Genetic Purity Testing

Prof. Kumari Rajani

(Seed Technology)
BPSAC, Purnea (BAU, Sabour)
 What is genetic purity?

Genetic purity refers to TRUENESS TO TYPE, or the degree of

contamination of seeds caused by undesired genetic varieties or
species.

The success of Hybrid Seed Production is dependent on the genetic

purity of parental lines.

Both outcrossing and the inadvertent mixing of seed can compromise

seed quality, therefore genetic purity tests are critical tools for seed
producers and plant breeders

Prof. Kumari Rajani, DSST, BAU, Sabour

 Minimum Standards for genetic purity
Class of Seed Standards for Minimum Genetic
Purity (%)
Breeder Seed 100.00

Foundation Seed 99.00

Certified Seed
(i) Varieties, composites,
98.00
synthetics & multilines

(ii) Hybrids 95.00

(iii) Hybrids of cotton, TPS,

90.00
muskmelon, brinjal & tomato

(vi) Hybrid castor 85.00

IMSCS, 2013
Prof. Kumari Rajani, DSST, BAU, Sabour
 Genetic Purity Testing

It is a test performed to determine the genuineness of cultivar.

Varieties being grown for seed production should periodically be
tested for genetic purity by this test to make sure that they are
being maintained in their true form.

Prof. Kumari Rajani, DSST, BAU, Sabour

 Objective of Genetic Purity Testing
❖The object of species and variety verification is to determine the
extent that the submitted sample conforms to the species or variety as
requested by the applicant (ISTA, 2015).

❖The determination of a species or variety is valid only if the species

or variety is stated by the applicant and an authentic standard sample
of the species or variety is available for comparison to ensure the
certainty of the determination.

❖Genuineness of a cultivar is tested by means of heritable characters

and the traits compared may be MORPHOLOGICAL,
PHYSIOLOGICAL, CYTOLOGICAL or CHEMICAL.
Prof. Kumari Rajani, DSST, BAU, Sabour
 Methods to assess Genetic Purity
1. Examination of seeds
2. Examination of seedlings (ISTA)

i. Coleoptile pigmentation in cereals

ii. Seedling color test in Beta spp.
iii. Color of cotyledons in Brassica spp.
iv. Fluorescence test in Lolium spp and Festuca spp.

3. Cytological test (Ploidy test)

4. Grow Out Test (Field plot test)
5. Growth Chamber test
6. Chemical test

Prof. Kumari Rajani, DSST, BAU, Sabour

 CHEMICAL TESTS

Phenol Test Rice, Wheat, Barley, Oat

Peroxidase Test Soybean, Groundnut
Potassium Hydroxide Bleach Test Sorghum, Rice, Sunflower
Ferrous Sulphate Test Oat, Sunflower
Hydrochloric Acid Test Oat
Vanillin Test Faba Bean
Alkaloid Test Lupin
Copper Sulphate Ammonia Test Sweet Clover
Sodium Hydroxide Test Rice, Sunflower, Wheat

Prof. Kumari Rajani, DSST, BAU, Sabour

Recommended by International Seed Testing Association (ISTA)
i. Phenol Test in Wheat
ii. Lugol’s Test for Lupin
iii. Electrophoretic techniques in Wheat, Oat, Barley, Peas,
Maize, Sunflower and Lolium

Recommended by Association of Official Seed Analysts (AOSA)

i. Copper Sulphate-Ammonia test for Sweet clover

ii. HCL Test for Oat
iii. Peroxidase Test for Soybean
iv. KOH Test for Rice and Sorghum
v. NaOH Test for Wheat

Prof. Kumari Rajani, DSST, BAU, Sabour

 MORPHOLOGICAL
✓Morphological feature have been a major component of
cultivar identification.

✓Therefore, Grow Out Test (GOT) has been considered for

genetic purity testing under Indian Minimum Seed
Certification System.

LIMITATIONS

✓Time consuming and expensive

✓Required large land area
✓Required highly skilled manpower
✓Many morphological features are multigenic
✓Quantitative characters affected or altered by environment

Prof. Kumari Rajani, DSST, BAU, Sabour

Seed shape

Brush hairs
Prof. Kumari Rajani, DSST, BAU, Sabour
 Grow-Out Test for Cultivar Purity

✓ Grow-out test is conducted to determine the genetic purity of a

given seed lot of a released cultivar and the extent to which the
submitted sample conforms to the prescribed standards (IMSCA).

