E
TG/276/1
ORIGINAL: English
DATE: 2012-03-28

INTERNATIONAL UNION FOR THE PROTECTION OF NEW VARIETIES OF PLANTSGENEVA*HEMPUPOV Code: CANNB_SAT
Cannabis sativa L.

GUIDELINESFOR THE CONDUCT OF TESTSFOR DISTINCTNESS, UNIFORMITY AND STABILITY

Alternative Names:*
Botanical nameEnglishFrenchGermanSpanishCannabis sativa L.

Hemp

ChanvreHanf

CáñamoThe purpose of these guidelines (“Test Guidelines”) is to elaborate the principles contained in the
General Introduction (document TG/1/3), and its associated TGP documents, into detailed practical guidance forthe harmonized examination of distinctness, uniformity and stability (DUS) and, in particular, to identify
appropriate characteristics for the examination of DUS and production of harmonized variety descriptions.

ASSOCIATED DOCUMENTS
These Test Guidelines should be read in conjunction with the General Introduction and its
associated TGP documents.

*

These names were correct at the time of the introduction of these Test Guidelines but may be revised or updated.
[Readers are advised to consult the UPOV Code, which can be found on the UPOV Website (www.upov.int), for thelatest information.]TG/276/1
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TABLE OF CONTENTS
1.
2.
3.

PAGE

SUBJECT OF THESE TEST GUIDELINES..................................................................................................3
MATERIAL REQUIRED ...............................................................................................................................3
METHOD OF EXAMINATION.....................................................................................................................3
3.1 Number of Growing Cycles ....................................................................................................................3
3.2 Testing Place ...........................................................................................................................................3
3.3 Conditions for Conducting the Examination...........................................................................................3
3.4 Test Design .............................................................................................................................................4
3.5 Additional Tests ......................................................................................................................................4
4. ASSESSMENT OF DISTINCTNESS, UNIFORMITY AND STABILITY...................................................4
4.1 Distinctness .............................................................................................................................................4
4.2 Uniformity...............................................................................................................................................5
4.3 Stability ...................................................................................................................................................6
5. GROUPING OF VARIETIES AND ORGANIZATION OF THE GROWING TRIAL.................................6
6. INTRODUCTION TO THE TABLE OF CHARACTERISTICS ...................................................................7
6.1 Categories of Characteristics...................................................................................................................7
6.2 States of Expression and Corresponding Notes.......................................................................................7
6.3 Types of Expression................................................................................................................................8
6.4 Example Varieties ...................................................................................................................................8
6.5 Legend.....................................................................................................................................................8
7. TABLE OF CHARACTERISTICS/TABLEAU DESCARACTÈRES/MERKMALSTABELLE/TABLA DE CARACTERES.......................................................9
8. EXPLANATIONS ON THE TABLE OF CHARACTERISTICS ................................................................15
8.1 Explanations covering several characteristics .......................................................................................15
8.2 Explanations for individual characteristics ...........................................................................................15
8.3 Growth stages for Hemp .......................................................................................................................18
9. LITERATURE ..............................................................................................................................................20
10. TECHNICAL QUESTIONNAIRE ...............................................................................................................21

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1.

Subject of these Test Guidelines
These Test Guidelines apply to all varieties of Cannabis sativa L.

2.

Material Required

2.1
The competent authorities decide on the quantity and quality of the plant material
required for testing the variety and when and where it is to be delivered. Applicants
submitting material from a State other than that in which the testing takes place must ensurethat all customs formalities and phytosanitary requirements are complied with.
2.2
The material is to be supplied in the form of seed or young, non-flowering plants in
pots, of sufficient size and with sufficient development to express all the characteristics of thevariety in the first growing cycle.
2.3

The minimum quantity of plant material, to be supplied by the applicant, should be:
Vegetatively propagated varieties: 50 young plants.
Seed-propagated varieties: 500 grams of seed.

In the case of seed, the seed should meet the minimum requirements for germination, species
and analytical purity, health and moisture content, specified by the competent authority.
2.4
The plant material supplied should be visibly healthy, not lacking in vigor, noraffected by any important pest or disease.
2.5
The plant material should not have undergone any treatment which would affect theexpression of the characteristics of the variety, unless the competent authorities allow orrequest such treatment. If it has been treated, full details of the treatment must be given.
3.
3.1

Method of Examination
Number of Growing Cycles
The minimum duration of tests should normally be two independent growing cycles.

3.2

Testing Place

Tests are normally conducted at one place. In the case of tests conducted at more than
one place, guidance is provided in TGP/9 “Examining Distinctness”.
3.3

Conditions for Conducting the Examination

3.3.1 The tests should be carried out under conditions ensuring satisfactory growth for the
expression of the relevant characteristics of the variety and for the conduct of theexamination.

