rice (Oryza sativa) (249 samples), mungbean (Vigna radiata)(126 samples), blackgram (Vigna mungo) (68 samples), foxtail millet (Setaria italica)(64 samples),sorghum (Sorghum bicolor) (59 samples),commonbean (Phaseolus
vulgaris)(58 samples) and pearl millet (Pennisetum americanum) (52 samples).
Rice cultivation
is mainly concentrated in four distinct agro-ecologicalzones
in Pakistan (Chaudhri,1986). The first zone consists of northern mountainous areas
including the North-West Frontier Province (NWFP). In the NWFP rice is cultivatedin the areas between 500 and 2000 meters. This area has great variationin air andwater temperatures (Rosh and Syed, 1986). The second zone lies in the broad strip of
irrigated land between the Rivers Ravi and Chenab in Punjab Province. The climate
is subtropicaland suitable for cultivating
fine aromatic varieties such as Basmati. The
third and fourth zones comprise the large tract of land on the west bank of the River
Indus and the Indus delta in the Sind Province.
Landraces of rice are still grown either in flat valleys or terraced valley sides
in northern Pakistan. Paddy fields in this area were covered by two different japonica
rice varieties,
Nali and Byene (Bayan),which are characterized
by round grains andtolerance
to cold injury. These two varieties differedfrom one another in cultural
practices(transplanting
or direct-seeding),
resistanceor susceptibility
to rice blast
fungus, positiveor negative reaction to phenol, and presence or absence of seed
dormancy (Katsuta

et al., 1996).

Genetic variability

in seed proteinof commonbean germplasm was analyzed

using sodium dodecyl sulfate polyacrylamide
gel electrophoresis(SDS-PAGE).
Based on patterns of phaseolin which is a major seed storage protein in commonbean,landraces collected were classified
into two types, A and B which correspond to the
T and S patterns describedby Gepts et al. (1986).
These two types were further
subdivided
into 6 patterns of type A (A1-A6) and 4 patterns of type B (B1-B4), basedon variation in proteins with higher molecular weight. A total of 10 patterns wereobserved in the accessions from the New World where commonbean originatedandis most diverse.
Seven out of the 10 different
patterns
detectedin the world
collectionsof commonbean occurred in Pakistanilandraces (Okuno et al., 1995).
They involved only 3 of 6 subgroups of type A and all subgroups of type B. Morethan 80% of Pakistani landraces showed type B which was more frequently
detectedin the accessions of the Middle America. Most Pakistanilandraces of commonbean
may have been introduced from the Middle America. The geographic distributionofpatterns of seed protein in common bean is shown (Fig.1).
Type B3 predominatedthroughout NWFP and Punjab Province.
There are 3 different types of endosperm starch in foxtail millet germplasm,
glutinous, low amylose and nonglutinous(Afzal et al., 1996). These starch propertiesare controlled
by multiple
alleles at the wx locus. All the samples of foxtail millet
collected
in Pakistan were nonglutinousand produced a higher level of Wx proteinwith a molecular weight of 60kDa which is responsible
for amylose synthesisin theendosperm and pollen grain (Echt and Schwartz, 1981). Landraces of foxtail millet
could be divided into 3 groups, the Chitral, the Baltistan
and the Dir groups, based on
several agronomic characters such as plant height, size of panicle and number oftillers(Ochiai
et al., 1994). Each group was similar to the landraces from different
regionsof Eurasia

surrounding

northern

Pakistan.

Plant

Genetic Diversity
in Central Asia and North Caucasia
The Central Asian countries
of Turkmenistan,
Uzbekistan,

Kazakhstan,

Kirgizstan
and Tajikistan
are a center of diversity
for such cultivatedplants as apple,
pear, broad bean, onion, garlic and spinach.
North Caucasia is located adjacenttothe primary center of genetic diversity
for wheat, barley, oats and their wild relatives.
Since 1992, collaborative
missions between NIAR and the N. I. Vavilov ResearchFig.1

Geographicalvulgaris)

Instituteand altitudal

distribution

colletctedof Plant

proteinelectrophoregramsof common bean

were undertaken

to investigate(Phaseolus

in Pakistan.

Industry

distributionof seed(VIR),of different

Russia,kinds of plant genetic

geographic

resources in these regions.

Geographic
variationand phylogenetic
differentiationof the genus Aegilops in theCentral Asia and north Caucasia as revealed RAPD analysis will be discussed here.
A team of Japanese scientists
undertook a scientificexpedition
to Pakistan,
Afghanistan
and Iran to collect Aegilops and Triticum germplasm (Kihara et al.,1965). Our collaborative
missions were undertaken to explore the genus Aegilops inCentral Asian republics, Turkmenistan, Uzbekistan and Kazakhstan in 1993 and north
Caucasia in 1994. Five species of the genus Aegilops, Ae. squarrosa, Ae. cylindrica,
Ae. crassa, Ae. juvenalis and Ae. triuncialis
were collected in the Central Asia. Four
species,Ae. squarrosa, Ae. cylindrica,Ae. biuncialis
and Ae. triuncialis
werecollected
in north Caucasia between the Black Sea and the Caspian Sea. Amongthem, Ae squarrosa,Ae. cylindricaand Ae. triuncialis
were widely distributedin bothregions and Ae. squarrosa includes 2 subspecies,
typica and meyeri.
Intraspecificand interspecificdiversityof Aegilops species, except for Ae.
juvenalis,
was analyzed using accessions
collected
in Central Asia and north
Caucasia. The accessions were grown in Tsukuba and DNA extracted from leaves was
subjected
to RAPD analysis. A total of 197 fragments generated by PCR using 22
primers of 10 bases were scored and data were analyzed by the UPGMA method. The
results revealed that accessions from Central Asia could be divided
into 2 majorclusters, A and B, corresponding
to the DD genome species (Ae. squarrosa: DD, Ae.
cylindrica:
CCDD, Ae. crassa: DDMM, commonwheat: AABBDD) and the UU
genome species (Ae. biuncialis:
UUMM,Ae. triuncialis:
UUCC), respectively(Fig.2).
These 2 clusters were also divided into sub-groups which corresponded
to the four
species of Aegilops and commonwheat. Intraspecificdiversity
in Ae. biuncialis
andAe. triuncialis
with the UU genome from north Caucasia was much greater than Ae.
squarrosa and Ae. cylindrica
with the DD genome (Fig.3).
Accessionsof Ae.
cylindrica
were clearly divided into two groups based on their natural habitat (Fig.4).
Such geographicalvariationof Aegilops also occurred in Ae. squarrosa and Ae.
triuncialis.
Accessions from Central Asia formed a separate cluster from those ofnorth Caucasia,
suggestingthat populations
differentiatedas a result of geographic
isolation.
in the genus Aegilops indicatedthat accessions
that those from north Caucasia.

of Aegilops
in these two regions
Comparative analysis of the diversity
from Central Asia were more diverse

Fig.2

A clustering

dendrogramof the genus Aegilops

collected

in central

Asia basedon RAPD

analysis.
Wh:common wheat, cy:Ae. cylindrical
tr:Ae. triuncialis,
Tur:Turkmenistan,
AFG:Afghanistan

st:Ae. squarrosa var. typica, cr:Ae. crassa,
Uzb:Uzbekistan,
Kaz:Kazakhstan,
TUR:Turkey,

Fig.3A clustering
analysis.

dendrogramof the genus Aegilops

collected

in north

Caucasia

basedon RAPD

Fig.4

A clustering

dendrogramof Aegilops

cylindrica

collected

in central

Asia and north

Caucasia.

PlantGenetic Diversity
in Vietnam
Since 1994, collaborative
missions between NIAR and Vietnam Agricultural
Science Institute(VASI) have been carried out to collect genetic resources of rice,
food legumes, vegetables, citrus and taro. Rice germplasm shows greater diversity
ina broad region from Nepal to northern Vietnam (Chang, 1976). Within Vietnam,
geographic and ethnic diversity,in addition to a long traditionof rice cultivation
anddiversified
farming practices,
have resulted in broad diversity
in Vietnamese rice
germplasm (Trinh et al., 1995). A major objective of the collaborative
missions was
to focus on exploration
for and collectionof rice landraces. Five missions exploredrice germplasm in northern Vietnam near the border with China and Laos and haveresulted in more than 600 indigenous
varieties being collected
since 1994.
About 450 landraces of rice collected in 18 provinces throughout Vietnam wereanalyzed
for esterase isozyme patterns,
SDS-PAGE patterns of seed protein,endosperm properties and reaction of seeds to phenol. On the basis of the genotypes
for 3 different
loci, Est1, Est2 and Est3, which are responsible
for variations
inesterase isozymes, rice varieties worldwide were classified
into 12 types (Nakagahra,
1976). Among them, 11 types were found in landraces collected in northwest Vietnam
compared to 12 types found in landracesfrom neighbouring
Yunnan province of
China(Fig.5).
Seven and 8 types existed in landraces
collected
in central andsouthern provinces of Vietnam. Type 3, which was widely spread in indica varieties,
predominatedamong landraces in the south. This result showed in agreement withthe results obtained
based on 5 isozyme loci, Pgi1, Pgi2, Amp1, Amp2 and Amp3
(Trinh et al., 1995).
Trinh et al. (1995)
reported
that 88.8% of landraces from
northern

Vietnam belonged to indica rice and 9.5% belonged to japonica rice.
There are three different
subunits of glutelin,which is a major seed storage
protein, in the rice endosperm. No variation was detected in -1 and -2 subunits among
Vietnamese rice landraces. Landraces were differentiatedinto 2 types of -3 subunit
comprisinghigher (type A) and lower (type B) molecular weight. About 70% of
landraces collectedin central and southern Vietnam showed type A, whereas only
36% of landraces
collected
in northwest Vietnam showed type A. One landracecollected
in central Vietnam lacked -1 subunit.
From data on the amount of amylose
and starch granule bound Wxprotein, landraces were divided into 3 types of starchrepresenting

glutinous,

intermediateand nonglutinous

endosperm

characteristics.

Fig 5 Variationof esterase

isozyme zymograms in Vietnameserice landraces.

More than 60% of landraces from northwest Vietnam were glutinous and 30% of
those from central and southern Vietnam were glutinous. About 90% of landraces inthe south reacted positively
to phenol, while 60% of those in the northwest did not(Fig.6).
Geographic
cline in Vietnamese rice landraces was clearly detected forglutelin
subunits, starch characteristics
and reaction of seeds to phenol.

Fig. 6 Geographicalclineof reactionof seeds to phenol

in Vietnameserice landraces.

Conclusion
For the past two decades, International
collaboration
with more than 40
countries and CGIAR centers has been carried out to conserve ex-situ plant genetic
resources worldwide. In 1985 the MAFF Genebank Project was developedtocontributeto Internationaland national
efforts to conserve genetic resources. In
addition,
exploration,collection
and research on plant genetic resources has beenenhanced with support from the International
Plant Genetic Resources Institute(IPGRI).
Collaborative
researches have contributed
not only to Improved conservationof genetic resources but also to deepening our knowledge of plant genetic diversity.
However, since the "Conventionof Biological
Diversity"
came Into effect In
December of 1993, some countries have requested new types of bilateralagreementon joint explorations
and transfer of plant materials collected.
This issue is under
on-going discussion
Internationallyand Japan Is developing
new collaborative
mechanisms to ensure conservation of plant genetic resources.
Further in depth research is required to fully understand what leads to genetic
erosion on the one hand and also genetic diversity
on the other. Greater emphasis isplaced on collaborative
researches both in the field and in the laboratory
to Improve
explorationand conservation
activities.
In particular,
laboratory
research now
increasingly
uses different
DNA markers. Among them, microsatellite
DNA and
AFLP markers are considered to be new technologies
for detecting
genetic variation

in plant

genetic

resources

efficiently.

References
Afzal, M., Kawase, M., Nakayama, H. and Okuno, K. 1996. Variation
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in foxtail

millet.

In Progressin electrophoregramsof total seedin New Crops, ed. J. Janick,

ASHS Press,

p.191-195.
Chang,

T.T., 1976.
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The origin,evolution,

rices. Euphytica

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elements

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C.G. and Schwartz,
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T.C., Rashka,

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K. and Bliss,

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Results

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and genetic

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resources
1986.

Rice

centersofdiversity

M., Nakagahra,M.,

in Pakistan.

Plant

Resources

cultivationin mountain

Trinh,

L.N., Tuan, D.T., Brar, D.S., de los Reyes,

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Breeding

M.S. and Ahmad, Z. 1995.

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Pakistan.

M., Takeya,

genetical

Scientificand distribution

in the mountainous

Nakano, H., Anwar, R.,Bhatti,

forms

of rice germplasm

Afghanistan

italicaOkuno, K., Katsuta,

in wild

for multiple

differentiation

sativa L.) by isozyme analysis.
Trop.Agric.Res.Ser.11:77-82.
Y., Kawase, M. and Sakamoto, S. 1994. Variation
P. Beauv.)

variability

JARQ 30:61-67.

and Triticum

relatives,the

99:275-284.

K. and Tanaka, M. 1965. Morphological,

studieswithin40:451-468.

M. and Anwar,

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and Hindu Khush, 1955, Vol.1,
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Ochiai,

Evidence

domestication.
collected
Kihara,D. 1981.

rice (Oryza
milletScience(Setaria

44:413-418.

Kawase, M., Nagamine,of Pakistan
Newsletter

T.,

and Japan

101:16-19.

of NWFP. Progressive

Farming

6:34-37.
B.G. and Khush,G.S.

rice germplasm

from Vietnam

based on isozyme

Riceed. Denning,

G.L.and Xuan, V.T. IRRI

Research,pattern.

1995.Classification

In Vietnam

and IRRI:

and MAFI, p.81-83.

of traditional
A partnershipinAppendix

List of overseas explorations

in which

staff members of the Laboratoryof Plant

Genetic

Diversity,

C o lla b o ra tin g c o lle c to r s

NIAR, have taken part in since

1984

C o u n tr y

C o lla b o r a t in g in s t it u t io n s

P e rio d o f e x p lo ra tio n

N ep al

RIn etes or nu arct ioe sn a l B o a rd f o r P l a n t G e n e t ic

1 N o v .‑ 1 1 D e c , 1 9 8 4

R e g io n

S a m p le s c o lle c te dw e st a n d e ast N e p a l

c e r e a ls , f ru it t re e s ,v e g e ta b le s , 1 1 5 0

N ep al

RIn etes or nu arct ioe sn a l B o a rd f o r P la n t G e n e t ic

8 O c t.‑ 2 6 N o v ., 1 9 8 5

B . K . B a n iy a , M . N . S u b e d i n o rt h w e s t N e p a lc e re a ls

c e r e a ls , f ru it t re e s ,v e g e ta b le s , 2 8 7 0

In d o n e s ia

C ro
e n pt sra l R e s e a rc h I n s t itu te f o r F o o d

2 0 F e b .‑ 1 4 M a r ., 1 9 8 9

S . K a rt o w i n o to

S u m a t ra

r ic e , 2 0 9

P a k is ta n

CP ao ku isn ctailn A g r ic u ltu r a l R e s e a rc h

1 1 O c t .‑ 2 4 N o v ., 1 9 8 9

R . A n w a r, M .S .B h a tt i,
Z . A h m ad , M . A fz al

N W F P , P u n j a b , B a ltis t a n ,B a lu c h is ta n , S in d ,K a s h m ir

m u lt i c ro p s ,7 0 5

P a k is ta n

CP ao ku isn tc ailn A g r ic u ltu r a l R e s e a rc h

1 1 S e p .‑2 O c t. 1 9 9 1
1 3 O c t .‑ 6 N o v . 1 9 9 1

R .A n w a r, M .S . B h a tt i
Z . A h m a d , M . A fz al

N W F P , P u njab N W F P ,G ilg itf o o d le g u m e s , 3 0 2m ille t s , 1 1 2I n d ia

RN ea stioo unrca le sB u r e a u o f P la n t G e n e t ic

2 7 S e p .‑ 1 2 O c t., 1 9 9 2

M .N .K oppar

M a h a r a s h tr a , K a rn a ta k a

se sa m e, 16 3

M a la y s ia

M a la y s ia A g r ic u ltu r a l U n iv e r s ity

8 F e b .‑ 1 9 F e b ., 1 9 9 3

I. B . B u j a n g
S . A n th o n y s a m m y

P e n in s u la r M a la y s ia

V ig n a , 1 0 5

rc eepn ut bra lic
l A s s ia n

V ad vu iloIn
s t ryv R e s e a rc h I n s tit u te o f P la n t

9 J u n .‑ 3 0 J u n ., 1 9 9 3

V . N o s u lc h a k

T u rKk am z eank ish statan n U z b e k is ta n

A e g ilo p s , H o r d e u m
A v en a , 1 2 3rc eepn ut rbalicl A s s ia n

V ad vu iloIn
s t ryv R e s e a rc h I n s tit u te o f P la n t

2 8 A u g .‑2 4 S e p ., 1 9 9 3

E . P o to k in a
K . I . B a im e t o v

U z b e k is t a n , K ir g iz s ta n

f o o d le g u m e s , 6 4

R u s s ia

V ad vu ilo
Ins trvy R e s e a rc h In s tit u te o f P la n t

4 J u n .‑ 2 J u l., 1 9 9 4

A . N . A fo n in
N .A . N av ru zb e k o v

n o r th C a u c a s ia

A e g ilo p s , H o r d e u m
A v en a , 1 2 3

V ie tn a m

V ieIns ttitun atme A g r ic u lt u ra l S c ie n c e

2 D e c ‑2 4 D e c , 1 9 9 4

L . N . T r in h
T . V . K in h

n o r th w e s t V ie t n a m

r ic e , 1 8 9S ri L an k a

P la n t G e n e t ic R e s o u rc e s C e n tre

8 F e b .‑ 2 2 F e b ., 1 9 9 5

W . M . W a s a a la
S . B a n d a ra
W . S . G . S am a rasing h
A . S . U . L iy a n a g e

w e s t e r n , c e n tr a l a n d
s o u t h e r n p a rts

f o o d le g u m e s , 1 1 9o th ers, 2 7

V ie t n a m

V ieIns ttitn ua tem A g ric u lt u ra l S c ie n c e

7 N o v .‑2 D e c , 1 9 9 5

N .T . Q u y h n
D . H . C h ie nn o r th w e s t V ie t n a m

r ic e , 1 5 4

V ie t n a m

IV n ies tit
t n ua tem A g ric u lt u ra l S c ie n c e

7 N o v .‑ 1 D e c , 1 9 9 6

L .T . T u n g , V . L . C h i
D .H .C oune

T h a n h H o a, N g h e A n

ric e , 1 5 3

riceIn-situ Conservationof Plant Communities:
Trends in Research
Genetic Variation and Differentiationof Plant Populations

into

KAZUHITO MATSUO
National

Instituteof Agro-Environmental

Sciences,

Kannondai,

Tsukuba,

Ibaraki

305, Japan

Abstract
Recent ecologicalresearch on the intraspecificvariation in plants has shown an
increasing use of genetic analysis. In order to clarify the relationships
between ecologicalfactors and genetic variationin plant populations,most studies
includeisozymeelectrophoresis,
calculationof standard genetic diversity
statistics
and an examination ofgenetic variation within and among populations in addition to field observations. These studies
show that the breeding system of species, kind of available
pollinatorsand ecologicalconditionsin habitats
are important factors to maintain the genetic diversityof plant
populations.
So, the integrationof basic informationfrom both phytogeographicalandecological studies to diversity analysis will be necessary for successful in-situ conservation of
species.
Introduction
Within populationsof most wild species, different
individuals
look quite
similar, but they are probably geneticallydistinctfrom all other individuals,
due tohabitatconditions
and breeding
system.
Ecologicalresearch into intraspecificvariationof plants over the last four or five years has increasingly
used genetic
analysis. There are several basic methods to analyze population
genetic diversity.
The
most widely employed technique
is allozyme electrophoresis.
A specific method,
called sequencing, involves directly
determining
the DNA base sequence making up
the code of genetic information.
Other methods which measure variation at the DNA
level
include,Restriction
Fragment
LengthPolymorphism(RFLP),
DNA
fingerprinting,and Random Amplificationof Polymorphic
DNA (RAPDs).
In this workshop, I discuss how these techniques
are used to analyze thegenetic variationin wild plant populations.
Then I will introduce
some recent
eco-genetic
studies on wild plants which provide basic and valuable information
relevant to in-situ conservationof plant communities.

Advances in analysis of genetic variationin plant populations
The number of publishedplant studies between 1991 to 1996 with genetic
variationor genetic diversity
as key words is shown (Table 1). These studies which
use four genetic methods -allozyme electrophoresis,
RAPDs, RFLPs and DNA
fingerprinting
The method"isozyme" electrophoresis
would probably
include"allozyme" electrophoresis
since "isozyme" is more general term than "allozyme".
Isozymes are multiple forms of a single enzymes. While the term allozyme is used torefer to allelicform of an enzyme. Therefore allozymes are genetic markers forquantifying
heterozygosity,
intra- and interpopulation
genetic variation.
Allozyme
methodology
is the most frequently
used of the four methods when both genetic
variationand genetic diversity
are used as key words.
Studies of allozyme variation in plants have several advantages over othermeasures of genetic variation. It is relatively
inexpensive and it can be applied to mostplant species. The same isozyme loci can be analyzed in all populations
or acrossrelated species, and estimates of the levels and distributionof genetic variation canbe directly compared. Allozyme loci have great utility as markers to describe patternsof genetic diversity
and it can also be a useful yardstick to measure the effectivenessof in situ and ex situ conservation
programs (Hamrick et al.,1991).
Most studies
includeallozyme electrophoresis
to clarify
the relationships
between ecologicalfactors and genetic variation in plant populations.
Allozyme variation is widely used
to study plant populationsin Japan.
Plant species which have been analyzed for intraspecificvariation by genetic
methods in recent years is shown (Table 2). Some of these species are native to Japan.
Other species are referred to in Japanese botanicaljournals-The
Journal of Plant
Research,
The Journalof Phytogeography
and Taxonomy and Plant Species
Biology-which
publish research on taxonomy, genetics, ecology. Species in thesestudies were analyzed by ecologicalgenetic and phytogeographic
methods rather thanby taxonomic methods. These studies have different
objectives,
but the data analysis
proceduresare very similar.
The general procedures for the ecologicalgenetic studies listed in table 2,
include data analysis
after resolutionof putative
loci, the detectionof variationatloci, the calculationof standard genetic parameters, the percentage of polymorphic
loci, number of alleles per locus, and genetic diversity(Fig.1).

Table1. Number of publishedin plant

speciesstudiesanalyzed

K e y w o rd s

using

"genetic

by four genetic

variation"
methods

and "genetic

diversity",

as key words,

from 1991 to 1996*.

