Kai Dnyandev Patil Charitable Trust , Chikhurda

COLLEGE OF AGRICULTURE BUSINESS MANAGEMENT

LATUR
AFFILIATED TO
VASANTRAO NAIK MARATHWADA KRISHI VIDYAPEETH PARBHANI

PRACTICAL MANUAL
OF

Course No: BOT-243

Course Title- Principle and Practices of seed Science and
technology

Name of student :
Registration no :
Semester : IV (New)
Course teacher : Prof. Pathan R S
Academic Year :
1
 COLLEGE OF AGRICULTURE BUSINESS
MANAGEMENT , LATUR
(Affiliated To Vasantrao Naik Marathwada Krishi Vidyapeeth Parbhani )

CERTIFICATE

This is Certified that Mr./Miss………………………………….… Reg.

No………………………… has successfully completed all the practical of the

Course No. BOT- 243 Course title- Principles and practices of seed science

and technology for the requirement of II year IV (New) semester

examination of the Academic session 20 - 20

Place: Latur
Date:

Course Teacher

Prof. Pathan R S
2
 INDEX

Sr. Name of Exercise Date Signature

No.
1 Identification of seeds of field and horticultural
crops.
2 Study of seed structure in monocot and dicot seeds.
3 Study of floral biology of important self pollinated
agriculture and horticulture crops.
4 Study of floral biology of important cross pollinated
agriculture and horticulture crops.
5 Study of floral biology of important often cross
pollinated agriculture and horticulture crops.
6 Types of isolation, determination of isolation
distance, study of isolation requirements in different
crops for foundation and certified seeds.
7 Study of emasculation and hybridization techniques.
8 Study of distinguishing morphological characters in
A, B & R lines.
9 Study of synchronization techniques for hybrid seed
production, planting ratio, supplementary pollination
techniques, border rows for hybrid seed production.
10 Study of seed cleaning and grading technique, seed
packing and seed treatment techniques.
11 Seed testing - seed sampling.
12 Seed testing - physical purity test and moisture test.
13 Seed testing - seed germination test.
14 Economics of seed production.
15 Visit to seed production plots of public and private
sector companies.
16 Visit to seed production organization.

3
 Exercise No. 1
Identification of seeds of field and horticulture crops

Q.1 Identify the given seed sample and write their common name, botanical name, family,

Chromosome number and mode of pollination.

4
 Exercise No. 2
Study of seed structure in monocot and dicot seeds.
Seeds: A seed is matured ovule that comprises an embryo or a miniature undeveloped plant
food reserves, all enclosed within a protective seed coat.

Structure of seed:

Seed structure of kidney bean

 Tesla: It is the outer coat of the seed that protects the embryonic plant.

 Micropyle: It is a tiny pore in the testa that lies on the opposite of the tip of the radicle. It
permits water to enter the embryo before active germination.

 Hilum: Is a scar left by the stalk which attached the ovule to the ovary wall before it became
a seed.

 Cotyledon: In some plants, this contains high quantities of starch and will provide a source
of food for the developing embryo prior to germination, in other plants this role is performed
by an endosperm. In monocotyledons, there is just one cotyledon whereas
in dicotyledons there are two. Depending on the type of germination (epigeous or
hypogeous) the cotyledons may remain below ground or be pulled above ground.

 Radicle: This is the embryonic root which will develop into the primary root of the plant. It
is usually the first part of the embryo to push its way out of the seed during germination.

 Plumule: This is the embryonic shoot. It appears as a bud which will give rise to the shoot
and the remaining structures in the plant.

 Endosperm: In many plants, a separate part for storage of starch develops and this is called
the endosperm. It is seen in maize and wheat.

Functions of Seeds:

They help in germination of the new plant.

The seeds contain food reservoirs in the form of cotyledons and endosperm.

The seed coat is protective in nature which protects the embryo inside.
5
Monocot seeds: Seeds that are made out of a single (mono) embryonic leaf or cotyledon are
called as monocot or monocotyledon seeds eg. Grains (wheat, corn, rice, millets, sugarcane,
banana and grasses.

Dicot seeds: Seeds having two embryonic cotyledons are called as dicot or dicotyledonous
seeds e.g. Legumes (pea, beans, lentils, peanuts) tomato, lettuce, mint, oak.

Structure of monocot seeds:

Embryos of monocot seeds possess only one large cotyledon called scutellum. The
scutellum is generally shield shaped and is located laterally towards a side of the embryo axis.
As in the dicot the embryo axis of monocot possess shoot tip, plemule, enclosed in a sheath
called coleoptile and root tip, radical, enclosed in coleorhiza. In a monocot seeds the endosperm
is covered by proteinous layer called aleurone layer.
Majority of the monocot seeds are albuminous seeds, i.e., they have hick, swollen
endosperm for nourishment. Endosperm is not completely consumed during embryo
development, and it is the nourishing tissues in seeds. However, some monocots like orchids
show exceptions. Other monocot seeds examples include ginger, onion, coconut and garlic.

Structure of monocot and dicot seeds

Structure of dicot seeds:

Dicotyledons are also known as dicots. Embryo consist of an
embryo axis and two cotyledons. Cotyledons generally have a swollen appearance as it acts as
food reserve for developing seeds. The embryo axis has two ends. The one which forms a shoot
tip is called plemule and the portion lower end which forms a root tip is called radical. The
whole content is enclosed within a protective cover called seed coat. Seed coat is made up of an

6
outer layer called testa and an inner layer called tagmen.moreover, the seed is attached to the
fruit through a structure called hillum. e.g. plum, almomnds, cashew.

Difference between monocot and dicot seeds:

Sr. Monocot seeds Dicot seeds

no.
1 The monocot embryos have a single The dicot embryos have a pair of
cotyledon. cotyledons.
2 Cotyledons are thin and small Cotyledons are fleshy and store food
material.
3 They have fibrous root system. They have tap root system.
4 Leaves in monocot have parallel venation. Leaves in dicots have reticulate or net
venation.
5 Endosperm store large amount of food for Endosperm is reduced or even absent.
the embryo.
6 Plemule occurs terminally. Plemule occure laterally.
7 Shapes of monocot seeds are variable, Shape of dicot seeds are variable, usually
usually less symmetrical and smaller in more symmetrical and larger in size.
size.
8 E.g. Grains (wheat, corn, rice, millets, E.g. Legumes (pea, beans, lentils, peanuts)
sugarcane, banana and grasses. tomato, lettuce, mint, oak.