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3.3.2 The optimum stage of development for the assessment of each characteristic is
indicated by a number in the second column of the Table of Characteristics. The stages ofdevelopment denoted by each number are described in Chapter 8.
3.4

Test Design

3.4.1 In the case of seed-propagated varieties, each test should be designed to result in a
total of at least 200 plants, which should be divided between at least 2 replicates.
3.4.2 In the case of vegetatively propagated varieties, each test should be designed to resultin a total of at least 40 plants.
3.4.3 The design of the tests should be such that plants or parts of plants may be removed
for measurement or counting without prejudice to the observations which must be made up to
the end of the growing cycle.
3.5

Additional Tests
Additional tests, for examining relevant characteristics, may be established.

4.
4.1

Assessment of Distinctness, Uniformity and Stability
Distinctness
4.1.1 General Recommendations

It is of particular importance for users of these Test Guidelines to consult the General
Introduction prior to making decisions regarding distinctness. However, the following points
are provided for elaboration or emphasis in these Test Guidelines.
4.1.2 Consistent Differences
The differences observed between varieties may be so clear that more than one
growing cycle is not necessary. In addition, in some circumstances, the influence of theenvironment is not such that more than a single growing cycle is required to provide
assurance that the differences observed between varieties are sufficiently consistent. One
means of ensuring that a difference in a characteristic, observed in a growing trial, issufficiently consistent is to examine the characteristic in at least two independent growing
cycles.
4.1.3 Clear Differences
Determining whether a difference between two varieties is clear depends on manyfactors, and should consider, in particular, the type of expression of the characteristic being
examined, i.e. whether it is expressed in a qualitative, quantitative, or pseudo-qualitative
manner. Therefore, it is important that users of these Test Guidelines are familiar with therecommendations contained in the General Introduction prior to making decisions regarding
distinctness.

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4.1.4 Number of Plants / Parts of Plants to be Examined
Unless otherwise indicated, for the purposes of distinctness, all observations on single
plants should be made on 20 plants or parts taken from each of 20 plants and any otherobservations made on all plants in the test, disregarding any off-type plants.
4.1.5 Method of Observation
The recommended method of observing the characteristic for the purposes of
distinctness is indicated by the following key in the second column of the Table ofCharacteristics (see document TGP/9 “Examining Distinctness”, Section 4 “Observation ofcharacteristics”):
MG: single measurement of a group of plants or parts of plants
MS: measurement of a number of individual plants or parts of plants
VG: visual assessment by a single observation of a group of plants or parts of plants
VS: visual assessment by observation of individual plants or parts of plants
Type of observation: visual (V) or measurement (M)“Visual” observation (V) is an observation made on the basis of the expert’s
judgment. For the purposes of this document, “visual” observation refers to thesensory observations of the experts and, therefore, also includes smell, taste andtouch. Visual observation includes observations where the expert uses reference
points (e.g. diagrams, example varieties, side-by-side comparison) or non-linear
charts (e.g. color charts). Measurement (M) is an objective observation against acalibrated, linear scale e.g. using a ruler, weighing scales, colorimeter, dates,
counts, etc.
Type of record: for a group of plants (G) or for single, individual plants (S)For the purposes of distinctness, observations may be recorded as a single record
for a group of plants or parts of plants (G), or may be recorded as records for anumber of single, individual plants or parts of plants (S). In most cases, “G”
provides a single record per variety and it is not possible or necessary to apply
statistical methods in a plant-by-plant analysis for the assessment of distinctness.
In cases where more than one method of observing the characteristic is indicated in the Tableof Characteristics (e.g. VG/MG), guidance on selecting an appropriate method is provided in
document TGP/9, Section 4.2.
4.2

Uniformity

4.2.1 It is of particular importance for users of these Test Guidelines to consult theGeneral Introduction prior to making decisions regarding uniformity. However, the following
points are provided for elaboration or emphasis in these Test Guidelines:

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4.2.2 Seed-propagated varieties: the assessment of uniformity of seed-propagated varieties
should be according to the recommendations for cross-pollinated varieties in the General
Introduction.
4.2.3 Vegetatively propagated varieties:
for the assessment of uniformity ofvegetatively propagated varieties, a population standard of 1 % and an acceptance probabilityof at least 95 % should be applied. In the case of a sample size of 40 plants, 2 off-types areallowed.
4.3

Stability

4.3.1 In practice, it is not usual to perform tests of stability that produce results as certain as
those of the testing of distinctness and uniformity. However, experience has demonstrated
that, for many types of variety, when a variety has been shown to be uniform, it can also be
considered to be stable.
4.3.2 Where appropriate, or in cases of doubt, stability may be further examined by testing a
new seed or plant stock to ensure that it exhibits the same characteristics as those shown by
the initial material supplied.
5.