A llo z y m e(Is o z y m e )RA PD sG e n e tic v a ria tio n

10 1(10 1)

6

5

15

G e n e tic d iv e rsity

64(8 5 )

4

14

24*Data obtainedTable

2. Species

selectedjounals*.

from a searchthroughanalyzed

for intraspecific

R FL Ps

D N A f in g e rp rin tin g

BIOSIS.
variationby allozyme

electrophoresis

in recent years fromS p e c ies

F a m ilyA u th o rs

A g a th is b o rn e e n s is
C a ly s te g ia so ld a n e lla

A ra u c a ria c e a e
C o n v o lv u la c e a e

K ita m u ra e t a l.( 19 9 2 )K im a n d C h u n g (1 9 9 5 )C a m e llia ja p o n ic a
C a m p a n u la p u n c ta ta

T h e a c e ae
C a m p a n u la ce ae

I n o u e a n d K a w a h a ra (1 9 9 0 )C a rp in u s lax ifl o r a

B e tu la c e a e

K ita m u ra e t a l.(1 9 9 2 )E u ry a em a r g in a ta

T h e a ce ae

C hun g and K ang (199 5)

E . j a p o n ic a

T h e a c e ae

C h u n g a n d K a n g ( 1 9 9 4 ) , O h e t a l (1 9 9 5 )F a g u s c re n a ta

F agaceae

K ita m u ra e t a l.(1 9 9 2 )F . j a p o n ic a
G ly c in e s o y a

F agaceae

K ita m u ra e t a l.(1 9 9 3 )K ia n g e t a l (1 9 9 2 ) , Y u an d K ia n g ( 1 9 9 3 ) , F u jita

H o s ta c a p ita ta

L ilia ce ae

C h u n g (1 9 9 4 )H . c la u sa
H . m in o r

L ilia ce a e
L ilia ce ae

C h u n g (1 9 9 4 )H . y in g er i

L iliac e a e

C h u n g (1 9 9 4 )C h u n g an d C h u n g (1 9 9 4 )M o n o c h o r ia k o r sa k o w ii

P o n te d e riac e a e

W an g e t a l.(1 9 9 6 )M . v a g in a lis

P o n te d e ria c e a e

W an g e t a l.(1 9 9 6 )P rim u r a c u n e if o lia

P rim u la c e a e

S h in d o e t a l.( 1 9 9 5 )S a lso la k o m a r o v i

C h e n o p o d ia c e a e

K im a n d C h u n g ( 1 9 9 5 )S a u ss u re a n ip p o n ic a

C o m p o s itae

Im ( 1 9 9 1 )T r iilliu m k a m ts c h a tic u m

L ilia c e a e

O h a ra e t a l.( 1 9 9 5 )V ite x ro tu n d ifo lia

V e rb e n a c e a e

L e g u m in o s a e

O h e t a l. ( 1 9 9 5 )e t a l. (1 9 9 7 , in p re ss )*The Journal
Biologyof Plant

Research,

The JournalY e e h n et a l (1 9 9 6 )of Phytogeography

and Taxonomy and Plant

Species

Hamrick and Godt (1989)
reviewed plant allozyme literature
at the species
level for 440 species of angiosperms. To calculate the distributionof genetic variation
within and among populations,many authors have used Nei's (1973)
genetic diversity
statistics,

-Ht
-Gst

Fig.1.

General procedure for ecologicalextraction and electrophoresis

genetic

analysisin plant

populations

Resolutionof putativeloci and detection

Calculationof standard

genetic

Comparison

with previous

data. Many authors

referredto the review by Hamrick

Calculationof Nei's

genetic

statistics

withinComputation(Nei,1972)of genetic(1973)

of variation

studiesparameters

distance

and/orat lociat the species

diversity

identity

Generating
dendrograms based on the genetic
analysis(Sneath and Sokal, 1973)

after enzyme

and within

betweendistances

populationlevelsand Gott (1989)

and among populations

populations

or identity

for pair-wisematrixcomparisonby the UPGMA cluster, total genetic diversity;-Hs, gene diversity
within populations;and, differentiation
among populations.
After computing the genetic distance
and/or the genetic identity
betweenpopulations
for the pair-wise comparison method (Nei, 1972), dendrograms generated
are often using the UPGMA cluster analysis method (Sneath & Sokal, 1973).
Analysisof the genetic variation
in a wild progenitor
of a crop
Breeding system is a major determinantof the genetic structure of population.
Considering
the maintenance mechanisms of genetic variationin wild progenitorsofcrops, reproductive
features from self-incompatible
to self-compatible
and from
outbreeding
to inbreeding,
are known affect genetic variationin crops. The wild
soybean, G. soja Sieb. et Zucc. is an annual herbaceous species and widely distributed
in north-eastern
Asia. This species is the most probableancestor of cultivatedsoybeans.

The cultivatedsoybean,Glycinemax (L.) Merr., has been cultivatedforthousands of years in China, Korea and Japan. It is an important crop because of its
high protein
and oil content. Wild soybeans consistently
shows a higher level ofgenetic variation than the cultigen. Many studies reporting the genetic variationof G.
soja based on isozymes and other genetic markers. Results of these studies show thatthe amount of variationis comparable to, or higher than that, in other self-fertilizedplant

species.
Comparative
estimate of outcrossing
rates between cultivatedand wild
soybean is shown (Table 3). The previous reports suggested that the rate of crosspollinationin cultivatedsoybean was very low, that is less than 3%, because it isself-fertileand self-pollinating.
However, the outcrossing
rate reported by Beard andKnowles (1971)
was increased
to 14% in artificially
increasing
the honeybee
population.
Although the wild soybean, G. soja, is also believed to be predominately
self-pollinating,a study in Japan showed a low outcrossing rate of 2.3%( Kiang et al.,1992) which is similar to cultivatedsoybeans.
Fujita et al. (1997,
in press) reportthat the outcrossing
rate for G.soja was about 13%, which is much higher than theoutcrossing rate estimatedby Kiang et al. (1992).
In the study of Fujita et al. basedon the genetic structure of G. soja populations
along the Omono River in Akita
Prefecture,
the authors noted the number of flowers visited by insects.
Previously
little attention
has been paid to the prevalence of pollinatinginsects on wild soybeans.
The frequency distributionof the visitors to flowers of G. soja is shown(Fig.2).
Honeybees were the most frequent visitors to the flowers and carpenter bees
the second most frequent. It seems that pollinators
are readily available
to pollinate
wild soybeans. The effect of habitat factors on genetic variation in G. soja populationsin this area is as follows. A relatively
undisturbed
habitat allows a larger and higher
density
population
to be established.
Consequently,
many insects frequently
visit
flowers to collect nectar, providing
a great potential
for cross-pollinationand higher
outcrossing
rate.
This study demonstrates
the importanceof insects and their
behavior
in visiting
flowers to maintain and/or increase genetic variationin wild
soybeans. A major conclusion of this study was that maintaining
large populationsof wild progenitorsof crops, which are important genetic resources, should be a
criticalconsiderationin the recurrent selectionof plants or the developmentof
cultivars.

Table 3. Outcrossing

rate of Glycine

maxand G. soja

C ultiv ated so yb ean
S p ecies

G . m ax

A utho r

G . m ax

Fig. 2 Frequency
individuals

capturedG . soja

B eard andK ian g et al.

F ujita et al.

stu dies*

K n ow les (197 1)(19 92)(in p ress)< 3%

14 % **:from Ahrent and Caviness 1994; Caviness 1966;**:the data obtained
from the experimentin whichthe natural

G . soja

P reviou sO utcrossing rate

beyondW ild soyb ean

2 .3%

13 %

Weber and Hanson 1961.
the honeybee populationis artificially

increased

levels.

distribution
in a squareof visitorsof flowersof Glycineof 4‡u in two hours (datasoja. Percentages

from Fujita

et al., 1997of visitors

to totalin press)

Clarificationof the relationshipbetween breeding system and genetic structure
Considering
differentiation
mechanisms
between plant
populations,differencesin genetic structure are associatedwith contrasting
breeding
systems
Ohara et al. (1996) clarified
the relationshipbetween the breeding system and genetic
variationof Trillium
kamtschaticum
based on morphologicalvariation
andHabitats hardly disturbed by shore protection and other human interventions
Preservation of larger G. soja populations and higher plant densities
Providing an attractive reward (nectar) for potential pollinators
Frequent visits by insects (honeybees and carpenter bees) to flowers
Ample opportunities for cross pollination
Higher outcrossing rate
Higher within-population genetic variation
Fig. 3. Effects

if habitat

situations

on genetic

variation

in G.soja population.

distribution
using pollination
ecology methodology.
T. kamtschaticum is an Asiatic
species of the genus Trillium.
This species is a herbaceous plant of temperature
woodland and is distributed
mainly in Hokkaido and northern Honshu, Japan. Large
populationsof this species
are found in eastern Hokkaido.
The habitatof T.
kamtschaticum
is broad-leaved
deciduous forests, dominatedmainly by Ulumus
davidiana
var. japonica, Fraxinus mandshurica var. japonica, Quercus mongolica
var. grosseserrata and Acer mono.There has been a decrease in size of populations
as a result of human activities.
Twenty three populations
were examined by the authors in Hokkaido. Floral
morphology of T. kamtschaticum in eastern populations
are characterizedby moreoval petals than northern and southern populations.
Petals of plants in the northernand southern populations
tend to be narrower. Previous comparative studies ofchromosomal variationin natural populationsof T. kamtschaticum(Kurabayashi,1957)were based on structural changes in chromosomes shown by differential
staining
at low temperatures. The results from this study revealed three, north, east and south,
geographicalgroups. To clarify
the differentiation
mechanisms between T.
kamtschaticum
populations,
Ohara et al. (1996)
discussed
the differentiation
mechanisms between allozyme characteristics
and breeding
system based on apollination

experimentwith bagged

flowers(Table4). Effects

were evaluatedbyTable4. Fourtreatmentsinpollination

experiments

on singled

floweredplantsinTrilliumkamtschaticum.
T re a tm e n t

E ffe c t *(A ) F lo w e rs u n tre a te d

O p e n p o llin a tio n (C o n tro l)(B ) B a g g in g f lo w e rs w ith n y lo n b a g p r io r to

P o wlline ra tio n w ith p o lle n fro m a n th e rs w ith in aflo

a n th e s is(C a)v Einmg aths ce um la intin ag n floo pw e en rs‑p po llinlerio ra tote da nc tho n eds itio
is a nn dbP yo llinth e a wtioinn d wo ith
r b yo thin es re cfl tso w e r 's p o lle n c a rrie da(Dn d) Eb am g ag scin ug law tioithn no ef tsflo( 1w m e mrs px rio1 m r mto ma ne thsh e)s is

oP no llinfro
mth eva iswtioitin
inn d gw a fithn lod wpo rethe rve er nflotiow n e or f's lap orglle
e r n inc sea rriec ts d*:calculationindividualof seed-setting
to total

rate per individual

number of ovules per individualfrom ratio(fromof total

number of seeds

producedper

Ohara et al., 1996).

seed-setting
rate (S/O ratio) which is the ratio of seed number per plant to total
number of ovules per plant.
The dendrogram derived by UPGMA clustering from a matrix of pair-wisecomparisons of Nei's genetic distancesfor 23 populations
revealed
two major
populationgroups. Most of the populationsin the eastern region with bagged flowers
did not produce any seeds in treatment (B), which suggests that self-fertilization
did
not occur in these populations.
The remaining major group consisted of northern andsouthern populations
and were characterized
by a low percentage of polymorphic
lociand lower genetic diversity
than eastern populations.
In all of the northern andsouthern populations,bagged individuals
produced mature seeds. This suggests thatthe plants in these populations
were self-compatible.
Some of the factors affecting
genetic
diversity
in T. kamtschaticum
populations
are shown (Table 5). The petal size is associatedwith the attractionofpollinators.
As mentioned earlier, petals of flowers in eastern populations
are thelargest and widest.
Consequently,
floral morphologyof self-incompatible
eastern
population
seem to exhibit
floralcharactersbetter suited to cross pollination.
Furthermore, larger population
size and higher density populations
tend to belong tohigher genetic diversity
groups. These results suggest that larger population
size andthe higher plant density should maintain self-incompatibility
and outcrossing
systems.
Both Fujita et al. (1997)
and Ohara et al. (1996)of plants characteristics
and plant ecology for conservation

pointed to the importanceof plant genetic resourcesTable 5. Comparison
kamtschaticumof floral,

ecologicaland reproductive

featuresaffectinggenetic

diversityin Trillium

populations.

P op ulation

E astern po pu latio ns

N o rth ern and sou thern p op ulatio ns

F lo ral featu res

L arger an d w id er p etals

S m aller an d n arro w er p etals

E co lo gical featu res

L arger p op ulation size

S m aller p op ulatio n size

R ep rod uctiv e featu res

poA ollinutb
lm oreed
st
atio
all
inn gseedarisins resug fro
lt mfromin sectoS ueedtb reeds froinm g manixdedin bsystem
reedingof

G enetic featu res

H ig her g enetic diversity

L ow er g en etic d iv ersity

and genetic

diversityof plants.

Conservation
biologyof sand dune species based on genetic variation
andpopulation
structure
Plant species, growing on beaches and sand dunes, are ecologicallyimportantbecause they initiate
and enhance formation and/or stabilizationof sand dunes. For
example, Salsola komarovi (Chenopodiaceae)
is a herbaceous
annual native tonorthern China, Japan, and Korea and Sakhalin island, Russia. The species grow only
on beaches and coastal sand dunes. After initial colonization
by annual plants, suchas, Atriplex gmelinii,
Polygonum polyneuron and Salsola komarovii, other dune
species such as, Carex kobomugi, Ischaemum anthephoroides,
Zoysia macrostachya,Calystegia
soldanella,
and Vitex rotundifolia
in succession on coastal sand dunes(Fig.4).
Aerial shoots of these species assist in the accumulationof sand, while theroots help to bind the sand deposited(Kim and Chung, 1995a).
These plants arevaluable not only in sand dune formation but also in establishmentof windbreak
forests. Both in Korea and Japan, despite the ecologicalimportance of sand dune
plants, the natural habitatsof coastal and sand dune plants are being destroyedbydunebank construction
and human disturbancein the summer season. Consequently,
size and genetic diversityof these plant populations
are decreasing.
Recently in Korea, Chung and his coworkers have conducted research which
focused on genetic effects
due to the destructionof natural habitatand habitat
fragmentationon the coastal plant species Eurya japonica (Chung and Kang, 1994),
E. emarginata(Chungand Kung, 1995),

Calystegia

soldanella(Kim

and Chung,Fig.4

Zonation

the seaside

on coastal

to the inlands.

75%, 4:75-50%,

1995b)

3:50-25%,sand dunes basedon vegetational

Six coveragegroupsare classified

2:25-10%,

1:10-1%and :1%)and Vitex rotundifolia(Yeehndata obtainedby vegetational

et al., 1996).

by belttransect

methodcover in a quadrat,

from
5:100-

Kim and Chuung (1995a)

conducted experiments on Salsola komarovi which focussed on the genetic erosion
resultingfrom isolationand decrease in population
size by human impact. The
objectivesof their studies were: i) to estimate how much total genetic diversityismaintainedin the species; ii) to describe how genetic variation is distributed
withinand among populations;iii) to compare genetic diversityof species with similar life
history
traits;and iv) to make a decision about which Korean populationsof thisspecies should be protected.
Throughout
these studies on allozyme variations
incoastal plants, they provide information
about the genetic resources of the species andmade suggestions

regardingin-situconservationof Korean coastal

plants.

Conclusion
Recently in Japan, there has been a gradual increase in research on biologicalconservationand publications
relatedto maintaining
biodiversity
and ecosystems

(Washitani

et al.,1991;

Shindoet al.,1995;

Washitaniof studies

on Lilium

lancifolium,

which

is an endemicand Hayashi(1992)

reported

the distributionof native

habitats

in Tsushima

conditions
horticultural

viewpoints.

trees,

rubra

Myricaal., 1989)

were studied
There

activities

will

resultingconservationof plant

application

of

Asia,

Noda

and the environmentalof vegetationof wild

fruitsin habitats

and Vaccinium

of in-situ

preservationin the fragmentationof plant

resources,of Eastspeciesandin destruction

analysislily

ecologicalbe an increase
genetic

In a series

from cytotaxonomical,

1988)

from viewpoint

diversity

phytogeographical& Sasaki,1996).

of populations

The management(Ohkuroand Yahara,ulginosum(Ohkuroof genetic

resources.

of natural
populations.

by

For successful

in-situ

habitatis essential.

The

integrationof basic

informationfromstudieshelp

in-situconservation.

willof natural

humanand further

and ecologicalprotection

habitats

etrational

Acknowledgements
I am grateful
Univ.)

for showing

to Dr. Shimamoto,me theirmanuscript

Mr. Fujita(Hokkaido

Univ.)and Dr. Ohara (Tokyoin pressand giving

me suggestions

leading

to thepresentationin this workshop.
I also would like to express my thanks to Dr. Hayashi (Tokyo
Univ.) for his valuable suggestion
and information
on Lilium lancifolium.
I thank Dr. Ohkuro,
my colleague,

for his comments on the managementof natural

genetic

resources..

References
Beard, B. H. and Knowles, C. G.. 1971. Frequency of cross-pollinationof soybeans after seed irradiation.
Crop Sci.1 1:489-492
Chung, M.G., and Kang, S.S. 1994. Genetic variation and population
structure in Korean populationsof Eurya japonica (Theaceae). Am.J. Bot. 81(8): 1077-1082.
Chung, M.G., and Kang, S.S. 1995. Allozyme diversity
and gene structure in Korean populationsofEurya emargita (Theaceae). Jpn. J. Genet. 70: 387-398.
Fujita, R., Ohara, M., Okazaki, K. and Shimamoto, Y. 1997. The extent of natural cross pollinationinwild soybean (Glycine soja). J. Heredity (in press).
Hamrick, J.L.and Godt, M.J.B. 1989. Allozyme diversity in plant species. In Brown.A.H.D. et al. (Eds.):
Plant Population
Genetics, Breeding, and Genetic Resoures, pp.43-63. Sinauer Associates,
Sunderland,MA.
Hamrick, J.L., Godt, M.J.B., Murawskii, D.A. and Loveless, M.D. 1991. Correlations
between species
traits and allozyme diversity:
implications
for conservation
biology.
In Falk, D.A. andHolsinger, K.E. (Eds.): Genetic and Conservation of Rare Plants, pp.3-30. Oxford University
Press, New York.
Kiang, Y.T., Chiang,Y.C. and Kaizuma, N. 1992. Genetic diversity
in natural populationsof wild
soybean in Iwate Prefecture,
Japan. J. Heredity 83: 325-329.

Kim, S.T. and Chung, M.G. 1995a. Genetic variationand population
structure
in Korean populationsofsand dune species Salsola komarovi (Chenopodiaceae).
J. Plant Res. 108: 195-203.
Kim, S.T. and Chung, M.G. 1995b. Genetic and clonal diversity
in Korean populationsof Calystegia
soldanella(Convolvulaceae).
Isr. J. Plant Sci. 43: 213-226.
Kurabayashi,
M. 1957. Evolution
and variationin Trillium
IV. Chromosome variationin natural
populationsof Trillium
kamtschaticum
Pall. Jpn. J. Bot., 16: 1-45.
Nei, M. 1972. Genetic distances
between populations.
Amer. Nat. 106:283-292
Nei, M. 1973. Analysis
of gene diversity
in subdivided
populations.
Proc. Nat. Acad. Sci. USA 70:
3321-3323.
Noda, S. and Hayashi, K. 1992. Environment and samplings in the natural habitatsof Lilium lancifoliumin Tsushima, Japan. Bull. Cul. Nat. Sci. Osaka Gakuin Univ. No.25: 19-53.
Ohara, M., Takeda,T., Ohno, Y. and Shimamoto,Y. 1996. Varationsin the breeding
system and the
population
genetic structure of Trillium
kamtscahticum(Liliaceae).
Heredity
76: 476-484.
Ohkuro, T. and Sasaki, H. 1988. Studies on the community structure and vegetation
management in the
habitatof Myrica rubra J. JILA 51(5):192-197.
Ohkura, T., Takeuchi, K., Ide, H., Yoshida, R, Imagawa, T. and Kajiura, I. 1989. Studies
on the habitat
distributionof wild fruit Vaccinium
uliginosum
in connection
with the forest destrcution
caused by volcanic eruption on Mt. Kusatsu-sirane,
central Japan. J.JILA 52(4):245-254.
Shindo,S., Zento, H., Watano, Y., Kinoshita,E., Ueda, K., Yonezawa, K. Nomura, T. and Shimizu,
T. 1995. Conservation
biology of Primula cuneifolia
var. hakusanensis:
Genetic variationanddifferentiationof populations.
J. Phytogeogr.
And Taxon. 43: 103-109.
(in Japanese)
Sneath,P.H. and Sokal, R.R.1973.
Numerical Taxonomy: The principles
and practiceof numericalclassification.
W.H. Freeman, San Francisco.
Washitani,I., Namai, H. Osawa, R and Niwa, M. 1991. Species
biologyof Primula sieboldii
for theconservationof its lowland-habitat
population:
I. Inter-clonalvariationsin the flowering
phenology,
pollen load and female fertility
components. Plant Species
Bio. 6: 27-37.
Washitani,I. and Yahara, T. 1996. An introduction
to conservation
biology:fromgene to landscape.
Bunichi-sogo-shyuppan,
Tokyo, (in Japanese)
Yeehn, Y., Kang, S.S., Chung, H.G., Chung, M.S. and Chung, M.G. 1996. Genetic and clonal diversity
in Korean populationsof Vitex rotundifolia(Verbenaceae).
J. Plant Res. 109:161-168

Questions

and Answers in Session

2

Questions to Dr. Strelchenko
Q. Will you explain the reason why in your RFLP analysis 2-rowed and 6 rowedbarley groups were separated in group B (occidental),
but not in group A (oriental).
(Morishima)
A. Wehad only a few accessions (22) in group A in this study. This may explain why
we failed
to find any order in clustering
in relation
to spike morphology.
(Strelchenko)
Questions to Dr. Li
Q. What proportionof wheat diversity
has already been collectedfrom Xinjiang
Province, and how much additional
diversity
is planned to be collected in 1997-99.
(Riley)A. Most of the local varieties
and wild relatives(about
70 species of Triticeae)ofwheat are distributedin Xinjiang.
The explorationand collection
for wheat genetic
resources has been carried out twice. So far almost all of the local varieties
havebeen collected
and conserved. In the wild relativesof wheat, however, about 10
species have not yet been found. In the species collected
only a few seeds were
harvested,
it is thus difficult
to study population
diversity.
In 1997-99 we areplanning to collect mainly those species not found in the previous explorations
andsamples growing in extreme environmental conditions. The seeds of those species
which were collected in the last explorations
were also harvested according to thedemands of population
diversity
analysis.
(Li)
Q. Do you have plans to introduce wheat germplasm from other countries.(Gupta)
A. It is very important to broaden wheat genetic basis and keep sustainably
increasing
production.
We are planning to introduce germplasm with desirable
characters,
especially
germplasm with high tolerance
to cold and resistanceto powdery
mildew.(Li)Questions

to Dr. Okuno

Q. Are the species you reported corresponding
to biological
species by Harlan and de
Wet? (Sano)
A. The species of Aegilops used in our experiments were identified
by morphologicalcharacteristics.
Therefore they corresponded
to taxonomic species. The Aegilops
species we used would correspond to species in the secondary genepool (Harlan

and de Wet)of wheat basedon information

on hybridizationof Triticum

andAegilops given by Kimber and Feldman (1987).
(Okuno)Q. What are the techniques that you use in exploration
to capture a larger proportionof genetic diversity?
(S.R. Gupta)
A. We have undertaken explorationand collectionof plant genetic resources
according to the manual issued by the Laboratoryof Plant Genetic Diversity,
NIAR. The manual describes ways to collect samples of cultivatedand wild
species. (Okuno)
Q. We can recognize center (or centers) of genetic diversityof crop species. Do you
think center of genetic diversity
exists in wild species? (Morishima)
A. Yes, I do. Based on the results obtained by RAPD analysis, we recognizedconsiderable
differencesin genetic diversityof wild relatives
collected
fromdifferent
locations. One of the difficulties
in clarifying
centers of genetic diversityof wild relatives is to obtain well identified
samples worldwide. We are focussing
on collectionof wild relatives as one of research topics in the 2nd phase of MAFF
Genebank project. (Okuno)
C. Centers of diversity
for wild species may well be very difficult
to determine in
relation to close relatives of crops where gene flow may occur. (Vaughan)
C. Dr. Okuno mentioned about a relationshipbetween cultured diversity
and genetic
diversity
analysis,with reference to the cultural diversity,and rice diversity
incollections
from Vietnam (Riley)
C. In plain areas in northern Vietnam, genetic diversityof cultivatedrice has been
rapidlyreplacedby a few improved varieties.
On the other hand, upland rice
grown in the mountainous areas still holds a wide range of diversity,
partly a resultof the taste and quality preferencesof the ethnic groups in this area. (Okuno)
Questions to Dr. Trinh
Q. In your classification,
Dr. Trinh,secondarygroup with respect

most of the introduced
to degree of genetic

germplasm belongs to thediversity?
What is thedifference
between the introducedgroup of crop species and the secondary groupfor degree of genetic diversity.(Hayashi)
A. In general, the introduced
crops have a lower degree of genetic diversity
than theendemic ones. I gave the terminology"primary degree of diversity"
to the cropshaving a high diversityand "secondary" degree of diversity
to those with lower
diversity.(Trinh)
C. I was surprised to find almost all farmers fields in Yenzian (a mountain area of
Yunnan), where many different
minoritygroups are living, were occupied byhybrid rice. Landraces were found only in remote, high altitude
areas. Genetic
diversityof major crops observed at present reflects the power of the government(or extension offices).(Morishima)
C. The management of in-situ areas (or GMZ=gene management zones) depends on
many factors, such as target species, annual or perennial, weedy plants or trees, thesize of area etc. You can protect conserved areas in many ways, it depends on yourbudget. However, the most important thing is how the area can be characterized
and evaluated, how often and what the benefits of the process are. (Sari)
Q. I understand that in the southern region of Vietnam the local rice varieties were
almost all replaced by modern varieties. I would like to know the present situation
regarding varietal replacement in northern Vietnam. (Kikuchi)
A. In general, throughout Vietnam in intensively
cropped areas where non-glutinous
rice is grown, almost all landraces
are replaced by modern varieties.
However,recently
as high quality
rice has been in demand some land races have beenreturning to production.
With regards glutinous rice, landraces still exist because
it is difficult
to find varieties
which have the required quality. (Trinh)
Q. I am interested in home gardens as a means of in-situ (on-farm) conservation. May
I know the diversityof crops maintained in home gardens in Vietnam. (Mariscal)
A. Twokinds of crops are widely cultivatedin home gardens in Vietnam:
Vegetables. The vegetables of temperate origin are cultivatedboth in the field
and in home gardens but vegetablesof tropicalorigin are mainly cultivatedinhome gardens.
Fruit crops. These are mainly grown in home gardens. Thus the home garden
is an important place for the in-situ conservation of these two crop groups. (Trinh)
Q. Dr. Okuno mentioned a relationshipbetween cultural
diversityand genetic
diversity
analysis,with reference to the cultural
diversityand rice diversity
incollections
from Vietnam. Does Dr. Trinh have any comment. (Riley)
A. Diversity
in ethnic groups creates diversity
in crop genetic resources. There are
two main factors which affect genetic diversity
in relation to ethnic groups (1) theagro-environmental conditions in which each ethnic group lives and (2) each ethnic
group has its own preferences with respect to food quality. (Trinh)
Q. Indo-China has a number of minorities
who are playing an important role in onfarm conservationof many crop species. Are you working with these farmer
communities or do you have project-typework between public institutions
andfarmer communities in your country? (Nakagahra)
A. Weare trying to work with farmer communities at the district level on the topic ofplant genetic resources conservation. We had a short term project with Crocevia
International
Center (CIC) from Italy for 2 years (1994-1995).
We got some results
and experience as a result of this project. (Trinh)
Questions to Dr. Matsuo
Q. In your paper you presented a very high level of outcrossingin G. soja (13%).
there any evidence of hybridization
between wild and cultivatedsoybeans
Japan? (Vaughan)
A. The paper from which thisfigurecame did not mentionIsinsuch hybridization.

