DUS Test Guidelines for Maize

DUS Testing Guidelines for Maize

Ph.D. (Genetics & Plant Breeding)Reg. No. : 04-00899-2011

Course Code: GP 691

Presented by: K. Satish

CONTENTS

Maize

• Introduction

• Production constraints

• Importance of maize

PPV & FR Act, 2001

• Objectives

• Criteria for protection

• Benefits

DUS Testing

• Assessment of DUS

• Characteristics and Symbols

• Technical questionnaire

• Table of characteristics of maize

Conclusions

Future thrust

Introduction

1

Taxonomically, Zea mays L. belongs to tribe Maydeae, family Poaceae and genus

Zea (Doebley et al., 2004). The center of origin of Zea mays L. (2n=20) is reported to be Mexico and Central

America (Chopra et al., 2002)

The main maize growing countries include U.S.A., China, Brazil, European Union,

Ukraine, Argentina, India and Mexico.

Maize is the third most important cereal crop in India after rice and wheat. It

accounts for 9 per cent of total food grain production in the country.

Nine states viz. Karnataka, Andhra Pradesh, Tamil Nadu, Rajasthan, Maharashtra,

Bihar, Uttar Pradesh, Madhya Pradesh and Gujarat account for 85 per cent of

India’s maize production and 80 per cent of area under cultivation (FAO, 2013)

Table: 1 Comparison of Area, Production and Yield of Maize, 2013-14

Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation, GOI (2013-14) USDA (2013-14)

State Area under cultivation

Million Hectare

ProductionMillion tonnes

YieldKg / Hectare

World 177.4 960.2 5508India 9.43 23.35 2583Gujarat 0.46 0.69 1501

2

• Maize is a monoecious plant.

• Male inflorescence called tassel

• Female inflorescence known as silk

• Generally protandrous

• Each floret contains three anthers and each plant produces about 3-5 million pollen grains.

• Wind borne nature of the pollen and protandry leads to cross pollination but, there may also be reported 5 % self pollination.

Plant Morphology of Maize

Chopra et al., 20023

Importance of Maize

Top ranking cereal in terms of production and productivity globally

Highest potential of carbohydrate production per unit/day (62.2 %)

Great significance as human food, animal feed and industrial products

Grown in unmatched diversity of environments

Queen of cereals

4

Maize Utilization for Various Purposes

Source of human food (24%), animal feed (11%), poultry feed (52%), starch (11%), brewery (1%) and seed (1%)

In addition more than 500 industrial products like starch, oil, protein, alcohol, acetic acid, sucrose, glucose, paper, rayon, dyes, synthetic rubber, resin etc. are being manufactured from it.

5 Ranum et al., 2014

PPV & FR Act, 2001

Government of India enacted the Plant variety protection & Farmer’s Rights Act on Oct. 30th 2001 Plant variety protection under PPV & FR Act, provides legal protection to the

plant variety

Article 27.3(b) – Member country to provide protection either by a patent or

Sui generis system or in combination

To a breeder in the form of Plant Breeder’s Rights

To a farmers in the form of Farmer’s Rights

To a Researcher in the form of Researcher’s Rights PPV & FR Act, 2001 was enacted by Indian Government based on UPOV Convention Act.

PPV & FR Authority, India6

Conti…

Objectives Registrable plant varieties

New Variety Extant Variety Farmer’s Variety Essentially Derived Varieties (EDV)

Criteria for protection Novelty– [(Article 15.3(a)] variety was not sold out before 1 year of registration Distinctness- [(Article 15.3(b)] at least one essential character Uniformity- [(Article 15.3(c)] sufficiently uniform Stablilty- [(Article 15.3(d)] not susceptible to change after repeated propagation

Who can apply?

Benefits7

DUS Test

Why? Essential requirement for registration of new variety(s) under PPV & FR Act, 2001 Variety be examined for compliance with Distinctiveness, Uniformity and Stability

criteria needed for registration.

Sujay et al., 20018

How?Involves comparison of new (candidate) variety against existing varieties for recording a number of morphological/physiological characters, by growing new and existing varieties side-by-side.

