CHAPTER-3

SEEDS AND SEED TESTING METHODS

 3. SEEDS AND SEED TESTING METHODS

3.1 INTRODUCTION
In order to have uniformity in seed testing it is important that the methods employed
for testing seeds by different laboratories are the same. Uniformity in seed testing
procedures is also necessary to play a statutory role under the Seeds Act (1966). In
case of dispute in a court of law the result cannot be checked and verified unless the
method adopted in analysis is uniform. Uniformity is also needed for reproducing results
in different laboratories. For this purpose, it is essential that rules are written and printed.

There are 38 major erops of the country comprising cereals, millets, pulses, oil
seeds, fiber crops, forage and sugar crops. For seeds testing two samples of each
cereal, pulses and oilseeds are selected. A brief introduction of each seed [66] and
seed testing methods [1] are given below which will be the bases for the project work o
progress.

3.2 TYPES OF SEEDS

3.2.1 Wheat ( Triticum Aestivum L)
Wheat is the worlds most widely food crop. It contains more protein than other
cereals. Wheat has relatively contented of niacil and thiamine. On account of the particular
physical and chemical qualities of the gluten of its grain, wheat makes more palatable
and better bread than any other cereal. India stands fourth in area and production (22.5
million hectares and 34.98 million tones respectively- 1989).

Weed Control
Weeds emerge with the emerging crop seedlings and if not controlled in the early
stages of the crop growth these may cause reduction in yield resulting from 10-40 %
depending upon the intensity and kind of weeds present in the area. Generally 2,4-D is
used to kill all the broad leaved weeds in wheat field.

Diseases
Wheat crop suffers from several diseases, which reduce its yield and quality. The
major diseases of wheat are rusts, alternaria leaf blight, loose smut, karnal bunt and
powdery mildew.

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Insect Pests
Wheat is attacked by number of insect pests in the field and in storage. Some of
the most important and the common ones are- termites, gujhia weevil, army worms,
brown wheat mites, aphids and jassids.

3.2.2 Sorghum (Sorghum BicolorL. Moench)

Sorghum (Jowar) is one of the four major food grains of the world. It contains
about 10 -12 % proteins, 3 % fat and 70 % carbohydrates. India ranks first in acreage
and second in production in the world.

Weed Control
During kharif season both grassy and broad-leaved weeds grow with sorghum
crop. In case weeds are not brought under control, there is 20 - 60 % reduction in
yield. Altrazine at the rate of 0.5 -1.0 kg active ingredients per hectare is used.

Diseases
Sorghum crop suffers from a number of diseases resulting in considerable loss in
yield. The important diseases are seed decay and seedling blight, downy mildew.

Insect Pests
Sorghum crop is subject to attack by number of insect pests. The major insect
pests are - sorghum shoot fly, stem borer, pink borer, sorghum midge, leafroller, sorghum
ear head bug, hairy caterpillar etc.

3.2.3 Green Gram (Vigna Radiata L. Witczek)

Green gram (moong bean) is an excellent source of high quality protein. It contains
about 25 % protein. In India moong bean is grown in almost all the states. It is grown
about 27.63 lakh hectares with a total production of 9.1 lakh tones of grain.

Weed Control
Two weedlings should be given to keep the crop free from handful weeds. First
weedlings should be done 20 -25 days after sowing and second 45 days after sowing.
Weeds do not pose a serious during summer. Use Basaline 1 kg a.i per hectare in 800
- 1000 liters of water as pre planting spray. It should be well incorporated in the soil
before sowing.

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Diseases
There are several important diseases of moong - yellow mosaic, mosaic mottle,
leaf crinkle, leaf curl, seed and seedling rot etc.

Insect Pests
The important insect pests are - hairy caterpillar, galerucid beetle, leaf hooper,
jassids etc.

3.2.4 Bean (Phaseolus Vulgaris)

It is mainly eaten as a green vegetable. It contains about 22% protein. Very little
fibre or oil is present, and the major of the seed is carbohydrate. The major exporter of
dried bean seed is the United States. Maharashtra state is another region were bean is
cultivated as a Kharif Crop.

3.2.5 Soybean (Glycine max L. Merrill)

Apart from 'he high yield potential, soybean possesses a very high nutritional value.
It contains about 20 % oil and 40 % high quality protein. Soybean protein is rich in
valuable amino acid lycine (5 %) in which most of the cereals are deficient.