(Matsuo)
Q. In relation to hybridization
between soyabean and wild Glycine soja, do you haveany data on the frequency with which honey bees visit soybean fields? (Sano)
A. Wehave no data on this. We presume it is not frequent.(Matsuo/Shimamoto)
Q. Given that the taxonomy of plants is not very consistent or stable as shown fromthe difficulties
in choosing character states and on-going revisions. Do you thinkthat ecologicalgenetic analysis
and phytogeographicalstudies would give a
consistentor stable classificationof plants? (Mujaju)
A. That is a difficult
question.
These data would give an insight
into plantclassification.
But for stable classification
a whole range of characteristics
need tobe taken into consideration.
(Matsuo)
C. Due to the natural dynamic nature of ecologicalsystems ecologicaland geographic
characteristics
would not be too helpful for classification.
However, such genetic
analysisand phytogeographicalstudiesof crop plants
do provide useful
information

to the farmer.(Kresovich)

Topic3:
Cooperative Mechanisms to Improve
Evaluation of Plant Genetic Resources

Chairpersons
A. G. Yunus
S. Miyazaki

Mechanisms for the Evaluationof Plant

Genetic

Resources in JapanHIDEFUMI SEKO
National

Instituteof Agrobiological

Resources,

Tsukuba,

Ibaraki

305, Japan

1. MAFF Genebank Project
The MAFF(Ministryof Agriculture,
Forestryand Fisheries
of Japan)
genebank project was initiatedin the fiscal year of 1983, and in 1986 the Genetic
Resources Center was establishedat the National
Instituteof Agrobiological
Resources, NIAR. At present
four out of six categories
of germplasm (plants,
microorganisms,
animals, DNA, forest trees, and aquatic organisms)
are concerned
at NIAR (Fig.1).
Plant genetic resources have the longest history of conservationinJapan. Systematic plant breeding started in 1920, and breeders maintained their own
genetic resources as the crossing materials for their breeding programs. As awarenessof the importance of the diversity
in genetic resources emerged, 3 laboratories
wereestablishedfor rice, wheat and barley, and soybean in the National
InstituteofAgricultural
Sciences, Central Agricultural
Experiment Station, National Tohoku
Agricultural
Experiment Station, respectively
in 1953.
2. Plant

Genetic Resources System in MAFF, JapanThe MAFF genebank for plants consists of the Central bank at NIAR and 15
sub-banks located from Hokkaido in the north to Okinawa, the southern most island
in Japan. Sub-banks belong to 13 National
Research Institutes.
From 2 to 16
laboratories
in each instituteand center participate
in the MAFF genebank project andconduct genetic
resources activities,
collecting,preservation,
multiplication,
evaluation
and use in research and plant breeding. Forty three designatedresearch
units in prefectural
agriculturalexperiment station are also participatingin this project(Fig.2).

The project

divides

agricultural

crops into 12 groups; rice, wheat/barley,

tuber

crops, legumes, small grains/industrial
crops, forage crops, fruit tree, vegetables,
ornamental plants, tea, mulberry tree and tropical crops. A curator is appointed
foreach plant group. Curators for rice, wheat/barley,tuber crops, legumes, small
grains/industrial
crops belong to the National Agriculture
Research Center, for forage
crops to National

Grassland

Research

Institute,for fruit

tree to Fruit

Tree ResearchFig. 1 MAFF Genebank

systemAFFRC: Agriculture,
Forestry and Fisheries
Research
NIAR: National
Instituteof Agrobiological
Resources
KFTBI: Kanto Forest Tree Breeding
Institute
NBIR: National Research Instituteof Aquaculture

Station,for vegetables,

ornamental

Vegetables,
Ornamental Plants
Sericulture
and EntomologicalResearch Center for Agriculturalin respective
crops.

plants,Counciland tea to National

Research

Instituteofand Tea, for mulberry to the National InstituteofSciences, for tropicalcrops to Japan International
Sciences. Most of curators are active plant breeders

Fig 2. MAFF Genebanks

network for PGR

NARC: National
Agriculture
Research Center, NIAR: National
Instituteof Agrobiological
Resources, NGRI: National Grassland
Research Institute,
FTRS: Fruit Tree Research Institute,
NIVOT: National
Instituteof Vegetables,
Ornamental
Plants and Tea, HNAES1: Hokkaido
National
Agriculture
Experiment
Station, TNAES: Tohoku National
Agriculture
Experiment
Station,
HNAES2: Hokuriku
National
Agriculture
Experiment
Station,
CNAES: Chuugoku
National
Agriculture
Experiment
Station,
SNAES: Shikoku
National
Agriculture
Experiment
Station,KNAES:
Kyushu National Agriculture
Experiment
Station,NISES: National
Instituteof Sericulture
and Entomological
Sciences,
JIRCAS: Japan International
Reserch Center for
Agricultural
Sciences,
NCSS: National
Center for Seed and Seedlings,
NLBC: National
Livestock
Breeding
Center

3. Evaluation

Mechanisms

In order to use germplasm stored in the genebank it is necessary to have as
much information as possible available to scientists.
The Center bank and sub-banks
collaborate
to characterize
and evaluate their germplasm collections
systematically.
Germplasm, including old varieties,
land laces, wild relatives,
breeding lines,
and materials introduced
from overseas are shared with participating
laboratories
related to crop groups and three levels evaluation;
primary, secondary, and tertiary,
and two categories in each level, compulsory and optional items are investigated.
The
descriptors
for compulsory items for primary evaluation
are limited in number toabout 10 essential
charactersfor identifying
strains, such as plant height, panicle
length. Secondary characters (compulsory)
include resistanceto pests and diseases
such as brown plant hopper, blast, preharvest
sprouting. The tertiary characters ofcompulsory items are, for example, productivity,
grain quality,
1000 seed weight.
Optional items are amylose content of cereal endosperm, electrophoretic
zymogram
patterns, DNA analysis. A textbook of guidelines
for evaluating PGR has been issued
for the 12 crop groups (Table 1).
A total of 110 crops are included in this evaluation
manual; 29 crops for thevegetables group and one for rice, tea, and mulberry groups, respectively.
The numberof accessions investigatedand data obtainedfor primary, secondary and tertiary
evaluation
over the past 3 years are shown (Table 2).
The MAFF Genebank Project has establishedmanagement systems forpassport data, stock control data, and evaluation
data. Evaluation
data recorded at
sub-banks can be entered into the database in the central bank by sub-banks throughthe MAFF on line network system (Fig. 3). Preparation
for providing
passport databy internet has progressed and it will be on line in 1997.
4. International
Cooperation
International
cooperation on plant genetic resources activities
in NIAR include
interaction
with IPGRI and the FAO, holding annually an international
workshop on
genetic
resources, conducting
a 6-months JICA training
course on PGR, andcollaborative
genetic resources projects with a number of developing
countries. Rice
germplasm, collectedduring the collaborative
IPGRI project, were characterized
for esterase

explorationin Vietnam within the
isozyme alleles. This was conductedTable 1. Charactersfor evaluationby the MAFF manual
N o . o f c h a ra cte rs re q u iredNcroo .op fs
C ro p g rou p

L ev e l 1
C om p0 1 :R ic e

L ev e l 2

L e v el 3

O p t1

C om pO p t1

C om pO p t1

T o ta l

1

R ice

13

18

12

8

8

5

64

2

W h ea t

9

19

10

9

7

12

66

0 3 :L e g u m es

3

S o yb e a n

12

5

3

9

3

6

38

0 4 :T u b er c ro p s

2

S w e et po tato

15

14

15

12

12

14

72

c0 ro
5 :Mp s ille t & In d u stria l

18

mF o illet
xtail

11

13

3

2

4

1

34

0 6 :F o rag e c ro p s

18

Ita lia n ry e

9

8

4

11

6

6

44

0 2 :W h e at &B a rle y

0 7 :F ru it tre e

22

A p p le

10

2

13

14

10

5

54

0 8 :V e g etab le s

29

M e lo n

12

44

8

16

10

24

1 14

0 9 :O rn a m en tal p lan ts

10

R o se

12

35

4

10

3

2

66

10 :T e a

1

T ea

11

16

8

5

1 1

7

58

1 1 :M u lb e rry1

M u lb erry

9

37

6

12

5

1

70

1 2 :T ro p ic a l p la n ts

3

P in e ap p le

7

15

2

2

4

4

Level 1 : Characteristics
essential
to identifying
strain.
Level 2 : Importantcharactersfor user's such as resistanceLevel 3 : Chemicallyanalyzed characters such as amylose,Fig 3. MAFF PGR Activities

34
1 10

T o tal c ro p s de sig n ate dto pests and diseases.
protein, DNA, and productivity.

Table 2. Characters
C ro pevaluatedby the MAFF manual in recent 3 years

Y ear

19 93

L ev el

199 4

19 95

1

2

3

1

2

3

1

2

3

0 1 :R ic e

22 5 6

3 78 0

16 08

185 6

15 08

33 80

190 7

33 16

2 14 40 2 :W h e a t & B a r le y2 7 127 30 360 86

2 6 10

69 73

57 20

20 04

7 9 125 96 50 3 :L e g u m e s

10 3 2

68 3

22 79

8 14

2 19

2 194

501

20 1

20 0

0 4 :T u b e r c r o p s

5 184

4 33 732 40

5 04 6

34 82

35 12

4 3 362 3 124 14 6

I0 n5 d: uM s tilrleia tl &c r o p s

14 3 85 69 74 2 56

143 8

60 97

42 56

14 97

20 8 23 7 120 6 :F o r a g e c r o p s

118 1

2 36 3

3 24

105 4

19 85

3 24

7 54

127 8

124 9

0 7 :F r u i t t r e e

837

2 57 03 6 33

73 7

19 65

24 45

4 74

1 5 13

2 22 20 8 :V e g e ta b le s

13 1 1

2 64 4

68 52

135 6

26 04

68 52

1 124

2 28 85 43 3p0 l9 a:On t sr n a m e n ta l

6 55

108 4

5 46

65 5

2 190

4 96

63 1

15 5 95 18

1 0 :T e a

1 137

125 4

2 185

198 6

10 20

20 68

175 0

12 18

720

1 1 :M u lb e r r y

351

84 4

50 0

351

8 44

5 00

10 6

9 60

5 84

1 2 :T r o p ic a l p la n ts
T o ta l c r o p s

0

0

0

0

0

0

13 5

0

0

2 26 72

3 3 265

3 130 9179 0330 7 59

2 98 75

16 2 19

2 46 3 9

2 68 93d e s ig n a t e d

Level
Level
Level

1 : Number
2 : Number
3 : Numberof accessions investigated.
of Data obtained.
of Data obtained.

by a Vietnamese scientist invited to Japan. It was found that germplasm collected innorthern districts
displayedwider diversity
in esterase isozyme pattern (11 out of 12
expected patterns)
than those collectedin southern districts(Okuno et al., 1996).
Recent collaboration
between Vietnam and Japanese scientists
has added much to ourunderstandingof variationin the most geneticallydiverse parts of Vietnam for rice.
NIAR has proposed a new project to IPGRI for the evaluationof rice and legumes
collaboratively
collected
in Nepal several years ago.
Primary evaluation has been carried out for introduced
rice germplasm and theresults
were publishedfrom the Laboratoryof Plant Diversity,
NIAR entitled"Primary
evaluationof induced
rice germplasm-Catalog
of accessions
inNIAR/MAFF" that were directly collectedby Japanese and collaborating
scientists
from 1979 to 1991.

5. Topics on the successful use of PGR
By using valuable but primitive
genetic resources, many parental lines andpre-breeding
materials have been bred such as brown plant hopper resistant lines, lowtemperature tolerant lines, wide compatible lines, blast resistant lines in rice, leaf rust
resistant lines in wheat, high-protein
and high-lysine
lines in barley, disease resistantlines in tomato.
An outbreak of rice stripe disease transmitted
by brown plant hopper was aserious problem in the western part of Japan in the 1960s and 70s. Highly resistantgenetic resources were found in Japanese upland rice and indica rice cultivars.
The
first stripe disease resistantJapanese paddy rice line 'St 1' was selected from
offsprings
backcrossed between an indica cultivar 'Modan' as a donor and 'Norin 8'
as the recurrent parent (Fig. 4). From various crosses with 'St 1', many stripe disease
resistantcultivar
were developed
such as 'Mineyutaka','Musashikogane'and'Hoshinohikari'(Toriyama,1992).
Barley yellow mosaic, a soil-borne virus disease, was epidemic in the malting
barley producing
areas of Japan. Barley genetic
resources were screened forresistanceto BYMV in infested
fields and 'Mokusekko 3' and 'Mihorihadaka'
arefound to be resistantby scientists
at Okayama University.
Extensive efforts forresistancebreeding
were conductedat the Tochigi
Prefectural
Agricultural
Experiment Station, designatedas a barley breeding unit of the National Government.
Various 2-rowed resistantparental lines were bred at the institute,and the first
BYMVresistant cultivar having acceptable
malting quality was released as 'Misato
golden. Many BYMV resistant cultivars
were bred after this cultivar which was aturning point in Japanese malting barley breeding (Seko, 1987).
Waxy cultivars have been identifiedin cereals such as rice, maize, barley, andsorghum. However, waxy cultivars of bread wheat have not been describedso far.
Amylose content was analyzed for wheat breeding lines of National
Agriculture
Research Center and Kanto 107 was found to have significantly
reduced amylose
content (Kuroda et al., 1989). The lack of Wxproteins involved the A and B genomesof Kanto 107. Reduced amylose content was determinedby two-dimension
electrophoreisis.
About 2,000 wheat genetic resources were analyzed to detect thelack of Wxprotein concerning D genome, and it was found in one line, 'Bai-huo' from
China. Using the haploid
breeding
method the first waxy bread wheat has beendeveloped from the cross between 'Kanto 107' and 'Bai-huo' (Hoshino et al., 1996).

PhotoPhoto

1. BYMV screening

2. Farmer's

field

infested

nursery

by BYMV

Fig. 4. Rice stripe

virus

resistantcultivars

derivedfrom Modan cross.

6. Future Perspectives
A prerequisite
to the efficientuse of PGR is a good collection
wellcharacterized
and evaluated.
Plant genetic
resources
efficiently
used for the
improvement of crops is a major reason for conservation
activities.
Developmentof
definiteand efficient
screening
method is necessary
to use genetic
resources
effectively
for plant breeding.
In this context, evaluationof various charactersisindispensable
and informationexchange should be strengthen.
In the MAFF
Genebank Project a certain amount of funds are allocated to pre-breeding,the creation

Photo 3. Breeding of waxy wheat
Cut surface of grains stainedof breeding
valuablematerials
characteristics.

collaboration
amongcollaboration

PGR

with

using

primitive,In

iodine(courtesyof Mr. Yoshikawa)

but

interesting,genetic

addition,furtherefforts

among

PGR researchers,

breeding

researchers

researchandandon evaluation

genomeand informationarebiotechnology
exchangeresources

necessarytoand breeders
research.

shouldalso

havingenhance
as wellasInternationalbe enhanced.

References
Hoshino, T., Ito, S., Hatta, K., Nakamura, T. and Yamamori, M. 1996. Development of waxy common
wheat by haploid
breeding.
Plant Breeding
46:185-188.
Kuroda, A., Oda, S., Miyagawa, S. and Seko, H. 1989. A method of measuring amylose content and its
variation
in Japanese wheat cultivars
and Kanto breedinglines. Japan J. Breed. 39(Suppl.
2):142-143.
Okuno K., Fukuoka, S., Tien, N.D. and Ha, N.P. 1996. Genetic variationin rice landraces
collected
inVietnam and its geographicalcline. Breeding
Science 46, Suppl.1:
Seko, H. 1987. Developmentof two-rowed malting barley cultivar
resistant
JARQ 21:162-165.
Toriyama, K. 1992. Disease and insect resistance,
in Utilizationof plant
improvement.
(JICA):12-15.

306
to barley
genetic

yellowmosaic.

resources

for cropEvaluation and Characterization: Present Situation
and Prospectsof Plant

Genetic

Resources

P.N. GUPTA, I. S. BISHT, MATHURARAI and K. P. S. CHANDEL
National Bureau of Plant Genetic Resources, Pusa Campus, NewDelhi-110

in India012, India

Abstract
The National Bureau of Plant Genetic Resources (NBPGR) is the nodal organizationin India for planning, conducting, promoting, coordinating
and leading all activities
concerning
collection,
introduction,exchange,evaluation,
documentation,safe conservation
andsustainable
management of diverse germplasm of agri-horticultural
crop plants and their wild
relatives.
Characterizationand evaluationof germplasm is carried out at the NBPGRHeadquarters
and its nine regional stations located in diverse agro-ecologicalregions of India
well over two decades on more than 75 major and minor crops with a current germplasm
holdingof about 119,000 accessions. NBPGR has strong linkages
with over 30 centres,
designatedas National Active Germplasm Sites and maintain about 173,000 accessions ofspecific crops/crop groups, on related activities.
The Bureau has published
65 catalogues and
38 inventories
on different crops and has already initiatedstudies on characterizationof plant
diversityusing modern molecular techniques. Establishmentof core collection,ecogeographic
studies and ethnobotany
are important activities
on several indigenous
crops. Pre-breeding
activities
are considered a major future
Plantsustain

Genetic

the present

Resources(PGRs)

are the basic

raw materials

that,

day crop improvement

programmes,

but willbe required

meet the needs of future generations
who may require
while
facingunforeseenchallenges.
Despitethisgermplasm

collections

is stilla collection

has been properly

it has little

practical

of genetic

resources,use.

evaluated

Germplasm
starting

or detailed

and its attributes

resources(Frankeland Bennett,

countries.

Until

become known to breeders,in a broadof the material

sense and in the context

in a collection.

It coversof the new samples

characterizationand preliminary

evaluation

The genetic

in the developing

evaluation,toaltogether
new sources of genes
wide recognition
the use offrom the receipt

these for seed increase,also for further
categories

particularlyis the description

whole range of activities
and growing

limited,also

not only

theby the curator
evaluationandand documentation.

of crop plants1970).

can be functionallydivided

into

four

1. Advanced varieties in current use and bred varieties
no longer in commercial use;
2. Primitive"folk" varieties
or "land races" of traditionalprescientificagriculture;
3. Wild or weed relatives of crop plants and wild species of actual or potentialuse
in crop breeding or as new crops; and4. Genetic stocks such as mutations, cytogenetic
stocks (translocation,
inversion andadditionlines),and linkage testers.
Until recently the major emphasis in genetic resources programme was on thelandrace varieties of the important food crops that could be conserved ex situ as driedfrozen seed. A major reason was the pivotal role that landrace varieties
have playedin the development of scientificagriculture.
They are the antecedents
of all modern
varieties.
Another reason was their potential
value as sources of variation for future
plant breeding and the fact that they were often under imminent threat of extinction.
IndianNational Plant Genetic Resources System
South Asian subcontinentis a major centre of crop diversityof more than 20
major agri-horticultural
crops. Nearly 160 domesticatedspeciesof economic
importance and over 325 species of their wild forms and close relatives
are native tothis region and constitutea reservoir of genes that can be used for developing
newvarieties.
India developeda system for the increased use of PGRs. In addition
India
is playing
a role in coordinatingsuch efforts for the Asia and the Pacific Region.
Indian initiatives
has also succeededin evolving
interestin this subject
among
SAARC (South Asian Association
on Regional Cooperation)
and G15 developing
countries who are now pooling their know-how as well as other resources to adopt a
regional strategy and coordinatedaction plan for conservation,
inventory, evaluationand sustainable
use of PGR (Rana, 1994).
The National
Bureau of Plant Genetic

Resources(NBPGR)is the nodal

organizationin India for planning,
conducting,promoting, coordinating
and leadingallactivities
concerning
collection,
introduction,exchange,evaluation,
documentation,safe conservation and sustainable
management of diverse germplasmof crop plants and their wild relatives
with a view to ensuring their continuous
availability
for use of breeders and other researchersin India and abroad (Fig 1,
Appendix I). One of the main objectivesof the NBPGR is to characterize
and evaluate
the available
germplasm and to coordinate
such activities
with other crop based

Fig. 1. Indian

National

Plant

Genetic

Resources

Systeminstitutes,
institutions,
resources.

coordinatedprojects,
state agricultural
universities
and international
and to help in preparing
inventories
and catalogues on available genetic
The work on characterizationand preliminary
evaluation
is carried out at

the Bureau's Headquarters
and its Regional
Stations.
Its regional
stations/centres
represent different
phytogeographicalregions with distinct
ecologicalconditions
andthese are located in the temperate region at Shimla; arid region at Jodhpur;semi-arid•Ž

region at Hyderabad,
Akola and Amravati; humid tropicalregion at Trichur andhumid subtropicalregion at Shillong.
It also has 10 explorationbase centres; 7 ofthese located in the existingregional
stations
and 3 located at Cuttack (Orissa),
Ranchi (Bihar)
and Srinagar (Jammu and Kashmir) (Fig.2).
NBPGR's Headquarters
and regional stations have defined crop responsibilities
for 75 major and minor crops
with a current germplasm holding of over 120,000 accessions (Appendix
II, III). Crop
curators for all major crops have been identified
within NBPGR and also in the ICAR
crop based institutes
and state agricultural
universities.
NBPGR is thus linked
effectively
with over 30 centres, designatedas National
Active Germplasm Sites(NAGS)
for specificcrops and has assigned them responsibility
for maintaining,
characterizing,evaluating
and supplying
germplasm out of its collectionsof different
crops which are also under long term storage at -20
in the National Genebank at
the Bureau's Headquarters(Fig3, Appendix IV).
In view of the wide range of genetic variability
in germplasm collections
ranging from wild and weedy types to high yielding
varieties,
specific
strategies
for
their evaluationand characterization
are necessary.
Also breeding aims change
rapidly.
By and large, for effective
evaluationof germplasm, a close organizational
and personal contact between curator and breeder is necessary
in the context ofbreeding

objectives

vis-a-vis

evaluation

program.