Grouping of Varieties and Organization of the Growing Trial

5.1
The selection of varieties of common knowledge to be grown in the trial with thecandidate varieties and the way in which these varieties are divided into groups to facilitate
the assessment of distinctness are aided by the use of grouping characteristics.
5.2
Grouping characteristics are those in which the documented states of expression, even
where produced at different locations, can be used, either individually or in combination withother such characteristics: (a) to select varieties of common knowledge that can be excluded
from the growing trial used for examination of distinctness; and (b) to organize the growing
trial so that similar varieties are grouped together.
The following have been agreed as useful grouping characteristics:
(a)
(b)
(c)
(d)
(e)
(f)

Time of male flowering (characteristic 11)
Inflorescence: THC content (characteristic 13)
Plant: proportion of hermaphrodite plants (characteristic 14)
Plant: proportion of female plants (characteristic 15)
Plant: proportion of male plants (characteristic 16)
Plant: natural height (characteristic 17)

5.4
Guidance for the use of grouping characteristics, in the process of examining
distinctness, is provided through the General Introduction and document TGP/9“Examining Distinctness”.

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6.
6.1

Introduction to the Table of Characteristics
Categories of Characteristics
6.1.1 Standard Test Guidelines Characteristics

Standard Test Guidelines characteristics are those which are approved by UPOV for
examination of DUS and from which members of the Union can select those suitable for their
particular circumstances.
6.1.2 Asterisked Characteristics
Asterisked characteristics (denoted by *) are those included in the Test Guidelines
which are important for the international harmonization of variety descriptions and should
always be examined for DUS and included in the variety description by all members of theUnion, except when the state of expression of a preceding characteristic or regional
environmental conditions render this inappropriate.
6.2

States of Expression and Corresponding Notes

6.2.1 States of expression are given for each characteristic to define the characteristic and toharmonize descriptions. Each state of expression is allocated a corresponding numerical notefor ease of recording of data and for the production and exchange of the description.
6.2.2 In the case of qualitative and pseudo-qualitative characteristics (see Chapter 6.3), allrelevant states of expression are presented in the characteristic. However, in the case ofquantitative characteristics with 5 or more states, an abbreviated scale may be used tominimize the size of the Table of Characteristics. For example, in the case of a quantitative
characteristic with 9 states, the presentation of states of expression in the Test Guidelines may
be abbreviated as follows:
State
small
medium
large

Note
3
5
7

However, it should be noted that all of the following 9 states of expression exist to describe
varieties and should be used as appropriate:Statevery small
very small to small
small
small to medium
mediummedium to large
large
large to very large
very large

Note
1
2
3
4
5
6
7
8
9

6.2.3 Further explanation of the presentation of states of expression and notes is provided in
document TGP/7 “Development of Test Guidelines”.

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6.3

Types of Expression

An explanation of the types of expression of characteristics (qualitative, quantitativeand pseudo-qualitative) is provided in the General Introduction.
6.4

Example Varieties

Where appropriate, example varieties are provided to clarify the states of expression ofeach characteristic.
6.5

Legend(*)

Asterisked characteristic

QL
QN
PQ

Qualitative characteristic– see Chapter 6.3
Quantitative characteristic– see Chapter 6.3
Pseudo-qualitative characteristic – see Chapter 6.3

MG, MS, VG, VSC– see Chapter 6.1.2– see Chapter 4.1.5

Additional test in greenhouse(a), (b)(+)
0003, etc.

See Explanations on the Table of Characteristics in Chapter 8.1
See Explanations on the Table of Characteristics in Chapter 8.2
Growth stage– see Chapter 8.3

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Table of Characteristics/Tableau des caractères/Merkmalstabelle/Tabla de caracteres

English1.

Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

françaisdeutschespañol

Cotylédon : forme

Keimblatt: Form

Cotiledón: forma

narrow obovate

obovale étroit

schmal verkehrt
eiförmigoboval estrecha

Fibrimon1

medium obovate

obovale moyen

mittel verkehrt
eiförmigoboval media

Epsilon 68

2

broad obovate

obovale large

breit verkehrt
eiförmigoboval ancha

Futura 75

30003 Cotyledon: shape
VG

Note/
Nota

(+)
QN

C

2.

0003 Cotyledon: color
VG

Cotylédon : couleur Keimblatt: Farbe

Cotiledón: color

PQ

Cyellow

jaune

gelb

amarillo

Chamaeleon

1light green

vert clair

hellgrün

verde claro

Fedora 17

2

medium green

vert moyen

mittelgrün

verde medio

Ferimon3

dark green

vert foncé

dunkelgrün

verde oscuro

Dioica 88

4

Hypocotyl:
Intensität der
Anthocyanfärbung

Hipocotilo:
intensidad de lapigmentación
antociánica

3.

0003 Hypocotyl: intensity Hypocotyle :VG of anthocyanin
intensité decoloration
la pigmentation
anthocyanique

QN

C

4.

QN

weak

faiblegeringdébil

Uso 31

3

mediummoyennemittelmedia

Dioica 88

5strong

forte

stark

fuere

KC Dora7

Pflanze:
Anthocyanfärbung
der Krone

Planta:pigmentación
antociánica de lacorona1006 Plant: anthocyanin Plante :VG coloration of crown pigmentation
anthocyanique dela couronne
absent or very weakabsente ou très faible fehlend oder sehrgeringausente o muy débil

1

weak

faiblegeringdébil

Felina 32

3

mediummoyennemittelmedia

Epsilon 68

5strong

forte

stark

fuerteFinola7

TG/276/1
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Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

Note/
Nota

5.