Weed Control
Soybean is very sensitive to early weed competition. A weed infestation in a
soybean field may reduce yield by 40 - 45 % depending upon the intensity, nature and
the duration of the weed competition.

To avoid competition during the early growth stages, soybean field should be kept
free of weeds for the first 30-40 days after sowing. Herbicides have been found very
effective in weed control.

Diseases
Some of the important diseases are- seed, seedling rot, forged eye leaf spot, pod
blight or anthracnose, purple seed stain, bacterial pustule, yellow mosaic, ring spot etc.

Insect Pests
The important insect pests are - stem fly, girdle beetle, Bihar hairy caterpillar,
tobacco caterpillar, semi looper, leaf roller, leaf miner etc.

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3.2.6 Sunflower (helianthus annuus L.)
Sunflower is familiar plant in India. It has got 45 - 50 % good quality oil and high
amount of quality protein in cake. Sunflower is rich source of (64%) linoleic acid, which
helps in washing out cholesterol deposition in the coronary arteries of the heart.

Weed Control
Intercultural operations are essential to minimize the competition of sunflower plant
with the weeds. Weed free conditions up to 60 days after sowing results in better
performance. Use of Sirmate at the rate of 4 kg / hectare applied as pre emergence has
been found effective in controlling weeds in sunflower crops.

Diseases
There are number of diseases which are responsible for reduction in yield both
quantitatively and qualitatively. Important diseases are seed rot, charcoal rot, altemaria
blight, sclerotium wilt, sclerotinia wilt and rot etc.

Insect Pest
There are number of insect pests associated with this crop but very few sporadic
pests cause serious damage to it. The important insect pests are - seedling pests, foliage
feeding pests, ear head pests etc.

3.3 LIFE CYCLE OF SEED INSECTS

The life cycle, mode of damage, distribution, etc. of major seed insect [63] are
described below in brief (Atwal, 1991; Sinha, 1999; Zakaladnoi and ratanova, 1987.)

3.3.1 Rice Weevil Sitophilus Oryzae Linn

Life Cycle
The female lays around 25 translucent and white eggs after making a small
excavation in the soft part of the grain. These hatch in about 4- 6 days depending upon
the weather. The tiny young grub is white with a yellow brown head. It bores into seed
and feeds on its starchy contents. Usually, only one grub is found inside a seed, though
sometimes there may be two or three depending on the seed size. The grub stage last
for 20-30 days and then it pupates inside the hollow grain for 3 - 6 days depending on
temperature. The adult weevil emerges from the infested seed and starts a new

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generation. The average period from egg to adult is 30 days. Usually there are five
generations in a year but the number of generations in a year depends on prevailing
temperature and humidity conditions.

The species is susceptible to temperature. Physical environmental factors play a

very vital role in population dynamics. It is unable to breed in the grain ifthe moisture is
less than 9 %.

The host is wheat maize, paddy, sorghum, cotton seed, linseed, etc.

3.3.2 Lesser Grain Borer, Rhyzopertha Dominica Fab

The pest breeds from March to November and in December it enters hibernation
as an adult or as a larva. The females mate several times during there life which may be
as long as eight months, and in favorable conditions lay about 300 - 400 eggs. When
the temperature drops below 21° C, development stops and the adults hibernate inside
the grains. The eggs hatch in 5 days at 35 0 C and 26 days at 21 °C. The larva moults
four times on an average and the change in appearance occurs progressive after the
second moult. The complete development cycle requires 27 days at 35 °C and 183
days at 21°C, the optimum being around 34°C (Atwal, 1991; Sinha, 1999; Zakaladnoi
and ratanova, 1987).

The insect has susceptibility to cold, which is compensated by a remarkable

resistance to dryness and heat. It is unable to breed in the grains if the moisture is less
than 9 %.

The host is wheat, maize, rice, millet, sorghum, flour, and biscuits. It is considered
the greatest enemy of grain in the world after the rice weevil, S.orygzae.

3.3.3 Khapra Beetle, Trogoderma Granarium Everts

The insect remains active from April to October. The female begins to lay eggs on
grain singly or sometimes in clusters of 2-5. The eggs are white, translucent, cylindrical,
rounded at one end and narrow at the other. The incubation period is 6-16 days
depending upon the prevailing temperature and humidity. The insect hibernates in the
larval stage from November to March in cracks and crevices of walls and floors or in
other sheltered places. The pupal stage last 4- 6 days. The adults are incapable of

24
flying. Normally there are 4- 5 generations in a year in favorable circumstances.

