INDIA

ITG/09

25-7-2003

National Guidelines

for the Conduct of Tests for

Distinctness, Uniformity and Stability

WHEAT

Triticum aestivum L
      

Contents
  I.           Subject

II.           Material required

III.          Conduct of tests

IV.         Methods and observations

V.          Grouping of varieties

VI.         Characteristics and symbols

VII.        Table of characteristics

VIII.        Explanations on the table of characteristics

IX.          Literature

X.           Technical questionnaire

XI.          Annexure

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I.   Subject

These test guidelines apply to all varieties, hybrids and parental lines of Wheat (Triticum aestivum L.)

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II.   Material required

  1. The Plant Variety Protection (PVP) Authority decides when, where and in what quantity and quality the seed material required for testing the variety is to be delivered.  Applicants submitting material from a country other than India must make sure that all customs formalities are complied with.  The minimum quantity of seed to be supplied by the applicant should be :

    Variety and hybrid     -           3000 gram
                                                                         (in one submission only)
    Parental lines           -         1500  gram  
      

  2. The seed should meet the minimum requirements for germination capacity, moisture content and physical purity prescribed for certified seed in India . Especially for storage, which requires a higher standard, the applicant should state, the actual germination capacity which should be as high as possible but not less than 90%. The seed supplied should be visibly healthy, not lacking in vigour or affected by any important pest or disease.  
     

  3.  In addition, at least 200 ears should also be submitted. The ears should be well developed and not obviously affected by any pest or disease.  They should contain a sufficient number of viable seeds to establish satisfactory row of plants for observation.  
     

  4. The seed material must not have undergone any treatment unless the competent authorities allow or request such treatment.  If it has been treated, full details of the treatment must be given.

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III.   Conduct of tests

  1. The minimum duration of tests should normally be two independent similar growing seasons with reference to the ecosystem of the variety submitted for DUS test.
      

  2. The test should normally be conducted at two test locations. If any important characteristics of the variety can not be seen at these places, the variety may be tested at an additional place.
      

  3. The field tests should be carried out under conditions ensuring normal growth. The size of the plots should be such that plants or parts of plants may be removed for measurement and counting without prejudice to the observations which must be made upto the end of the growing period. Each test should include about 2000 plants, which should be divided among three replications. Separate plots for observation and for measurement can only be used if they have been subjected to similar environmental conditions.  
      

  4. Test plot design  

    Number of rows : 06  
    Row length : 06
    Row to row distance : 30
    Plant to plant distance : 05
    Number of replications : 03

     

  5.   Observations should not be recorded on plants in border rows.  
      

  6. Additional tests for special purpose may be established.

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IV.   Methods and observations

  1. The characteristics described in the table of characteristics should be used for the testing of varieties, inbred lines and hybrids for DUS.  
      

  2. For the assessment of distinctness and stability observations should be made on 15 plants or parts of 15 plants which should be divided among 3 replications (5 plants per replication).  
      

  3. For the assessment of uniformity of characteristics on the plot as a whole (visual assessment by a single observation of a group of plants or parts of plant), the number of aberrant plants or parts of plant should not exceed 5 in 2000.
      

  4. For the assessment of uniformity of characteristics on single ear-rows, plants or parts of plant (visual assessment by observation of a number of individual ear-rows, plants or parts of plants) the number of aberrant ear-rows, plants or parts of plants should not exceed 3 in 100.
      

  5.   For the assessment of colour characteristics, it is recommended that Royal Horticultural Society (RHS) colour chart should be used.
      

  6. Additional tests for special purposes may be established

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V.   Grouping of varieties

  1. The collection to be grown should be divided into groups to facilitate the assessment of distinctness. Characteristics that are suitable for grouping purposes are those which are known from experience not to vary, or to vary only slightly, within a variety and which in their various states are fairly evenly distributed within the collection.  
      