VG Leaf: intensity ofgreen color

Feuille : intensité de Blatt: Intensität der Hoja: intensidad della couleur verte
Grünfärbung
color verde

QN(a) light

clairehellligeroChamaeleon

1

mediummoyennemittelmedio

Fedora 17

2

dark

foncéedunkeloscuroEpsilon 68

3

6.

MS Leaf: length ofpetioleFeuille : longueurdu pétiole

Blatt: Länge desBlattstiels

Hoja: longitud delpecíoloQN(a) short

court

kurz

corto

Santhica 27

1(b) medium

moyen

mittelmedio

Fedora 17

2

long

long

lang

largo

Ermes

3

Blatt:
Anthocyanfärbung
des Blattstiels

Hoja: pigmentación
antociánica delpecíolo7.
(*)

VG Leaf: anthocyanin Feuille :coloration of petiole pigmentation
anthocyaniquedu pétiole

QN(a) absent or very weak absente ou très faible fehlend oder sehrausente o muy débil

Fibrol1(b) weakgeringfaiblegering

débil

Ruby

2

mediummoyennemittelmedia

Dioica 88

3strong

forte

stark

fuerteEpsilon 68

4very strong

très forte

sehr stark

muy fuerte

Finola

5

8.
(*)
(+)

MS/ Leaf: number of
VG leaflets

Feuille : nombre de Blatt: Anzahl
foliolesBlattfiedern

Hoja: número defolíolosQN(a) few

faiblegering

bajo

Ermes

1

(b) medium

moyen

mittelmedio

Epsilon 68

2

many

élevé

groß

alto

Kompolti

3

9.

MS Central leaflet:
lengthFoliole centrale :longueurMittlere Blattfieder: Folíolo central:
Länge
longitudQN(a) shortcourte

kurz

corto

Santhica 27

3(b) medium

moyennemittelmedio

Epsilon 68

5

long

longue

lang

largo

Kompolti

7

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Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

Note/
Nota

10.

MS Central leaflet:
width

Foliole centrale :largeur

Mittlere Blattfieder: Folíolo central:
BreiteanchuraQN(a) narrowétroiteschmalestrechoSanthica 27

3(b) medium

moyennemittelmedio

Dioica 88

5

large

breit

ancho

Kompolti7

broad

Époque de floraison Zeitpunkt dermâle
männlichen Blüte

Época de floración
masculina

very early

très précoce

sehr früh

muy temprana

Finola1

earlyprécocefrüh

temprana

Santhica 27

3mediummoyennemittelmedia

Dioica 88

5

latetardivespät

tardíaFutura 75

7very latetrès tardive

sehr spät

muy tardía

Kompolti

9

11.
(*)
(+)

MG Time of male
floweringQN

12.

2102 Inflorescence:
2304 anthocyanin
VG coloration of male
flowers

Inflorescence :pigmentation
anthocyaniquedes fleurs mâles

Blütenstand:
Anthocyanfärbung
der männlichen
BlütenInflorescencia:
pigmentación
antociánica de lasflores masculinas

QN

absent or very weaknulle ou très faible

fehlend oder sehrgeringausente o muy débil

Kompolti

1

weak

faiblegeringdébil

Beniko3

mediummoyennemittelmedia

Uso 315strong

forte

stark

fuerteErmes

7very strong

très forte

sehr stark

muy fuerte

Finola9

Inflorescence :teneur en THC

Blütenstand:
THC-Gehalt

Inflorescencia:
contenido en THC

absent or very low

nulle ou très faible

fehlend oder sehrgeringausente o muy bajo

Santhica 23

1mediummoyennemittelmedio

Uso 313very hightrès élevéesehr hochmuy alto

Medisins

5

13.
(*)
(+)
QN

MG Inflorescence:
THC content

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14.
(*)
(+)

QN

15.
(*)
(+)

QN

16.
(*)
(+)

QN

17.
(*)
(+)

QN

françaisdeutsch2102 Plant: proportion of Plante : proportion Planze: Anteil
2202 hermaphrodite
de plantes
zwittriger Pflanzen
2302 plants
hermaphrodites
2304
MS/
VG

español

Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

Note/
Nota

Planta: proporciónde plantas
hermafroditas

lowfaiblegeringbaja

1mediummoyennemittel

media

3

high

élevée

hoch

alta

5

Planta: proporción
2102 Plant: proportion of Plante : proportion Planze: Anteil
2202 female plants
de plantes femelles weiblicher Pflanzen de plantas femeninas
2302
2304
MS/
VG
low

faiblegeringbaja

1mediummoyennemittel

media

3

high

élevée

hoch

alta

5

Planta: proporción
2102 Plant: proportion of Plante : proportion Planze: Anteil
de plantes mâles
männlicher Pflanzen de plantas
2202 male plants
masculinas
2302
2304
MS/
VG
low

faiblegeringbaja

1mediummoyennemittel

media

3

high

élevée

hoch

alta

5

Plante : hauteur
naturelle

Pflanze: natürliche
Höhe

Planta: altura
natural

short

basse

niedrigbaja

Finola3

mediummoyennemittelmedia

Uso 31

5

long

haute

hoch

alta

Ferimon7

2202 Plant: natural
2302 height
VG/
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Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