Components of Germplasm Evaluation
After collectionof germplasm, there is a need for its systematic
evaluationin order to know its various morphological,
physiologicaland developmental
characters including
some special features, such as stress tolerance,pest and disease
resistance.
The germplasm accessions are usually evaluated for two consecutive years
for documentationand preparationof crop catalogues.
The following
steps andcomponents
1.

of germplasm

evaluation

can be distinguished.

Selectionof germplasm accessions for characterization
The following
categories
of germplasm may be included
New collections
through explorations
New exotic introductions
New accessions generated from parasexual

methods/vegetative:

propagules

Fig.

2. National

Quarantine

Stations/tissue

Bureauof Plant

and Satellite

cultureraisedGenetic

Resources(Headquarters,

Regional

Stations,

Base Centers,

Stationpropagules

etc.

Samples redrawn from genebank after long intervals
to monitor the changes
in expression
to stable (characterization)
traits may also be included,andSamples procured from other genebanks as duplicate
sets to monitor thechanges due to the location effect in character expression.

Fig. 3. Indian

National

Actve

Germplasm

Sites.

1 DWR, Karnal; 2 CRRI, Cuttack;3
AICRP on Maize, New Delhi; 4 AICRP on barley, Karnal;
5 NRC
for sorghum, Hyderabad;
6 AICRP on pearlmillet,
Pune; 7 AICRP on small millet,
Bangalore;
8 IIPR,
Kanpur; 9 NRC for soybean,
Indore;10 DOR, Hyderabad;
11 AICRP on rapeseed
and mustard, Hisar;
12 NRC for groundnut,
Junagarh;
13 SBI, Coimbatore;
14 CICR, Nagpur;
15 CIJAF, Barrackpore;
16
DVR, Varanasi;
17 CPRI, Shimla;
IGFRI, Jhansi;
19 NRC for spices, Calicut;
20 CTRI, Rajamundri;
21
CPCRI, Kassargod;
22 NRC for M&AP, Anand; 23 NRC for agroforestry,
Jhansi;24 AICRP on semiarid fruits, Hisar; 25 NBPGR Regional
Station,Shimla;26 IIHR, Bangalore;
27 NRC for citrus, Nagpur;
28 CIHNP, Lucknow; 29 CTCRI, Trivandrum;
30 NBPGR Reg. Stn. Shimla.

2.

Seed increase
Initialseed increase needs care as it involves
the risk of losing a particularaccession due to poor adaptation,disease and pest damage, introducing
admixtures
through contamination
or error and altering
the genetic compositionof the original
genetic make-up through conscious (human) and unconscious (natural)
selection.
Therefore, it is essential
to increase seed stocks sufficiently
in one cycle so that theharvested seeds can be used for evaluation,differentiationand storage. On receiving
the samples, a portion of the seeds is saved for another planting, in case the first effort
fails, besides serving as a reference sample. During initial
seed increase,data onmany morphologicaltraits and other traits of interest
are recorded.
Duplicateaccessions are also identified
at this stage and promisingones are identified
forintensive evaluation.
The plant quarantine needs can be met during this stage as well.
After germplasm is collectedfrom nature or from farmer's field and placed in a
genebank or regenerated,
loss of genetic variationor change in the genetic structureof the collection
may occur. One of the most important duties of curators is, therefore,
to minimize such genetic changes. In order to do so, a sufficiently
large effective
population
size is preserved
and, whenever the populationis regenerated,asufficiently
large number of plants are grown and enough pollinations
are made or
facilitatedto maintain large effective
populations.
possible

The environmental conditionsof the multiplicationsite(s)are kept as near as
to those under which the accession evolved or was cultivatedfor a long

period.
is likely

Since the distribution
ranges of accessions of all major crops vary greatly, itthat two or more multiplicationsites are necessary to determine adaptability

and site X genotype
interaction.
The advantage
of choosing
such a range lies in
reducing the evaluation
period, because the complete range of climatic
factors may
be encountered over a shorter period of time.
A pure line is to be multipliedby growing only a few plants and the actual
number will depend on the multiplication
rate and the seed quantity
required,
whereas, a heterozygous
population
would need to be multiplied
from a much larger
population
sample and much care is taken to ensure the maintenanceof genetic
integrity.
The need for multiplication/rejuvenationof germplasm is a function of sizeof the initial sample, user demand and seed longevity
under the conditionof storage.

The aim during rejuvenation
is to retain the essential
genetic characteristicsof theaccession and obtain sufficientquantities
of high quantity seed to satisfy requirement
for storage and user demand. During the regeneration
process care is taken to reducechanges due to contamination
through mutation,
foreign pollen or seed, and tominimize genetic drift or shift by ensuring sufficient
population
size and reducing
opportunities
for natural selection.
3. Preparationof descriptor
list
The process of characterizationand evaluation
begins with the adoption ofdescriptor
lists.
The IPGRI descriptor
lists are widely used. NBPGR has also
developed
suitablelists of descriptors
and descriptor
states for a number of crops
suited to Indian conditions
which are advocated for uniform documentationin theNational PGR system (Gupta et al., 1995).
4. The design of experiment
The germplasm accessions are invariably
grown in an augmented block
design. The number of checks used may be 3-5 which are replicatedand randomized
in each block of 10, 15 or 20 accessions, depending
upon the size of the experiment.
Single row (3-5 m) plot or small plots of more than one row, depending
quantityof seeds available
and the nature of plant species, are generally

upon thegrown forgermplasm evaluation.
Even space is kept between plants to permit them to express
their differences
and avoid competition.
Accessions belonging
to the same maturity
group are planted
togetheron one date of sowing. Accessions suspected
to beduplicatesare grown side by side to facilitatecomparison while evaluatingin thefield.
During the process of growing, attention
is given to minimize natural crosspollination,
contaminationand erroneous labeling.
5. Type of charactersand measurement data
Observableor qualitative
characters
are identified

in single

plantswhereasnon-observable characters or quantitative
traits are generally
recorded on 5 plant basis
at the time of harvest.
The choice of check lines depends very much on circumstances.
For
preliminary
evaluation,
locallyadaptedcultivars
familiarto breeders,provideunderstandable

comparisonsand a dependable

way of monitoring

trial-to-trial(oftenyear to year) variation.
For further evaluation,
which usually addresses one trait ata time, there will often be a well recognized
set of checks that cover the likely rangeof scores (e.g. known resistantand susceptible
cultivars
or accessions
for disease
screening).
Appendix V lists the characters of specific significanceduring evaluationof the germplasm in some of the agri-horticultural
crops.
6. Documentationand cataloguing

Both evaluationand documentation
are seen as pre-requisite
for the use of
germplasm collections.
The passportand characterization
data should be readily
available
to the users in order to select the desired germplasm. Hence, information
management and manipulation
with plant genetic resources.
documentation
greatlywhich are indispensableof information
are essential parts of all practicalwork
The use of (personal)
computers in modern genebank

facilitatessorting,
retrieval,analysis,to the potential
users of germplasm

collationcollection.

etc. of data

Based on evaluation data, over the years, several crop catalogues/inventories
have been prepared (Appendix
VI, Anonymous, 1994). These crop catalogues
aredistributed
to concerned plant breeders for identifying
the useful germplasm for use
in their breeding
programmes.
Molecularusing

Characterizationof Plant Genetic Resources
NBPGR conducts research on biosystematics
and characterizationof PGRs
modern biotechnologicaltools.
New opportunities
to assess the extent ofgenetic variation
among accessions
in germplasm collections,
thereby helping
todecide which accessions are essentially
duplicatesand which should be includedina core collection
are now available.
Electrophoretic
isozyme survey and Random
Amplified
Polymorphic
DNA analysis, RFLP, SSR etc. have been performed to assess
the genetic diversity
in Solanum and Musa species. Similar studies are underway inother crop species such as sesame, Cucumis, okra (Bhat et al., 1992a, 1992 b; Bhat
and Jerret,

1995;

Establishment

Karihalooand Gottlieb,

1995;

Karihaloo

et al., 1995).

of Core Collections

Core collections,representative

sub sets of a basecollection,

have beenrecently advocated by some workers to cope with the difficultyof dealing with larger
genetic
resources collections.
About 10% of the accessions
may be drawn bydifferent
sampling techniques to form a core set which facilitate
initialevaluationor
study.
Preliminary
findings
can then be used to determine which eco-geographic
sectors of the base collection
can be studied more intensively
for specifictargets(Frankel,
1984, 1986; Frankel and Brown, 1984; Brown , 1989).
The NBPGR has initiatedstudies on establishmentof core collectionsin okraand sesame with an objective to have a core collection
with the widest possible rangeof variability
available
for breeders and other users. These core collections
represent
the genetic diversity
in the collection
and its selection
requires quality passport andcharacterization
data (Bisht et al., 1995; Mahajan et al., 1996).
Germplasm Enhancement/
Pre-breeding
Pre-breeding'or 'germplasm enhancement' is the early phase of any breeding
programme. Many improvement programmes concerned with the use of plant
germplasm include the process of pre-breedingas part of the total project.
Thoughthe end products of pre-breeding
are usually deficientin certain desirable
characters,
they are attractive
to plant breeders due to their greater potential
for direct use in abreeding
programme than the original
unadapted
exotic sources. Where several
sources with resistanceto biotic
and abiotic
stressescan be incorporated
into
improved populations
they can be used in breeding programs.
NBPGR feels pre-breeding
is an activity
level and considers it to be a major future thrust.
Cucumis, Solanum, Abelmoschus, Asiatic Vigna.
NBPGR's National
Informationlevels. Conservation

to be undertaken at the curatorial
The priority
crop species include

Information
Systemdatabase system is very importantof genetic resources,
not onlyat national,
regional and global
for immediateuse of alreadyconserved and evaluated/characterized
germplasm in the ongoing plant breeding
programmes, but also for future use. The National
databasesystem gathers all
relevant data from diverse sources that are needed by user scientists
belongingtodifferent
cooperation

disciplines.

NBPGR proposes

and international

linkages.

to expand its databasethroughstrongregionalGermplasm Advisory Committees
The National PGR System has been strengthenedby the constitutionof Crop

Advisory Committees, which have been set up recently for specific crops or groupsof crops. They advise the Bureau regarding the status of current holdingsof different
crops, shortcomings in storage and management system as well as gaps in exploration,collectionand evaluationof indigenous
genetic variabilityof native crops and alsosuggest the countries to be approached for introductionof new genetic variabilitytosustainour crop improvement

programmes.

International
Collaborations
i. Collaboration
with CGIAR crop based institutes
Besides operation of the above mentioned network of active germplasm sites
in the country, the Bureau also actively collaborates
with International
Agricultural
Research Centres in India and abroad. International
Plant Genetic Resources Institutehas contributed
significantly
to the Bureau's efforts offeringexpertise,training
andfundingfor research. NBPGR has active collaboration
with ICRISAT
on joint
explorationand multi-location
evaluation
programme on five ICRISAT mandate
crops. This has helped in documentationof germplasm collections
in pearl millet,
sorghum, pigeonpea, chickpea and groundnut. Considerable
exchange of germplasm
takes place between NBPGR and ICARDA (Syria),
IRRI (Philippines),
CIMMYT(Mexico)and IJO (Bangladesh).

ii. Bilateralcollaboration
Many countries have well developed
systems for assemblage,
enrichment,
documentationand conservationof plant
genetic
resources
and also havecomputerized database network. ICAR has memorandum of understanding
as well as
bilateralagreementswith several
internationalorganizations
and national
programmes. NBPGR is involved with over 80 countries on various plant genetic
resources
i i. Global

activities.

responsibilityof PGR
Followinga criticalassessment of the infrastructural
facilities
and trained
manpower available
at NBPGR, the Bureau has been designatedas responsibleforglobalandregional

base

Okra

workshopInternational

collections

genetic

resourcesof IPGRI(IBPGR,

1991;

Arora

PGR

Project

Indo-USAID
The Indian

the systemsof othercurrently

operatingPlantnations

and has adapted

a 7 years

is being

implemented

enhanceits

at the international

in 1993

and Riley,

National

to enhancea dozenandcrops.

an international

at NBPGR

The

first

workshopunder

the sponsorship1994).

Genetic

Indo-USthanin 1990

was organizedproject

role

more

was organizedon sesameiv.

ofResources
these
Project

Programmeto the Indianon Plant

NBPGR'ssituation.

Genetic

national

has learned

from

NBPGR

Resources.

capability

and

is
This

also

to

level.

References
Anonymous. 1994. NPBGR Publications
1976-1993.
National Bureau of Plant Genetic Resources, New
Delhi-110012, 20p.
Arora, R.K. and Riley, K.W. (Eds.). 1994. Sesame Biodiversity
in Asia:Conservation,
Evaluation
andImprovement. Proceedings of IBPGR-ICAR/NBPGRAsian Regional Workshop on "Sesame
Evaluation
and Improvement"
held at Nagpur and Akola, India, 28-30 September, 1993.
Bhat, K.V., Bhat, S.R. and Chandel,
K.P.S. 1992a. Survey of isozyme polymorphism
for clonal
identificationin Musa. I. Esterase, Acid phosphatase
and Catalase. J. Hort. Sci. 67:501-507.
Bhat, K.V., Bhat, S.R. and Chandel,
K.P.S. 1992b. Survey of isozyme polymorphism
for clonal
identificationin Musa. II. Peroxidase,
Superoxide
dismutase,
Shikimatedehydrogenase
Bhat,

and Malate dehydrogenase.
K.V. and Jerret, R.L. 1995.

J. Hort. Sci. 67:737-744.
Random amplified
polymorphic

Bisht,

Indian Musa germplasm. Genetic Resources and Crop Evolution,
I.S., Mahajan, R.K. and Rana, R.S. 1995. Genetic diversity
in South

DNA and genetic

diversityin

42:107-118.
Asian okra (Abelmoschus

esculentus)
collection.
Annals of Applied Biology,
126: 539-550.
Brown, A.H.D. 1989. The case for core collection,
pp. 136-156. In: The Use of Plant Genetic Resources,
Editedby Brown, A.H.D., Frankel,O.H.,
Marshal,D.R.
and Williams,J.T.,
Cambridge
University
Press, Cambridge.
Frankel,O.H. 1984. Genetic perspectiveof germplasm conservation,
pp. 161-170.
In: Genetic
Manipulation:Impact
on Man and Society, Edited by Arber,W.K., Llimenusee,K., Peacock,Frankel,W.J.and Starlinger,
O.H. 1986. Genetic

P. Cambridge University
resources: The founding

Frankel,internationalboard. Diversity
9:30-33.
O.H. and E. Bennett. 1970. Genetic Resources

Press, Cambridge.
years. III. The long road to the internationalin Plants-

Their

Exploration

and Conservation.

Blackwell,

Oxfordand Edinburgh.

Frankel,O.H. and A.H.D. Brown. 1984.

Gupta,

P.N.,

pp. 1-11.

In: Genetics:

MathuraRai

and descriptor

statesCurrent

New Frontiers,
andCrops-Management,

Plant

Vol. IV. Oxford

S. Kochhar.

1995.

for vegetable

crops,and Utilization.

pp. 77-90.

and Kochhar, S. NBPGR, New Delhi.
1991. Report of an international
workshop

Karihaloo,

Delhi, India,
8-12 October,
1990, 133p.
J.L., Brauner, S. and Gottlieb,
L. D. 1995.
eggplant,

Solanum

MathuraResourcesof Vegetable

byGupta,

P.N.,

resourcesheldat the NBPGR, New

Random amplified

polymorphic

Rai

DNA variation

Solanum

melongena

on South

L.D. 1995.

Methodology

Allozyme

variationfor establishinga representative

Resources and Crop Evolution,national
plant genetic
resources

Theoretical

Mathuraeggplant,

Mahajan,

exploration,

descriptor

Theoreticaland Applied
Genetics,
90:578-583.
I. S., Agrawal,
R.C. and Rana, R.S. 1996. Studies

L. (Solanaceae).
R.K., Bisht,

data. Genetic
Rana, R.S. 1994. Indianevaluation

Genetics

Karihaloo,

Collection:

and

In: Genetic

L. (Solanaceae).

appraisal,

and IBH, New Delhi.

Editedon okra genetic-a criticaland Applied

90:767-770.
J.L. and Gottlieb,

melongena

Resources

Characterization

Conservation

IBPGR.

theGenetic

in thecore set using

43:249-255.
system, pp. 1-19, In: Plant

evaluationand maintenance,

EditedRai, Kochhar,

S. and Duhoon,

S. S. NBPGR, New Delhi.

by Rana, R.S., Bhag Singh,

Asian

Okra

characterization
Genetic

Resources

Koppar,M. N.,

Appendix
NationalImandate of NBPGR

To plan, conduct and coordinate
plant explorations
germplasm of cultivatedplants, their wild relativesof economic importance.

for collectionof diversity
inand naturally
occurring species

To undertake introductionand exchange of plant germplasm for research purpose.
To examine seed and plant propagules
under exchange for the presence of
associatedpests and pathogens
and also to salvage healthymaterials
from theinfected/infested/
contaminatedsamples.
To undertake and promote characterization,
evaluationand documentationof plant
germplasm collections
and their distribution
to user scientists.
To undertaken
and promote conservationof plant genetic resources on a long termbasis employing
in vivo, in vitro and cryopreservation
techniques
and also to assist
in situ conservation
efforts.
To develop
and operate the National
Databasefor storage and retrievalof
information
on plant genetic resources.
To conduct basic researches for providing
a sound scientificback up to its services.
To develop and operate the National
Herbariumof Crop Plants and their Wild
Relatives.
To organize suitable training
programmes at the national,
regional
and international
levels.
To develop and implement
workplans based on memoranda of understandingandbilateralagreements.

Appendix

ActiveGermplasm holdings

S ta tio n/C e ntre
D e lh i

at Various

H o ld in g s
3 3 ,2 25

NBPGR Centres

M ajo r cro p s/c ro p s g ro u p s
W h ea t, B a rley , M a iz e , C lu sterb e a n (D ), C o w p e a,
B lac k g ram , P ea , C h ick p e a(D ), L e n til(D ), S u n flo w er,
P ea rl m ille t(D ), S o rg h u m , F o rag es, R a p ese ed ‑m u sta rd ,B rinja l, T o m ato , O n io n , G a rlic , C u c u rb its, C o ria n d er,
L e afy a nd ro o t v eg e tab les, M & A P , M in o r fru its e tc.

A k o la

3 0 ,6 6 0sm
C h ick
all pm eillets,a, P igeS oo ny pb eeaa,n S, oS rg
afflo
h u mw ,e Gr,roL inu nseed n ud t,, SMesaille
m ts
e ,a n d

N ig e r(D ), A m a ran th (D ), H o rseg ram , O k ra .
A m rav ati

4 ,2 0 0

G ree n g ra m , B lack g ram (D ), L ab la b b e a n , S w eet p o tato ,C h illies, P ap a y a, T ro p ica l fru its .

S h im la

1 2 ,3 8 1

F re n ch b e an , R ic e b e an , S o yb ea n , P e a , H o rse g ra m ,M in o r M illets, A m a ra n th , B u c kw h ea t, O ilsee d s,
T e m p e rate fru its, O rn a m e nta ls .

Jodhpur

1 2 ,3 8 0

Gm uillet,ar, MC oo th
w pbeeaa(Dn ,),
M Cu asto
n g bre aannd(DF ),ru Sits.
e sam e(D ), P e arl

T h rissu r

1 3 ,2 0 7

P ad d y , H o rse g ra m (D ), C o w p ea , F in g er m ille t, S e sam e,
B itte rg o u rd , G in g e r, C u rcu m a , C o lo ca sia , O k ra,
C assav a , A m o rp h o p h a llu s, M usa .