  2. It is recommended that the competent authorities use the following characteristics for grouping varieties.

    i.                    Flag leaf : anthocyanin colouration of auricle (characteristic 4)

    ii.                  Time of ear emergence (characteristic 7)

    iii.                Plant : length (including awns/scurs) (characteristic 14)

    iv.                 Straw : pith in cross section (half way between base of ear and stem node below) (characteristic 15)

    v.                   Awns or scurs : presence (characteristic 19)

    vi.                 Outer glume : pubescence (characteristic 23)

    vii.               Ear : colour (characteristic 24)

    viii.             Season type (characteristic 36)

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VI. Characteristics and symbols

  1. To assess distinctness, uniformity and stability, the characteristics and their states as given in the Table of Characteristics should be used.
      

  2. Notes (1 to 9), for the purposes of electronic data processing, are given opposite the states of each characteristics.
      

  3. Legend :

    (*)        Characteristics that should be used every growing period for the examinations of all varieties and should always be included in the description of the variety, except when the state of expression of a preceding characteristic or regional environmental conditions render this impossible.

    (+)       See Explanations on the table of characteristics in Chapter VIII.  
      

  4. The optimum stage of development of plant growth for the assessment of each characteristics is indicated by a number in the sixth column of table of characteristics. The stages of development denoted by each number are described below : 

    Stage Code

    General Description

    Germination

    09

    Leaf just at coleoptile tip

    10

    First leaf through coleoptile

    11

    First leaf unfloded

    Tillering

    25

    Main shoot and 5 tillers

    26

    Main shoot and 6 tillers

    27

    Main shoot and 7 tillers

    28

    Main shoot and 8 tillers

    29

    Main shoot and 9 tillers

    Booting 

    40

    Early boot stage

    41

    Flag leaf sheath extending

    43

    Boots just visibly swollen

    47

    Flag leaf sheath opening

    49

    First awns visible  [in awned forms only]

    Inflorescence

    50-51

    First spikelet of inflorescence just visible

    52

    1/4 of inflorescence emerged

    Anthesis

    60-61

    Beginning of anthesis

    64-65

    Anthesis half-way

    68-69

    Anthesis complete

    Milk development

    73

    Early milk

    75

    Medium milk

    77

    Late milk

    Dough development

    83

    Early dough

    85

    Soft dough

    87

    Hard dough

    Ripening

    91

    Caryopsis hard (difficult to divide by thumb-nail)

    92

    Caryopsis hard (can no longer be dented by thumb-nail)

    93

    Caryopsis loosening in daytime

    94

    Over-ripe, straw dead and collapsing

     

  5. Type of assessment of characteristics indicated in column seven of table of  characteristics is as follows :

    MG : Measurement by a single observation 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 observations of individual plants or parts of plants
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VII. Table of characteristics

Sl.No.

Characteristics

States

Note

Example Varieties

Stage of observation

Type of assessment

1.
(*)
(+)

Coleoptile :
Anthocyanin colouration

absent
present

1
9

DDK 1001
HD 2009

09-11

 

VS

2.
(*)   (+)

Plant : growth habit

erect
semi-erect
intermediate
semi-prostrate
prostrate

1
3
5
7
9

PBW 175
PBW 343
K 8027
SK 184 EC367999
EC 364998

25-29

 

VG

3.
(*)

Foliage : colour

pale green

green

dark green

1

5

9

Motia, C 306
WL 711, DWR 162
WL 410, DL 153-2

40-45

VG

4.
(*)

Flag leaf : anthocyanin colouration of auricles

absent
medium
very strong

1
5
9

K 9644
GW 89
CPAN 1922  

49-51

 

VG

5.
(*)

Flag leaf : hairs on auricle

absent

medium

strong

3

5

7

K9162,K 9006
NIAW 34, PBW 343
PBW 54, WH 711

49-51

 

VG

6.
(+)

Plant : flag leaf attitude

erect
semi-erect
drooping

1
3
5

PBW 343
HD 2687

47-51

 

VG

7.
(*)

Time of ear emergence (first spikelet visible on 50% of ears)

very early
early
medium
late
very late

1
3
5
7
9

 