The insect is susceptible in its egg and pupal stages and also to reduction in oxygen
content. Generally its infestation occurs on the top layer of the seed because the insect
fails to penetrate beyond a certain depth below the grain surface. An airtight storage
structure is quite dependable against damage by these pests.

The host is wheat, rice, maize, sorghum, gram, pulses, etc.

3.3.4 Pulse Beetles, Callosobruchus Chinenis Linn And C. Maculates Fab

The pests breed actively from March to the end of November. The females lay
eggs either on maturing pods in the field or on the seeds in the godowns. Eggs are glued
on the surface of seed. A female lays around 50-100 eggs in her lifetime. Under favorable
environmental conditions, eggs hatch in 4- 5 days. The grub bores into grain immediately
after hatching and spends it life inside the seed. It moults 4 times and completely develops
in 2-3 weeks. It then transforms into a pupa. The adults normally emerge in four days
but in winter it may take up to 28 days. Complete lifecycle from egg to adult takes
around 25-30 days in summer and 50-60 days in winter. The adult normally survives for
about 10 days. In a year 7-8 generations are recorded (anonymous, 1976; Sinha, 1999).

In dry seeds the adult cannot feed it and its longevity is reduced. The host is
gram, moong bean, cowpea, pigeon pea, etc.

3.3.5 Grain Moth, Sitogroga Cerealella Oliv

Normally, breeding takes place from April to October. The female lays around
400 eggs and larva emerges in about 4 days during summer and 7 days in winter. It
feeds inside the seed and spends its life there, moulting five times and developing in 14-
20 days. It pupates inside the seeds in cocoon. The moth emerges in 7-10 days. It
takes about 1 month to complete one-generation in favorable conditions.

This pest is unable to complete its development in dry seeds. The host is paddy,
maize, sorghum, barley and other cereals.

3.3.6 The Rice Moth, Corcyra Cephalonica (Stainton)

The rice moth is active from March to November. The adults remain active at

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night and the female lays eggs singly or in groups of 3 - 5 on the grains. The fully grown
larva is 10-12 mm long and is whitish and sometimes slightly grey. It produces abandoned
webs and when distributed it lets itself down to the ground on the end of its thread.
Pupation takes place in a thick spindle shaped cocoon. In summer, development can be
completed in 6 weeks depending on the temperature and the humidity conditions. There
are 2-4 generations per year depending on the climate.

This pest is unable to complete its development in dry seeds.The host is rice,
gram, sorghum, maize, groundnut, soybean, etc.

3.4 SEED TESTING METHODS

3.4.1 Seed Sampling
Seed sampling is aimed at obtaining a sample of the required size and consisting of
the same components as the whole lot of seeds. The quantity of seed tested in the
laboratory is small, compared with the size of the seed lot, which it is intended to
represent. No matter how accurately the laboratory work is done, the results can only
show the quality of the sample submitted for analysis. Consequently, every effort must
be made to ensure that the sample sent to the seed-testing laboratory accurately
represents the seed lot in question. Seed testing laboratory personnels are not necessarily
engaged in the sampling of seeds. But, nevertheless they should be well acquainted with
the principles of seed sampling and should also be able to guide properly the persons
engaged in this job.

3.4.2 types of Seed Samples

There are three types of seed samples received by a seed-testing laboratory. They
are (1) Service. (2) Certification (3) Official samples. [1]

Service Samples
Service sample means a sample submitted to the central seed laboratory or to a
state laboratory for testing the results to be used as information for seeding, selling or
labeling purposes. These samples must be tested promptly to provide the sender with
immediate information about the quality of seed with which he is dealing.

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Certification Sample
Certification sample means a sample of seed drawn by a certification agency or
by a duly authorized representative of a certification agency established under section 8
or recognized under section 18 of the Act.

Official Samples
Official sample means a sample of seed drawn by a seed inspector to ascertain
that the seeds meet minimum limit of specified quality.

All possible efforts are made to start testing a sample on the day ofreceipt. However
under unavoidable circumstances the sample shall be stored in dry cool, ventilated room
so that changes in the quality of seed are minimized.