B h o w a li

5 ,0 6 6

Wc h heillin
at,g Bfruarle
itsy., L en til, B e an s, A llium , C h illie s(D ), L o w

C u ttack2 ,2 9 4

P ad d y (D ), W ild R ic e

S h illo n g

1 ,8 4 0

H ill p ad d y , M a iz e(D ), R ic e b e an , R o o t cro p s, F ru its

R an c h i

3 ,6 0 6

P ig eo n pe a(D ), B ra ss ic a , A rto ca rp u s , N ig er,
L a thy ru s(D ) , L in se ed (D ), P a pa y a(D )H y d e rab a d
T o talNote: 'D' denotes

1,55 7C hillie s(D ), B rinjal(D ), B la ck g ra m (D ), S im a ro u b a

1 2 0 ,4 16

duplicateholdings

IIAppendix

Research

Projects

on Evaluation

C e n tre s

PNrojectso .o fD e lh i

20

of Plant

Genetic

III

Resources

C ro p s/c ro p g ro u p s

C e rea ls (4 ), O ilse ed s (1), L eg u m es (1 ), V eg e ta b les (1 ),F o ra g e cro p s (1), B io ch e m ic al a n d p h y to c he m ic al
ev alu atio n (6 ), U n d eru tiliz ed cro p s (1 ), A rid leg u m es (1),E v alu atio n ag a in st b io tic stre sses (2 ), C o re co llectio n (1),D o cu m en tatio n a n d in fo rm atio n m an a g em en t (1)A k o la6

L e g u m e s(1), O ilse ed s(1), S o yb e an an d lin seed (1), M illets(1 ), O k ra an d o th e r m isce llan eo u s cro p s (2 )A m rav ati

1

M un g b e an an d o th er m isc ellan eo u s ag ri‑h o rticu ltu ra lc ro p s

B h o w alli

6

C e rea ls ,tem p era te le g u m e s a n d v e g etab le s (3), W ild
relativ es o f c ro p p la n ts(1 ), H o rtic u ltu ral c ro p s(1),M isc ella n eo u s cro p s (1)C u ttac k

1

R ice

H y d erab a d

4

M isce lla n eo u s ag ri‑ho rtic u ltu ral cro p s : p ig e o n p e a,c h illies, b rinja l, b lac k g ra m etc . (4 )Jo d h p u r

3

A g ricu ltu ral c ro p s o f arid reg io n (1 ), A rid z o n e
h o rtic u ltu ra l c ro p s(1), M isce lla n eo u s eco n o m ic p lan ts(1)R a n ch i2

R ice a n d o th er m isc ellan e o u s c ro p s o f th e re g io n (2 )S h illo n g

2

A g ri‑h o rticu ltu ral cro p s o f n o rth ‑ea st re g io n (2 )S h im la

6

P seu d o c ere als(1), F ren c h b ea n (1), T em p e rate fru its(2),O rn am en tal cro p s (1), G rain leg u m e s(1)T h rissu r

3

In d ig e no u s ag ri‑h o rticu ltu ra l cro p s o f so u th ern In d ia (2 ),N ew c ro p s to th e reg io n(1 )Appendix

Directory
S .N oof National
C ro p

ActiveIV

Germplasm Sites
N A C S ite

N o .o f ac c e ss io n s

1

W heatD ire c to ra te o f W h e a t R e se a rc h , K a rn a l‑1 3 2 0 0 1 (H a ry a n a )1 8 ,0 0 0

9

R ic eC e n tra l R ic e R e se arc h In stitu te , C u tta c k ‑7 5 3 0 0 6 (O r is sa )4 2 ,0 0 0

3

M a iz e

A ll In d ia C o o rd in a te d M a iz e Im p ro v e m e n t P ro je c t, In d ia n

2 ,5 0 0

A g ric u ltu ra l R e se a rc h In stitu te , N e w D e lh i 1 1 0 0 1 2
4

B a rle y

A ll In d ia C o o rd in a te d B a rle y Im p ro v e m e n t P ro je c t IA R I
R e g io n a l S ta tio n , K a rn a l 1 3 2 0 0 1 (H a ry a n a )5

S o rg h u m

N atio n a l R e s e a rc h C e n tre fo r S o rg h u m ,5 ,1 6 0

R aje n d ran a g a r ,H y d e ra b a d (A n d h ra P ra d e s h )‑5 0 0 0 3 06

P e a rl

A ll In d ia C o o rd in a te d P e a rl m ille t Im p ro v e m e n t P roj e c t, C o lle g e

m ille t

o f A g ric u ltu re , S h iv aj i N a g a r , P u n e (M S ) 4 1 1 0 0 5

S m a ll

A ll In d ia C o o rd in a te d S m a ll M ille t Im p ro v e m e n t P ro je c t,

M ille ts

U n iv e rs ity o f A g ril. S c ie n c e s B a n g a lo re (K a rn a ta k a) 5 6 0 0 6 58

P u lse s

D ire c to ra te o f P u lse s R e se a rc h (IC A R ) , K a n p u r‑2 0 8 0 2 4 (U P )9 ,3 1 0

9

S oy bean

N a tio n a l R e s e a rc h C e n tre fo r so y b e a n , In d o re ‑ 4 2 5 0 0 1 (M P )2 ,5 0 0

10

O ils e e d s

D ire c to ra te o f O ilse e d s R e se a rc h (IC A R ) , R aj e n d ra n a g a r,

1 5 ,6 2 9

7

8 ,5 7 2

H y d e rab a d ‑5 0 0 0 3 0 (A P )11

RM aupsetas erde d &A JI In d ia C o o rd in a te d C ro p Im p ro v e m e n t P ro je c t R a p e se e d &8 ,0 8 2

M u sta rd ), H A U , H isa r 1 2 5 0 0 4
12G ro u n d n u t

N a tio n a l R e se a rc h C e n tre fo r G ro u n d n u t, T im b a w a d i J u n a g a rh

6 ,4 3 2

3 6 2 0 1 5 (G uj a rat)
13

S u g a rc a n e

S u g a rc a n e B re e d in g In stitu te , C o im b a to re (T N )3 ,9 7 9

14

C o tto n

C e n tra l In stitu te fo r C o tto n R es e a rc h , P .B . N o . 1 2 5 ,

6 ,8 9 6

N a g p u r‑4 4 0 0 0 1 (M a h a ra sh tra )15

AJulliete &d

C e n tra l In stitu te o f Ju te & A llie d F ib re , B a rra c k p o re

3 ,2 2 6

7 4 3 1 0 1 (W e st B e n g a l)
F ib re s
16

V e g e ta b le sD ire c to ra te o f V e g e ta b le R e s e a rc h , V a ra n a s i‑2 2 1 0 0 5 (U P )1 6 ,1 3 917

P o ta to

C e n tra l P o ta to R e s e a rc h In s titu te , S h im la ‑ 1 7 1 0 0 1 (H P )2 ,3 4 2

18

F o rages

In dian G rasslan d & F o dd er R ese arch In stitute, Jh an si‑2 84 0 03

6,2 6 9(U P )
19

S pice s

N ation al R esearch C entre for S pices, M arik un nu , C alicut

2 ,84 7

(K erala) 6 73 0 12
20

T ob acco

C en tral to b acco R esearch In stitu te, R ajahm u n dry (A P ) 53 3 10 5

1 ,50 021

P lan tatio n

C en tral P lantation C ro ps R esearch In stitute, K asarg od 6 7 1 0 24

30 7

C ro p s(K erala)

AM ro
edici
m ati
n cal &

N ation al R esearch C entre fo r M & A P A n an d (G ujarat)

22

37 5

P lan ts
23

24

25

A g rofo restry

N ation al R esearch C entre fo r A g ro‑fo restry, In d ian G rasslan d &P lan ts

F o d der R esearch in stitute Jhan si 2 84 00 3 (U P )F ru its

A ll In dia C o ord in ated P roject (S em i A rid F ruits) H aryan a A g ril.

(S em i A rid)

U n iversity, H isar 12 5 0 0 4 H aryan a

F ru its

N B P G R R egion al S tatio n , P h ag li, S h im la 17 1 0 04 (H P )

40

54 1

6 67

(T em pe rate)26

lH Co rti
ropcu
s lturaInd ian In stitute of H o rticultu ral R esearch, 25 5 , U pp er p alace

13 ,118

O rch ard s, B an galore 5 60 0 80 K arnatak a27

C itru s

N atio nal R esearch C en tre fo r C itru s, Sem ina ry H ills, N agp u r

90

44 0 0 0 6 (M ah arashtra)
28

M ang oC en tral In stitute fo r H o rticu lture fo r N o rthern P lain s, L uck no w

7 27

22 6 0 16 (U P )29

30

T u be r

C en tral T ub er C ro ps R esearch In stitute, S ree‑k ariy am ,C ro psT riv and ru m (K erala )‑69 5 0 17

P se ud o

N B P G R R eg io nal S tatio n , P hagli, S h im la 17 1 00 4

cereals

3 .6 43

3 ,6 82

Specific

Characters

C ro p g ro u p s/c ro p

Recorded during

Evaluation

for Identification

Appendix V
of Promising Lines

C h a ra c te rs

V e g e ta b les
B rin ja l

PH higo hm oypiesis
ld bpligo teh nt tia
a n l,
d sletessm asenedd efru
d ,itebaorlin
re re s s , le s s s p in in e ss , re s ista n c e to

O k raHm igo sah icy iev iru
ld ,seaanrlin
d fru
e s its, anlo dw step umb ebsoc reen rc e o f fru its , re s is ta n c e to y e llo w v e in

C h illie s

Ha nigd hlepaufncguerl
n c y , ea rlin e s s, lo n g sh elf life , h ig h y ie ld , re s ista n c e to a n th rac n o se

L a b la b b e a n

Pfohliao tor dinissee na sesitiv
s e ty p e s , e a rlin e ss , fle s h y p o d s , lo n g s h e lf life , re sista n c e to

T o m a to

aH nigd ho thy iee rldfo, dliae teg ermd iniseaates etys p e , h e a t to le ra n t, h ig h T S S , re sista n c e to le a f c u rl

L eg u m e s
G re e n g ram /E re c t h a b it, s y n c h ro n o u s flo w e rin g p e rio d , e a rly m a tu rity , h ig h H . I.,b la c k g ra m

re sis ta n c e to y e llo w m o sa ic , le a f c rin k le , o th e r fo liag e d is e a se s an d b ru c h id s

C e r ea ls
R ic eH ig h y ie ld , g ra in q u a lity , re s is ta n c e to b io tic an d a b io tic stre s se s

O ils e e d s&R aMp ue sestae rdd

Y ie ld p o te n tia l, lo w e ru c ic a c id , h ig h o il c o n te n t, re s is ta n c e to A lte rn a ria
b lig h t, w h ite ru st a n d ap h id s.

F r u it C ro p s
Z izy p h u s

P u lp /sto n e ra tio , c risp n e s s, ju ic in e s s a n d h ig h T S S , e a rly m a tu rity

A eg leS c u ll th ic k n e ss , h ig h T S S , p u lp c o lo u r, le s s se e d a n d m u c ila g e in th e p u lp

E m b lic a

F ru it s iz e , sk in c o lo u r, fib re c o n te n t, q u a lity a n d v ita m in C c o n te n t

B a n an akS eu ecpkinin gg qh ua ab lityit, b u n c h a n d fin g e r c h a ra c te rs, fru it q u a lity , p e e l th ic k n e ss a n d

M ang oSinecxidratio
e n c e, bo ie
f nm naia
n gl/ao nmnaulfo
a l rmb e aarin
tio gn, fru it q u a lity , s to n e /p u lp ra tio , a ro m a a n d

L itc h icP ounlpd /seitioend s ra tio , fla v o u r, s w e e tn e s s, to le ra n c e to fru it c ra c k in g u n d e r d ry

Ja c k f ru it

S h a p e a n d s iz e , p u lp /se e d ra tio , sw e e tn e s s , fla v o u r, fib re c o n te n t

mM ea/len d ma rin
o n /liFrerus isittasen t/d
ce ro
top v, iru
ju icsina ne ss
d ,M leLss
O ssedeisd e da ,sepsh y s io lo g ic a l d iso rd e rs o f th e fru it a n d

Appendix
List of Catalogues
S r.N o .

PublishedVI

by NBPGR

C ro pB o ta n ic a l n am e

p uY b lic
e a ra tioofn

a c Nc eos .sioo fn sd e sNc ripo . tioof n sA m a ra n th

A m a r a n th u s s p p .

198 1

40 0

31

2

B a n an a

M usa

199 3

19 1

60

3

B a rle y

H o rd e u m v u lg a r e

198 3

25 9

35

‑d o ‑‑d o ‑

198 3

1 155

27

4

‑d o ‑‑d o ‑

198 4

74 2

15

5

‑d o ‑‑d o ‑

198 5

2 17

15

6

‑d o ‑‑d o ‑

198 6

28 0

8

7

C h ic k p e a

C ic e r a rie tin u m

199 3

12 06

15

8

C lu s te rb e a n

C y a m o p s is te tra g o n o lo b a

19 81

1 150

22

9

‑d o ‑‑d o ‑

198 3

83 0

26

10

‑d o ‑‑d o ‑

198 5

15 40

24

11

‑do ‑

‑d o ‑

19 89

15 78

21

12

‑d o ‑‑d o ‑

199 5

52 0

23

13

C ow pe a

V ig n a u n g u ic u la ta

198 1

70 7

34

14

‑d o ‑‑d o ‑

198 2

6 83

24

15

E g g p la n t ‑ I

S o la n u m m e lo n g e n a

199 5

1 188

52

16

F o x ta il M ille t

S e ta r ia ita lic a

19 87

73 6

52

17

F re n c h b e a n

P h a s e o lu s v u lg a r is

198 1

17 73

16

18

G re e n g ra m

V ig n a ra d ia ta

198 3

30 2

19

19

‑d o ‑‑d o ‑

199 6

15 32

61

20

H o rse g ra m

M a c ro ty lo m a u n iflo ru m

199 5

92 0

12

21

K o d o m ille t

P a sp a lu m s c ro b ic u la tu m

198 7

20 6

33

‑d o ‑‑d o ‑

198 7

18 6

33

22

L e n til

L en s c u lin a ris

1 9 8 2 ‑8 3

24 0

14

23

L in s e e d

L in u m u s ita tiss im u m

199 3

621

19

24

M a iz e

Z ea m a y s

198 4

38 0

25

25

‑d o ‑

‑do ‑

198 5

7 68

12

26

‑d o ‑‑d o ‑

1 9 8 6 ‑8 7

4 62

21

‑d o ‑‑d o ‑

1 9 8 6 ‑8 7

14 4

19

‑d o ‑

‑do ‑

199 1

63 5

26
19
19

1

27

‑d o ‑‑d o ‑

199 1

30 4

‑d o ‑‑d o ‑

199 1

58 1

‑d o ‑‑d o ‑

199 1

23 0

19

28

‑d o ‑‑d o ‑

199 5

19 7 1

26

29

M o th b e a n

V ig n a a c o n itifo lia

198 0

28 5

17

30

‑d o ‑‑d o ‑

198 1

84 8

17

31

‑d o ‑

‑do ‑

198 3

8 29

32

M ustard

B rassica sp p.

198 6

5 55

20
7

100 0

31
45

33

O ats

A ven a sp p .

199 0

34

O k ra P art 1

A belm osch us escu len tu s

199 0

558

35

O k ra P art 2

‑do ‑

199 1

3 94 319

36

O k ra P art 3

‑do ‑

199 3

5 80

42

37

O k ra (W ild )A belm osch us sp p .

199 5

24 1

35

38

O p iu m P o pp y

P ap aver so m n iferu m

198 0

14 5

19

39

P earl m illet‑1

P enn isetum g la ucu m

199 3

193 8

20

40

P earl m illet‑2

‑d o‑

199 3

2 45 8

18

41

R ice

O ry za sativa

198 8

10 2

56

42

S afflow er

C a rth a m u s tin cto rius

198 2

48 1

31

43

‑do ‑

‑d o‑

199 5

85 1

27

44

S esam e

S esam um ind icu m

198 2

29 7

22

45

‑do ‑

‑d o‑

198 3

13 93

29

46

‑do ‑

‑d o‑

199 3

2 06 839

47

S esb an ia

S esb an ia spp .

198 2

54

31

48

S o rg hu m

P art‑ 1

S orgh um b ico lo r

199 1

1 10

34

49

S org hu m

P art‑ 2

‑d o‑

199 2

150 0

26

50

S o ybean

G ly cine m ax

198 3

2 00 9

18

51

‑do ‑

‑d o‑

199 3

2 53 923

52

S un flo w er

H elian th us an n uu s

198 2

35 2

13

53

T o m ato

L ycop ersicon escu len tu m

198 2

80

21

54

T rigo nella

Trig on ella sp p

198 0

17 1

27

55

W h eat & T ritic ale

Triticu m aestivu m , T ritica le

19 82 ‑83

17 18

25

56

‑d o‑

‑do ‑

198 4

19 79

14

57

‑d o‑

‑do ‑

198 4

2 14 3

14

58

‑d o‑

‑do ‑

198 6

15 29

8

17 18

8

59

‑d o‑

‑do ‑

19 86 ‑87

60

‑do ‑

‑do ‑

19 87 ‑88

2 7 97

8

61

‑d o‑

‑do ‑

19 88 ‑89

3 5 92

8

62

‑do ‑

‑do ‑

19 89 ‑90

3 33 9

8

56 8

25

143 9

31

63

‑d o‑

Triticu m spp .

19 83

64

W ing ed b ean

P sop h oca rp us tetrag on o lo bu s

198 3

65

C atalog ue o n C ro p G enetic R esou rces
C ow pea

V ig na u ng uicula ta

198 4

88

31

R edg ram

C aja nu s cajan

198 4

25 9

23

H o rseg ram

M a cro ty lom a un iflo rum

198 4

39 9

14

C h illies

C ap sicum spp .

198 4

40 3

12

T u rm ericC u rcum a sp p .

198 4

10 2

9

D iosco rea sp p .

198 4

1 12

22

Y am

Internationalizationof Elite Germplasm for Farmers : Collaborative
Mechanisms to Enhance Evaluationof Rice Genetic Resources
R. C. CHAUDHARY
INGER Global
International

Coordinator,
Rice

Research

GRC
Institute,

P. O. Box 933, Manila,

Philippines

Abstract
Ancestorsof rice evolved in South and SE Asia, and Niger basin of Africa.
Domesticatedover 10,000 years, these evolved into land races, and bred over last 100 years
into elite germplasm.
A collaborative
network mechanism called International
Network for
Genetic Evaluation of Rice (INGER)
facilitatedevaluation
and utilizationof rice germplasm
since 1975, through 1000 scientists
located at 700 locationsin 95 countries.
Out of 40,000
elite breedinglines evaluatedglobally,
577 were released as varieties
in 63 countries
away
from their origin. Several thousand lines were used in local hybridizationtransferring
superior
charactersand diversifying

Introduction
Genetic
human race.
added by plant
process, plantand economic

diversity

farmers

varieties.

But PVR and IPR may endanger

is the basic raw material

for the growth

INGER.

and sustenanceof

The genetic diversity
created by nature and genetic recombination
breeders form the basis of varietal improvement globally.
In the whole
breeders try to adjust the genotype of the plant to agricultural,socialenvironment where these are expected to be grown. Oryza sativa (ricecultivatedworld over) originatedin the humid tropics of South and South East Asia,and Oryza glaberrima(ricecultivatedin parts of Africa)
originatedin Niger basin.
Under domestication
over 10,000 years, it evolved into various ecotypes and landraces under influenceof natural and farmer selection(Oka, 1988; Vaughan, 1994).
The available genetic variability
needs to be selected for a particularagro-climate
toachieve high and stable yield (Swaminathan,1993).
In this process germplasm
sharing
followedby testing
and acclimatization
play key role.
International
institutions
like International
Rice Research Institute(IRRI),
International
Instituteof Tropical Agriculture(IITA),
Centro Internacionalde Agricultura
Tropical (CIAT),West Africa Rice Development Association(WARDA) and most of 115 national
agriculturalresearch system (NARS) have rice breeding
program on going at somelevel.
Increasingsustainable
yields and broadening
the genetic
base of farmers'varieties can only be obtained through international
exchange, evaluation,and use ofdiverse germplasm (Alluri et al., 1995; Nguyen et al., 1994). Unrestrictedsharing andexchange of germplasm across geographicaland politicalboundaries
requires soundnetwork and commitment of the members.
Collaborative
mechanism in International
germplasm testing
Networks are inexpensive yet effective
catalysts
for research. They provide
opportunities
for isolated scientists
to form structured working partnershipsthat boost
research efficiency,
jump start a country program, save on time, and reduce costs.
The networks also help spread useful research results among regions with similaragro-ecologies
despitecontrasting
political,
religiousor socialbackgrounds(Chaudhary,1994).
Baum (1986)
defined3 types of networking
in agriculturalresearch, based on coordination
unit, network members, and communication among
them. Plucknett and Smith (1984)
proposed 7 principles
for the success of a network,
to which Greenland et al. (1987)
added 2 more. May not be for final but Seshu(1988)
added 4 more to that list. But the sum and substance of a successful networkis the joint ownership, individual
benefit,mutual trust, and free-flow of germplasm.
Ignore any of the four and the network lands on the shore. Networks have been
establishedto test crop germplasm over a broad range of environments, explore waysof boosting the efficiencyof the scientists,scientificinstitutions
and thereby improve
the lots of farmers and consumers in a shortest possible
time. Networks can assume
various forms (Baum, 1986).
International
Network for Genetic Evaluationof Rice(INGER)establishedat IRRI assumed one such form.
Evolutionof INGERInternational
cooperationin agriculturalresearch is rapidly
increasingwiththe tighteningof funds and realizationof the benefitsof collaboration.
Networking
among agriculturalscientists
is not new but the current level of collaborationisunprecedented
where scientists
in over 95 countries
forged partnershipon a global
scale for mutual support, trim costs, avoid duplication,shorten time frame of varietal
development
boost efficiency
and acceleratetransferof technology
to farmers(Chaudhary,
unparalleled1994; Chaudharyand Ahn, 1994).
But the form assumed by INGER is
where 2 way flow of breeding
material
and information,and entails

commitment of resources from participating
nations.
The International
Rice Research Institute(IRRI)
was founded in 1960 withthe aim of improving
the rice production
technology
and sharing this with therice-growing
countries of the world. The exchange of breeding material started in
1963, though informallywith a few interested
breeders.
The necessity was realized
to start an organized
and formalized
forum through which the genetic
materialdeveloped
at IRRI as well as by the national
agriculturalresearch system (NARS)
should be pooled and evaluated for sharing.
This gave birth to the International
Rice
Testing Program (IRTP) in 1975. The IRTP as a project was first approved by the
UNDP in January 1975, for a period of 5 years with a funding support for US$2.0
million.
In its second phase INGER was extended for 1980-84 with a grant of US$7.8
million from the UNDP, and renamed as International
Rice Testing and Improvement
Program (IRTIP).
IRTIP had greatly
expanded workplan including
germplasm
collection,
cooperative research networks on innovative techniques
for rice breeding,and biologicalnitrogenfixation.
The next phase of the project
1985-90
was also
funded by the UNDP. During 1991 to 1996 the UNDP continued its funding ascomponent III of the Global Program entitled"Development of technology
whichhave less dependence on synthetic
fertilizers
and agro-chemicals".
INGER came into being to replace IRTP in 1989 with the following
objectives:
To make the world elite rice germplasm available
to all rice scientists
fordirect use or in crosses within breeding programs.
To provide rice scientists
with an opportunity
to assess the performance of
their own advanced breeding lines over a wide range of climatic,
cultural,soil, disease, and insect-pest
conditions.
To identify
genetic
sources of resistanceto major biotic
stressesandtolerance
to abiotic stresses.
To monitor and evaluate the genetic variationof pathogens
and insect-pests.
To serve as a center for informationexchange on how varietal characteristics
interact with diverse rice growing environments.
To promote cooperation
Globaland interaction

thinkingwith regional
focus.
The germplasm are shared and evaluatedamong rice improvement

scientists.

through

Basically

INGER nurseries.

there are two types of nurseries (AppendixI); ecosystem oriented and stress oriented.
Ecosystem oriented nurseries are focused towards identifying
germplasm, suitable for
irrigated,upland lowland and flood prone ecosystem. Stress oriented nurseries
arefocused towards identifying
donors for resistanceand tolerance to various abiotic andbiotic stresses like, disease,pest, temperatureand soil stress. For each ecosystemoriented nurseries there are observational
and yield nurseries.
About 1000 scientists
located at 700 locationsin 95 countries (AppendixII) receive, evaluate and utilize
these nurseries.
INGER has regional
focus such that to focus special problems
specific nurseries are composed for Africa (INGER-Africa),and Latin America andCaribbean(INGER-LAC).
The INGER-LAC locatedat CIAT Colombia,
and
INGER-Africa
located at IITA Nigeria (to be shiftedto WARDA, Cote d'lvoire)supervise these nurseries.
Modus operandi of INGER
INGER operates from IRRI Los Banos in the Philippines
to coordinate
theactivities.
Its flow chart of activities
are depicted
in Fig.1, and operations
described
below.
A. Field operations
1. Introductionof the promising
germplasm.
Promisingelite germplasm
are contributedby all research
institutions
belongingto NARS and IARC, though some contribute more than the others (Fig. 2).
Information
are sent to most NARS (Appendix II), and all IARC for nominating
their
best breedinglines for specificnurseries(Appendix
I). Scientists
respond by
nominating entries for specific nursery, and provide seeds. In case seed are sufficientfor the nurseries,
these are directly
accepted for testing or else are multiplied
at LosBanos, Philippines.
2. Multiplicationof seed.
The introduced
seed are checked thoroughly
by Seed Health Unit if they are"clean" and meet the Philippines
Plant Quarantine
requirements.
Upon clearance,these
are multiplied
in isolationarea, and are observed
for any unwanted
contaminationof pests, diseases and weeds. The second stage multiplication
under normal field conditions,and limited
observations
are recorded
suitabilityof the proposed entry for a particularnursery.

is doneto see theFig.