What is DUS Test?Plant variety protection can only be granted in respect of a new plant variety after examination, for that UPOV Convention Acts laid down criteria viz. novelty (N), distinctness (D), uniformity (U) and stablity (S) ‘DUS’ or ‘NDUS’ in short.

Must be clearly distinguishable for at least one essential

character from the varieties of common knowledge in

India and out side.

Essential character is a heritable trait which is determined

by one or more genes or other heritable determinants that

contribute to the principal features, performance or value of

the plant variety.

DISTINCTNESS

9

Uniform, when subjected to the variation that

may be expected and allowed due to the

specific nature of the reproduction of the crop

such as vegetative, self- and cross-pollinated.

UNIFORMITY

STABILITY

Stable when all essential characters remain

unchanged after repeated propagation or a

specified cycle of propagation.

10

Guidelines for Conduct of Test for Distinctiveness, Uniformity and StabilityOnMaize

UPOV Species Code: ZEAAA_MAY(Zea mays L.)

Protection of Plant Varieties and Farmer’s Rights Authority (PPV & FRA)Department of Agriculture & Co-operationMinistry of Agriculture, Government of India11

Requirements for DUS Testing

Planting material Candidate Variety Reference Collection

Staff Scientific/Technical/Secretarial/Field

Facilities Land/Gardens(flowers)/Orchard(trees) DUS Test Guidelines DUS test centres Laboratory with equipments RHS colour Chart Data base of Extant/Notified varieties

(NORV and IINDUS) Software for database / analysis

(OOS, PLOD_PLAN, ANALIST 3.1, DUSTNT)

12 source: www.plantauthority.gov.in

Rice Wheat Maize Sorghum Pearlmillet

DRR, Hyderabad DWR, Karnal DMR, New Delhi (Kharif)

NRCS, Hyderabad

MPKV, Rahuri

CRRI, Cuttack IARI, Indore ANGRAU, Hyderabad (Kharif and Rabi)

MPKV, Rahuri

Mandore

IARI, Karnal CSUAT, Kanpur RAU, Dholi (Kharif and Rabi)

GBPUA&T, Pantnagar

AAU, Jorhat UAS, Dharwad VPKAS, Almora, Uttarakhand (Hill region Kharif )

TNAU, Coimbatore

PAU, Ludhiana

DUS testing centre in India for cereals

source: www.plantauthority.gov.in

Table: 2 DUS test centres

13

Planting material for DUS testing

Crop Quantity (g)Maize 3000(varieties & hybrids)

1500 (parental lines)Wheat 3000(varieties & hybrid)Rice 3000 (varieties & hybrids)

1500 (parental lines)Sorghum 1500 (varieties & hybrids)

500 (parental lines)Pearl millet 1000 (varieties & hybrids)

500 (parental lines)

Annonymous, 2007

A. Germination capacity

i Inbred line & single cross hybrids

80% (minimum)

ii Varieties & double cross hybrids

90% (minimum)

B. Moisture content 10-12% (maximum)

C. Physical purity 98%(minimum)

Table - 3: Seed Quantity for Test Table - 4: Seed Standards, Moisture and Physical purity

14

Table - 5: Test plot design for Maize

● Number of rows i) Inbred line & single cross hybrids 4 ii) Varieties & other hybrids 8

● Row length 6 m● Row to row distance 75 cm ● Plant to plant distance 20 cm● Number of replication 3

Number of growing cycleTest locationCondition favoring normal growth and expression.Observations should not be recorded on plants in border rows.Additional test for special purpose may be established.

New Delhi PPV & FR Authority

Conduct of Test

15

Assessment of DUS

Distinctness Distinctness is established by comparing the candidate variety with existing varieties/reference

collections. Consistent differencesClear differences

If similar state of expression - variety is not distinct

UniformityPopulation standard of 3 % and an acceptance probability of at least 95 % should be

applied. In sample size of 100 plants, 3 off-types are allowed.Same should applied to clear case of out-crossed plants in inbred lines as well as single cross

hybrids.For visually assessed characteristics, the uniformity of the candidate variety is checked against

known comparable varieties.