7
 Exercise No. 3
Study of floral biology of important self pollinated agriculture and
horticulture crops
1) Agriculture crops

A) Wheat

Botanical Name : Triticum monococcum (2n=2x=14)

Triticum durum (2n=2x=28)
Triticum aestivum ( 2n=2x=42)

Family : Gramineae/Poaceae
Chromosome Number : 2n = 42

Mode of pollination : Self of pollination

Floral biology: Inflorescence is a terminal compound, distichous spike, with rachis,

awned or awnless, glabrous or hairy. Self pollinated.

Spikelets: Sessile, and borne singly at the nodes on alternate sides of the zigzag rachis.
Each spikelet consists of two to five florets attached alternately on opposite sides of a
short central axis called rachilla, and is covered by two sterile or empty glumes.
Lemma broad with an acute tip or awn, palea thin. The lower lemma are fertile
while the top one or two sterile. Lodicules are two in number, stamens three with thin
filaments and large anthers, superior ovary, styles two, single ovule, bifid feathery
stigma.

B) Rice:

Botanical Name : Oryza sativa L.

Family : Poaceae

Chromosome No. : 2n =24

Mode of pollination: Self pollination

The inflorescence of rice is called panicle. The spikelet is the unit of

inflorescence.The spikelets are borne on the branched panicle.

8
Spikelet Morphology: The spikelet consists of two short sterile lemma, called glume 1
and glume 2, the grain a normal fertile

Lemma and palea. The fertile lemma and palea enclose the spikelet. The sexual
organs viz., six stamens arranged in two whorls and a pistil at the centre. The
stamen consists of bilobed anthers borne on slender filaments, while the pistil consists
of ovary, style and feathery bifid stigma.

C) Soybean:

Botanical Name : Glycine max L.Merr.

Family : Fabaceae

Chromosome Number : 2n =40

Mode of pollination : Self pollination

Inflorescence: Axillary racemes. Self pollinated crop.

Flowers: Bracteate, bracteolate, bracteoles two, shortly pedicellate, bisexual, hypogynous,

zygomorphic, complete, pentamerous, cyclic.

Calyx: Five gamosepalous, tubular and five unequal lobes.

Corolla: Papilionaceous corolla, posterior standard petal two lateral wing petals and two
anterior keel petals.

Androecium: The ten stamens diadelphous Pattern (9+1).

Gynoecium: The single pistil is unicarpellate and has one to four campylotropous ovules.
The style is about half the length of ovary and curves backward towards the free posterior
stamen. The stigma is capitate. Hairs are present on the pistil, and the outer surface of the
calyx tube.

D) Green gram/Black gram

1. Botanical name : Vigna radiate/Vigna mungo

2. Family : Fabaceae
3. Chromosome number : 2n= 22

4. Mode of pollination Self pollination

 Flowers are in an axillary or terminal receme, peduncle up to 13 cm in

length with clusters of 10to 20 flowers.
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  Corolla is yellow in colour and papilionaceous, sometimes curved 5-10 cm
long. Small flowers are borne in capitates clusters on the end of long hairy
peduncles.
 Petals are five in numbers, three kinds of petals, 1 standard, 2 wings and 2
keels.
 Androecium: male reproductive part stamen has got two parts anther and

filament.
 Gynoecium: Female reproductive part made up of stigma, style and ovary.
Gynoecium is monocarpellary with a superior unilocular ovary.
 The stigma is hairy and placentation is marginal.
 Keel encloses reproductive organs, 10 stamens and one gynoecium.

E) Groundnut:

Botanical name : Arachis hypogaea L.

Family : Fabaceae
Chromosome number : 2n =40
Mode of pollination Self pollination

The flowers are borne in the axils of the leaves, mostly near the base of the
plant.Sepals are 5and form a green colored tube

Petals are 5 and yellow in color, one standard, two wings and two keels.
The stamens are 10 monoadelphous with the stamina column surrounding
the ovary. Two of them are usually sterile.

Long style, club shaped stigma with hairy end, which is enclosed in keel.
Stalk of the ovary elongate to form a peg and curves towards soil.

Flowers open between 6.00 to 8.00 am. Anthers dehisce about 1-2 hrs before
opening of the flowers and fertilization completes before mid-day (6 hours
after pollination)

10
5

2) Horticulture crop:
A) Grape
Botanical Name: Vitis vinifera
Family :Vitiaceae
Floral biology: Flowers are small, green, sweetly scented and are borne on panicles
on current season growth. Three types of flowers viz., male, female and
hermaphoridite occur in grapes. Varieties of V. vinifera are mostly hermaphrodite
Perfect Flowers : Pistil is functional; stamens are erect and produced fertile pollen
Female flowers: Pistil is well developed; stamens are refluxed and may produce
abundant pollen, but remain sterile owing to the absence of germpores.
Male flowers: Stamens are erect and anthers produced well-developed fertile pollen
but pistil is only rudimentary without stigma and style with only a small ovary
containing incompletely developed ovules. The number of stamens varies from two
to seven but majority of flowers have five stamens viz., 1. Stamens having upright
filament and 2. Those in which the filaments are bent backwards and downwards
soon after the cap fall.

Petals and sepals are (five in number) fused and during anthesis the petals detach
from the base forming a cap like structure called ‘calyptra’.

Anthesis starts early in the morning and continues beyond 5.00 p.m., the peak
between 6.00 to 10.00 a.m. The time taken for completion of anthesis varies from
half a minute to one day, depending upon the variety, temperature etc. Stigmatic
receptivity has been characterized by the presence of sugary secretion on the stigma,
appearance. Once the stigmatic surface dries, it becomes black, indicating the loss
of receptivity. Stigma becomes receptive one day prior to anthesis and remains so, a
day after, with maximum receptivity on the day of anthesis.

5
 Exercise No. 4
Study of floral biology of important cross pollinated agriculture and horticulture crops
1) Agriculture crop

A) Maize:
Botanical Name: Zea mays
Family: Gramineae/Poaceae
Chromosome No: 20
Mode of pollination: cross pollination

Floral biology:
Maize bears monoecious flowers.
Staminate flowers are produced in tassel and Pistillate flowers
are on the shoot in the axil of leaf.

