ASSIGNMENT

Course: Varietal Development and Maintenance Breeding

(GPB-504)
DUS Test Guidelines for Maize

Submitted to Dr. Tosh Garg

Submitted by Sandeep Singh

Admission No. L-2022-A-277-M
Department Plant Breeding and Genetics
 Introduction:
 Taxonomically, Zea mays L. Belongs to tribe Maydeae, family Poaceae.
 The center of origin of Zea mays L. (2n=20) is reported to be Mexico and Central America.
 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 % of
total food grain production in the country.
 Nine states: Karnataka, Andhra Pradesh, Tamil Nadu, Rajasthan, Maharashtra, Bihar, Uttar Pradesh,
Madhya Pradesh and Gujarat accounts for 85% of India’s maize production and 80% of area under
cultivation (FAO, 2013).
Plant Morphology of Maize:
 Maize is a monoecious plant.
 Male inflorescene called tassel.
 Female inflorescene 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.
PPV & FR Act 2001:
 Government of india enacted the plant variety protection and farmers’s rights act on Oct 30, 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 system or in combination
 To a breeder in the form of Plant Breeder’s Rights
 To a farmers in the forms 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.
What is DUS testing ?
DUS means Distinctness-Uniformity-Stability.
Through DUS testing, one can ascertain whether a recently bred variety is distinct from other varieties
within the same species whether the characteristics that were used to establish the uniformity of the
trait are expressed consistently and whether the stability part of the trait is maintained over successive
generations.
Why?
 DUS tests are in place to ensure that new varieties can enter the market lawfully through the National
List and to grant plant breeders rights, a type of intellectual property rights intended to protect the
significant financial outlay associated with contemporary plant breeding.
How?
A DUS test is typically carried out over the course of two growing seasons in a glasshouse or the
field. Many primarily morphological traits are noted during this time on the new variety as well as on
related varieties, contributing to what is referred to as "Common Knowledge".
Guidelines for conduct of test DUS on Maize:
NOTE: The following test guidelines shall apply to all varieties, hybrids and parental lines of Maize
(Zea mays L.).
Seed material required:
 Minimum quantity of seed to be provided for the candidate variety or hybrid = 3000 gm.
 Minimum quantity of seed to be provided for the parental line of hybrid = 1500 gm.
Each of these seed lots shall be packed and sealed in ten equal weighing packets and submitted in one
lot.
 The submitted seed material by applicants shall have the following standards:
Table no. 1
Germination capacity
Inbreds and single cross hybrids 80% (Minimum)
Varieties and double cross hybrids
90% (Minimum)
Moisture content 8-10% (Maximum)
Physical purity 98% (Minimum)

 Together with the seed, the applicant must submit certified germination test results that were obtained
no more than one month before the submission deadline. It must also meet the strictest requirements
for sanitary, phytosanitary, genetic purity, and uniformity.
 The plant material cannot have undergone any biophysical or chemical processing.
Conduct of test:
 Typically, the test needs to be carried out at least twice (for two seasons). A different suitable test site
or special test conditions may be used to examine the variety if any of its essential characteristics are
not expressed for visual observation at these locations.
 The field tests must be conducted in an environment that supports each test characteristic's normal
development and expression. Plots must be large enough so that individual plants or plant portions
can be taken out for measurement and observation without affecting subsequent observations on the
standing plants until the end of the growing season.
 Each test will have a plot size of roughly 250 plants. Only when subjected to comparable
environmental conditions can separate plots be used for measurement and observation.
Design of test plot:
Table no. 2
Number of rows
Inbred lines and single cross hybrids 4
Varieties and other hybrids 8
Row length 6m
Row to row distance 75cm
Plant to plant distance 20 cm
Number of replications 3

Methods and observations:

 The qualities listed in the Table of Characteristics will be applied when evaluating hybrids, inbred
lines, and varieties for their DUS.
 At least 30 plants for inbreds and single cross hybrids and 60 plants for varieties and other hybrids
must be observed in order to determine the distinctiveness and stability of the hybrids.
 Inbred lines and single-cross hybrids will be assessed for uniformity using a population standard of
1% and a 95% acceptance probability. The maximum number of variants permitted in a sample of 100
plants would be three for inbreds and single cross hybrids, and six for other varieties and hybrids.
 The upper, well-developed ear is where all ear observations should be made, and the most recent
Royal Horticultural Society (RHS) color chart should be used to evaluate all color characteristics.
Grouping of varieties:
To make the Distinctiveness assessment easier, the candidate varieties for DUS testing will be
grouped. Features that, in their various states, are fairly evenly distributed across all varieties in the
collection and that are known from experience to either not vary at all within a variety or to vary very
slightly within a variety are appropriate for grouping purposes.
The following characteristics are proposed to be used for grouping maize varieties:
Table no. 3
Tassel Time of anthesis
Ear Time of silk emergence (50% plants)
Ear Anthocyanin colouration of silks
Plant Length
Ear Type of grain

Characteristics and Symbols:

 To assess distinctness, uniformity and stability, the characteristics and their states should be used.
 Notes (scales 1 to 9) should be use for the purpose of electronic data processing.
 Legend :
 (*) characteristics that should be observed during every growing season on all varieties, included in
the variety description, 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, 2007)
Table no. 4: 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 8
(41- as per UPOV) (16)
Onion 27 12
The optimum stage of plant growth assessment of each characteristics is indicated by decimal code.
Table no. 5: Decimal code for the growth stage
Stage code General description
00 Dry seed
12 2 leaf unfolded
14 4 leaves unfolded
51 Inflorescene 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