50-52

 

VG

8.  
(*)

Flag leaf: waxiness of sheath

absent
weak  
medium  
strong  
very strong

1  
3  
5  
7  
9

NW 1014  
K 9107  
DT 46

60-65

 

VG

9.
(*)

Flag leaf: waxiness of blade

absent
weak  
medium  
strong  
very strong  

1  
3  
5  
7  
9

HI 7483

60-65

VG

10.  
(*)

Ear: waxiness

absent
weak  
medium  
strong  
very strong  

1  
3  
5  
7  
9

 

60-69

 

VG

11.  
(*)

Culm: waxiness of neck (Peduncle)

absent  
weak  
medium  
strong  
very strong

1  
3  
5  
7  
9

 

60-69

 

VG

12.

Flag leaf : length

short  
medium  
long

1  
5  
9

HD 1949  
C 306  
GW 496

70-80

MS

13.

Flag leaf : width

narrow  
medium  
broad

1  
5  
9

NP 824  
PBW 343  
H 961

70-80

MS

14.
(*)

Plant : length (including awns/scurs)

very short  

short  
medium  
long  
very long

1  

3  
5  
7  
9

Lal Bahadur  
HD 2009  
PBW 343  
C 306  
IC 138905

75-92

 

MS

15. 
(*)  
(+)

Straw : pith in cross section (halfway between base of ear and stem node below)

thin
medium  
thick

3  
5  
7

 

80-92

 

VS

16.  
(*)  
(+)

Ear : shape in profile

tapering  

parallel sided  

semi-clavate  
clavate  

fusiform

1  

2  

3  
4  

5

K 88, VL 804  
NW 1014, HI 1500  

SK 13, SK 43  
PI 164160

92

 

VS

17.  
(*)  
(+)

Ear : density

very lax  
lax  
medium  
dense  
very dense

1  
3  
5  
7  
9


Sonalika  
DL 153-2  
GW 192  
Raj 215

80-92

 

VS-MS

18.
(*)

Ear : length (excluding awns and scurs)

very short  
short  

medium  
long  
very long

1  
3

5
7  
9

 
PBW 34, NI 917  
C 306  
DL 803-3  
K 9006, VL 616

80-92

 

MS

19.  
(*)  
(+)

Awns or scurs: presence

both absent  
scurs present  
awns present

1  
2  
3

NP 4  
Ridley  
WH 147

80-92

 

VG

20. 
(*)

Awns : length

very short  
short  
medium  

long  

very long

1  
3  
5

7  

9

 
HD 2009  
KRL 1-4, NIAW 34  
MACS 2938  
PDW 215

80-92

 

VG or MS

21.

Awn : color

white

brown  
black

1

2  
3

UP 2425, WH 147  
WL 711
K 9644

80-92

 

VS

22.

Awn : attitude

appressed  
spreading

1  
2

 

80-92

VS

23.
(*)

Outer glume :  
 pubescence

absent  
medium  
very strong

3  
5  
7

WH 542  
K 9644

90-92

VS

24. 
(*)

Ear: colour

white  
light brown  
dark brown  
black  

1  
2  
3  
4

PBW 343  

HD 2329

90-92

VG

25.  
(+)

Lower glume: shoulder width (spikelets in mid-third of ear)

absent or very narrow  
narrow  
medium  
broad  
very broad  

1  

3  
5  
7  
9

80-92

 

VS

26. 
(*)  
(+)

Lower glume: shoulder shape (as for 25)

sloping  
round  

straight  

elevated  

strongly elevated with 2nd point present

1
3  

5  

7

9

Kalyansona  
C 306, HD 1982  
HD 2643, K68  
HYB 65, JOB 666

80-92

 

VS

27.