Staminate flower (tassel):

• The main stem of the maize plant terminates in a tassel, bearing two flowered
staminate spikelets.
• Each staminate flower having lemma, palea and three stamens.
• As the tassel flower opens, the anthers are pushed-out by the elongating
filaments and pollen grains are come out from the extruded anthers.

Pistillate flowers (silk / spadix):

• The shoots arise as branches from nodes about mid-way of the stalk. Each shoot is
composed of a shank from which the husk arises and terminates in the ear on which the
pistillate flowers are borne .

• The spikelets are borne in pairs. Each spikelet normally contains one fertile and one
sterile ovule. This results in an even number of rows of kernels on the ear.

• Fresh silks functions both as a stigma and style, being receptive to fresh pollen
throughout their entire length

•
Fertilization of the ovule usually occurs within 12-28 hours after the silk have been
pollinated.
B) Sunflower:

Botanical name- Helianthus annus

Family – Compositae
Chromosome No. = 2n=36
Mode of pollination- Cross pollination
Inflorescence: Head or capitulum which contains two types of florets
a) Disc floret b) Ray floret.
Disc floret: Bracteate, sessile, bisexual, complete, actinomorphic and
6
epigynous.
Calyx: calyx is reduced to pappus hairs, gamosepalous.
Corolla: United to form a tube.

Androecium: Stamens five, syngenesious anthers, epipetalous stamens, ovary inferior,

monocarpellary with single ovule and the style passes through syngenesious anthers
with bifid stigma.

Ray floret: With ligulate corolla, yellow in colour zygomorphic and calyx is
reduced to pappus hairs. Normally florets are sterile or sometimes pistillate.
Highly cross pollinated crop.
Fruit: Achene (or) cypsela.

2) Horticulture:
A) Mango
Botanical Name: Mangefera indica
Family : Anacardiaceae
Floral biology: Flower starts opening early in the morning from 4-7 a.m. and maximum flowers
open between 9.30-10.30 a.m. and complete 11.a.m. Dehiscence of anthers takes place at 11.30 a.m.
and it continues up to 3.45 p.m. The pollen grains are oval, or triangular or oblong. Stigma becomes
receptive even 18 hours before flower opening. Mode of pollination is entomophily; nectar is present
to attract the insects. The flowering duration is usually of short i.e. 2 to 3 weeks. The mango
inflorescence or panicle bears mainly two types of flowers – male and perfect. The number of
flowers per panicle varies between 1000 to 6000 depending upon the variety and climatic factors.
The percentage of perfect flowers varies between 0.74 per cent in Rumani, 16.41 to 55.7 per cent in
Neelum and up to 69.8 per cent in Langra.
B) Citrus:
Botanical Name: Citrus sp.
Family: Rutaceae
Floral biology: Flowers are produced on current season growth in cymes, both axillary and terminal
position. Two types of flowers, perfect and imperfect are found. The flowers are white in colour in
most of the species except lemon and citron where they are purplish on the outside. Flower opening,
starts from morning and extends up to evening but maximum anthesis is between 11.00 a.m. to 12.00
noon. The viability of pollen grains varies from 45-80% depending upon the season. The dehiscence
of anthers takes place there 45 minutes before anthesis or within 45 minutes after anthesis. It varies
up to 5 hours after anthesis. The receptivity of stigma starts either 15 minutes to 2 hours before
anthesis or within 35 minutes to 5 horus after anthesis depending upon weather. The receptivity lasts
for 4-8 days after anthesis.
7
C) Banana:
Botanical Name: Musa sp.
Family: Musaceae
Floral biology: Flowers are placed in the axils of the bracts, arranged biseriately and number
about 12 to 20 per node. Basal flowers behave as pistillate flowers while the terminal ones as
staminate. At the bottom end, they form a bulbous male bud. Pistillate flowers are large in size and
have well developed ovaries. Stamens (5) are reduced to staminodes, ovary inferior and trilocular.
Sytle stiff and long, stigma club shaped and sticky. Staminate flowers have long stamens 5, filaments
filiform, free, anthers two lobed. The female and male flowers opne by 6.30-8.00 a.m.

D) Sapota:
Botanical Name: Achras sapota

Family : Sapotaceae.

It is a wind pollinated one. Flowers are protogyny and the stigma grows out of the bud about two
days before anthesis. Flowers open between 4-4.30 a.m. Anthers dehisce between 8-10 p.m. The
flowers keep fresh for nearly two days. The stigma is found to be receptive two days before
opening and continues to be like that up to 12 hours after opening. Peak receptivity is between 8-
10 a.m. The total time taken from fruit set to maturity is 10-12 months under North Indian
conditions but in Tamil Nadu it takes only 4-5 months.

Flowers are emasculated and bagged before 4-5 p.m. and well before the stigma protrudes out of
the bud. The actual procedure consists of making a circular incision around the flower bud with
sharp knife or blade, so that 2/3 of the upper floral cup is removed including the portions of
calyx, corolla and epipetalous stamens. The style is left in position in remaining 1/3rd of the
floral cup. Stamens from male parent, which should shed their pollen in the early hours of next
day, are collected in the previous day evening and kept over night in a petridish. These are used
to pollinate the receptive stigma of the emasculated flower between 8-10 a.m. in the next day.

E) Guava:
Botanical Name: Psidium guajava
Family : Myrtaceae
Floral biology: The guava flower has a superior calyx with 5 lobes and the corolla of 6-10
petals arranged in one and two whorls. The androecium consists of 160 to 400 thin filaments
carrying bilobed anthers closely packed together. The gynoecium consists of an inferior
ovary, syncarpous with axile placentation and subulate terminal style. The style is smooth
and red at the summit. It is larger than filaments but bent over stamens in bud stage .
Three flowering seasons were reported in the peninsular regions of India, namely, ambe
bahar, mrig bahar and hatti or hastha bahar. In north Indian subtropics, there are only two
flowering seasons, however, have reported three distinct flowering and fruiting periods in
spring, rainy and winter seasons in Delhi. The peak anthesis was found to be between 5.00
and 6.30 AM in most of the varieties. However, in Chittidar and Lucknow Round, it was
observed between 6.30 and 7.00 AM The dehiscence of anthers starts 15 to 30 minutes after
anthesis in all the varieties and continues up to 2 hours. After dehiscence, the anthers assume
a whitish colour caused by the pollen. No fixed relation has been observed between the
8
 atmospheric temperature and humidity and the time of anthesis and dehiscence.Pollen
fertility has been found to be high in all the cultivars of guava.