Table no. 6: Characteristics for leaf: Angle between leaf blade and stem
Sr. No. Characteristics States Notes Examples Stage Assessment
 variety/
(1 to line
9)
1. (+) Leaf: angle between Narrow 1 HHM 2 61 VG (visually in
leaf blade and stem (<45°) beggining group)
(on leaf just above of anthesis
upper ear)
Wide 9 Vivek 5
(>45°)
2. (+) Leaf: attitude of leaf Straight 1 HHM 2 61 VG (visually in
blade (on leaf just beginning a group)
above upper ear) of anthesis

Droopy 9 HKI 323

3. Stem: Anthocyanins Absent 1 HKI163 65-75 VS (visually in
colouration of brace anthesis a single)
roots halfway

Present 9 HKI 3271 Caryopsis

watery
ripe,
medium
milk
4. (*) Tassel: time of Very 1 HKI 335 65 Anthesis VG (visually in
anthesis (on middle early (<45 halfway a group)
third of main axis, D)
50% of plant)
Early (45- 3 HKI 1025
50)

Medium 5 HKI 323

(50-55)

Late 7 HKI 1126

(>55 D)
5. (+) Tassel : Anthocyanin Absent 1 HKI 1344 65 Anthesis VS (visually in
 colouration at base of halfway a group)
glume HKI 161
Present 9
6. Tassel : Anthocyanin Absent 1 HKI 209 65 Anthesis VS (visually in
colouration of glumes halfway a single)
excluding base
Present 9 CM 300
7. Tassel : Anthocyanin Absent 1 HKI 209 65 Anthesis VS (visually in
colouration of anthers halfway a group)

Present 9 CM 145
8. Tassel : density of Sparse 3 HKI 1126 65 Anthesis VS (visually in
spikelets halfway a group)
HKI 288-
Dense 7 2

9. (*) (+) Tassel : angle between Normal 1 CM 145 65 Anthesis VS (visually in

main axis and lateral (<45°) halfway a group)
branches

Wide 9 CM 140
(>45°)
10. (*) (+) Tassel : Attitude of Straight 1 HKI 193- 65 Anthesis VS (visually in
lateral branches 1 halfway a group)
Curved 5 HKI 323

Strongly 9 HKI 163

curved
11. Ear : time of silk Very 1 HKI 335 65 Anthesis VS (visually in
emergence early (<48 halfway a group)
D)

Early (48- 3 HKI 1025

53 D)

Medium HKI 323

 (53-58) 5

Late HKI 1126

(>58) 7

12. (*) Ear : Anthocyanin Absent 1 HKI 1025 65 Anthesis VS (visually in

colouration of silks halfway a group)

Present 9 CM 323
13. Leaf : Anthocyanin Absent 1 HKI 163 71 VS (visually in
coloration of leaf Caryopsis a single)
sheath watery ripe

Present 9 Cm 300
14. Tassel : length of main Short 3 HKI 1128 71 MS
axis above lowest side (<20cm) Caryopsis (measurements
branch watery ripe in a single)

Medium 5 HKI 327T

(20-30)

Long 7 V25
(>30cm)
15. 1 (*) Inbred line only: plant Short 3 HKI 1348 75 medium MS
length (<120) milk (measurements
in a single)
Medium 5 HKI 323
(120-150)

Long
(>150) 7 HKI 1128
15.2 (*) Hybrids and open Short 3 Vivek 75 medium MS
pollinated varieties (<150) milk (measurements
only: Plant length in a single)

Medium 5 Prakash
 (150-180)

Long
(180-210) 7 HQPM1

Very long 8 African

(>210) tall
16. Plant : ear placement Low 3 HKI 323 75 medium MS
milk (measurements
Medium 5 HKI 295 in a single)

High 7 HQPM1
17. Leaf : width of leaf Narrow 3 HKI 323 75 medium MS
blade milk (measurements
in a single)
Medium 5 HKI 295

Broad 7 HKI 1126

18. Ear : length without Short 3 HKI 536 92 MS
husk caryopsis
hard
Medium 5 HKI 163

Long 7 HQPM1
19. Ear : diameter without Small 3 HKI 323 92 MS
husk caryopsis
Medium 5 HKI 327 hard

Long 7 THQPM1
20. Ear shape Conical 1 HKI 1344 92 MS
caryopsis
Conical- 2 HKI 295 hard
cylindrica
l
 cylindrica 3
l HKI 1105
21. Ear: no of row of Few 3 - 92 MS
grains caryopsis
hard
Medium 5 HKI 163

Large 7 HM5
22. (*) Ear: type of grain Flint 1 HKI 1105 92 MS
caryopsis
Semi flint 2 HKI 1344 hard

Dent 3 HM5

23. Ear : color of top of White 1 Cm 300 92 MS

grain caryopsis
hard
White 2 HKI 1344
with cap

Yellow 3 HKI 1050

Yellow 4 HKI 209

with cap

Orange 5 HKI 323

Red 6

Variegate 7
d
24. (*) Ear :Anthocyanin White 1 HKI 163 92 VG
 color of glumes of cob caryopsis
hard
Light
purple 2 SUWAN

Dark
purple 3 HKI 161
25. Kernel : row Straight 1 HM 2 92 VG
arrangement caryopsis
hard
Spiral 2 HM 5

irregular 3 HKI 1344

26-29. Ear: type of grain Field corn 1 CMI 128 92 VG
caryopsis
Pop corn hard
2 VL
popcorn
Waxy
3 Vivek hy-
Opaque 4
4 Vivek hy-
Sweet 5
corn 5 Madhuri
30. (+) Kernel shape Shrunken 1 Madhuri 92 VG
caryopsis
Round loosening
2 HKI 1342 day time

Indented HM 5
3
Toothed HKI
4 1348-6-2
Pointed
 5
31. Kernel size Very 1 Madhuri
small

Small 3 HKI 1025

Medium 5 Prabhat

Large 7 [Link]-9

Conclusion :
 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 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.