Note/
Nota

18.
(*)

2202 Main stem: color
2302
VG

Tige principale :couleur

Haupttrieb: Farbe

Tallo principal:
color

PQ

(c) yellow

jaune

gelb

amarillo

Chamaeleon

1

medium green

vert moyen

mittelgrün

verde medio

Epsilon 68

2

dark green

vert foncé

dunkelgrün

verde oscuro

Kompolti

3

purplepourprepurpurnpúrpuraFibranova

4

19.

2202 Main stem: length of Tige principale :2302 internodelongueur deMSl’entre-nœud

Haupttrieb:
Internodienlänge

Tallo: longitud delentrenudoQN(c) short

court

kurz

corto

Ferimon3

mediummoyen

mittelmedio

Uso 31

5

long

long

lang

largo

KC Dora720.

2202 Main stem:
2302 thickness
MS/
VG

Tige principale :épaisseur

Haupttrieb: Dicke

Tallo principal:
grosorQN(c) thin

mince

dünn

delgadoFinola1

mediummoyennemittelmedio

Epsilon 68

2

thick

épaissedickgruesoKompolti

321.

2202 Main stem: depth of Tige principale :2302 grooves
profondeur
VG
des cannelures

Haupttrieb: Tiefe
der Furchen

Tallo principal:
profundidad de lossurcosQN(c) shallow

peu profondes

flachpoco profundos

Finola1

mediummoyennes

mittelmediosFerimon2

deep

profondes

tief

profundos

Dioica 88

3(+)

2204 Main stem: pith in
2306 cross-section
VG

Tige principale :moelle en section
transversale

Haupttrieb: Füllung Tallo principal:
im Querschnitt
médula en sección
transversal

QN(c) absent or thinabsente ou finefehlend oder dünn

ausente o delgada

Ermes

1

mediummoyennemittelmedia

Santhica 27

2

thick

épaissedickgruesaChamaeleon

322.

TG/276/1
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1 000 graines

Samen: 1 000Korngewicht

Semilla: pesode 1000 semillas

Example Varieties/
Exemples/Beispielssorten/
Variedades ejemplo

Note/
Nota

23.

2205 Seed: 1,000 seed
2307 weightMG

QN

very lowtrès faible

sehr gering

muy bajo

Finola1

low

faiblegeringbajo

Chamaeleon

2

mediummoyen

mittelmedio

Uso 31

3

high

élevé

hoch

alto

Fedora 17

4very hightrès élevé

sehr hochmuy alto

Epsilon 68

5

Graine : couleur
des téguments

Samen: Farbe derSamenschale

Semilla: color deltegumento

light grey

gris clair

hellgrau

gris ligero

Fibrol1

medium grey

gris moyen

mittelgrau

gris medio

Finola2

grey brownbrun grisgraubraun

marrón gris

Futura 75

3yellowish brownbrun jaunâtre

gelblichbraun

marrón amarillento

Santhica 27

4

brown

bruns

braun

marrónErmes

5Graine : marbrure

Samen:
Marmorierung

Semilla: veteado

weak

faiblegeringdébil

Finola1

mediummoyennemittelmedio

Kompolti

2strong

forte

stark

fuerteFutura 75

3

24.

PQ

25.
(+)
QN

2205 Seed: color of testa
2307
VG

2205 Seed: marbling2307
VG

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8.
8.1

Explanations on the Table of Characteristics
Explanations covering several characteristics

Characteristics containing the following key in the second column of the Table ofCharacteristics should be examined as indicated below:
(a)
(b)
(c)

8.2

Observations should be done in the period between the beginning offlowering (growth stage 2101, 2201 or 2301, whichever is earliest) and the
beginning of seed maturity.
Observations should be done on the last opposite, fully expanded leaves
Observations should be done on the internode below the last opposite leavesof female and/or hermaphrodite plants only.

Explanations for individual characteristics

Ad. 1: Cotyledon: shape1
narrow obovate

2
medium obovate

3
broad obovate

Ad. 8: Leaf: number of leaflets
Few is less than 7 leaflets.
Medium number of leaflets is 7 (predominant number of leaflets).
Many is more than 7 leaflets.

Ad. 11: Time of male flowering
Monoecious varieties: 50 % of all plants with first male flower open.
Other varieties: 50 % of all male plants with first male flower open.
First male flowers mostly appear from the axils of the leaves on the main stem. Male
flowers usually appear about 2 weeks before the styles of female flowers are visible.