F) Annual moringo:
Botanical Name: Moringa pterygosperma
Family : Moringaceae.

Anthesis has been reported to commence as early as 4.30 AM and continue till 6.30 lAM peak.
Observed at 5.30 A.M In another report the anthesis was found to be form 5 to 9 hrs. In association
with temperature range of 27.3 to 29.20C and RH 68 to 78%. At Horticultural College and Research
Institute, Periyakulam the anthesis was reported to commence as early as 2.30 AM and continue till
7.00 AM peak being 5.40 AM.
Anthesis was irregular. However no flower was found to open after 7.00AM. The anther
dehiscence starts around 4.00 AM and continues upto 6.30 AM the peak at 5.30 to 5.45 AM. In
anther of longest stamen dehisces first followed by the stamens in the descending order of filamental
length. At full maturity the anthers are greenish yellow and after dehiscence they turn to pale colour.
On an average each anther has 7400 pollen and the diameter of each pollen measures 5-4 microns.
The stigma becomes receptive a day prior to opening of flowers and continues to be receptive on the
day of opening. The receptivity is Breeding of Field & Horticultural Crops www.AgriMoon.CoM
lost on the next day of anthesis too.
The flowers are good source of nectar and hence the pollination was predominantly by
honeybees. Cross-pollination yields good fruit set and seed set than self-pollination. The pollen
viability is 72% at anther dehiscence. Pollens stored beyond 66 hours were unable to germinate. By
hand pollination using fertile pollens there was even 100 percent set. But under natural condition the
fruit setting ranges from 11-15% depending upon the seasons.

9
 Exercise No. 5
Study of floral biology of important often cross pollinated agriculture and horticulture
crops
A) Sorghum:

Botanical Name : Sorghum bicolor L.

Family : Poaceae
Chromosome No. : 2n =2x =20
Mode of pollination : Often cross pollinated

Floral biology:
Inflorescence:
The inflorescence is a panicle (called arrow) with a central rachis from which primary branches
arise. They give rise to secondary and sometimes tertiary branches which carry the racemes of
spikelet.
Panicle:
The central axis of panicle the rachis may be striated and it may be hairy or glabrous.
Raceme:
A raceme always consists of one or several spikelet .Raceme may vary in length according to number
of nodes and length of internodes.
The spikelet generally occurs in pairs, one being sessile which is bisexual perfect and fertile,
these can born on short pedicel which maybe sterile or staminate.
Flower:
 Perfect flower consist of two glumes, one hairy lemma, a small palea, small awn, three
stamens, two lodicules and one pistil.
 Ovary has two styles with feathery stigmas.
 Sorghumisshortdayplantandfloweringishastenedbyshortdaylengthandhigher temperature.
Anthesis:
 The sorghum flowers bloom just prior to sunrise.
 Blooming starts from apex and moves downward.
 Anthers and stigma pushed out as the glumes open.
 Pollen remain viable for 3-6hrs. while stigma remain receptive for 7 days but are most receptive up
to 3days after emergence.

B) Cotton:
Botanical name: Gossypium hirsutum
Family : Malvaceae

Chromosome No: Gossypium hirsutum (2n = 52)

Cultivated spp. G. arborium (2n = 26)

10
 G. herbaceum (2n = 26)

G. barbadense (2n = 52)

Mode of pollination: Often cross pollination

 The base of cotton flower is surrounded by leaf like triangular bracts that is commonly
known as squares.
 On the day before pollination the twisted corolla emerges from the bracts.
 When the corolla first opens the petals may be white, cream, yellow or purple in the
different varieties. The following day the corolla turns pink gradually changes to red
and finally falls from the plant.

 The stamens are numerous, forming a tube like staminal column around the style
which is united with the inside base of the corolla.
 Pistil is formed 3-5 carpels corresponding to the number of locks in the ball.
 Pollen is shed directly on the stigma when the anthers open or it may be carried to the
stigma by insects may in considerable (5-25%) cross pollination.
c) Pigeon pea:

Botanical Name : Cajanus cajan L.

Family : Fabaceae
Chromosome Number : 2n =22
Mode of pollination : Often cross pollination

Inflorescence: Flowers are borne in axillary corymbose racemose, often forming a terminal
panicles. Self pollinated crop.

Calyx: Five tubular campanulate.

Corolla: Papilionaceus, five petals, one standard, two wing and two keel, petals,

Polypetalous.

Androecium: Stamens 10, diadelphous. The stamens show dimorphism of the 10 stamens,
four have short filaments and six including the old posterior one, have long filaments.

Gynoecium: The ovary superior, sub sessile, densely pubescent and glandular punctate
with 2-9 ovules. The style is long filiform, and glabrous. The stigma is capitate.

11
 Exercise No. 6
Types of isolation, determination of isolation distance study of isolation requirements
in different crops for foundation and certified seeds.

Isolation distance: It is a minimum separation required between two or more varieties of the same
species for the purpose of keeping seed pure.
The crop raised for seed production should be separated from other fields of the same crop
species by a minimum distance, which varies from one crop to other. It is essential to prevent pollination
from unwanted pollens in case of cross pollinated species and often cross pollinated species and to avoid
mechanical mixture, chance of cross pollination in self pollinated species.
The minimum isolation distance required for a foundation seed crop is markedly greater than that
of certified seed crop in case of all those crops where pollen contamination is feared.
Types of isolation:
1) Spatial isolation
2) Temporal isolation
3) Physical barrier
4) Physical isolation

1) Spatial isolation: The spatial (space) separation required between a seed field and other sources
of genetic and mechanical contamination especially between verities of cross pollination.
More the extent of out crossing wider the distance.
Higher the class of seed wider the distance.
In hybrid seed production wider the distance than that of the variety.

2) Temporal isolation/ Time isolation : crop of seed production should be sown early or late by a
margin of 15-20 days than neighboring fields of same or other variety to prevent entry of foreign
pollens in the field of seed production.