Lower glume: beak length (as for 25)

very short  
short  
medium  
long  
very long

1
3  
5  
7  
9

CPAN1922  
JU 12  
HUW 234  
HYB 65  
CPAN1676

80-92

 

VS

28.  
(+)

Lower glume: beak shape (as for 25)

straight  
moderately curved  
stronglycurved
geniculate

1  
2  

3  
4

HB 208  
NP 4  

HUW 234  
HI 617, HP 1744

80-92

 

VS

29.
(*)

Peduncle : length

short  
medium  
long

1  
5  
9



Kharchia local

80-92

MS

30. (*)

Grain: color

white

amber
red

1  

2  
3

Safed Lerma  
Sonalika  
Kharchia 65

92

 

VG

31.
(*)  
(+)

Grain: coloration with phenol

none

light  
medium  
dark  
very dark

1

3  
5  
7  
9  

Gulab, PDW 215  
VL 719, HUW 234  
WL 711, PBN 51

92

 

VS

32.  
(*)  
(+)

Grain : shape

round  
ovate  
oblong  

elliptical

1  
2  
3  

4

SK 7,SK56  
Lerma Rojo  
LOK 1, Kalyansona
Meghdoot

92

VG

33.

Grain : crease

shallow  
medium  
deep

3  
5  
7

HD 2277
CPAN3004  
HB 208  

92

VG

34.  
(*)  
(+)

Brush hair : length

absent/short  
medium  
long

3  
5  
7

HD 2402  
HD 2610  
Girija

92

VG

35.  
(*)

Seed : size (weight of 1000 grains)

small  
medium  
large  
very large

3  
5  
7  
9

CPAN1676  
HS 277  
NP 4  
Bawaji

92

VG or M

36.
(*)

Seasonal type

winter type  
alternative type  
spring type

 

1
2  

3

 

 

VG

 

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VIII. Explanations on the table of characteristics

 Characteristic 1. Coleoptile : anthocyanin colouration

Method for Determination of Anthocyanin Colouration of coleoptile
      

Number of grains per test: 20 grains for distinctness, 100 grains for homogeneity
   
Preparation of grains: Set up non-dormant grains on moistened filter paper covered with a petri dish lid during germination
    
Place: Laboratory or greenhouse
   
Light: After the coleoptiles have reached a length of about 1 cm in darkness, they are placed in artificial light (daylight equivalent), at 15,000 lux continuously for 3-4 days
   
Temperature: 15 to 200C
    
Time of recording: Coleoptiles fully developed (about 1 week) at stage 09-11
    
Scale of recording: See characteristics 1
    
Note: At least, two of the example varieties should be included as a control when testing for distinctness.
    

Characteristic 2. Plant : growth habit 

The growth habit should be assessed visually from the attitude of the leaves and tillers. The angle formed by the outer leaves and the tillers with an imaginary vertical axis should be used

Characteristic 6. Plant : flag leaf attitude

1
erect

3
semi-erect

5
Drooping


Characteristic 15. Straw : pith in cross section (half way between base of ear and stem node below)

3
thin

5
medium

7
thick

All stems of the plant should be checked and the strongest expression per plant recorded  

Characteristic 16. Ear : shape in profile

1
tapering

2
parallel sided

3
semi-clavate

4
clavate

5
fusiform

 

Characteristic 17. Ear : density

 1
very lax

3
lax

5
medium

7
dense

9
very dense


Characteristic 19. Awns or scurs : presence

 1
both absent		 2
scurs present		 3
awns present

 

Characteristic 25. Lower glume : shoulder width (spikelet in mid-third of ear)

1
absent or very narrow

3
 narrow

5
 medium

7
 broad

9
 very broad

 

Characteristic 26. Lower glume : shoulder shape (spikelet in mid-third of ear)

1
 sloping

3
 round

5
 straight

7
elevated

9
 strongly elevated with 2nd point present

 

Characteristic 28. Lower glume : beak shape (spikelet in mid-third of ear)

1
 straight

2
 moderately curved

3
 strongly curved

4
 geniculate

 

Characteristic 31. Grain : colouration with phenol

Method for Determination of Phenol Reaction
    
Number of grains per test: 20 grains for distinctness, 100 grains for homogeneity. The grains should not have been treated chemically
  
Preparation of grains: Soak in tap water for 16 to 20 hours, drain and remove surface water, place the grains with crease downwards, cover dish with lid
 
Concentration of solution: 1 per cent Phenol-solution (freshly made up)
  
Amount of solution: The grains should be about 3/4 covered
  
Place: Laboratory
  
Light: Daylight - out of direct sunshine
 
Temperature: 18 to 200C
   
Time of recording: 4 hours (after adding solution)
   
Scale of recording: See characteristics 31 in the Table of characteristics
   
Note: At least, two of the example varieties should be included as a control.