UPOV, 200916

Conti…

Stability Require 2-3 years to perform a test of stability which leads to the same certainty as

the testing of the distinctness and homogeneity.

Submitted sample to be homogenous during the test, the material can also be considered to be stable.

In case of doubt inbred and open pollinated varieties may tested by growing a further generation, or by testing a new seed stock.

All observation on ear should be made on the upper well-developed ear.

Latest Royal Horticultural Society (RHS) colour chart should be used.

17Selvi et al.,2013

1. Reference collection Similar to Candidate variety

2. Grouping of varieties To facilitate the assessment of distinctness, varieties to be divided into groups

based on grouping characteristics. It is recommended that the testing organizations / centers use the following

characteristics for grouping varieties (Maize):

a) Tassel: Time of anthesis (cha # 4)

b) Ear: Time of silk emergence (cha # 11)

c) Ear: Anthocyanin coloration of silks (cha # 12)

d) Plant: Length (cha # 15.1/15.2)

e) Ear: Type of grain (cha # 22)

UPOV, 200918

Characteristics and Symbols

To assess distinctness, uniformity and stability, the characteristics and their states should be used.

1. Notes (scale 1 to 9) should be use for the purpose of electronic data processing.

2. Legend : (*) Characteristics that should be observed during every growing season

on all varieties, included in the variety descriptions, except when the state of expression of a preceding characteristics show this inappropriate.

(+) Characteristics is illustrated by explanation or drawing.

(S) Possible segregation in three way cross and double cross hybrids.

PPV & FRA, 200719

Table - 6: Range of characters as per PPV & FR Act

Crop No. of characters Characters with * mark

Wheat 33 20

Rice 30 5

Sunflower 40 13

Peas 58 27

Maize 31 (41- as per UPOV)

8 (16)

Onion 27 12

* Mandatory characters to record.

20

Table - 7: Decimal code for the growth stage

Stage code General Description

00 Dry seed

12 2 leaves unfolded

14 4 leaves unfolded

51 Inflorescence just visible

61 Beginning of anthesis

65 Anthesis halfway

71 Caryopsis watery ripe

75 Medium milk

85 Soft dough

92 Caryopsis hard ( can no longer be dented by thumbed nail)

93 Caryopsis loosening day time.

New Delhi (Anonymous 2007)

The optimum stage of plant growth assessment of each characteristics is indicated by decimal code.

21

Technical Questionnaire

A Performa containing technical questionnaire about the candidate variety is to be submitted by the applicant seeking plant variety protection. The applicant is asked:

1. Name of species

2. Applicant (name and address)

3. Proposed denomination or breeder’s reference

4. Information on origin, maintenance and reproduction

5. Type of material

6. State of expression for each of the grouping characteristics

7. Most similar varieties and the difference of candidate variety from these varieties

8. Any additional information which can help to distinguish the variety

9. Resistance to pests and disease

10. Any special condition required for examination of the variety

11. Any other useful information

Cont….22

Cont……

IX. Technical Questionnaire

Reference Number (Not to be filled in by the

applicant.)

To be completed with every application made on candidate variety in accordance with specific Indian dust test guidelines for registration under PPVFR&FR Act, 2001.

1. Species Zea mays L.

Maize2. Applicant (Name and address): ( In concurrence with column 3 of

form no 1.)3. Proposed denomination of the

(candidate variety as given in form no 1)

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

4.1 Type of variety Inbreed line . [ ] Single cross hybrid. [ ] Three way cross hybrid [ ] Double cross hybrid [ ] Open pollinated variety [ ] 4.2 Variety details Field corn [ ] Pop corn [ ] Baby corn [ ] Waxy corn [ ] Quality Protein Maize [ ] High oil corn [ ] Fodder corn [ ] 4.3 Pedigree of proposed variety/line 4.4 Other information

5. Characteristics of the candidate variety to be given (The number in note column refers to the different states of given characteristics described in column 3. Tick out within the brackets, which based corresponds to the character expression of the candidate variety):

23

6. Similar varieties and difference from these varieties:

Characteristics in which similar variety is different*

Denomination of similar variety

State of expression of candidate variety

*: in the case of identical states of expression of both varieties, indicate the difference in grade of expression.