3) Physical barrier: In surrounding of crop of seed production particularly on bunds crop of well
plant height and densely planted should be grown to prevent entry of foreign pollens in the field
of seed production.

4) Physical isolation: It can be practiced by covering the floral part or whole plant we can avoid the
physical contaminants e.g. Paper bags

Factors affecting on isolation distance:

 Pollination behavior of the crop: Plants that primarily self pollinate require less isolation
distance, while plants that primarily cross pollinate require greater distance.
 Pollinating agent: On the basis of pollination agent pollination categorized as:
A) Abiotic pollination:
Wind pollination
Water and rain pollination
B) Biotic pollination:
Domesticated bees (honey bees)
12
 Butterflies
Moths
Flies
Wasps
 Pollen trials:
Pollen viability
Pollen production capacity
Pollen mate
Time of anthesis
 Wide pollinator
Inter varietal crossing
Intra generic crossing
Inter generic crossing
 Designated diseases
It refers to the disease specified for the certification of seeds and for which certification standards must
be met with.
Crop designated disease Causal organism
Wheat Loose smut Ustilago tritici
Sorghum Grain smut/kernel smut Spacelotheca sorghii
Mustard Alternaria blight Alternaria sp.
Pearl millet Green ear Sclerospora graminicola
ergot Claviceps microcephala
Sesame Leaf spot Alternaria sp.
Brinjal Little leaf Datura virus 2
Chillis Anthracnose leaf blight Gloesporium piperatum
Cucurbits Mosaic Cucumis virus
Cow pea Anthracnose Colletotricum species
Bhendi Yellow vein mosaic Hibiscus virus
Tomato Early blight Alternaria solani
Leaf spot Xanthomonas vesicatoria

 Type of seed production: In hybrid seed production wider the distance than that of varietal
improvement (inbred pure line)

 Class of seed: Higher the class of seed wider the distance i. e. breeder seed production requires
relatively large isolation distance compared to foundation seed production.

Importance of maintaining isolation distance

Isolation prevents unwanted cross pollination
It is primary practice required to keep varieties true to type
Isolation begins with an understanding of a distance required to limit or eliminate chances of cross
pollination between two varieties of the same species
Isolation distance managed through several methods:
By distance
By timing flowering
By contaminant
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Isolation by distance is the most reliable method, which involves providing enough distance between
variety and any source of contaminating pollen to ensure that the variety remains true to type.

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16
 Exercise No. 7

Study of emasculation and hybridization techniques

EMASCULATION:
Removal of stamens or anthers or killing the pollen of a flower without the female
reproductive organ is known as emasculation. In bisexual flowers, emasculation is essential to
prevent of self-pollination. In monoecious plants, male flowers are removed. (castor, coconut)
or male inflorescence is removed (maize). In species with large flowers e.g. (cotton, pulses)
hand emasculation is accurate and it is adequate.

Methods of Emasculation:

1. Hand Emasculation:
In species with large flowers, removal of anthers is possible with the help of forceps. It is
done before anther dehiscence. It is generally done between 4 and 6 PM one day before
anthers dehisce. It is always desirable to remove other young flowers located close to the
emasculated flower to avoid confusion. The corolla of the selected flower is opened with the
help of forceps and the anthers are carefully removed with the help of forceps. Sometimes
corolla may be totally removed along with epipetalous stamens e.g. gingelly.

In cereals, one third of the empty glumes will be clipped off with scissors to expose
anthers. In wheat and oats, the florets are retained after removing the anthers without
damaging the spikelets. In all cases, gynoecium should not be injured. An efficient
emasculation technique should prevent self pollination and produce high percentage of seed
set on cross pollination.

2. Suction Method:
It is useful in species with small flowers. Emasculation is done in the morning
immediately after the flowers open. A thin rubber or a glass tube attached to a suction hose is
used to suck the anthers from the flowers. The amount of suction used is very important
which should be sufficient to suck the pollen and anthers but not gynoecium. In this method
considerable self-pollination, up to 10% is like to occur. Washing the stigma with a jet of
water may help in reducing self-pollination; however self pollination cannot be eliminated in
this method.

3. Hot Water Treatment:

Pollen grains are more sensitive than female reproductive organs to both genetic and
environmental factors. In case of hot water emasculation, the temperature of water and
duration of treatment vary from crop to crop. It is determined for every species. For sorghum
42-48OC for 10 minutes is found to be suitable. In the case of rice, 10 minutes treatments
with 40-44OC is adequate. Treatment is given before the anthers dehiscence and prior to the
opening of the flower. Hot water is generally carried in thermos flask and whole inflorescence
is immersed in hot water.

4. Alcohol Treatment:
It is not commonly used. The method consists of immersing the inflorescence in alcohol
of suitable concentration for a brief period followed by rinsing with water. In Lucerne the
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 inflorescence immersed in 57% alcohol for10 second was highly effective. It is better method
of emasculation than suction method.

5. Cold Treatment:
Cold treatment like hot water treatment kills the pollen grains without damaging
gynoecium. In the case of rice, treatment with cold water 0.6OC kills the pollen grains
without affecting the gynoecium. This is less effective than hot water treatment.
6. Genetic Emasculation:
Genetic/ cytoplasmic male sterility may be used to eliminate the process of emasculation.
This is useful in the commercial production of hybrids in maize, sorghum pearlmillet, onion, cotton,
and rice, etc.
In many species of self-incompatible cases, also emasculation is not necessary, because
self-fertilization will not take place. Protogyny will also facilitate crossing without
emasculation (e.g.) Cumbu.

7. Use of Gametocide:
Also known as chemical hybridizing agents (CHA) chemicals which selectively kills the male
gamete without affecting the female gamete. eg. Ethrel, Sodium methylarsenate, Zinc methyl
arsenate in rice, Maleic hydrazide for cotton and wheat.
Bagging
Immediately after emasculation the flower or inflorescence enclosed withsuitable bags
of appropriate size to prevent random cross-pollination.

Hybridization: Crossing between two different species of the same genus or

two different genera of the same family is called distant hybridization. Such
crosses are called as wide crosses or distant crosses. Wide hybridization is an
effective mechanism in transferring desirable genes into cultivated plants from
related species and genera.
It mainly depends on chromosome number, crossability, fertility and
homology between the two species. Distant hybridization is of two types
(a). interspecific hybridization and (b). intergeneric hybridization.