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Hemp, 2012-03-28- 16 -

Ad. 13: Inflorescence: THC content
The method to determine the THC content is based on a quantitative determination of
Δ9-tetrahydrocannabinol by gas chromatography after extraction with a suitable solvent.
Sampling
The sample (mixture of 20 plants) should be taken from the upper 30 cm of the main stem,
containing the female inflorescence. Sampling should be carried out in the period from 20
days after the beginning of female flowering up to the end of flowering. The sample should bedried as soon as possible (within 48 hours) at a temperature below 60º C. Samples should bedried to a constant weight and to a moisture content of 8 – 13 %. After drying samples can be
stored (without crushing) at below 25º C in a dark place.
Determination of THC content (see also Cole, 2003).
1. Preparation of the test sample
Remove stems and seeds over 2 mm in size from the dried samples.
Grind the dried samples to obtain a semi-fine powder (passing through a 1 mm mesh sieve).
The powder may be stored for 10 weeks at below 25º C in a dark dry place.
2. Reagents and extraction solution
Reagents- Δ9-tetrahydrocannabinol, pure for chromatographic purposes.
- squalane, pure for chromatographic purposes, as an internal standard.
Extraction solution- 35 mg of squalane per 100 ml hexane.
3. Extraction of Δ9-tetrahydrocannabinol
Weigh 100 mg of the powdered test sample, place in a centrifuge tube and add 5 ml of
extraction solution containing the internal standard.
Place in an ultrasound bath and leave for 20 minutes. Centrifuge for 5 minutes at 3,000 r.p.m.
and then remove the supernatant THC solution. Inject the solution into the chromatograph andcarry out a quantitative analysis.
4. Gas chromatography(a) Apparatus- gas chromatograph with a flame ionization detector and a split/splitless injector- column allowing good separation of cannabinoids, for example a glass capillary column
25 m long and 0.22 mm in diameter impregnated with a 5 % non-polar phenyl-methylsiloxane phase.
(b) Calibration ranges
At least three points including points 0.04 and 0.50 mg/ml Δ9-THC in extraction solution.
(c) Experimental conditions
The following conditions are given as an example for the column referred to in a).
oven temperature
260º C
injector temperature
300º C
detector temperature
300º C(d) Injection volume: 1 µl

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Results
THC should be determined to two decimals in grams of Δ9-THC per 100 grams of analyticalsample dried to constant weight. A tolerance of 0.03 g per 100 grams applies. The results are
expressed in % dry weight.
Although varietal differences for THC content are consistent, absolute levels of THC content
are sensitive to environmental variation. States of expression need to be calibrated by
Example varieties.

Ad. 14, 15 and 16: Plant: proportion of hermaphrodite plants, female plants and male plants
resp.
Cannabis sativa L. is dioecious by nature, containing approximately equal proportionsof male and female plants. Hermaphrodite plants (male and female flowers on one plant)
occasionally occur naturally but are specially created by breeding activity (Bócsa, 1998).
Several intersexual forms exist and sex expression can be modified by environmental factors.
Hermaphrodite plants: plants with both male and female flowers
Female plants: plants with female flowers only
Male plants: plants with male flowers only
Proportion
low
low to medium
mediummedium to high
high

Note
1
2
3
4
5

Ranges (percentage)<= 5 %
6-35 %
36-65 %
66-95 %>= 96 %

Proportion should be based on at least 200 plants for seed propagated varieties and at least 40
plants for vegetatively propagated varieties (numbers are rounded to whole numbers).

Ad. 17: Plant: natural height
Natural height should be observed on female and/or hermaphrodite plants including
inflorescence.

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Ad. 22: Main stem: pith in cross-section1
absent or thin2
medium3
thick

Ad. 25: Seed: marbling
Marbling of testa: black mosaic patterns.

1
weak

8.3

2
medium

3
strongGrowth stages for Hemp

All characteristics should be recorded at the appropriate time for the plant concerned.
Growth stages of hemp are recorded by a four-digit code describing the principal growthstages, depending on the sex of the plant followed by detailed developmental stages(Mediavilla, Vito et al., 1998):
Principal growth stages
Four principal stages describe the life cycle of a plant and are coded by their first digit of the
four-digit code.
First-digit of code0
1
2
3

Definition
Germination and emergence
Vegetative stage
Flowering and seed formation
SenescenceTG/276/1
Hemp, 2012-03-28- 19 -