1. INGER's

flowchartof activities/linkages

Fig. 2. Percentage

of entries

contributedby NARS in INGER nurseries(1975-1995)

3. Nursery composition.
Entries are composed into specificnurseriesof nursery and number of sets required(Appendix

whose number depend on typeI). The whole process of thereceiptof nomination
for testingand indent for the nursery
evaluation
and report preparation
follows a strict time line.
B. In-house operations
1. Incoming seed.
The incoming seed has in-house aspects
cooperators of the seed status, clearance through
IRTP number to each seed lot, computerization
lot is divided into three parts, one for seed file,of it, which
Seed Healthtilldispatch,data

involves informing theUnit (SHU), assigningof the information.
Each cleared seedsecond for multiplicationand third asremnant.
2. Seed conditioning.
Seedmultiplication
has associatedin-house
operation
calledseed
conditioning
which involves air and screen cleaning, washing and floatation,
grading,
removal of genetic impurities,
specialprocessing,and seed treatment (hot water,
chemical)and germinationtest.
3. Decision on test locationsof the nurseries.
Typical of the philosophyof INGER, tests locations
are voluntary.
A circular
is sent around June every year to a number of cooperatorsin every NARS informing
them the type of nursery available
during the following
years. They can decide thetype of nursery they want to request for evaluation.
These requests are compiled andform the basis of the number of sets for each nursery to be composed.
4. Plant quarantine
and seed health.
Every country has its own plant quarantine,seed health,and seed importregulation.
INGER is firm believer of safe exchange of seed and thus over the last 20
years of its operation,no case has come where a pest or disease was imported orexported.
Every seed lot imported
must be accompanied
by a Phytosanitary
Certificateof the originating
country and Philippine
Import permit. Similarly
every
exported seed lot contains the Import Permit of the importing
country (if applicable),
Plant

Quarantine

certificate

and the seed list.

5. Dispatchof nurserysets.

Nursery sets to the countries north of the equator are done mostly during
February to May. Similarly
seeds are dispatchedto south of the equator countries
during June to September.
Seeds of some specificnurseries like boro nursery aredispatchedduring June to September for seeding any time after October.
6. Data receipt and verification.
After the trials are conducted and observations
recorded, three sets of the data
get recorded automaticallyin the field books. One set is mean for INGER, anotherfor the national
coordinatoran the third for the cooperator.
The last date for thereceipt of the data is set for July 15.
7. Analysis
and preparationof the report.
Data are analyzed appropriately
and nursery reports are prepared.
The
printed reports are made available to cooperators before the end of the year, for study
and use. Through the information
thus shared, cooperators
get the performance of
their breeding lines across the variable environment of the globe. They can also see
the stabilityof agronomic or resistancecharactersin various locations.
This does
provided information
not only on the stabilityof the resistancebut also on the racesof pathogen and biotypes of the insect-pests.
New analytical
tools of G X E analysis
make the data more useful,training.

which will be appreciatedmore at the end of the present

C. Monitoring
and Coordination
operations
1. Monitoringteam visits.
On case basis, monitoring
tours to some specific nurseries and locations
aremounted annually.
Members of the team include scientists
from NARS and IARC.
Monitoring
tours provide opportunity
to review a specificnurseries
or locations
critically.
It is also good opportunity
for consultations
and lateral learning.
2. Site visits and general supervision.
On individual
basis, a close contact is maintained
locatedat various sites. Routine visits
are maintained
information.
3. Project
SupportA Projectwith various
to exchange

cooperators
views andTeam meetings.
SupportTeam comprisingof seniorscientists

at IRRI

providestechnicalbackup and support to INGER. Members represent
divisions,consortia
and programs.
Meetings are held twice
problems and seek opinion on issues of importance.
4. INGER Steering
Committee meeting.

variousannually

disciplines,
to sort out

INGER Steering Committee comprising of scientists
and administrators
frommajor rice growing countries and international
institutions
concerned with rice. This
committee meets annually to discuss the results of the previous year, plan for future
nurseries and activities
and provide policy guidelines.
Impactof INGER

Genetic

resources sharing and evaluation.
From 1975 to 1995, over 40,000 test entries were evaluated through INGER(Fig. 3, Appendix
I). These lines originatedfrom the breeding
programs of 95
countries and IRRI, IITA, CIAT. While the contributionof NARS and IARCs mayvary (Fig. 2, Table 1), the sharing is un-restricted.
Political
neutralityof INGER helps
override those barriers,and our operations
are the same be it U.S.A., India, Iran or
Iraq.
Evenson and Gollin (1996)
concluded that INGER nurseries stimulated
more
international
search for genetic resources.
More than 3,000 breedinglines andvarieties
distributed
through INGER have been used in hybridization
by national
programs to improve the productivityof local varieties.
Similarly,
IRRI and other
IARCs working with rice had easy access to NARS' breeding material.
Of 1790
modern varieties released, 390 were borrowed- developed in one country and released
in another. IRRI provided 75 percent of the borrowed varieties,
most of which weremade available
through INGER.
Varietalreleases.
Varietalrelease stemming out of the exchanged germplasm made significant
contributions
to production
increases in several rice growing countries.
Over the last
20 years, 577 INGER provided lines have released varieties
in 62 countries
of Asia,
Africa and Latin America (Table 2). In Vietnam, China, Indonesia
over 60% of thetotal rice area is covered by INGER distributedlines. More than 10 million
hectareof rice area in China are planted to materials taken directlyfrom INGER nurseries arederivedfrom crosses

made with

INGER entries.

About

65 million

hectaresareTable1. Volume of INGER nurseriesn Nu ros e.o rfie se nN t roie.os f

Y ear

and the source of germplasm,

1975 to 1995.

S o u r c e o f e n t r ie s ( % )N A R SI A R C 's

IR G *

cNo ou .on trf i e s

t r iNa lo s.oe tsf

19 75

12

19 07

5 2

4 6

1

3 6

4 6 1

19 76

14

20 10

5 9

3 9

2

3 8

5 73

19 77

15

23 14

65

3 3

2

3 9

5 77

19 7 8

15

24 83

54

4 4

2

5 0

8 62

15

24 53

50

48

2

5 8

9 56

1 98 0

17

2 108

52

44

5

5 8

10 8 1

1 98 1

3 2

20 26

48

49

3

5 2

12 15

1 98 2

2 8

26 05

58

40

1

6 0

13 67

198 3

2 3

25 78

50

46

4

6 2

1 195

198 4

2 3

27 48

47

48

5

5 3

13 0 1

198 5

2 5

2 895

45

50

4

4 9

17 0 7198 6

2 5

15 4 3

42

55

3

5 1

14 64

198 7

2 5

12 9 3

47

50

3

50

15 5 046

12 17

19 7 9

198 8

2 6

169 1

54

43

2

198 9

2 5

1280

54

44

2

4 1

9 5 9

199 0

2 3

174 6

59

3 7

4

35

92 1

199 1

2 5

198 8

58

35

7

49

86 7

199 2

19

1560

50

4 1

9

35

63 8

199 3

15

89 6

59

3 7

4

42

77 7

199 4

14

115 4

47

4 6

7

37

76 4

199 5

15

135 5

50

4 4

6

40

67 4

T o ta l

4 0

4 0 63 3

53

4 3

4

95

2 108 6

International

plantedRice Genebank

to these varieties

annuallyin the world.

Increaseddiversityand complexityof pedigree.
With the increased availabilityof diverse germplasm, the number of parents
entering
into a released variety increased(Table 3). Pedigree
analyses of 1,709
varieties
released from 1975 to 1991 and available
at IRRI data bank, revealed thata total of 11,592 ancestors were used in developing
these varieties.
Only 3 varieties
released before 1965 contained more than 4 ancestors, 22 varieties
released during
1986-1991
could be traced to 5 or more ancestors, and 72 had more than 15 ancestors.
Growing complexity of pedigrees is said to have a definite
advantage for stabilityofperformanceand resistance.

Table2. Rice varieties

C o u n try o f re le a sereleasedin 62 countries

V a rie ty N o .

E A S T A S IA
C h in a29

out of INGER nurseriestestedduring

1975-1995

O rig in a tin g c o u n try o r o rg a n iz a tio n
B an g la d e sh , C o te d 'lv o ire , In d ia , IR R I, K o re a , P a k ista n ,P e ru , S ri L a n k a , T a iw a n (C h in a) , U .S .A .

S O U T H E A S T A S IA
B ru n e i2

IR R IC a m b o d ia

9

In d ia , IIT A , IR R I

In d o n e sia

21
4
28

In d o n e s ia , IR R I , T h a ila n d

M a la y s ia
M y anm ar

C IA T , IR R I, M a la y sia
A u stra lia , B a n g la d e s h , In d ia , In d o n e s ia , IR R I,
P h ilip p in e s , S ie rra L e o n e , S ri L a n k a , T h a ila n d

P h ilip p in e s

5

T h a ila n d
V ie tn am

1
42

IR R IT h a ila n d
B a n g la d e sh , C o te d 'lv o ire , In d ia , In d o n e s ia , IR R I, IR A T ,P h ilip p in e s , T a iw a n (C h in a ), V ie tn a m , T h a ilan d

S O U T H A S IA
B a n g la d e sh

10

B a n g la d e sh , In d o n e sia , IR R I

B h u ta n
In d ia

840

B a n g la d e sh , In d ia , IR R I, J a p an , K o re a , S ri L a n k a
B a n g la d e sh , In d ia , In d o n e s ia , IR R I, P h ilip p in e s ,S ri L a n k a

N epal10

B a n g la d e sh , In d ia , IR R I, N e p al, S ri L a n k a

P ak istan

3

In d ia , IR R I

W E S T A S IA & N O R T H A F R IC A
E gy pt

4

IR R II ra n4

In d ia , IR R I, J a p a n

S u d anT u rk e y

1
3

IR R IB u lg ar ia , Ita ly , U S S R

15
15

C IA T , IIT A , In d ia , IR R I, L ib e ria , S ri L a n k a
B a n g la d e sh , B u rk in a F a so , C o te d 'lv o ire , In d ia , IIT A ,S U B ‑S A H A R A N A F R IC A
B e n inB u rk in a F a so

IR R I, L ib e ria , P h ilip p in e s
B u ru n d i

9

B a n g la d e sh , B u rk in a F a so , C o te d 'lv o ire , In d ia , IIT A ,In d o n e s ia

C a m e ro o n
C e n tra l A fric a n

15
1

C o lo m b ia , In d o n e s ia , IIT A , IR R I, IR A T , T a iw a n (C h in a )IR R I10

B a n g la d e sh , F re n c h G u in e a , IR R I, S ie rra L e o n e ,GR ea pm ubb ialic
S ri L a n k a , T a iw a n (C h in a)Table 2 (continued)
C o u n try o f re le a se
G h an aV a riety N o .
16

O rig in a tin g c o u n try o r o re an iz atio n o u n trv o f re le a s e
B a n g la d e sh , C o te d 'lv o ire , IIT A , In d ia , IR R I, N ig e r ia ,S ri L a n k a , P h ilip p in e s

G u in e a

3

C o te d 'lv o ire , IR R I, T a iw a n (C h in a )G u in e a B iss a u

519

F . G u in e a , S ie rr a L e o n e , S ri L a n k a , T h a ila n d
B ra z il, B u rk in a F a so , C o te d 'lv o ire , H a iti, IR R I, In d ia ,C o te d 'lv o ire

In d o n e sia , S e n e g al, S ri L a n k a , Z a ire
K en ya8

B a n g la d e sh , B ra z il, In d ia , IR R I , S ri L a n k a

L ib e ria

8

IR R I, IIT A , L ib e ria , M ala y s ia

M a la w i

2

In d ia , IR R I

M a li5

C o te d 'lv o ire , In d ia , IR R I

M a u r itan ia

6

M o z a m b iq u e
N ig e r

10

IR R I.
IIT A , IR R I, P h ilip p in e s2
24

IR R IB u rk in a F a s o , C o te d 'lv o ire , IIT A , IR R I, N ig e ria ,R w an da

1

IR R LS eneg al
S ie rra L e o n e

1220

C o te d 'lv o ire , In d ia , IR R I, S e n e g a l, S ri L a n k a
B a n g la d e sh , C o te d 'lv o ire , C u b a , G u y a n a , IIT A , IR R I ,N ig e ria

L ib e ria , M a la y s ia , N ig e ria , S ri L a n k a .
T a n z a n ia

19

IIT A , In d ia , IR R I , S ri L a n k a , T a iw a n (C h in a ) , T a n z a n ia

T o goU g and a

52

C o te d 'lv o ire , In d ia , IIT A , IR R I, L ib e r ia

Z a m b ia

7

In d ia , IIT A , S ri L a n k a , T a iw a n (C h in a )Z im b a b w e

4

IIT A , IR R I

L A T IN A M E R IC A
B e liz e

2

C o lo m b ia , C IA T

B o liv ia
B raz il

11
35

B o liv ia , C IA T , C u b a , IR R I
B ra z il, C IA T , C o lo m b ia , C o te d 'lv o ire , IR R I, M a la y s ia ,B u rk in a F a s o , IR R I

S ri L a n k a , T a iw a n (C h in a) , T h a ila n d .
C o lo m b ia

5

C IA TC o s ta R ic a

5

C IA TC ub aRD eo pm u inb lic
ic a n4
3

C u b a , IR R I
C IA T , D o m in ic a n R e p u b lic , IR R I

E cu ado r

3

C IA T , IR R I

E l S a lv a d o r

2

C IA TG u a te m a la

5

C IA T , C o lo m b ia , K o re aG uy ana

2

C IA T , IR R I

H o n d u ra s

5

C IA T , C o lo m b ia

Table 2 (continued)
C o u n try o f re le a s e

V a rie ty N o .

M e x ic o

8

C IA T , IR R I, M ex ic o , T h a ilan d

N ic a ra g u a

4

C IA TP an am a2

C o lo m b ia , P a n am a

P e ru6

C IA T , IR R I, P e ru

V e n e z u e la

5

C IA T , In d ia , T a iw a n (C h in a )T o ta l

5 77

Fig.

3. Intra-and inter-continental

O rig in atin g co u n try o r o re a n iz a tio n o u n trv o f re le a se

movementof eliterice

germplasm

through

INGER

during

1975

to

1995.

Donors for resistanceand tolerance to stresses.
The INGER observational
nurseries on resistanceand tolerance

to biotic

andabiotic stresses have facilitatedthe evaluationof resistance/tolerance
in elite breedinglines against various stresses, and also identify
donors. As a result, a large numberof donors for resistanceagainstdiseasesand pests, and tolerance against various

Table 3. Average number of ancestors
1965

to 1991(data

source:in a released

Evenson and Gollin

varietyand the source of ancestralmaterial

during1993).

N u m b e r o f a n c e s to r s u s e d i n th e v a r ie t y
P e rio d

A v e rag e N o .

f ro mn o n ‑IR R I

%

fro m

IR R Is o u rc e ss o u rc e s
2 .7

P re ‑ 1 9 6 5

2 .5 5

2 .4 81 9 6 6 ‑7 0

4 .0 1

1 .8 95 2 .91 9 7 1 ‑7 5

5 .2 92 .1 55 9 .4

1 97 6‑80

8 .1 5

1 .6 97 9 .3

198 1‑85

7 .4 9

1 .9 5

7 4 .0198 6‑91

7 .2 3

2 .1 86 9 .8

abioticstresses are available
now (Tables 4, 5, 6). These have been used by plant
breeders in the hybridization
program to make the local varieties
resistantto thesestresses. The exact value of these donors is hard to estimate as they protect the crop,
stabilize
their yield, save on pesticides
and other agrochemical,
and thus protect theenvironment ahd human health.
Impacton global production.
Varietal release stemming out of the exchanged germplasm made significant
contributions
to production
increases in several rice growing countries.
In Vietnam,
China, Indonesia
over 60% of the total rice area is covered by INGER distributedlines. For example, more than 10 million ha of the rice area in China are planted tomaterials
taken directlyfrom INGER nurseries or derived from crosses made with
INGER entries
In the past, two deficientcountries
namely Vietnam and Myanmar
became the exporters of rice, the third and fourth respectively.
Evenson and Gollin(1993,
1996) analyzed that the economic value of each released variety is 2.5 million
US$. The economic value of modern varieties in the indica rice region was estimatedto be 3.5 billion
US$ in 1990.
Sustainability
The widened

genetic

base of farmers'

varieties

reducesvulnerability

to theattacksof pest and diseases
and its wide scale impact.
This improves
thesustainabilityof the production
technology(Chaudhary,
1995; Nguyen et al., 1994).
The fact the origin of 1709 modern varieties mentioned by Evenson and Gollin (1993)
can be traced to 11,592 cultivars
used in developing
those. Varieties
with superior

Table 4. Best varieties

for resistanceO r ig inagainstdiseases

screenedin INGER nurseries

during

1975-1995.

E n tr ie s

B la s t:
B r a z il

T r e s M a r ia s

B u r k in a F a s o

IR A T 1 4 4

C h in a

M G ‑3C o l o m b ia

C IA T I C A 5

C o t e d ' lv o ir e

IR A T 1 3

IR R II R 1 9 0 5 ‑ 8 1 ‑ 3 ‑ 1 , I R 1 4 1 6 ‑ 1 2 8 ‑ 5 ‑ 8 , IR 2 7 9 3 ‑ 8 0 ‑ 1 ,I R 1 4 1 6 ‑ 1 ‑ 4 2 ‑ 2 ‑3 ‑ 3 ,IR 1 9 0 5 ‑P P 1 1 ‑2 9 ‑4 ‑6 1 , IR 4 5 4 7 ‑6 ‑3 ‑2 , IR 5 5 3 3 ‑P P 8 5 0 ‑ 1 ,I R 3 2 4 2 9 ‑ 4 7 ‑ 3 ‑ 2 ‑ 2 , W H D ‑ I S ‑ 7 5 ‑ 1 , I R 5 9 6 0 6 ‑ 1 1 9 ‑3

K o re a

IR I 3 8 7

P h il ip p i n e s

C a rreo n , T a d u k a n

T a iw a n ( C h in a )T a ‑ p o o ‑ c h o ‑ z , H u a n ‑ s e n ‑g o o

V i e tn a m

T e te p

B a c t e r i a l b li g h t :B a n g la d e s h

A C 1 9 ‑ 1 ‑ 1 , B R 1 7 1 ‑2 B ‑ 8 , B R 3 1 9 ‑ 1 ‑ H R 2 8 , D V 8 5 , K a l im e k r i 7 7 ‑ 5(A C C 6 6 1 3 ) , B R 2 5 6 4 ‑ 2 B ‑ 6 ‑ 1

In d ia

B J 1 , K A U 1 7 2 7 , R P 6 3 3 ‑5 1 9 ‑ 1 ‑3 ‑8 ‑1 , R P 6 3 3 ‑7 6 ‑ 1 ,R P 2 1 5 1 ‑ 1 9 2 ‑1 , R P 2 1 5 1 ‑ 1 9 2 ‑2 ‑5 , R P 2 1 5 1 ‑2 2 4 ‑4 , R P 2 1 5 1 ‑3 3 ‑2 ,R P 2 1 5 1 ‑4 0 ‑ 1

I n d o n e s ia

C is a d a n e

IR R IIR 2 0 , IR 4 4 4 2 ‑4 6 ‑3 ‑3 ‑3 , IR 1 3 4 2 3 ‑1 7 ‑1 ‑2 ‑1 , IR 5 4,

IR 1 7 4 9 4 ‑3 2 ‑1 ‑ 1 ‑3 ‑2 , IR 4 0 , IR 2 2 0 8 2 ‑4 1 ‑2 , IR 2 5 5 8 7 ‑ 1 3 3 ‑2 ‑2 ‑2 ,I R 3 2 8 2 2 ‑ 9 4 ‑3 ‑ 3 ‑ 2 ‑ 2 , I R 3 5 4 5 4 ‑ 1 8 ‑ 1 ‑ 2 ‑ 2 , I R B B 7 , I R B B 5 , I R 4 8 ,IR 5 4 , IR 3 2 7 2 0 ‑ 1 3 8 ‑2 ‑ 1 ‑1 ‑2

T u n g ro :B a n g la d e s h

D W A 8 , H a b ig a n j D W 8

In d iaA m bem o har 15 9,A R C 7 14 0, A R C 10 34 2 ,A R C 10 4 95 ,A R C
1 1 3 5 3 , A R C 1 1 5 5 4 , A C 4 2 3 6 , K a t a r ib h o g , P T B 1 8 , P a n k h a r i 2 0 3

I n d o n e s ia

U t r i M e r a h ( A C C 1 6 6 8 2 ) , U tr i R a j a p a n ( A C C 1 6 6 8 4 )T h a il a n d

B K N B R 10 3 1 ‑4 1 ‑2 ‑6 , B K N B R 1 0 3 1 ‑7 ‑5 ‑4 , G a m P a i 3 0 ‑1 2 ‑1 5degree of resistanceto pests and diseases

avoid the use of harmful

pesticides,

and thus

insuring the sustainabilityof human health and environment.
Impact on less-developed
countries.
Less developed
countries
have been the
maximum beneficiaryof INGER.
Countries
such as Cambodia, Myanmar andVietnam where research infrastructure
were lacking or even non-existent took the best
advantageof the breedinglines

developed

in otherinstitutions(Chaudhary,

1990;

Table 5.Best varieties

for resistanceagainstO r ig inW

insect-pests

screenedin INGER nurseries

during

1975-1995.