7. Additional information , which may help to distinguish the variety

a. Resistance to pests diseases (indicate the race etc.)

b. Produce quality

c. Special conditions for the examination of the variety

d. Other Information

8. Authorization for release

State of expression of similar variety

UPOV 2009

Sr. No.

Characteristics States Note(1 to 9)

Example variety/ line

Stage Assessment

7. Tassel: anthocyanin coloration of anthers (in middle third of main axis on fresh anther )

Absent 1 CM 400 65 Anthesis halfway

VG (visually in a group)Present 9 CM 145

HKI 161

24

Sr. No.


Example variety / line

Stage Assessment

1.(+)

Leaf: angle between leaf blade and stem (on leaf just above upper ear)

Narrow(<45o )

1 HHM 2 61Beginning of anthesis

VG (visually in a group)

Wide (>45o )

9 Vivek 5

Table -8: Characteristics for Leaf: Angle between leaf blade and stem (on leaf just above upper ear)

Narrow

25

Wide

1 9

Sr. No.



Stage Assessment

2.(+)

Leaf: Attitude of leaf blade (on leaf just above upper ear)

Straight 1 HHM 2 61Beginning of anthesis

VG (visually in a group)Droopy 9 HKI 323

Table - 9: Characteristics for Leaf: Attitude of leaf blade (on leaf just above upper ear)

Straight

Droopy

26

1 9

Sr. No.



Stage Assessment

3. Stem: Anthocyanins colouration of brace roots

Absent 1 HKI 163 65-75Anthesis halfway

Caryopsis watery ripe, Medium milk

VS (visually in a single)

Present 9 HKI 327T

Table -10: Characteristics for Stem: Anthocyanins colouration of brace roots

27

1 9

Sr. No.



Stage Assessment

4.(*)

Tassel: Time of anthesis (on middle third of main axis, 50 % of plant)

Very early(<45 D)

1 HKI 335 65

Anthesis halfway


Early(45-50 D)

3 HKI 1025

Medium (50-55 D)

5 HKI 323

Late (>55 D)

7 HKI 1126

Table -11: Characteristics for Tassel: Time of anthesis (on middle third of main axis, 50 % of plant)

28

Sr. No.


Example variety/line

Stage Assessment

5.(+)

Tassel : Anthocyanin colouration at base of glume

Absent 1 HKI 1344 65Anthesis halfway

VS (visually in a single)

Present 9 HKI 161

Table -12: Characteristics for Tassel: Anthocyanins colouration at base of glume

29

1 9

Sr. No



Stage Assessment

6. Tassel: Anthocyanin colouration of glumes excluding base (in middle third of main axis )

Absent 1 HKI 209 65 Anthesis halfway

VS (visually in a single)Present 9 CM 300

HKI 161

Table -13: Characteristics for Tassel: Anthocyanin colouration of glumes excluding base (in middle third of main axis )

9

30

1 9

Sr. No.



Stage Assessment

7. Tassel: Anthocyanin colouration of anthers (in middle third of main axis on fresh anther )

Absent 1 HKI 209 65 Anthesis halfway

VG (visually in a group)Present 9 CM 145

HKI 161

Table -14: Characteristics for Tassel : Anthocyanin colouration of anthers (in middle third of main axis on fresh anther)

91

31

1 9

Sr. No.



Stage Assessment

8. Tassel: Density of spikelet (in middle third of main axis )

Sparse 3 HKI 1126 65Anthesis halfway

VG (visually in a group)Dense 7 HKI 288-2

Table -15: Characteristics for Tassel: Density of spikelet (in middle third of main axis)

91

32

3 7

Sr. No.



Stage Assessment

9.(*)(+)

Tassel: Angle between main axis and lateral branches (in lower third of tassel)

Normal (<45o )

1 CM 145 65Anthesis halfway


Wide (>45o )

9 CM 140

Table -16: Characteristics for Tassel : Angle between main axis and lateral branches (in lower third of tassel)

33

1 9

Sr. No.