Secondary growth stages
The secondary growth stages are described by the second digit, which indicates the sex of theplant, the third and fourth digits indicating the developmental stage of the plant.
CodeDefinition
Remarks
Germination and emergence0000
Dry seed0003
Cotyledons unfolded
Vegetative stage refers to main stem. Leaves are considered unfolded when leaflets are at least one cm long1002
1st leaf pair
1 leafletnd1004
2 leaf pair3 leafletsrd1006
3 leaf pair5 leaflets
10xx
Last opposite leaf pairxx = 2 times nth leaf pair
Flowering and seed formation refers to the main stem including branches
2000
GV point (i.e. induction of Change of phyllotaxis on the main stem fromflowering)
opposite to alternate. Distance between
petioles of alternate leaves at least 0.5 cm
2001
Flower primordia
Sex nearly indistinguishable
Male Plant
2100
Flower formation
First closed staminate flowers
2101
Beginning of floweringFirst opened staminate flowers
2102
Flowering
50 % opened staminate flowers
2103
End of flowering
95 % of staminate flowers opened or withered
Female Plant
2200
Flower formation
First pistillate flowers
Bract with no styles
2201
Beginning of flowering
Styles on first female flowers
2202
Flowering
50 % of bracts formed
2203
Beginning of seed maturity
First seeds hard
2204
Seed maturity
50 % of seeds hard
2205
End of seed maturity
95 % of seeds hard or shattered
Hermaphrodite plant
2300
Female flower formation
First pistillate flowers
Perigonal bracts with no styles
2301
Beginning of female flowering First styles visible
2302
Female flowering
50 % of bracts formed
2303
Male flower formation
First closed staminate flowers
2304
Male flowering
50 % opened staminate flowers
2305
Beginning of seed maturity
First seeds hard
2306
Seed maturity
50 % of seeds hard2307
End of seed maturity
95 % of seeds hard or shattered
Senescence
3001
Leaf dessication
Leaves dry
3002
Stem dessication
Leaves dropped
3003
Stem decomposition
Bast fibers freeTG/276/1
Hemp, 2012-03-28- 20 -

9.

Literature

Bócsa, I., 1998: Genetic Improvement : Conventional Approaches. In: Advances in Hemp
Research. Paolo Ranalli (Ed.). Haworth Food Products Press, New York. 272 pp.
Bredemann, G., 1922 : Die Bestimmung des Fasergehaltes in Bastfaserpflanzen bei
züchterischen Untersuchungen. Faserforschung 2. Leipzig : Hirzel Verlag. S. 239-258.
Clarke, R.C., 1998: Botany of the Genus Cannabis. In: Advances in Hemp Research. Paolo
Ranalli (Ed.). Haworth Food Products Press, New York. 272 pp.
Cole, M.D., 2003. The analysis of controlled substances – a systematic approach. John Wileyand Sons Ltd., Chichester, UK. ISBN 0-471-49252-3.
Mediavilla, V., Jonquera, M., Schmid-Slembrouck, I., Soldati, A., 1998. Decimal code for
growth stages of hemp (Cannabis sativa L.). Journal of the International Hemp Association
5(2) : 67-72.
Meijer de, E., 1995: Fibre hemp cultivars : A survey of origin, ancestry, availability and briefagronomic characteristics. Journal of the International Hemp Association 2(2) : 66-73
Meijer de, E., 1998: Cannabis Germplasm Resources. In: Advances in Hemp Research. Paolo
Ranalli (Ed.). Haworth Food Products Press, New York. 272 pp.

TG/276/1
Hemp, 2012-03-28- 21 -

10.

Technical Questionnaire

TECHNICAL QUESTIONNAIRE

Page {x} of {y}Reference Number:
Application date:(not to be filled in by the applicant)

TECHNICAL QUESTIONNAIRE
to be completed in connection with an application for plant breeders’ rights
1.

2.

Subject of the Technical Questionnaire
1.1

Botanical name

Cannabis sativa L.

1.2

Common name

Hemp

ApplicantName
Address

Telephone No.
Fax No.
E-mail address
Breeder (if different from applicant)3.

Proposed denomination and breeder’s reference
Proposed denomination(if available)
Breeder’s referenceTG/276/1
Hemp, 2012-03-28- 22 -TECHNICAL QUESTIONNAIRE#

Page {x} of {y}Reference Number:

4. Information on the breeding scheme and propagation of the variety
4.1 Breeding scheme
Variety resulting from:
4.1.1

Crossing(a)controlled cross(please state parent varieties)(…………………..……………..…)
female parent(b)#(……………..…………………..…)
male parent

partially known cross(please state known parent variety(ies))(…………………..……………..…)
female parent
(c)

x

x

[ ]

[ ]

(……………..…………………..…)
male parent

unknown cross

[ ]

4.1.2

Mutation(please state parent variety)

[ ]

4.1.3

Discovery and development[ ](please state where and when discovered and how developed)

4.1.4

Other
(please provide details)[ ]Authorities may allow certain of this information to be provided in a confidential section of the Technical
Questionnaire.