E n t r ie sh it e b a c k e d P la n t h o p p e r :In d ia

W C 1 2 4 0 (A C C 1 3 7 4 2 )IR R IIR 1 3 4 7 5 ‑7 ‑ 3 ‑ 2 , IR 2 0 3 5 ‑ 1 1 7 ‑ 3 , I R 1 3 4 5 8 ‑ 1 1 7 ‑2 ‑ 3 ‑ 2 ‑3 , IR 1 5 5 2 7 ‑ 2 1 ‑2 ‑3 , IR 2 7 3 1 6 ‑ 6 ‑ 2 ‑2 ,IR 1 5 5 2 9 ‑ 2 5 3 ‑ 3 ‑ 2 ‑2 ‑ 2 , I R 1 5 7 9 5 ‑ 1 5 1 ‑ 2 ‑ 3 ‑2 ‑ 2 , I R 1 5 7 9 7 ‑7 4 ‑ 1 ‑ 3 ‑ 2 , IR 1 7 3 0 7 ‑ 1 1 ‑ 2 ‑ 3 ‑ 2

S ri L an k a

R ath u H ee n ati

B r o w n P la n th o p p e r :In d i a

C R 9 4 ‑1 3 , M u d g o (A C C 6 6 6 3 ) , P T B 3 3 , P T B

19 (A C C 5 3 4 3 1 ), P T B

1 9 (A t h ik r a y a ,A C C 6 1 0 7 ) , P T B 2 1 (T e k k o n , A C C 6 1 1 3 ), R P 1 5 7 9 ‑ 1 8 6 4 ‑7 0 ‑3 3 ‑ 5 4 , R P 1 7 5 6 ‑ 1 2 1 ,R P 1 5 7 9 ‑5 2 , R P 1 7 5 6 ‑3 9
IR R IIR 1 3 5 4 3 ‑6 6 , IR 1 3 5 3 8 ‑4 8 ‑ 2 ‑3 ‑2 , IR 1 5 3 2 3 ‑2 6 ‑3 ‑ 2 , I R 1 7 4 9 2 ‑ 1 8 ‑ 6 ‑ 1 ‑ 1 ‑3 ‑3 ,IR 1 7 4 9 4 ‑3 2 ‑ 1 ‑ 1 ‑3 ‑2 , I R 1 9 6 5 7 ‑ 8 4 ‑ 3 ‑ 2 ‑2 , IR 1 9 6 6 1 ‑2 3 ‑3 ‑ 2 ‑ 2 , IR 2 1 9 1 2 ‑ 1 3 1 ‑3 ‑ 3 ,IR 2 5 6 0 3 ‑2 0 ‑2 ‑ 1 ‑3 ‑2 , I R 5 2 , I R 5 6 , IR 9 7 8 2 ‑ 1 1 1 ‑ 2 ‑ 1 ‑2 , IR 9 8 5 2 ‑ 2 2 ‑3 , IR 2 5 5 8 8 ‑ 3 2 ‑ 2 ,IR 1 9 6 6 0 ‑4 6 ‑ 1 ‑ 3 ‑ 2 ‑2 , IR 1 9 6 7 0 ‑ 5 7 ‑ 1 ‑ 1 ‑3 , IR 3 2 8 2 2 ‑2 ‑2 ‑ 3 ‑ 2 , IR 3 5 3 2 3 ‑ 9 3 ‑ 1 ‑ 3 ‑ 1 ,IR 1 7 4 9 4 ‑3 2 ‑2 ‑ 2 ‑ 1 ‑3 , IR 2 7 3 1 6 ‑ 7 8 ‑ 3 ‑ 3 , IR 3 2 8 2 9 ‑5 ‑2 ‑2 , IR 2 9 7 2 5 ‑3 ‑ 1 ‑ 3 ‑ 2 , I R 3 6 ,IR 1 3 4 1 5 ‑9 ‑3 , I R 1 5 8 4 7 ‑ 1 3 5 ‑ 1 ‑ 1 , IR 1 5 8 6 9 ‑ 1 1 3 ‑ 1 , IR 9 7 5 2 ‑2 2 2 ‑3 ‑2 ‑ 2 , I R 1 1 4 1 8 ‑ 1 5 ‑2 ,IR 9 7 6 1 ‑4 0 ‑3 ‑ 2 , IR 9 8 4 6 ‑2 3 ‑2 , I R 9 8 5 2 ‑9 3 ‑ 2 ‑ 2 ‑2 ‑3 , IR 3 2 8 3 0 ‑2 3 ‑2 ‑2 , I R 2 9 6 9 2 ‑9 9 ‑ 3 ‑2 ‑ 1
IR 1 3 2 4 0 ‑8 2 ‑2 ‑ 3 ‑ 2 ‑3 ‑ 1 , I R 1 3 4 2 7 ‑ 4 0 ‑ 2 ‑3 ‑ 3 , I R 1 3 4 2 7 ‑4 0 ‑ 2 ‑ 3 ‑ 3 ‑3 ‑3 , I R 1 3 5 4 0 ‑5 6 ‑3 ‑2 ‑ 1 ,IR 2 5 5 8 7 ‑ 1 3 3 ‑3 ‑ 2 ‑ 2 ‑2 , IR 3 5 3 5 3 ‑9 4 ‑ 2 ‑ 1 ‑3 , IR

S ri L an k a

B a b a w e e (A C C 8 9 7 8 ) , B G

3 6 7 ‑2 , B G

7 4 , IR 1 1 4 1 8 ‑ 1 9 ‑ 2 ‑3 , IR 2 8 1 5 0 ‑ 8 4 ‑ 3 ‑3 ‑2

3 7 9 ‑2 , B G

3 6 7 ‑ 4 , B a l a m a w e e (A C C 8 9 1 9 ), B G

3 7 9 ‑1 ,G a n g a la ( A C C 7 7 3 3 ) , H o n d a ra w a la , K u ru h o n d o ra w a la (A C C 7 7 3 1 ), H o n d a r a w a la .
K u r u h o n d o r a w a la ( A C C 7 7 3 1 ) , R a t h u H e e n a t i (A C C 1 1 7 3 0 ) , S in n a S iv a p p u (A C C 1 5 4 4 4 ) ,S u d u r u S a m b a (A C C 3 6 8 5 1 ) , S u d u H o n d o ra w a la , (A C C 1 5 5 4 1 ), M a w e e ( A C C 3 1 4 8 2 )G a ll M id g e :In d ia

C R 9 5 ‑J R 7 2 1 ‑ 3 , C R

1 99 ‑1, R P W

A R C 1 0 6 6 0 (A C C 2 1 0 2 6 ), A R C

S te m6 ‑ 1 7 , P T B 2 1 (A C C 6 1 1 3 ) , W

IR R IIR 1 3 4 2 9 ‑ 1 5 0 ‑3 ‑ 2 ‑ 1 ‑2 , IR 4 2 3 4 2 ‑4 0 ‑ 3 ‑ 3 ‑2 ‑3

T h a ila n d

R D

S ri L a n k a

B G 4 0 2 ‑2 , O B 6 7 7 , 7 5 ‑ 1 5 9

9

B o r e r (D e a d H e a r t) :IR R I

IR 8 6 0 8 ‑7 5 ‑ 3 ‑ 1 ‑3 , IR 9 8 2 8 ‑2 3 ‑ 1 , I R 1 5 7 2 3 ‑ 4 5 ‑ 3 ‑ 2

In d ia

R P 6 ‑ 1 8 9 9 ‑ 2 5 ‑ 4 , R P 8 8 7 ‑4 6 ‑ 1 , R 3 4 ‑ 7 3 ‑2 0 0 , W

1 2 6 3 (A C C 1 1 0 5 7 ) , T K M 6 (A C C 2 3 7 ) ,R P 1 0 5 7 ‑3 9 4 ‑ 1 , R P 2 1 6 7 ‑ 3 5 3 ‑ 3 ‑2 ‑ 1
IR R IS te m

1 2 6 3 (A C C 1 1 05 7 ),5 9 8 4 ( A C C 2 0 2 9 7 ) , M R 1 5 2 3 (A C C 4 6 4 0 1 )B o r e r (WIR 5 9 6 0 6 ‑ 1 1 4 ‑3 S
h ite H e a d ) :In d iaW

1 2 6 3 ( A C C 1 1 0 5 7 ), T K M 6 (A C C 2 3 7 ) , R P 1 0 1 7 ‑ 7 6 ‑ 1 ‑3 ‑2In d o n e s ia

G H 1 4 7 ( M ) 4 0 K ra d 8 9 , B 5 2 7 8 ‑ 1 3 0 ‑ M R ‑ 5 ‑ 4

Table5. (Continued)

S t e m B o r e r (W h it e H e a d ) :IR R I

IR 8 6 0 8 ‑7 5 ‑3 ‑ 1 ‑3 , IR 1 5 3 1 4 ‑4 3 ‑2 ‑ 3 ‑3 , IR 9 8 2 8 ‑2 3 ‑ 1 , I R 5 6 , I R 1 7 4 9 4 ‑3 2 ‑ 1 ‑ 1 ‑3 ‑ 2 ,I R 9 2 8 8 ‑B ‑ B ‑5 2 ‑ 1 , I R 1 5 7 9 5 ‑ 1 5 1 ‑ 2 ‑3 ‑ 2 ‑ 2 , IR 1 3 4 7 5 ‑ 7 ‑ 3 ‑2 , IR 2 5 5 8 7 ‑ 1 3 3 ‑ 3 ‑ 2 ‑2 ‑2 ,I R 3 9 3 8 5 ‑ 1 2 4 ‑3 ‑ 3 ‑ 2 ‑3 , IR 1 8 2 0 ‑5 2 ‑ 2 , IR 1 9 7 4 3 ‑ 4 6 ‑ 2 ‑3 ‑ 3 ‑ 2 , IR 4 6 1 9 ‑5 7 ‑ 1 ‑ 1 ‑ 2 ‑ 1

T a iw a n

Table 6. Overall

T a ic h u n g s e n 1 0

best tolerant

entriesO r ig in

to abiotic

stressscreenedin INGER nurseries(1975-1995).

E n tr ie s

F o r c o ld to le r a n c e : :C h in a

C h in g H si 1 5 (A C C 3 6 8 5 2 ), Y u n le n 1 7 , Y u n le n 1 8 , C h u ra i, G e n d ra o 3
Y u n le n 1 2

In d ia

JK A U (K l‑4 5 0 ‑1 2 6 ‑2 , K 3 1 ‑ 1 6 3 ‑3 ( K h u d w a n i), K 3 9 ‑9 6 ‑ 1 ‑1 ‑ 1 ‑2 , K
7 8 ‑1 3 (B a rk at)

IR R IIR 1 9 7 4 6 ‑2 6 ‑2 ‑3 ‑3 , IR 2 4 3 1 2 ‑R ‑R ‑1 9 ‑3 ‑B

J a p an

F uj i 1 0 2 , E ik o (A C C 9 4 1 7 ) , T a tsu m i M o c h i

K o re a

D e o g ‑Je o g ‑J o d o

R u s sia

S te ja ree 4 5

T a iw a n (C h in a)

C h in a 1 0 3 9

F or D rou g h t:
B ra z il

IA C 4 7 , C a rijo , A g u lh a (A C C 3 8 9 7 6 )In d o n e s ia

B 2 9 9 7 C ‑T B ‑4 ‑2 ‑ 1

IR R IIR 4 7 6 8 6 ‑6 ‑2 ‑ 1 ‑1 , IR 4 7 6 8 6 ‑9 ‑4 , IR 5 5 4 1 1 ‑5 3 , IR 3 0 7 1 6 ‑B ‑1 ‑B ‑ 1 ‑2 .

S ri L a n k a

B W 3 1 1 ‑9

IR 2 7 0 6 9 ‑ B 5 3 ‑B ‑B ‑1 ‑4 ‑4F o r A lk a lin ity :In d iaG e tu , C S R 1 , P o k k a li

In d o n e sia

S 8 1 8 B ‑1 0 ‑2

IR R IIR 2 0 5 3 ‑4 3 6 ‑ 1‑2 , IR 4 2 2 7 ‑1 0 9 ‑ 1 ‑3 ‑3 , IR 4 2 2 7 ‑2 8 ‑3 ‑2 , IR 2 9 7 2 3 ‑ 1 4 3 ‑3 ‑2 ‑ 1 ,IR 5 5 1 7 8 ‑B ‑B ‑B ‑9 ‑3 , IR 6 3 7 3 1 ‑ 1 ‑1 ‑ 1 ‑1 ‑4

F o r S a lin ity :In d iaG e tu , N o n a B o k ra , B h u ra ra ta 4 ‑ 1 0 , P o k k a li

IR R IIR 4 6 3 0 ‑2 2 ‑2 ‑1 7 , IR 4 6 3 0 ‑2 2 ‑2 ‑5 ‑l ‑3

S ri L an k a

A 6 9‑1

Chaudhary
1991; Chaudharyand Fujisaka,
1992; Nguyen et al., 1994). Cambodia's
research infrastructure
was completely
ruined in the ongoing civil war since 1960's,
and no scientificcapability

existed(Chaudharyand Mishra,

1993;

Chaudhary

et al.,

1995).
Almost all cultivatedrice varieties
have been unimproved.
IR 8 the "green
revolution"
rice variety did not reach Cambodia.
But during 1988 to 1993 a total of
12 varieties
were released out of which 10 came directlyfrom INGER nurseries(Chaudhary,
1995;
Chaudharyand Mishra,1993).
This would have been an
impossible
task without INGER. The newly released varieties
have spread to over
100,000 ha in a short time and the country is moving towards export from the state of
deficit.
Human resource development.
Germplasm evaluation
networks address human resource development formaking the germplasm evaluation
and utilization
more effective.
This is done byformal training,
post docs, and organized monitoring
visits etc. INGER supported
more than 80 research scholars, post docs, and visiting
scientists,and 356 trainees.
The joint monitoring
visits,and field workshops have been avenues of informal
human resource development.
In 31 such activities
more than 750 scientists
participated,a number of land mark recommendations
were made which influenced
not only INGER activities
but also the national
programs and their research
prioritization.
Innovations
Possible
The collaborative
mechanisms of germplasm evaluation
globallyprovidesexcellent
opportunity
to introduce
a number of innovations
for operations
and datainterpretation:
Multi-media
based operations.
The data base and the operations
at the coordinating
center IRRI, are already fully supported
by computerized
system. Now it is feasible
to use electronic field book and data management system, as the INTERNET expands.
This will add speed and save on manual time. The test environment
at individual
locationcan be recorded and stored using multi-media,
for immediate
and futureinterpretationof the results.
G x E interaction
analysis.
Fixed genotypes once tested in variable
environments
provide multilocationtest data set which could best be analyzed by new tools in G
x E interaction
analysis.
The results could be used to stratify
the test locations,stratify
the test genotypes,
identify
stablegenotypes,
deploymentof genotypes,
extrapolation
by modelingetc. (Chaudhary
1994, Chaudharyand Movillon,
1995;
Gauch, 1992).

Biologicalmapping.
Through the use of "probe" genotypes, that are selected for their ability
todiscriminate
the environment, it is possible
to match gene diversity
to the needs offarmers in the heterogeneousand variable rice ecosystems. The differential
responseof the probe genotypes
is used to "biologically"
characterize
the diverse
environments. Thus the plant, not the geographer,is the sensor of the environment.
With multi-variate
analysisand GIS, the map for rice adaptation
can be drawn as
sensed by the rice plant. The development of this research will facilitate
the selectionof parental types for breeding.
Characterizationof key sites.
The sites selected
for representationof "domains of adaptation"requirecarefulcharacterizationof their physicaland biologicalcomponents, since thisinformationis needed to calibrate
the models now available
for further assessmentand extrapolationof genotype performance. This should help reducing the test sites
and saving on material, time and money.
Germplasm deployment.
Using the test data, modelingand extrapolation
techniques,it should bepossibleto deploy genotypes
even in the untested locations.
From the study of thepathogenic
variability
and stabilityof the resistance,
it should also be possible
todeploy and rotate the resistancegenotypes across the growing environments.
Why INGER succeeded?
INGER, over the last 20 years of operation has shared over 40,000 varieties
and breeding lines. This has resulted in the release of 577 varieties
in 62 countries.
Thousands of lines have been used in the crossing programs to further diversify
thegenetic base of farmers' varieties
and sustain yield.
Fair enough then that INGER has
been called the flag ship of IRRI and its most successful program. INGER mechanismof germplasm exchange and evaluation
and utilization
is a successfulmodel and has
several in-built
points of success. INGER has unprecedented
worldwide scope inwhich 1000 scientists
participate
and feel that they are the owners. It is jointly
owned
and operated by NARS and IRRI, and cost is also shared almost equally (Table 7).
The technicalprogram is suggested
and modified
continuously
by NARS and the
INGER Steering
Committee, and not by IRRI alone, making it truly democratic
anddynamic.

The freedom

to for all concerned

NARS and IARCs to joinor out makesTable 7. Annual expenses (US$)

incurredC o u n try

by some NARS in participating

to evaluate

INGER nurseries.

C o st o f e v a lu atin g v a rio u s IN G E R n u rse rie s
Y ie ld

O b se rv a tio n a l

A rg e n tin a

R e s is tan c e

N o . o f s ite s

T o ta l C o st5

1 2 ,5 0 0

B a n g la d e sh

4 10

2 ,5 2 0
165

80

18

1 2 ,8 0 2

B h u tan

33 1

140

41

4

1 ,2 0 0

B ra z il

19 0

115

15 0

15

2 ,2 5 0

50

50

12

60 0

C h in a

1 ,6 0 0

6 30

64 0

36

5 2 ,0 0

C o sta R ic a

38 2

3 00

2 10

3

90 0

6 19

3

1 ,8 5 7

C a m b o d ia

G u yan a
In d o n e sia

97 5

5 50

375

48

3 8 ,4 0 0 *In d ia

500

3 00

200

12 1

4 8 ,4 0 0 *Ita ly350

5

1 ,7 5 0

Ja p a n

4 13

13

7 ,3 6 9 * *1 ,6 9 0

2 ,0 0 0

2 1

3 7 ,8 0 0

23 9

165

3

78 1

K o re aM a d a g as c a r

377

M a la w i

60 4

1

60 4

14

2 2 ,9 6 0

4 25

26

2 1 ,7 5 8

M a la y s ia

1 ,6 4 0

M yanm ar

1 ,3 0 0

77 5

4 00

25

1 0 ,0 0 0

P a k istan

4 00

19 0

100

18

1 1 ,2 0 0 * *P h ilip p in e s

1 ,0 0 0

90 0

7 00

5 5 (6 )5 ,4 0 0

S o u th A fric a

1 ,2 0 02

1 ,2 0 0 *S en e g a l

661

38 0

2 00

8

3 ,3 1 2 * *S ie rra L e o n e

2 26

81

16

4

549

S ri L a n k a

2 80

18 5

145

19

3 ,8 5 7

T a iw a n

3 ,0 0 0

1 ,7 4 3

1 ,8 0 0

10

2 1 ,8 1 0

T u rk e y

55 0

00

100

4

1 ,4 0 0

U gan da

53 9

17 7

40

3

7 56

V ie tn a m

56 8

266

1 15

52

1 6 ,5 3 2

N e p al2

T o ta l* Senior staff

3 3 1, 1 4 7salaries** No staff salaries

not included.
included.

The cost does not include

the cost of breeding

lines

sharedthem more attached to the success of the network. It provides
a feedback on theperformance of the test entry to the nominating
plant breeders, and at the same timeleaves any scientists
to free use the entry as released
variety or parent. Weakest

NARS derivestrengthmechanismto evaluate

FutureProspects

from the strongest
germplasm

feeding

It is highly

the world.

It has its basedeep

imbeddedof the rice genome to match the eco-geographic

and edaphic

globe.

Under

years.

evolutionit has evolved

where reproductive

much diverse

directions,ratherbeen

it hasbarriers

thoughsteppedover 10,000

no longer

less thanup.

exist,

100 years

The createdold.

genetic

varieties.

VarietyRight

are coming up and are being

investmentsof elitein agricultural
germplasm

developingof IRRI?
withresearch.
is notIs future

How can INGER'sor "Perennial

the same enthusiasm
Common Wealthand inhas not stopped

Rightspatentingfasterhas to be sharedadvocated(Barton,the poor farmer
efforts

Bush Rices"would like the mechanism
question

to imagine

Evolution

Proprietary

What effectsof the(IPR)

and Plant

to promoteprivatewould bring1993).

The

and the pooresttoon the flow
wholeof theof the poor - the poorof "poor rice grower and consumer" secure in the leadershipsustainof IRRI

and openness,

NARS and IARCs is great.
Asia,

hard

world would suffer,rice consumer.
Rices"

issuesIntellectual

diversity

evolvediversity

the base of farmers

in genetic

Under man-guided

it would

diversify(PVR)

effective

Collaboration

diversity

domestication

cost

internationally.

of International

Rice is a cereal

institutions.

rice

productivity?

are readyfor sharing,will

as we shared

IR 8?

The enthusiasm

INGER would like to expandof Independent

Sates

countries

to continue

beyondthe life

marks on the availabilityof funds,

When the "Superof theits activities

in West-Central

and Eastern

Europe.

of IARCs.

and scenario

we be sharing

createdBut there

NARS

are serious

by IPR and PVR.

References
Alluri, K., Chaudhary, R. C and Akinsola, E. A. 1995. Genetic Evaluation and Utilization:
Gateway
to rice improvement in Africa. American Society of Agronomy, St. Louis, Missouri, U. S. A.,
Abs. No. C-1009, p. 44.
Barton, J. H. 1993. How will intellectual
property protection and plant patents influence germplasm
collection,enhancement, exchange and use? International
Crop Science, CSSA, Madison, U.
S.A. pp. 855-858.
Baum,W.C. 1986. Partners against hunger: Consultative Group on International
Agricultural
Research.
The World Bank, Washington, D. C. 337 p.
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P55.

Chaudhary,R. C. 1991. The Cambodian
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No. 5 p 4. February 1991.
Chaudhary,R. C. 1994. Global evaluationof rice genotypes
through
INGER: Expectations
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Netherlands- IRRI Philippines;
pp. 23-30
Chaudhary,R. C. 1995. Linking
the centers of diversity,
evolutionand utilizationof rice to sustain
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throughthe International
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D. W. 1995. Improvementof deepwater
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Appendix

I. Types of INGER nurseries

and entries

evaluated

globallyE c o lo g y /N u r s e rv

during1975

to 1995.

1 9 7 5 ‑1 9 8 4

1 9 8 5 ‑9 5

T o ta l

3 ,3 1 7

2 ,1 8 7

5 ,5 0 4755

810

1 ,5 6 5

749

0

749

I r r ig a te d
IR O N / IIR O N( O b s e rv a t io n a l)

II R Y N ‑ V E /E /M
IR A R O N(Y ie ld , v . e a r ly /e a r ly /m e d iu m )( A r id re g io n o b s e rv a tio n a l)

IR B O N(B o ro r ic e o b s e r v a t io n a l)

0

153

153

IR H O N( H y b r id ric e o b s e r v a t io n a l )

0

159

159

IR F A O N( F i n e g r a i n a r o m a tic o b s e rv a tio n a l)
1 ,6 7 4

1 ,3 5 4

3 ,0 2 8

243

25 1

494

1 ,3 3 0

1 ,2 2 797388

2 ,5 5 7
485

R a in fe d U p l a n d
IU R O N( O b s e r v a t io n a l)

IU R Y N / ‑ E /‑ M

( Y ie ld , e a rly / m e d i u m )R a in fe d L o w la n d
IR L O N( O b s e r v a t io n a l)

IR L Y N‑E / ‑ M(Y ie ld , e a r ly /m e d iu m )D ee p w a te r &F lo o d p r o n e

ID R O N( O b s e r v a t io n a l)

565

830

1 ,3 9 5

IF R O N(F l o a t in g r ic e )

16 7

78

245

IT R O N(T id a l w e tla n d s )

226

472

698

ID R Y N( Y ie ld )

0

57

57

3 ,7 7 4

2 ,4 3 4

6 ,2 0 8542

1 ,0 6 8

1 ,6 1 0

1 ,8 6 9

2 ,2 2 5

4 ,0 9 4

460

0

460

1 ,3 0 8

1 ,0 6 5

2 ,3 7 3

B io tic r e s is ta n c e
IR B N( B la s t)

IR B B N
IR T N( B a c t e r ia l b l ig h t)
(T u n g ro v ir u s )IR S H B N( S h e a t h b lig h t )IR B P H N( B r o w n p la n t h o p p e r )IR W B P H N

19 1

498

689

IR S B N( S te m b o re r)(W

h ite b a c k e d p la n th o p p e r)

349

369

718

IR G M N( G a ll m id g e )

539

172

711

IR U N( U f r a n e m a to d e )

0

165

165

78 1

690

1 ,4 7 1

0

14

14

2 ,1 5 1934

3 ,0 8 5

0

272

272

A b io t ic S t r e s s
IR S T O N / IR S S T N( P r o b le m s o ils )IR S T Y N( S a li n ity )IR C T N(C o ld to le r a n c e )IR D T N( D r o u g h t t o le r a n c e )S p e c ia l s c r e e n in g s e t s
IR G O N

0

82

82

O th ers(G r a in q u a lity )1 ,1 5 4586

1 ,7 4 0

T o tal2 2 ,2 4 1

1 8 ,5 4 0

4 0 ,7 8 1

IIRON:Interntnl

Irrigated

IRHON:Interntnl

Rice Hybrid

Rice Observational

IRTON:

Interntl.

Rice

IRLON:

Interntl.

Rnfd. Lowland

IDRON:

Interntl.

Deepwater

IRBN:

International

Rice Blast

IRBBN:

International

Rice

IRCTN:

International

Rice Cold

IRSBN:

International

IRWBPHN:

Interntl.

Nursery;

IIRYN-E,M:

Observational

Nursery;

IRBON:

International

Rice

Observtnl.

Nursery;

IRFAON:Interntl.

Rice Finegrain

Temperate

Rice Observtnl.

Observtnl.

Nursery;

Nursery;

Nursery;

Bacterial

IRLYN:

International

Internatnl

IURON:International
IRDTN:Interntl.

BlightNursery;

IRBPHN.

Tolerance

Nursery,

Rice

Stem Borer Nursery;

Rice

Whitebacked

Planthopper

Rice

Irrigated

Rice

Rnfd.

Aromatic

LowlandUplandRice Observational

Drought

Tolerance

Obsv. Nsery;
Nursery;
Nursery;

Nursery;

Rice

Brown Planthopper

IRTN:

International

Rice

TungroIRGMN:

International

Rice

Gall

IRSSTN:International

Nursery;
Nursery;

Rice Yield

Interntl.

