The Plant Variety Protection (PVP) Authority decides when, where and in what quantity and quality the plant 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 :

3000 grams (in only one submission)
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.
The plant 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

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.
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.

The test should be carried out under conditions ensuring normal growth. The size of the plot should be such that plants or parts of plant may be removed for measuring and counting without prejudice to the observation which must be made upto the end of the growing period. Each test should include a minimum of 300 plants which should be divided among 3 replications. Separate plots for observation and for measuring can only be used if they have been subjected to similar environmental conditions.

Test plot details

No. of rows	: 5
Row length	: 5m
Row to row distance	: 45cm(60 cm at hilly location)
Plant to plant distance	: 10cm
No. of replications	: 3

Observations should not be recorded on plants in border rows.
Additional tests for special purpose may be established.

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IV. Methods and observations

The characteristics described in the table of characteristics should be used for the testing of varieties for DUS.
For the assessment of distinctness and stability, observations should be made on 30 plants or parts of plants which should be divided among 3 replications (10 plants per replication). The number of aberrant plant should not exceed 4 in 30.
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 plants), a population standard of 0.5% with an acceptance probability of atleast 95% should be applied. In the case of a sample size of 300 plants, the number of off-types should not exceed 4.
For the assessment of colour characteristics, it is recommended that Royal Horticultural Society (RHS) colour chart be used.

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V. Grouping of varieties

The collection of varieties to be grown should be divided into groups to facilitate the assessment of distinctness. Characteristics, which are suitable for grouping purposes, are those, which are known from experience not to vary, or to vary only slightly, within a variety. Their various states of expression should be fairly evenly distributed throughout the collection.
It is recommended that the competent authorities use the following characteristics for grouping varieties:
(i) Flower: color (characteristic 8)
(ii) Pod: presence of hairs (characteristic 9)
(iii) Pod : color of hairs (characteristic 10)
(iv) Seed : hilum color (characteristic 18)
(v) Plant: time of maturity (characteristic 20)

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VI. Characteristics and symbols

To assess distinctness, uniformity and stability, the characteristics and their states as given in the Table of Characteristics should be used.
Notes (1 to 9), for the purposes of electronic data processing, are given opposite the states of the different characteristics.
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.

The optimum stage of plant growth for assessment of each characteristic is indicated in the sixth column of table of characteristics. These are explained below:

Code	Description
10	Cotyledons completely unfolded
65	Full flowering: about 50% of flowers openMain period of flowering
66	About 60% of flowers open
77	About 70% of pods have reached final length (15-20mm) Advanced pod filling
85	Advanced ripening: about 50% of pods are ripe; beans final colour, dry and hard. Main period of pod and seed ripening
89	Fully maturity: approximately all pods are ripe; beans final colour, dry and hard (= Harvest maturity)Majority of pods are ripe; beans final colour, dry and hard

Type of assessment of characteristics indicated in column 7 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

S.No.	Characteristics	States	Note	Example Varieties	Stage	Type of assessment
1. (*)	Hypocotyl:antho-cyanin colouration	absent present	1 9	NRC 37, PK 472 JS 335, NRC 12	10	VS
2. (*) (+)	Plant: Growth Type	determinate semi-determinate indeterminate	1 2 3	JS 71-05, NRC 7 NRC 37, MACS 58 Type 49, Improved Pelican		VG
3. (*)	Plant: time of flowering (50% plants with at least one flower open)	early medium late	3 5 7	JS 71-05, NRC 7 JS 335, NRC 37, JS 80-21 Hardee, MACS 124		VG
4. (+)	Leaf: Shape of lateral leaflet	lanceolate triangular pointed ovate rounded ovate	1 2 3 4	JS 90-41, PK 308, PK 1024 NRC 37,Type 49 JS 71-05	65	VG
5.	Leaf: size of lateral leaflet	small medium large	3 5 7	JS 71-05 JS 335, NRC 37 MACS 124, MACS 450	65	VG
6.	Leaf: intensity of green color	light medium dark	3 5 7	NRC 37, Indira Soya 9, Type 49 Hara Soya, JS 90-41 JS 71-05, JS 335, LSb 1	65	VG
7. (*) (+)	Plant: growth habit	erect semi-erect semi-erect to horizontal	1 3 5	NRC 2, Punjab 1,NRC 12 PK 472, JS 335, NRC 37 Type 49
8. (*)	Flower: colour	white violet	1 2	NRC 37, PK 472 JS 335, NRC 12	66	VG
9. (*)	Pod: presence of hairs	absent present	1 2	JS 71-05, JS 335 NRC 37, PK 472, NRC 12	77	VG
10. (*)	Pod: colour of hairs	grey tawny	1 2	PK 472, JS 75-46 NRC 37, MACS 58	77	VS
11 (*)	Plant : height	short medium tall	3 5 7	JS 71-05, JS 2, LSb 1 JS 335, NRC 12 JS 75-46, MACS 58, NRC 37	85	MS
12.	Pod: intensity of brown color	light medium dark	3 5 7	PK 472, JS 75-46, Hardee MACS 58, PK 416, JS 71-05 MACS 13, PS 1029	85	VS
13.	Seed: size (100 seed weight)	small (< 10 g) medium (10-13 g) large (> 13 g)	3 5 7	Type 49, Punjab 1 JS 335, NRC 2 JS 71-05, NRC 12	89	MG
14.	Seed : shape	spherical spherical flattened elongated elongated flattened	1 2 3 4	JS 71-05, PK 262, Monetta Type 49 MACS 13 MAUS 32, Indira Soya 9	89	VG
15. (*)	Seed: ground color of testa (excluding hilum)	yellow yellow green green brown black	1 2 3 4 5	JS 335, PK 472 JS 90-41 Hara Soya VLS 1, Kalitur, JS 76-205	89	VS
16.	Seed coat: lusture	shiny intermediate dull	3 5 7	MACS 450, VLS 47 Monetta, MAUS 32 PK 327, PS 1029, PK 472	89	VG
17. (+)	Seed: colouration due to peroxidase activity in seed coat	absent present	1 2	NRC 12, NRC 7 MACS 450, PK 416	89	MG
18. (*)	Seed: hilum color	grey yellow brown imperfect black black	1 2 3 4 5	NRC 2 Shilajeet NRC 37, PK 416, Hardee Pusa 16 PS 1029, KHSb 2	89	VS
19.	Seed: colour of hilum funicle	same as testa different to testa	1 2	MACS 58, Pusa 22 NRC 7, NRC 12	89	VS
20. (*)	Plant: time of maturity	early medium late	3 5 7	LSb 1, KHSb 2 JS 335, NRC 37 Hardee, Birsa Soy 1, Type 49	89	VG