A. INTERSPECIFIC HYBRIDIZATION
Crossing two different species of the same genus is called as
interspecific hybridization. It gives rise to three types of crosses namely, fully
fertile cross, partially fertile cross and fully sterile cross.
1. FULLY FERTILE CROSS
When the two parents in a cross, have complete chromosomal
homology then normal pairing of chromosomes during meiosis results in fully
fertile hybrids. Examples of fully fertile crosses in various crops are as
follows:

Cotton: G. hirsutum x G. barbadense

G. arboretum x G. herbaceum

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Wheat: Triticum aestivum x T. compactum

Oats: Avena sativa x Avena byzantiana

Soybean: Glycine max x Glycine soja

Sugarcane: Wakker made the first reported interspecific cross between

S.officinarum and S.spontaneum for incorporating disease resistance into
nobel canes.
Pearlmillet (Pennisetum sp): In, Pennisetum the primary gene pool includes
the wild, grassy, and weedy P.americanum subspecies (2n = 14) monodii and
stenostachyum that readily cross with pearlmillet and produce fertile hybrids.
The subspecies monodii is a rich source of germplasm that can be used to
improve pearlmillet.

2. PARTIALLY FERTILE CROSSES

When the two species involved in a cross differs in chromosome
number then the hybrids become partially fertile and partially sterile. Examples
Wheat.

Triticum aestivum x Triticum durum partially fertile F1

2n = 42 (AA BB DD) 2n=28 (AA BB) 2n = 35 (AABBD)

3. FULLY STERILE CROSSES

When the two species involved in a cross have no chromosomal

homology then the hybrids become sterility. The lack of homology does not
permit pairing
of chromosomes during meiosis. Such hybrids are made fertile by doubling
their chromosomes by colchicine treatment. Example

Nicotiana sylvestris x Nicotiana

tomentosa (2n=24) (2n=24)

F1 (Fully sterile)

Chromosome doubling with colchicine

Fully fertile tetraploid (2n=4x=48)

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B. INTERGENERIC HYBRIDIZATION
It refers to cross a between plants from two different genera
of the same family. Examples are as follows:
(i). Wheat x Rye cross: A cross between wheat (Triticum aestivum)
(2n=42) and Rye (Secale cereale) (2n=14) resulted in Secale cereal
(2n=56). The F1s were sterile and became fertile upon
colchicine treatment resulting in an amphidiploid called triticale.

DRAWBACK IN WIDE HYBRIDAZATION

A key observation on utilizing wild species germplasm for

crop improvement is the predominance of sterility in F 1 hybrids.
Even when partially fertile interspecific hybrids are produced,
linkages with undesirable genes many times limit the usefulness of
these hybrids. The most common fertility restoration method is
colchicine treatment of sterile F1 plants, but naturally occurring
unreduced gametes have also played a role in fertility restoration for
a few groups. Many examples could be cited when the initial
hybrids were made, fertility was restored, and the hybrids were
abandoned after a few generations, due to addition of sterility
barriers.

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 Exercise no. 8

Study of distinguishing morphological characters in A, B and R lines of

released hybrids.
A line: A line is male sterile line considered as female parent in hybrid seed production.

B line: Male fertile line Known as maintainer line.

A and B are isogenic lines. ( lines that posses same genotypic make up with one gene in
difference.

R line: This is fertility restorer line used as pollinator to the female parent.

Characteristics of A line: Stable sterility

Well developed floral traits

Easily, widespectrum strongly restored.

Characteristics of B line: Maintainable line.

Well developed floral traits.

Good combining ability

A and B nucleus should be same only cytoplasm will vary.

Characteristics of R line: Able to restore/ fertile the male fertility character.

Good combining ability

Taller than A line

Large pollen load, normal flowering traits.

No extra parents need for R line multiplication.

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 Exercise No. 9

Study of synchronization techniques for hybrid seed production, planting

ratio, supplementary pollination techniques, border rows for hybrid seed
production.

Synchronization: simultaneous flowering of male and female parent in seed production is

known as synchronization. The synchrony in flowering male and female parent is termed as
nicking. It is essential for commercial large scale production of hybrid seeds in different crops.
Synchronization in flowering between parental lines assumes greater importance as the seed set
on female parent depends on amount of pollen supplied from the male parent during flowering
period.

In producing hybrid seed considerable difficulties are experienced when two

parental lines in crossing plot do not flower at the same time. These difficulties could be
removed if a method were enabling as to synchronize the flowering periods. In hybrid seed
production seed yield and its quality attributes can be increased through proper synchronization
of flowering of parental lines and optimum female to male parental rows. The hybrid seed is
produced generally by planting the parental alternating each other at a definite row proportion.
This planting row proportion decides the amount of hybrid seed produced and is influenced by
vigour of the parents and especially the pollen production ability of male parent. To get more
hybrid seed, one may be tempted to put more number of female rows without taking care to
supply adequate pollen to all seed parent rows.

Synchronization techniques:

1) Staggered sowing of male parents:

2) By water management
3) By fertilizer management
4) Supplementary spraying

Synchronization in rice:

1) Staggered sowing of male parents:

seedling date is usually determined by leaf age, effective accumulated temperature (EAT)
and growth duration.
In general the period from initial to full heading of a CMS line is 4-6 days longer than for
a restorer line.
First sowing of male parent establishes the dates for second and third sowing.
The second sowing is done when the leaf emergence on the first sowing is 1:1 and third
sowing when the leaf emergence is 2:1
The planting ratio for sowing at different dates is 1:2:1

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 2) By water management:
Irrigation during later stages of panicle differentiation
Draining water from the field will delay the male parent panicle development
Higher standing water will speed panicle development.
3) By fertilizer management:
Beginning about 30 days before heading.
During first three stages of panicle differentiation, treat earlier developing parent with
quick releasing N fertilizer.
Urea spray @ 2% enhances flowering.
Spray the later developing parent with potassium dihydrogen phosphate @ 1.5%.
Foliar spray of 400 ppm maleic hydrazide (MH) to female parent at panicle initiation
delayed the flowering by 3.5 days.
4) By GA3 application: Spray GA3 @ 75g/ha with knapsack sprayer in two split doses, i.e.
first spray on 15-20% ear head emergence and second spray in the next day for enhanced
seed set.

Synchronization in sorghum:

Adopt staggered sowing.