Nursery;

Yield

Boro Observational

Nursery;

Nursery;
Midge
Rice

Nursery;
SoilStressNurseryAppendix

II. Number of regularly

activeINGER test locations

N u m ber o f

N um b er o f
R e g io n /c o u n try

lo c atio n s

and cooperators.

c o o p e ra to rs

R e g io n /c o u n try

lo c a tio n s

c o o p e ra to rs

W e st A s ia & N . A fr ic a (c o n t.)

E a s t A sia
C h in a

1273

M o ro c c o

1

1

Ja p a n22

S a u d i A ra b ia

1

1

K o re a831

S u dan1

1

T a iw a n

4

4

T u rk e y

1

2

S u b ‑S a h a ra A f ric a

S o u th e a st A s ia
C a m b o d ia

78

Iv o ry C o a s t

2

3

In d o n e s ia

16

23

M o z a m b iq u e3

2

L ao s1

1

N ig e ria

23

M a la y sia

45

S eneg al

2

4

M yan m ar

13

32

S o u th A fric a

1

2

P h ilip p in e s

7

10

T an z a n ia

2

4

T h a ila n d

2180

Z a ire1

1

V ie tn a m

18

36

Z a m b ia

1

1

L a tin A m e ric a

S o u th A sia
B a n g la d e sh

1237

A rg e n tin a2

2

B h u ta n

3

1

B ra z il

3

6

In d ia65

13 6

C o lo m b ia

35

N ep al

4

12

C o sta R ic a

1

1

P a k ista n

716

G uyan a1

1

S ri L a n k a414

N ic a ra g u a

1

1

W e s t A s ia & N . A fric a

E u ro p e

A fg h a n ista n

1

2

E g yp t

3

8

Ira n

5

17

Ira q

1

1

Ita ly1

1

3

1

O c e a n ia
P . N . G u in e aQuestions

and answers in Session

Questions to Dr. Seko
Q. To make the MAFF genebank

3

activity

more international,

standardizationofevaluation
methods would be useful, for example amylose content of cereals.
Standardization
also makes routine work more useful across centers. Do you haveany comment? (Hayashi)
A. To help internationalise
the MAFF genebank by the end of 1996 passport data will
be available on the internet. However, it may still be necessary to have catalogues
for those people that don't have computers and Internet connections.
(Seko)
C. For efficientcollaboration,exchange of informationand opinions
are very
important. Advanced information
systems will help this greatly.
All speakers insession 3 mentioned the availabilityof information
related to their activities
on
the Internet.
May I propose that all speakers/participants
furnish their e-mailaddresses and URL of home-page showing his or her activities.
I would like thetopic of information
management, also related activities
like DIP and SINGER
which Dr. Riley referred to yesterday
to be discussed elsewhere in this workshop.
(Suzuki)Q. Could you comment on pre-breedingas an activityof MAFF? How successful
have you found it? (Riley)
A. Pre-breedingis part of the MAFF genebank project and a certain amount ofbudget is allocatedto this. Laboratories
involved in pre-breeding
are for the mostpart also responsible
for breeding. Numerous good results have been obtained andmaterials generated have been used as parents. Thus pre-breeding
is a good way
to generate parental material. (Seko)
Questions to Dr.Gupta
Q. I would like to know the present situation
regardingin-vitro
India?(Kikuchi)
A. In vitro conservation
is being carried out at the "national
Facility
Repository"
at NBPGR. About 60 species are being multiplied

conservation

inof Plant Tissue
and conserved

through tissue culture.
The recalcitrant
species are being conserved by cryopreservation.(Gupta)
Q. To conserve original
seeds from farmers fields is quite difficult.
We need apractical
way of storage without eliminationof genetic diversity.
Any comments?

(Nakagahra)
A. In India we have no difficultyin collecting
farmers seed and conserving ex-situ in
our genebank. There is not much danger of eliminationof genetic diversity.
However, we are considering"on-farm conservation"
as a future strategy. A law
on "farmers rights" is in its final stage (Gupta).
Questions to Dr. Chaudhary
Q. Does INGER require pedigree data on material in its nurseries- i.e. can ultimate
landraces be determined?
(Vaughan)A. INGER requires and keeps in its database,the data on pedigree designation,
parentage and origin of each test entry. The ultimate parents and the pedigreearenot recorded but can be searched. (Chaudhary)Q. Should collaboration,
plant genetic resources and exchange networks, focus onelite material
or on genebank accessions?(Riley)
A. So far, INGER evaluated entries consist 98% elite breeding lines and 2% genebank
material. Plant breeders from NARS and IARC's would also be interested
in thegenebank materials
if found to have useful characteristics
as donor parents.
(Chaudhary)

Workshop SummaryGroup Discussion Summary

Chairpersons
T.Oishi
K.KatoWorkshop Summary
KEN RILEY
Regional

Director,

IPGRI-APO,

P.O.Box 236, UPM post Office 43400

Serdang,

Selangor

Darul Ehsan,

Malaysia

together

The 4th MAFF International
Workshop on Genetic Resources
89 participantsfrom 20 countries
to address"Characterization

brought
andEvaluationof Plant Genetic Resources for Improved Use of Plant Genetic Resources".
One of the features of the workshop is that, by bringing
scientists
togetherfrom different parts of the world, new linkages can be forged between scientist having
similar interests.
For example, during the workshop we heard from Dr. Sano on newevidence regarding species of the rice genus (Oryza) in Latin America. We also heard
briefly from Dr. Kresovich on collaboration
between his laboratory
and CENARGEN
in Brazil on different aspect of the same species. The bringing together of like mindedscientists,
in not such big groups, within an intimate atmosphere like this, can indeed
have beneficial
and synergist
results. Collaboration
was the theme of Dr. Okuno'spresentationand he gave many examples of successful two way collaborationin thefield and in the laboratory
involving
his active team in NIAR.
However, this workshop while bringing together
like minded scientists
did
not bring together scientistof all the same discipline.
Dr. Tosa presented
fascinating
results
on plant-pathogen
interaction
providing
us with the invaluable
precisionof a plant pathologist.
He helped greatly
in providing
perspectives
on co-evolutionary
relationships
between organisms.
Dr. Matsuo gave us an enlightening
paper of the relevance of detailed
ecologicalresearch and how it can lead to valuable understandingof genetic diversityof particular
relevance to in-situ conservation.
While Dr Seko and Dr. Chaudhary gave us breeders perspective
coupled with
their deep knowledge of genetic resources.
The network approaches Dr. Chaudhary explainedin his paper was followed
up during the discussion
when the need for strong within country networks in relation
to genetic resources was very apparent.
Dr. Morishima, in her keynote address, raised the alarm of genetic erosion.
Her long term monitoring experiments have given her unique authority
to warn of theconsequencesof neglecting

conservationin the fieldand she rightly

extendedherconcerns to erosion in the genebank.
Dr. Trinh provided an overview of primary and secondary centers of crop
diversity
in Indochina, while Dr. Li described the high level of wheat genetic diversity
in north-western
China's Xinjiang
Province. This paper was well linked with Dr.
Okunos' presentation
on Aegilops in Central Asia. Again with judicious
use of tools
for genetic diversity
analysis Dr. Strelchenko
identified
2 genepools in Central Asia
and Russia for barley.
Dr. Yunus and Dr. Morishima both compared various methods of genetic
diversity
analysis including isozyme and morphometric techniques.
It is important toconsider the techniques appropriate
to a given objective for analysis, as Dr. Kresovich
also pointed out in his presentation.
Dr. Vaughan illustrated
how in-situ conservation research can help answer
many basic and applied questions, particularly,
in relation to processes of evolution.
The topic of in-situ conservation generated great interest, particularly
theinfluence of human cultivators that affect the structure of cultivateddiversity.
In situconservation may require additional
interdisciplinary
approaches,
includinglinkages
between biologicalsciences and social sciences.
The analysis of rice diversity in Vietnam provided an example of the potentialof understanding
the relationshipbetween ethnic diversity,
local taxonomies andgenetic diversity.
The importanceof finding
economical and efficientmethods to gain
improved understandingof diversity,and how to conserve, it were repeatedlyemphasised.
Collaboration
by building
on strengthsof different
institutions
andcountries may be a very effective way to achieve this goal.
Representing
IPGRI, Dr. Riley in presenting
one of the keynote addresses
reviewed characterizationand evaluation approaches and raised a number of issues.
Of particularinterest
was the rapid developmentof information
tools including
exchange of non-standardized
data through tools such as SINGER
Information Systems on Genetic Resources) and DIP (Data Interchange(System Wide
Protocol).
The

importance of standardizing
information,such that it can be readily exchanged andunderstood, was emphasised by Dr. Hayashi and Dr. Chaudhary. Dr. Riley mentioned
IPGRI descriptor
lists which are designed for this but allow flexibility
in characterstaken.

Both Dr. Seko and Dr. Gupta highlighted

the importanceof strong integration

between genebanks and breeders in the large national programs in Japan and India.
It was agreed that similar linkages are necessary in all countries for effective
use ofplant genetic resources.
The workshop benefitted
from the active participationof the JICA trainees
including Mr. Ali Osman Sari from Turkey, Mr. Gupta from Nepal , Mr. Ekanayake
from Sri Lanka and Mr. Mujaju from Zimbabwe. Questions and comments from otherparticipantsfrom Japan and other countries stimulated
discussion - such that the time
was maximally used. In fact, discussions
continued well beyond the set time for theworkshop.
Three active discussion
groups debated needs and opportunities
related totechniques,
genetic diversity
and networking. The summaries of these discussion
groups is reported below.
Finally,
there was universal agreement that the workshop provided an
opportunity
for participants
to identify
areas of mutual scientificinterest
to beidentified
and developed. This process undoubtedly
will continue well beyond thisworkshop.

Group Discussions
A. Techniques/technologies
Discussion group
Leader : S. Kresovich, Rapporteur:
D. A. Vaughan
Four questions were raised:
1. What do we need technologies
for in PGR work?
2. What do we see as the main constraints to work at present?
3. What do we see in the future as technologicalneeds?
4. Collaboration-Vision for the future?
1. What do we need technologies
for in PGR work?
Identity
Structure
Relatedness
Inheritance,
gene function regulation
Evaluation
Chemical - quick kits for screening
Vector tags/Generation
tags
Differentiation/domestication
2. What do we see as the main constraints
Arranged as a priorities
Time/money
Materials/samples
Knowledge and expertise
Humancooperation and evaluation
Electronic networking
Data Analysis and Handling
Equipment and chemicals(Good) Unique idea

to at present

3. What do we see as future technologicalneeds?
Easier, faster, better and safer
More thoughtfulquestions asked and solved

scientificobjectives?

Easier interpretation
Not destructive
Multidimensional
analysis
Biologicalalienation
Genepool irrelevance
4. Vision - dreamed for collaboration?
Global network building on strengths
Chromosome Image Internet database
In-situ analysis- in field host/plants
Regional/Global
interdisciplinary
study - plant,
anthropology

animal, ethnology,

micro-organism,

B. Diversity
Discussion group
Leader : K. Okuno, Rapporteur: P. Strelchenko
The group had a wide ranging discussion on the topic of genetic diversity.
Many points were raised. Genetic erosion was a major topic of the group and clearlyscientifically

based early warning systems are needed.
Genetic resources are being threatenedby extinction
due to various factors
such as rapid urbanizationand introductionof improved varieties. It was realized that
germplasm of different
crops and their wild relatives must be collected before they
become extinct.
The following
recommendations
were made to safeguard
the genetic
resources from erosion and for their characterization.
* A keen watch must be kept on areas where genetic resources of particularspeciesare endangered. Such areas should be explored and germplasm be collected as quickly
as possible.
* In cases where genetic resources are disappearing
quickly,a proposal may besubmitted to IPGRI for collectionof germplasm. It was noted that in accordance with
Agenda 21 of the meeting held in Brazil, endangered species have already been listed.
Efforts are required to protect these species from erosion.
* Priorities
for germplasm collectionof different
crops should be fixed, because not
all crops need urgent explorationand collection.
Emphasis should be given tocollectionof criticallyimportant or threatened
germplasm.
* Germplasm should be multiplied,
rejuvenatedand characterisedin the areas ofcollection.
Multiplication
and rejuvenation
can also be undertaken in the greenhouse
under controlled
conditions
so that during this process genetic diversity
is not lost.
* Storage conditions in genebanks must be kept optimal, otherwise there is the dangerof lose of a considerable
amount of genetic diversity
in the genebank.
* Attempts should be made to conserve the germplasm in-situ, wherever it is possible.

C. Networks

Discussion

group

Leader : R. C. Chaudhary, Rapporteur:
A.G.Yunus
The group realized
the shrinking
resources and increasing
and thereby increasing
importance in networking.

interests

in PGR

A. Organization
1. International
networks for main crops with all centers including
the sharing,evaluation
and use, and increase of awareness on PGR.
2. A regional network on PGR with sub-networks focussed on specific crops andissues.
3. National networks which link all groups involved with PGR together.
4. Internationaland national PGR database on Internet for exchange of information
on gene bank accessions, PGR technologies
to retrieve information
on evaluation
and use.
B. Funding.
1. Country fund for their
2. For regional
network
countries etc.
3. International
fund foridentified.
4. Company or individual

own PGR network.
the countries
involvedmobilizationof international

who has interests

providethe fundnetworks.

Donors may bein the PGR project.

C. Operational
1. Development of informationexchange system.
2. Problem solving research related to conservation
3. Addressing
policy questions
such as IPR/PVR

and use of PGR.

bring in breedingcompanies to the network and sharing "rights"
mode of operating in PGR work will be different
4. Agreement among members on mechanism of germplasm exchange.
5. Operation with local expertiseand autonomy in fund use.
6. Use of accessions stored in international
research institutes.
7. Standardizationof testingprocedure.

e.g. ASEAN

8. Shorten the year of varietal recommendation and release.
9. Use of evaluation
data on genotype X environment interaction
studies.

and simulation

CLOSING REMARKSClosing

Remarks

HIDEFUMI SEKO
Genetic

Resources

Coordinator,

NIAR, JapanThank

you very much Dr.Riley

for thatvery

excellentsummary of theworkshop.
Wehave come to the end of the workshop and on behalf of the organizing
committee of the workshop, I should deliver a few words.
First of all I would like to thank all the chairpersons,speakers and otherparticipants
for their kind contributions
to this workshop. I would also like to thankthe Agriculture,
Forestry and Fisheries
Research Council and sister institutes
inTsukuba for their support and help.
The MAFF International
Workshop on Genetic Resources aims to promoteexchange of research ideas and collaborationon the development of technologies
andglobal strategies
for conservation
and use of genetic resources in national
programs
and research institutions.
Our deliberations
over the last few days have addressedcharacterizationand evaluationof plant genetic resources. I believe
we have alllearned a great deal from the speakers and ensuing discussions.
Thank you all for
helping
make this last three days so productive.
I will close this workshop by wishing you all a safe journey home.
Thank you very much.

"The 4th
MAFF
22-24
OctoberInternational
Workshop
1996,
Tsukuba,
Japan.

on

Genetic

Resources

Photographof workshop

participants:

Front row left to right:*.Ohmura, H. Seko, Y. Shimamoto,
Y. Tosa, L. N. Trinh, P.
Strelchenko,
M. Nakagahra,
R.C. Chaudhary,P. N. Gupta, Y. Kotaka, K. Riley, A. G.
Yunus, S. Kresovich,L. H. Li, M. D. Zhou, K. Hayashi.
Second row: S. Nakayama,
H. Yamane, A. Ghafoor,
Mariscal,F. Kikuchi,
H. N. Regmi, M. Afzal, C. Mujaju,
M. Ekanayake,
Y. Sano, K. Ebana, K. Okuno.
Third row. S. Fukuoka, J. Takahashi,
V. Y. Molodkin,
T. Oishi, Y. I. Chin,

T. Goto, H. Namai, A. M.
A. O. Sari, S. R. Gupta, E.

T. D. Hoang, T. Sato, N. T. Quynh, N. Katsura,
O. Welker, A. Yamamoto, S.Suzuki,
N. Mase.

Forth row: K. Shirata,
H. Nakayama, T. Nagamine, Y. Kunihiro,
A.S. Liyanage,
N.
Tomooka, T. Nishikawa,A. M. Melhim, K. Komaki, Y. Tsurumi, M. Shoda, D. A.
VaughanFifth
row: S. Miyashita,
Yamamoto, K. Shimizu,

K. Matsuo, T. Chibana,T. Shiina,
T. Sanada, K. Kato, M. Yamamori.

S. W. Prihatanti,

M.

LIST OF PARTICIPANTS

List of Participants
to the 4th MAFF Workshop
held at NIAR, Japan,22-24
October, 1996
Afzal,

on Genetic

Resource to be

Muhammad
Plant Genetic Resources Institute(PGRI),
Pakistan
Chaudhary,
Ram C. (Topic 3)International
Rice Research Institute(IRRI),
Philippines
Chen, Yi-Shin
Chia-Yi Agricultural
Experiment Station,Taiwan
Chibana,
TakashiNational
Instituteof Agrobiological
Resources (NIAR),Japan
Ebana, Kaoru (Steering
Committee)
NIAR, Japan
Ekanayake, E.M.D.S.Nalin
Plant Genetic Resources Center, Sri Lanka
Fukuoka, Shuichi
NIAR, Japan
Ghafoor, Abdul
PGRI, Pakistan
Goto, Torao
Agriculture,
Forestry and Fisheries
Technical
Information
Society(AFFTIS),Japan
Gupta, P. N. (Topic 3)
National
Bureau of Plant Genetic Resources (NBPGR),
India
Gupta, Salik Ram
Nepal Agricultural
Research Council, Nepal
Hasebe, Akira
NIAR, Japan
Hayashi,
Kenichi
Advisory committee of NIAR, Japan
Higo, Kenichi
NIAR, Japan
Hoang, Tran Due
Root and Tuber Crop Research Center, Vietnam
Horita, Mitsuo
NIAR, Japan
Hoshino, Takafumi(Organizing
Committee)
National
Agriculture
Research Center (NARC),Japan
Ideno, Aika
NIAR, Japan
Ishikawa,
Masaya
NIAR, Japan

Iwamoto, Masao
NIAR, Japan
Kaku, Hisatoshi
NIAR, Japan
Kato, Kunihiko(Organizing
Committee)
NIAR, Japan
Katsura, Naoki (Organizing
Committee)
NIAR, Japan
Katsuta-Seki,
Masumi
NIAR, Japan
Kikuchi,
Fumio (Organizing
Committee)
Tokyo Universityof Agriculture,Japan
Kimura, Tetsuya
National
Center for Seeds and Seedlings,
Japan
Komaki, Katsumi (Steering
Committee)
NARC, Japan
Kotaka, Yoshihiko(Welcome address)
Agriculture,
Forestry and Fisheries
Research Council Secretariat(AFFRC),Japan
Kresovich,
Steven(Topic 1)
USDA-ARS, USA
Kunihiro,
Yasufumi (Steering
Committee)
NIAR, JapanLe, Viet Dung
Hokkaido University
student from Vietnam
Li, Li Hui (Topic 2)
Instituteof Crop Germplasm Resources, China
Liyanage,
Athula S. U.
Plant Genetic Resources Center, Sri Lanka
Luu, Ngoc Trinh (Topic 2)
Vietnam Agricultural
Science Institute(VASI),Vietnam
Mariscal,
AlgericoM.
Philippine
Root Crop Research and Training
Center, Philippines
Mase, Nobuko
National
Instituteof Fruit Tree Science (NIFTS),
Japan
Matsuo, Kazuhito(Topic 2)
National
Instituteof Agro-environmental
Sciences (NIAES),
Japan
Melhim, Al-Muhamad
Directorateof Agricultural
Scientific
Research,
Syria
Miyashita,
Susumu
NIAR, JapanMiyazaki,

Shoji (Organizing
Committee)
Japan International
Research Center for Agricultural
Sciences (JIRCAS),
Japan, (presently.NIAR)
Molodkin,
Vadim Y.
N. I.Vavilov
Research Instituteof Plant Industry(VIR),Russia
Morishima,
Hiroko (Organizing
Committee, Keynote address)
National
Instituteof Genetics (NIG), Japan
Mujaju, ClaidNational
Herbarium and Botanic Garden, Zimbabwe
Nagamine, Tsukasa (Steering
Committee)
NIAR, Japan
Nagamine, Yoshitaka
NIAR, Japan
Nagai, Toshiro
NIAR, Japan
Nakagahra, Masahiro (Opening
address)
NIAR, Japan
Nakamura, Masaru
AFFRC, Japan
Nakayama, Hiroki
NIAR, Japan
Nakayama, Shigeki
NIAR, Japan
Namai, Hyoji
Universityof Tsukuba, Japan
Nguen, Thi Quynh
VASI,Vietnam
Nirasawa, Keijiro
NIAR, Japan
Nishikawa,
Tomotaro
NIAR, Japan
Oishi, Takao (Organizing
Committee)
NIAR, Japan (presently:
National
Instituteof Animal Industry)
Okuno, Kazutoshi(SteeringCommittee, Topic 2)
NIAR, Japan
Osono, Masanori
National
Center for Seeds and Seedlings,
Japan
Prihatanti,
Sri WinarniInternational
Potato Center (CIP),Indonesia
Regmi, HornNath
Nepal Agricultural
Research Council, Nepal

Riley,Kenneth W. (Organizing
Committee, Keynote address, Workshop summary)International
Plant Genetic Resources Institute(IPGRI),
Malaysia
Sanada, Tetsuro (Steering
Committee)
NIFTS, Japan
Sano, Yoshio (Topic 1)
Hokkaido University,Japan
Sari, Ali Osman
Aegean Agricultural
Research Institute,
TurkeySato, Takanori
National Instituteof Vegetables,
Ornamental Plants and Tea (NIVOT),Japan
Seko, Hidefumi (Chairman of Organizing
Committee, Chief of Steering Committee,
Topic 3, Closing remarks)
NIAR, Japan (presently:Yamaguchi
University)
Shiina, Tsugio
NIAR, Japan
Shimamoto, Yoshiya (Organizing
Committee)
Hokkaido University,Japan
Shimizu, Kunihiro
NIAES, Japan
Shirata,
Kazuto (Steering
Committee)
NIAR, Japan
Shoda, Moriyuki
Okinawa Prefectural
Agriculture
Experiment
Station,Japan
Strelchenko,
Pjotor (Topic 2)
VIR, Russia
Suzuki, Shigeru
AFFTIS, Japan
Takahashi,
Junji
Japan International
Cooperation
Agency (JICA),Japan
Takashima,
Satoshi
AFFRC, Japan
Takeya, Masaru
NIAR, Japan
Tanaka, Yoshiho(SteeringCommittee)
NIAR, Japan
Tomooka, Norihiko
NIAR, Japan
Tosa, Yukio(Topic
2)
Kobe University,
Japan
Tsuchiya,
Kenichi
NIAR, Japan

Tsurumi,

YoshiroNational
Instituteof Grassland
Research (NIGR),
Vaughan, Duncan A. (Steering
Committee, Topic 1)
NIAR, Japan
Welker, Ottomar
JIRCAS fellow from Germany
Yamamori, Makoto
NIAR, Japan
Yamamoto, Akio (Steering
Committee)
AFFRC, Japan
Yamamoto, Masashi
NIFTS, Japan
Yamane, Hiroyasu(Organizing
Committee)
NIFTS, Japan
Yunus, Abdul G. (Topic 3)
Universiti
Pertanian
Malaysia,Malaysia
Zhou, Ming-De
IPGRI ,China

JapanEditors
Editor in chief
Managing editorsC

onsultingeditors

Seko, Hidefumi
Vaughan, Duncan A.
Okuno, Kazutoshi
Shirata, Kazuto
Ebana, Kaworu
Miyazaki, ShojiPublishedJuly, 1998

Research Council
National InstituteKannondai

Secretariat of MAFF andof Agrobiological
Resources2-1-2, Tsukuba,ISBN 4-9900110-9-0

Ibaraki

305-8602,

Japan