TG/276/1
Hemp, 2012-03-28- 23 -TECHNICAL QUESTIONNAIRE

Page {x} of {y}Reference Number:

4.2 Method of propagating the variety
4.2.1 Seed-propagated varieties(a)Self-pollination

[ ]

(b)

Cross-pollination(i) population(ii) synthetic variety

[ ]
[ ]

(c)

Other
(please provide details)[ ]4.2.2 Vegetatively propagated varieties(a)cuttings[ ](b)in vitro propagation

[ ]

(c)

other (state method)

4.2.3 Other(please provide details)[ ]TG/276/1
Hemp, 2012-03-28- 24 -TECHNICAL QUESTIONNAIRE

Page {x} of {y}Reference Number:

5.
Characteristics of the variety to be indicated (the number in brackets refers to thecorresponding characteristic in Test Guidelines; please mark the note which best
corresponds).
Characteristics
5.1
(11)

Example Varieties

Note

Finola1[ ]Time of male flowering
very early2[ ]very early to early
earlySanthica 27

4[ ]early to medium
mediumDioica 88

Futura 75

5.2
(13)

7[ ]
8[ ]

late to very latevery late

5[ ]
6[ ]

medium to late
late

3[ ]

Kompolti

9[ ]Santhica 23

1[ ]Inflorescence: THC content
absent or very low

2[ ]

low
medium

Uso 314[ ]highvery high3[ ]Medisins

5[ ]TG/276/1
Hemp, 2012-03-28- 25 -TECHNICAL QUESTIONNAIRE

Page {x} of {y}Characteristics
5.3
(14)

5.4
(15)

5.5
(16)

Reference Number:
Example Varieties

NotePlant: proportion of hermaphrodite plants
low

1[ ]

low to medium

2[ ]medium3[ ]medium to high

4[ ]

high

5[ ]

Plant: proportion of female plants
low

1[ ]

low to medium

2[ ]medium3[ ]medium to high

4[ ]

high

5[ ]

Plant: proportion of male plants
low

1[ ]

low to medium

2[ ]medium3[ ]medium to high

4[ ]

high

5[ ]

TG/276/1
Hemp, 2012-03-28- 26 -TECHNICAL QUESTIONNAIRE
Characteristics
5.6
(17)

Page {x} of {y}Reference Number:
Example Varieties

NotePlant: natural heightvery short

1[ ]very short to short

2[ ]

short

Finola4[ ]short to medium
medium

Uso 315[ ]6[ ]medium to long
long

3[ ]

Ferimon7[ ]long to very long8[ ]very long9[ ]TG/276/1
Hemp, 2012-03-28- 27 -TECHNICAL QUESTIONNAIRE
6.

Page {x} of {y}Reference Number:

Similar varieties and differences from these varieties

Please use the following table and box for comments to provide information on how yourcandidate variety differs from the variety (or varieties) which, to the best of your knowledge,
is (or are) most similar. This information may help the examination authority to conduct its
examination of distinctness in a more efficient way.
Denomination(s) of
Characteristic(s) in Describe the expression
Describe thevariety(ies) similar to which your candidate of the characteristic(s)expression of the
your candidate variety variety differs from thefor the similar
characteristic(s) for
similar variety(ies)variety(ies)your candidate variety
ExamplePlant: natural height
short
long

Comments:

TG/276/1
Hemp, 2012-03-28- 28 -TECHNICAL QUESTIONNAIRE#

Page {x} of {y}Reference Number:

7.

Additional information which may help in the examination of the variety

7.1

In addition to the information provided in sections 5 and 6, are there any additional
characteristics which may help to distinguish the variety?
Yes

[ ]

No

[ ]

(If yes, please provide details)
7.2

Are there any special conditions for growing the variety or conducting the examination?
Yes

[ ]

No

[ ]

(If yes, please provide details)
7.3

Other information
Main use
(a)
(b)
(c)
(d)

8.

bast fibre and woody coreoil seedpharmaceuticals
other(please provide details)

[
[
[
[

]
]
]
]

Authorization for release(a) Does the variety require prior authorization for release under legislation concerning
the protection of the environment, human and animal health?
Yes
(b)

[ ]

No

[ ]

Has such authorization been obtained?
Yes

[ ]

No

[ ]

If the answer to (b) is yes, please attach a copy of the authorization.

#

Authorities may allow certain of this information to be provided in a confidential section of the Technical
Questionnaire.

TG/276/1
Hemp, 2012-03-28- 29 -TECHNICAL QUESTIONNAIRE
9.

Page {x} of {y}Reference Number:Information on plant material to be examined or submitted for examination.

9.1 The expression of a characteristic or several characteristics of a variety may be affectedby factors, such as pests and disease, chemical treatment (e.g. growth retardants or pesticides),effects of tissue culture, different rootstocks, scions taken from different growth phases of atree, etc.
9.2 The plant material should not have undergone any treatment which would affect theexpression of the characteristics of the variety, unless the competent authorities allow orrequest such treatment. If the plant material has undergone such treatment, full details of thetreatment must be given. In this respect, please indicate below, to the best of your knowledge,
if the plant material to be examined has been subjected to:(a)

Microorganisms (e.g. virus, bacteria, phytoplasma)

Yes [ ]No [ ](b)

Chemical treatment (e.g. growth retardant, pesticide)

Yes [ ]No [ ](c)Tissue culture

Yes [ ]No [ ](d)

Other factors

Yes [ ]No [ ]

Please provide details for where you have indicated “yes”.
……………………………………………………………

10. I hereby declare that, to the best of my knowledge, the information provided in thisform is correct:
Applicant’s nameSignature

Date[End of document]