The advancing parent has to be sprayed with 500 mg of maleic hydrazide in 1 litre of
Water, 45 days after sowing.
1% urea solution can be sprayed on late parent.
One irrigation may be skipped for the advancing parent.
Flowering can be delayed, by spraying CCC (Choro chlorine chloride )@ 300 ppm.
Synchronization in pearl millet:
Adopt staggered sowing of parental lines which are early in flowering.
1% urea sugar solution can be sprayed on lagging parental line.
Jerking of early parent delay the flowering.
To enhance the growth of lines lagging behind, extra nitrogen fertilization or foliar spray
may be used.
Synchronization in sunflower:
Flowering behavior of parental lines should be ascertained in the hybrid seed production.
There should not be more than 3 to 4 days difference in the flowering to avoid stagger
Problems.
In case of BSH-1, DRSH-1, and PKVSH-27 hybrid seed production, simultaneous sowing
of both parents is recommended.
However, R lines has to be sown 8days early in case of KBSH-1 and 4 days later in case of
KBSH-1 and 4 days later in case of APSH-11.

Planting ratio: It refers to number of rows of male parent of that of the female parent in hybrid
seed production plot.

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 - It depends on nature of pollination
- Amount of pollen shed
- Mode of pollination
- Pollen viability
- Stigma receptivity
- Wind velocity
- Density and movement of vector
- Height of the pollinator
- Agronomical management.
It differs from crop to crop, genotype to genotype and place to place.
First and last row should be preferably being of pollinator.
Row should be across the wind direction.

Supplementary pollination: For effective pollen dispersal during peak flowering stage for
Specific period (days & timing) for maximizing outcrossing for proper seed setting.
- Rope pulling or shaking of pollen parent with the help of two
bamboo sticks. 30-40% of spikelet opening stage. This process repeated 3-4 times during
day time (10 am to 1pm ) at an interval of 30 min. this repeated for 7-10 days during
flowering period.
- Rubbing the heads of two neighboring plants with each other. It is done during mid
flowering stage between 7-11 am for 2 weeks.

Border rows: Four rows of male parental lines to be grown all around the field called border
rows. Border rows will provide pollen grains to female plants.

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 Exercise No. 10 Study of seed cleaning and grading technique, seed packing and seed treatment techniques.

Type of upgrading operation and types of Principles of operation of the machine Use of machine / General Specific usage
machines usage
1 2 3 4
1. Sizing and grading These machines make extremely sensitive and a) Removal of splits from Sizing hybrid corn.
(Dimensional sizing) precise separations on the basis of differences in soybeans, peanuts, etc. Rice seed industry
(a) Width and thickness— sizing and width and thickness. The seeds are sized for b) The removal of chips
grading width by using round-hole screen openings, and and splits from sorghum
1. Horizontal flat screen separators, for thickness by using slotted screen openings. "seeds.
e.g., clipper corn sizer, superior The separators employ gravity, centrifugal force, c) Removal of cheat from
rock-it-corn grader product pressure, or a combination of these wheat.
2. Vertical ribbed screen separator, forces, to make width and thickness separations d) Removal" of cockleburs
e.g., Dockins seed grader. on screens. from cotton seed, wild
3. Cylindrical screen separators, e.g., onion from fescue, and wild
Carterrday precision grader, oats from barley.
superior high capacity sizer.
(b) Length sizing and grading Seeds are separated on a pure length basis. 1) Removal of weed seeds. Small grains,
1. Disc Separator In a disc separator, the discs lift uniformly 2) Cross broken crop seeds. Corn, soybeans.
shaped and sized and undersize particles, out of a 3) To upgrade general
mass of seed. The separation is not affected by appearance.
seed coat texture, weight per bushel or moisture 4) Size grade for precision
content. plantings

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 2. Cylinder The Cylinder separator operates on a centrifugal force principle
Separator in which the speed of the cylinder holds seed in the indents,
lifting them out of the mas until the indents are inverted to the
point where gravity causes the particles to fall. Shape and size
of the indents and seed, seed coat texture, moisture content and
weight of seed all combine to
affect seed separation.

2. Gravity or The gravity separator employs a flotation principle. In this 1. Removal of badly damaged,
weigh separation, seeds are vertically stratified in layers on the deck decriorated. Diseased seed and
separation according to density. Seeds of same size are stratified and insect damages crop seed.
s Gravity separated by differences in their specific gravity. Seeds of 2. Removal of empty, blind or
Separator same size are stratified and separated by differences in their sterile seed.
size. The oscillating movement of the table walks the heavy 3. Remove of abode balls, soil
seeds in contact with the deck uphill, while the air floats the particles and gravel and sand
light seeds downhill. The seeds travelling to the edge of the t mixed with certain kinds of seeds.
able range from light, at the lower end to heavy at the upper 4. Removal of contami -nating
end. The discharge can be divided into any number of density crop or weed seed.
fractions.

Stoners The stoner is essentially a modified gravity separator designed Removal of heavy inert material
to make two part separation by differences in specific gravity. from a larger volume of seed.
Its operationdepends upon vertical stratification of the seed
mixture according to weight, followed by

26
 separation of different strata on an oscillating

deck.

3. Air Separations pneumatic Pneumatic and aspirator separators use the movement 1. General cleaning. Precise separation e.g.,
Separators of air to divide seeds according to their terminal lifting seeds of meadow
velocities. Terminal velocity refers to the velocity of air 2. Close grading. foxtail from seeds of
required to suspend particles in a rising air current. 3. Specific Alta fescue.
Many factos such as density, shape and surface and separations
texture affect resistance of a particle to air flow. When
a seed mixture is introduced into a confined rising air
stream, all particles with a terminal velocity less than
the air velocity will be lifted; seeds with higher
terminal velocities will -fall below, against the air flow.
In a Penumatic

separator, the fan forces the air through the machine by

creating a pressure greater than.

27
 atmospheric. When the seed mixture encounters the air
stream, those seed and contaminants with a terminal
velocity, less than the air velocity (light seed), are lifted
and rise through the column to the top where they are
deflected into a discharge spout. Seed with a terminal
velocity greater than the air velocity (heavy seed), fall
through the column of air unitl they reach an inclined
screen positioned across the column. The seeds are then

deflected by the screen into the heavy seed discharge

spout.