3.4.3 Moisture Determination

Seed moisture content is one ofthe most important factor influencing seed quality
and storability. Therefore, its estimation during seed quality determination is important.
Seed moisture content can be expressed either on wet weight basis or on dry weight
basis. In seed testing, it is always expressed on a wet weight basis. Seed moisture
content can be determined either by air oven or moisture meter. However, if prescribed
standard for moisture content is less than 8 %, air oven method shall be used.

In this method, seed moisture is removed by drying at a specified temperature for

specified duration (Table 3.1). The moisture content is expressed as a percentage of
the original weight (wet weight basis), i.e.

M2 - M3 Loss in weight
M = ------------- X 100 = -----------------------------X 100
M2 - Ml Initial weight of seed

Where M = Seed moisture content.

M1 = Weight of the empty container with its cover.

M2 = Weight of the container with its cover and seeds before drying.

M3 = Weight of the container with its cover and seeds after drying.

The duplicate result of the determination may not differ by more than 0.2 %
otherwise the analysis should be repeated in duplicate.

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 If the material is predried, the moisture content is calculated from the results
obtained in the predried and dried stages using the following formula.

M = Si + S2 - ..
100

M = Moisture content

51 = Moisture percentage lost in predrying stage.

52 = Moisture percentage lost in drying state.

This is the most common and standard method for seed moisture determination.

Table 3.1 Method for moisture determination

Crop Botanical name Grinding Drying Predrying

Temp. Time necessary
(°C) (Hrs.) above the
stated moisture (%)
1 2 3 4 5 6

Sorghum Sorghum Bicolor FG 130 2 17

Wheat Triticumaestivum FG 130 2 17

Bean Lab Lab purpureus CG 130 1 17

Greengram Vignaradiata CG 130 1 17

Soybean Glycine max CG 103 17 10

Sunflower Hdianhus annus - 103 17 -

3.4.4 Germination Testing

Germination tests shall be made with seeds from the pure seed fraction of a purity
test. A minimum of four hundred seeds are required in four replicates of 100 seeds each
or eight replicates of 50 seeds each or 16 replications of 25 seeds each depending on
the size of seeds and size of containers of substrate.

The seeds shall receive no pretreatments excepting those recommended in table

3.2. If additional tests are undertaken after any other pretreatment, the result and pre­
treatment must be reported under other determinations on the certificate.

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 The seeds, arranged in replicates, are tested under favorable moisture conditions
and in accordance with the methods prescribed in table 3.2 After the period indicated
in table 3.2 the replicates are examined and counts made of the seedlings and seeds in
various categories required for reporting as presented elsewhere.

Procedures
Four hundred seeds are counted at random from the well-mixed pure seed.
Replicates of 100 seeds are normally used, spaced sufficiently far apart on the seedbed
to minimize the effect of adjacent seeds on seedling development. To ensure adequate
spacing, split replicates of 50 or even 25 seeds may be necessary, particularly where
there is seed borne disease. When the seeds are heavily infected it may also be necessary
with a paper substrate to change the substrate at an intermediate count.

Testing four hundred seeds is recommended on seed law enforcement, seed

certification and service samples.

Table 3.2 Germination methods (field crops)

Crop Botanical Prescription for Addition directions

name Substrata Temp. First Second including recomm-
°C count count endtaions for
(Days) (Days) breaking dormancy

Sorghum Sorghum Bicolor TP, BP 20-30, 4 10 Prechill

25

Wheat Triticumaestivum TP, BP,S 20 4 8 Prehet

Bean Lablab Purpureus S, BP 20-30, 4 10 -

20

Greengram Vignaradiata BP, S 20-30, 5 8 -

25

Soybean Glycine max BP, S 20-30, 5 8 -

25

Sunflower Helianhus annus BP, S 20-30, 4 10 Ethrel(25 ppm)

25 48Hrs.

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3.4.5 Seed Vigour Testing
Seed vigour is an important quality parameter which needs to be assessed to
supplement germination and viability tests to gain insight into the performance of a seed
lot in the field or in storage. ISTA congress in 1977 adopted the definition of seed
vigour as “the sum total of those properties of the seed which determine the level of
activity and performance of the seed or seed lot during germination and seedling
emergence”.

Procedure
1. First Count
The test is done along with the regular germination test. The numbers of normal
seedlings germinated on the first count day, as specified in the germination test for
each species, are counted. The number ofnormal seedlings gives an idea of the level
of seed vigour in the sample. Higher the number of normal seedlings greater is the
seed vigour.