 

Characteristic 32. Grain : shape

1
 round

2
 ovate

3
 oblong

4
 elliptical

 

Characteristic 34. Brush hair : length

3
 short

5
 medium

7
 long

 

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IX. Literature

PAYNE, P.I., LAWRENCE , G.J., 1983: "Catalogue of Alleles for the Complex Gene Loci, Glu-A1, Glu-B1, Glu-D1, Which Code for high Molecular Weight Subunits of Glutenin in Hexaploid Wheat". Cereal Research Communications 11, p. 29-35.

PAYNE, P.I., 1987: "Genetics of Wheat Storage Proteins and the Effect of Allelic Variation on Bread-Making Quality". Annual Review of Plant Physiology 38, p.141-153.

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X. Technical questionnaire 

 

 

Reference Number
(not to be filled in by the applicant)
 

Technical questionnaire  
to be completed in connection with an application for plant breeders' rights  

 
1. Species:                                                            Triticum aestivum L.
                                                                                        Wheat

 

 
2.      Applicant (Name and address) :  

 

 

 
3.  Proposed denomination or breeder's reference

  

 

  

4.  Information on origin, maintenance and reproduction of the variety

5. Characteristics of the variety to be indicated (the number in brackets refers to the corresponding characteristic in Test Guidelines; please mark the state of expression which best corresponds).  
S.No. Characteristics States Example varieties  Notes

5.1
(4)

 Flag leaf : anthocyanin colouration of auricle  

absent
medium
very strong  

K 9644
GW 89
CPAN 1922  

1 [ ]
5 [ ]
9 [ ]

5.2
(7)

 Time of ear emergence (first spikelet visible on 50% of ears)  

very early  
early  
medium  
late  
very late

 

1 [ ]  
3 [ ]  
5 [ ]  
7 [ ]  
9 [ ]

5.3
(14)		 Plant : length (including awns/scurs)

very short  
short  
medium  
long  
very long

Lal Bahadur
 
HD 2009  
PBW 343  
C 306  
IC 138905

1[ ]  
3[ ]  
5[ ]  
7[ ]  
9[ ]  

5.4.
(15)

Straw : pith in cross section (halfway between base of ear and stem node below)

thin  
medium  
thick

  

3[ ]  
5[ ]  
7[ ]  

5.5.
(19)		 Awns or scurs: presence

both absent  
scurs present  
awns present

NP 4  
Redley  
WH 147  

1[ ]  
2[ ]  
3[ ]  

5.6
(23)		 Outer glume : pubescence

labsent  
medium
very strong  

WH 542  
K 9644  

3 [ ]  
5 [ ]  
7 [ ]  

5.7
(24)		 Ear: colour  

white  
light brown  
dark brown  
black  

PBW 342

HD 2329

1[ ]  
2[ ]  
3[ ]  
4[ ]  

5.8.
(36)		 Seasonal type winter type  
alternative type  
spring type 		   1[ ]  
2[ ]  
3[ ]  
6. Similar varieties and differences from these varieties  

Denomination of
Similar variety

Characteristic in
which the similar variety is different 0)

State of expression of similar Variety

 State of expression of candidate variety
                          
0)  In the case of identical states of expressions of both varieties, please indicate the size of the difference.  

7. Additional information which may help to distinguish the variety

 

      7.1 Resistance to pests and diseases (indicate races, if any)

 

 

      7.2 Special conditions for the examination of the variety, if any

 

      
        7.3 Other information

 
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