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VIII. Explanation on the table of characteristics

Characteristic: 2. Plant: growth type

Layout: This characteristic should preferably be assessed in a special trial with 3 or 4 replicates of 20 plants each with about 9 cm between plants in the row. Any border effect must be avoided.

Plant material: Candidate and example varieties must be grown in groups according to their earliness at maturity (characteristic 20)

Observations: At the beginning of flowering time (1 flower at any level of the main stem), the apex of the plant must be identified with a mark.

At maturity (free kernels in the pod), the number of nodes between the mark and the top of the plant is counted. The average numbers per variety gives-in comparison with standard varieties-the state of expression of the characteristics.

In addition, the characteristic “Size of the terminal leaf” could also be considered to separate more clearly the state of expression “determinate” (Note 1) from other states. The terminal leaf on the main stem of determinate varieties is more or less equal to other leaves at lower levels. For other types, the terminal leaf is clearly smaller

Characteristic 4. Leaf: shape of lateral leaflet

Characteristic 7. Plant: growth habit

Characteristic 17. Seed: coloration due to peroxidase activity in seed coat

20 seeds per variety should be tested.

The seed coat of the seed should be removed carefully so that no piece of cotyledon remains. To facilitate this procedure, the seed should be placed in water for 2 hours.

The seed coat should be placed in a cell box or in tubes (one seed per tube) and 3 to 4 cm³ of 0.5% Guayacol solution should be added. The 0.5% Guayacol solution should be stored in the refrigerator for a period of not longer than 2 months. After having left it at room temperature for one day or more, it can no longer be used.

After 10 minutes waiting time, one drop of 0.1% H₂O₂ solution should be added.

The solution changes to dark red/brown color for a positive reaction or remains without color for a negative reaction. In order to check the 0.5% Guaycacol solution, it is advisable to include some seeds of a reference variety with a positive reaction. The recording of this reaction must be done not longer than 10 seconds after the H₂O₂ was added. It is very important that the observation must not be done longer than 60 seconds because it could lead to wrong results.

The cell box or the tubes could be softly shaken for a better reaction. For a better recording of the observation, the tubes or the cell box should be placed over a white surface

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IX.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: Glycine max (L.) Merrill SOYBEAN
2. Applicant (Name and address) :
3. Proposed denomination or breeder's reference
4. Information on origin, maintenance and reproduction of the variety 4.1 Genetic origin and breeding method Is the variety a GMO and does it require prior authorization for release under legislation concerning the protection of the environment, human and animal health? Yes [ ] No [ ] Has such an authorization has been obtained from the nodal agency (DBT, GOI/Others)? Yes [ ] No [ ] If the answer to that question is yes, please attach a copy of such an authorization. 4.2 Other information
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 (8)	Flower: color (at full flowering)		White Violet		NRC 37, PK 472 JS 335, NRC 12		1 [ ] 2 [ ]
5.2 (9)	Pod: presence of hairs (at advanced pod filling		absent present		JS 71-05, JS 335 NRC 37, PK 472, NRC 12		1 [ ] 2 [ ]
5.3 (10)	Pod: color of hairs (at advanced pod filling		Grey Tawny		PK 472, JS 75-46 NRC 37, MACS 58		1 [ ] 2 [ ]
5.4. (18)	Seed: hilum color (full maturity)		Grey Yellow Brown Imperfect black Black		NRC 2 Shilajeet NRC 37, PK 416, Hardee Pusa 16 PS 1029, KHSb 2		1[ ] 2[ ] 3[ ] 4[ ] 5[ ]
5.5. (20)	Plant: time to maturity (full maturity)		Early Medium Late		LSb 1, KHSb 2 JS 335, NRC 37 Hardee, Birsa Soybean 1, Type 49		3 [ ] 5 [ ] 7 [ ]
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 (specific biotype, if any) 7.2 Special conditions for the examination of the variety, if any 7.3 Other information
8. 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 { } No { } (b) Has such authorization been obtained? Yes { } No { } If the answer to that question is ‘Yes’, please attach a copy of such an authorization

Comments on the draft of National DUS Test Guidelines of Soybean

1. Minor modifications have been made from chapter I to VI to bring uniformity of format among the DUS test guidelines of various crops.

2. In the Table of characteristics (chapter VII), characteristics and their states of expressions have been assigned separate columns and an additional column of type of assessment has been incorporated at the extreme right.

3. In the fifth column of this table the stage of observation recording of characters has been assigned the code and the description of the code has been provided in chapter VI.

4. Kindly provide the range of measurement (days, cm etc.) for each state of expressions of characteristics 3 (plant : time of flowering), 11 (plant : height) and 20 (plant : time of maturity).

Kindly go through the edited draft critically and give your suggestions. May I request your input by May 25, 2003

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