2. Seedling Growth Rate And Dry Weight

The seedlings are grown either in laboratory, green house or field. In laboratory, in
between rolled towel paper method should be followed. Ten seeds are planted in
the centre of the moist towel papers in such away that the mieropyles are oriented
towards bottom to avoid root twisting. The rolled towel papers are kept in the
germinator maintained at a temperature recommended for crop in reference. After a
specified period of time (5-10 days) towel papers are removed and five seedlings
are selected, their lengths are measured and mean seedling length is calculated. Seeds
producing the taller seedlings are considered more vigorous than the seed lots producing
shorter seedlings. For dry weight determination, the seedlings are removed and dried
in an air oven at 100 °C temperatures for 24 hours. The seedling dry weight provides
additional information for assessing seed vigour.

3. Speed of Germination
One hundred seeds each in four replications are planted in recommended substratum
for germination. The substratum is kept in a germinator maintained at recommended
temperature for the crop in reference (table 3.2). Numbers of seedlings emerging

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 daily are counted from day ofplanting the seeds in the medium till the time germination
is complete. Thereafter a germination index (G.I.) is computed by using the following
formula.

Where n = number of seedlings emerging on day ‘d’

D= day after plating

The seed lot having greater germination index is considered to be more vigorous.

4. Seed Vigour Index (S.V.I.)

This is calculated by determining the germination percentage and seedling length of
the same seed lot. Fifty seeds each in four replications are germinated in towel
papers as prescribed for the crop species in germination test. While evaluating the
number ofnormal seedlings at the time offinal count, the seedling length of 5 randomly
selected seedlings are also measured. Seed vigour index is calculated by multiplying
germination (%) and seedling length (mm). The seed lot showing the higher seed vigour
index is considered to be more vigorous (Abdul-Baki and Anderson, 1973).

3.4.6 Seed Health Test (Detection of Insects)

Insect damaged seed in purity analysis are classified according to the so-called
“One half seed rule”’ If one half or more of the seed is damaged or consumed by the
insect it is considered inert matter. Standard for insect damaged seeds under certification,
however, does not allow apparent or visible evidence of damage by insect for both
foundation and certified seed in excess of 1.0% for the seeds of maize and legumes and
0.5% for the other crop seeds unless otherwise prescribed. The methods of insect
infestation are

Detection of External Infestation: Direct Examination

i) Seeds are examined in dry state with the help of magnifying glass (1 OX) or
stereoscopic microscope aided with light.
ii) Adult weevil, beetle, moth larvae, grubs etc, are separated and counted.

iii) The result is reported as number of insets (including its stages) per weight of the
sample.

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 Presence of insects in the seed sample, so detected, confirms that the seed lot
was infested. Their absence does not guarantee that the seed lot is free from insect
infestation i.e. insect injury to the seeds or internal (hidden) infestation. Thus, it is
imperative to carry out further tests to detect external and internal infestation.

Detection Of Internal (Hidden) Infestation

Some of the stored seed pests especially weevils and beetles are internal feeder.
Detection of their infestation is essential to determine actual infestation. The number of
infested seeds, thus obtained is added to the number of seeds found externally damaged
by the insects for final calculation.

Procedure
1. Seeds are taken in a beaker (100ml capacity) and then sodium hydroxide (NaOH)
10 % solution (Dissolve 1 Ogm NaOH pellets in 100 ml water) is poured in it until
the seeds are submerged.

2. The seeds are boiled in sodium hydroxide solution for 10 minutes or more
depending upon the type of seeds.

3. After decanting solution translucent seeds are washed with water and then examined
with the help of magnifying glass.

4. Seeds with visible internal infestation are separated, cut opened to confirm the
presence of insect or its stage and counted.

5. The result is reported after adding the count of damaged seeds as percentage (by
number).

3.5 CONCLUDING REMARKS

Weevil (Sond Kida) is one of the important and common insects pests which
attack cereal crops such as wheat, sorghum etc. It is unable to breed in the grains if the
moisture is less than 9 %. Collosobruchus (Bhunga) is the important pests for host
grains such as green gram, bean, cowpea etc. In dry seeds the adult cannot feed it and
its longevity is reduced. The seed testing methods play a statutory role under the Seeds
Act (1966). Seed quality evaluation is done in seed testing laboratories as per above
standards.

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