✓ The samples for grow-out test are to be drawn simultaneously with

the samples for other quality tests and the standard procedure shall
be followed.

✓ The grow out test shall be conducted in specified areas

recommended for the cultivar / hybrid or in off-season nurseries.

✓ The standard sample of a cultivar (control) is the official standard

against which all other samples of the seed of the cultivar will be
judged.

Prof. Kumari Rajani, DSST, BAU, Sabour

 Submitted sample size

Submitted sample size

Crops
(g)
Maize, cotton, groundnut, soybean and
1,000 species of other genera with seeds of a
similar size
Sorghum, wheat, paddy and species of other
500
genera with seeds of similar size
Beta and species of other genera with seeds
250
of similar size
100 Bajra, jute and species of all other genera
250 tubers/ planting Seed potato, sweet potato and other
stakes/ roots/corns vegetatively propagating crops

Prof. Kumari Rajani, DSST, BAU, Sabour

 Specification for grow out test
Space
Row Plant to plant Space between
between No. of
SN Crop length distance rows
plots replications
(meters) (cm) (cm)
(cm)
1. Wheat, barley, 6 2 25 50 2
oats
2. Pea, cowpea 6 10 45 90 2
3. Chickpea, green 6 10 30 60 2
gram, black
gram
4. Maize 10 25 60 90 2
5. Hybrid cotton 5 10 45 45 2
6. Paddy
(a) very early to 6 15 20 45 2
medium
(b) late and very 6 25 30 60 2
late
7. Pearlmillet 6 10 60 90 2
8. Sorghum 6 10 45 60 2
Prof. Kumari Rajani, DSST, BAU, Sabour
 Method of grow out test
Raising of desired population by following recommended cultural practices

Provide equal opportunity to each and every plant for full expression of
genetically controlled characters

Sow the various samples of the same variety/cultivar in succession and standard
sample at suitable intervals

Seed rate must be adjusted to produce approximately the same number of plants
in the plots

When absolutely necessary, thinning or transplanting of seedlings from elsewhere

into the plot is permitted

The tests is preferably conducted in area for which it is recommended

The minimum 200 plants from control samples should be raised along with the
test crop

The number of plants found to be of other cultivars, other species or aberrant

shall be calculated as a percentage of the number of plant examined
✓Crop should be so grown that the genetical difference
expresses them as clearly as possible.

✓In self-fertilizing species the individual of a cultivar may be

theoretically identical whereas the individual of a cultivar in
cross-fertilizing species may not be genetically similar, but
comprise a number of types.

✓Therefore, it is easier to determine the cultivar purity in self-

fertilizing species than in cross-fertilizing species.

✓Hence, it is essential to sow the various samples of the same

cultivar in succession and standard samples are sown at suitable
intervals (for example, one standard sample for every ten
samples to be tested.

Prof. Kumari Rajani, DSST, BAU, Sabour

 Observations

✓Observations are made during the full growing period, or for a

period specified by originating breeding institute and deviations
from the standard sample of the same variety are recorded.

✓At suitable development stage the plots are examined carefully

and plants which are obviously of other cultivar are counted and
recorded.

Prof. Kumari Rajani, DSST, BAU, Sabour

 Calculation, Interpretation and reporting of the result

✓Percentage of other cultivars, other species or aberrant found may

be calculated upto first place of decimal. While interpreting the result,
use of tolerance may be applied by using the reject.

Prof. Kumari Rajani, DSST, BAU, Sabour

A very limited number of discriminatory traits can be observed directly
on seed or seedlings, a limited number can be assessed on plants

Limitation of Grow Out Tests (GOT)

•Long duration and costs of field trials
•Effect of environment on phenotype
•Evaluation sometimes subjective

Prof. Kumari Rajani, DSST, BAU, Sabour

 Benefits of Genetic Purity Testing

Benefits for Seed Production & Distribution

✓Confirmation that seed meets genetic purity standards

✓Identification of selfing and outcrossing in hybrid seed lots

✓Determination of variants, segregation and seed mixes in hybrid or

inbred seed production lots

✓Variety verification, ensuring that producers and customers receive

the variety they expect

Prof. Kumari Rajani, DSST, BAU, Sabour