Stage Assessment

10.(*)(+)

Tassel: Attitude of lateral branches (in lower third of tassel )

Straight 1 HKI 193-1 65Anthesis halfway


Curved 5 HKI 323,46Strongly curved

9 HKI 163

Table -17: Characteristics for Tassel : Attitude of lateral branches (in lower third of tassel)

34

1 95

Sr. No.



Stage Assessment

11. Ear: Time of silk emergence (50 % plants)

Very early (<48 D)

1 HKI 335 65Anthesis halfway


Early (48-53 D)

3 HKI 1025

Medium (53-58 D )

5 HKI 323

Late (>58)

7 HKI 1126

Table - 18: Characteristics for Ear: Time of silk emergence (50% plants)

35

Sr. No.



Stage Assessment

12.(*)

Ear: Anthocyanin coloration of silks (on day of emergence)

Absent 1 HKI 1025 65Anthesis halfway

VG (visually in a group)Present 9 HKI 323

Table - 19: Characteristics for Ear: Anthocyanin coloration of silks (on day of emergence)

36

1 9

Sr. No.



Stage Assessment

13. Leaf: Anthocyanin coloration of leaf sheath (below ear)

Absent 1 HKI 163 71Caryopsis watery ripe

VS (visually in a single)Present 9 CM 300

Table - 20: Characteristics for Leaf: Anthocyanin coloration of leaf sheath (below ear)

37

1 9

Sr. No.



Stage Assessment

14. Tassel: Length of main axis above lowest side branch

Short (< 20 cm) 3 HKI 1128 71

Caryopsis watery ripe

MS (measurements in single)Medium (20-30 cm) 5 HKI 327 T

Long (>30 cm ) 7 V 25

Table - 21: Characteristics for Tassel: Length of main axis above lowest side branch

38

Sr. No.


Example Variety / line

Stage Assessment

15.1(*)

Inbred line only :Plant Length (up to flag leaf )

Short (<120cm) 3 HKI 1348-6-2 75Medium milk

MS (measurements in single)Medium (120-150 cm ) 5 HKI 323

Long (>150 cm) 7 HKI 1128

15.2(*)

Hybrids and open pollinated varieties only : Plant length (up to flag leaf)

Short (<150cm) 3 Vivek 75Medium milk

MS (measurements in single)Medium (150-180 cm ) 5 Prakash

Long (180-210 cm) 7 HQPM 1

Very long (>210 cm) 9 African Tall

Table - 22: Characteristics for Plant Length (up to flag leaf)

39

Sr.No.

Characteristics States Note(3 to 7 )


Stage Assessment

16. Plant: Ear placement

Low 3 HKI 1011 75Medium milk

MS (measurements in single)

Medium 5 HM 4High 7 HQPM 1

Table - 23: Characteristics for Plant: Ear placement

40

Sr. No.

Characteristics States Note(3 to7)


Stage Assessment

17. Leaf : Width of leaf blade (leaf of upper ear)

Narrow(<8cm) 3 HKI 323 75Medium milk

MS (measurements in single)

Medium(8-9cm) 5 HKI 295Broad (>9 cm) 7 HKI 1126

Table - 24: Characteristics for Leaf : Width of leaf blade (leaf of upper ear)

41

Sr. No. Characteristics States Note(3 to 7)


Stage Assessment

18.(*)

Ear: Length without husk

Short (<10cm) 3 HKI 536 92Caryopsis hard

MS (measurements in single)Medium (10-15 cm) 5 HKI 163

Long(>15 cm) 7 HQPM 1

19. Ear: Diameter without husk (in middle)

Small (<4 cm) 3 HKI 323 92Caryopsis hard

MS (measurements in single)Medium (4-5 cm) 5 HKI 327

Large (>5 cm) 7 THQPM 1

Table - 25: Characteristics for: Ear length without husk and Ear diameter without husk (in middle)

42

Sr. No.


Example Variety / line

Stage Assessment

20.(+)

Ear shape Conical 1 HKI 1344 92Caryopsis hard

VG (visually in a group )Conical-

Cylindrical2 HKI 295

Cylindrical 3 HKI 1105

Table - 26: Characteristics for Ear Shape

43

13

2

Sr. No.