Aspirators In an aspirator, the fan is at discharge end and induces a

vacuum which allows the atmospheric pressure to force
air through the separator.

In a scalping aspirator, the rough separation is made

when seed mixture is dropped into a rising air column
that has a velocity slightly below the terminal velocity
Scalping Aspirator of heavy, plump seeds. The leaves, trash and light seeds
rise with the air and are deposited in an enlarged
settling chamber. The denser plumper seeds fall
through the incoming air into a receiving bin below.

In a fractioning aspirator, when a seed mixture is

introduced into the lower end of an expanding air
column, heavy seeds fall against the air flow and light
seeds are lifted. Air velocity through the expanding
columns lessens and gradually drops out seeds with
lower terminal velocities. Each outlet along the column

28
 receives a lighter fraction of seeds; the mixture thereby
is separated into several grades.

Fractioning Aspirator

4. Surface Texture The velvet roll mill classifies seeds according to a Separation of smooth
Separation Roll Mill difference in texture of seed coat. clover seeds, e.g. from
rough seed coat, seeds
with irregular shape,
wrinkled or shriveled
seeds, broke.

29
 Seed packaging: After processing and treating are completed, seeds are packaged into
containers of specified net weight Packaging or bagging is essentially the last operation in
which seeds are handled in bulk flow. The packaging consists of the following operations:

1. Filling of seed bags to an exact weight.

2. Placing leaflets in the seed bags, regarding improved cultivation practices.
3. Attaching labels, certification tags on the seed bags, and sewing of the bags.
4. Storage/shipment of seed bags.
Equipment Used for Packaging of Seed
The bagger weigher: These are small machines which, when properly mounted beneath abin,
will fill and weigh a bag accurately in a single operation. Operational steps include:

a) The empty bag is suspended on the bagger weigher by a bag clamp.

b) Seed flow into the bag is begun, usually by a trip lever.
c) As seed flows into the suspended bag, a scale-type counter-balance mechanism is
actuated, so that when the proper weight of seed is in the bag, the seed flow lever is
tripped and seed flow is automatically stopped.
d) The bag now filled with the exact weight of seed is removed from the bagger
weigher and is closed. Bagger weigher and bagging scales used in. seed packaging
may be manual, semi-automatic or automatic.
Manual weighing: This type of scale, usually a portable plat- form, is considered
inefficient for volume weighing operations because of high labour requirements and
relatively low capacity, in terms of bags filled per minute. With this scale, bags are filled
to approximate weight, placed on the scale and then 'even weighed' with a hand scoop.
These scales are useful in following conditions:
(a) Weighing bags of non-free flowing seeds.
(b) A bagging bin is not available.
(c) Labour costs are minimal.
Semi-automatic: This is the most widely used scale. The scale is attached to the bottom
of a bagging bin, and the bag is clamped to the bottom of the scale. The feed gate is
opened manually and may be closed either manually or automatically when the proper
weight is attained. The scales have a capacity to weigh four to eight bags of 50 kg per
minute, depending upon the seed being packaged and the skill of the operators.
When selecting a scale of this type, the circumference and composition of bags or
containers must be considered. The orifice, or the bag clamp,, must be smaller than the
open end of the bag. However, too small an orifice and clamp will result in seed spilling
around the edge of the bag. The bag clamps hold bag materials of specific finish and
thickness, therefore, the composition of the bagging material, that is, jute, cloth, plastic,
paper, etc., should be stated when ordering the scale.
Automatic scales: Scales of this type are used primarily for small packages, e.g., vegetable
and lawn seeds. In these machines the entire weighing and filling process is done
30
 automatically. Installation is similar to the semi-automatic bagger. Some completely
automated systems pick up the empty bag; place it on a bagger, fill the bag and releases
the filled bag which then moves by conveyer to a bag closer.
Regardless of the types of scales used, they should be checked regularly to determine their
accuracy, particularly if they are portable. Frequent and careful cleaning of the weighing mechanism
will decrease the number of inaccurate weighings and extend the life of the scale.
Bag sewing machine : After an open-mouth bag is filled, the bag top must be sewed with
a bag sewing machine. Bag sewing machines are precision, high speed machines and must
be operated and maintained properly to prevent frequen* break-down and a short
operating life. For proper operation, the bag sewing machine must be:

Seed treatment: It is defined as application of fungicide, insecticide, bio-fertilizer or any other

Growth regulator either to control pathogens or insects or to improve the germination potential.

Objectives of seed treatment:

 Its main role is to protect seeds from seed borne diseases and pest attacks.

 To revive a seed that has been dormant for a long time.

 Drought tolerance is induced.

 Early emergence is used to increase the percentage of seeds that germinate.

 To keep birds and vermin out.

 Using X-rays, Gamma rays, and colchicines, obtain polyploides (genetic variety).

Different methods of seed treatment:

 Dry treatment: Mixing the seed with pesticides/nutrients in powder form.

 Wet treatment: Soaking the seeds in a pesticide/nutrient solution in liquid form.

 Slurry treatment: Seeds/seedlings are dipped in slurry. Rice seedlings, for example, are
immersed in phosphate slurry.

Pelleting: It is the process of coating seed with enough seed ingredients to make the seeds larger,

heavier, and consistent in size for sowing using seed drills. Pesticide pelleting is used to protect

soil organisms and pests, as well as to repel birds, ant.

31
Pre storage seed treatment

Fungicide, insecticide, or a combination of both, as well as any other chemical or plant product, are
applied to seeds prior to storage. The goal is to keep seeds fresh for longer by disinfecting them
against seed-borne or seed-storage diseases and storage insects, as well as minimizing seed
deterioration directly or indirectly.

Characteristics of ideal chemical seed treatment:

 It must be extremely effective in the face of harmful organisms.

 Seeds must be somewhat unaffected.

 Even if overused, it should be safe for humans, animals, and cattle.

32
Exercise No. 11
Seed testing – seed sampling

33
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35
36
37
38
 Exercise No. 12

Seed testing – Physical purity test and moisture test

39
40
41
Moisture test

42
43
 Exercise No. 13

Seed testing – seed germination test

44
45
46
47
Exercise No. 14

48
49
50
 Exercise No. 15

Visit to seed production plots of public and private sector companies.

51
 Exercise No. 16

Visit to seed production organization.

52