Stage Assessment

21. Ear : Number of row of grains

Few (≤ 8) 3 - 92Caryopsis hard

MS (measurements in single)Medium (10-12) 5 HKI 163

Large (≥ 14) 7 HM 5

Table - 27: Characteristics for Ear : Number of row of grains

44

L-16 row, R-14 row

Sr. No.



Stage Assessment

22.(*)

Ear : type of grain (in middle third of ear)

Flint 1 HKI 1105 92 Caryopsis hard

VG (visually in a group )Semi flint/

Semi dent2 HKI 1344

Dent 3 HM 5

Table - 28: Characteristics for Ear : Type of grain (in middle third of ear)

45

1 32

Table - 29: Characteristics for Ear : Colour of top of grain

Sr. No.



Stage Assessment

23. (*)

Ear: Colour of top of grain

White 1 CM 300 92Caryopsis hard

VG (visually in a group )White with cap 2 HKI 1344

Yellow 3 HKI 1050

Yellow with cap 4 HKI 209

Orange 5 HKI 323

Red 6

Variegated 7

23

5

6

46

41

Table - 30: Characteristics for Ear: Anthocyanin colouration of glumes of cob

Sr. No.



Stage Assessment

24.(*)

Ear: Anthocyanin colouration of glumes of cob

White 1 HKI 163 93Caryopsis loosening day time

VG (visually in a group )Light purple 2 SUWAN

Dark purple 3 HKI 161

1 2 3

47

Table - 31: Characteristics for Kernel : Row arrangement (middle of corn)

Sr. No. Characteristics States Note(1 to 3)


Stage Assessment

25.(*)

Kernel : Row arrangement (middle of corn)

Straight 1 HM 2 93Caryopsis loosening daytime

VG( visually in a group )

Spiral 2 HM 5

Irregular 3 HKI 1344

48

1 2 3

Table - 32: Characteristics for Ear : Type of grain

Sr. No.



Stage Assessment

26-29. Ear : Type of grain Field corn 1 CM128 92

Caryopsis hard

VG (visually in a group )Pop corn 2 VL. Popcorn

Waxy / opaque 3 Vivek Hy.4

Opaque tinge 4 Vivek Hy.5

Sweet corn 5 Madhuri

1

5

49

1

2

3

4

Sr. No.

Characteristics States Note (1 to 5)


Stage Assessment

30.(+)

Kernel shape Shrunken 1 Madhuri 93

Caryopsis loosening day time

VG (visually in a group )Round 2 HKI 1342

Indented 3 HM 5Toothed 4 HKI 1348-6-2Pointed 5 -

Table - 33: Characteristics for Kernel shape

PPV & FRA, 2007550

3 421

Table - 34: Characteristics for Kernel size (1000 kernel weight)

Sr. No.

Characteristics States Note (1 to 7)


Stage Assessment

31. Kernel size (1000 kernel weight )

Very small (<100g)

1 Madhuri 93Caryopsis loosening day time

MG (measurements in a group )

Small (100-200 g)

3 HKI 1025

Medium (200-300g)

5 Prabhat

Large (>300g)

7 V.HY.-9

51

Conclusions

India has the largest reservoir of the genetic material and accounts for the largest

skilled population in the world, it gives several opportunities to the Indian

agricultural research and development.

To validate the claims distinctness, uniformity and stability (DUS) tests will have to

be conducted as per the National Test Guidelines.

PPV & FR Act protects breeders varieties from unauthorized infringement, ensure

farmers rights on seeds and encourage growth of seed industries by making seeds of

protected varieties available to the farmers.

DUS Testing is one of the key feature of plant variety protection.

For easy identification and preservation DUS test is very important .

52

Future thrust

• DUS test should be modified according to need, cropping pattern and

environmental conditions.

• More emphasis should be given on DUS testing by scientists / technical qualified

persons .

• The institutes/ research organization engaged in plant breeding should stream line

their maintenance breeding programme so as to make available pure seed, which

shall be used for DUS tests and kept as reference sample for at least 7 years.

• A well conceived programme of training of staff for conduct of DUS tests, data

compilation, analysis and its interpretation should be organized.

53

54 Thank You

DUS Test Guidelines for Maize

Education