GPB 366 Practical Manual
GPB 366 Practical Manual
GPB 366 Practical Manual
Practical Manual
Crop Improvement –II(Rabi Crops)
EXERCISE NO. 1
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN WHEAT, OAT AND BARLEY
WHEAT
(Triticum species)
Family : Gramineae
Genus : Triticum
Species : aestivum
Chromosome Number : 2n = 6x = 42
Cultivated Species
There are two cultivated species of wheat, viz. common wheat (Triticum aestivum)
and durum wheat (T. turgidum L.). Common wheat is hexaploid (2n = 6x = 42), whereas
durum wheat is tetraploid (2n = 4x = 28). The former is more widely adapted than the later.
Common wheat is used for bread, cakes, noodles, cookies, chapatti etc., whereas the durum
wheat is used mainly for macroni and some flat breads. There are 16 wild species of wheat,
out of which six are diploids (2n = 2x = 14), seven tetraploids (2n = 4x = 28), and three
hexaploids (2n = 6x = 42). Wild species are used in hybridization programme for transfer of
resistance to biotic and abiotic factors, adaptation and other desirable characters into
cultivated species.
Wild Species:
T. timopheevii
T. dicoccoides
Aegilops speltoides,
A. squarrosa.
Origin :
Near East is the center of origin of bread wheat. It is believed that evolution of
common wheat in nature took place in two important steps. First, an amphidiploid tetraploid
species originated from a cross between two diploid species, one with A genome and other
with B genome. The amphidiploid after crossing with a diploid species with D genome gave
birth to the common wheat as given below:
The chromosome doubling took place in nature in above crosses. The other two forms
of hexaploid wheat, viz. T. compachim and T. spherococcum originated through spontaneous
mutations of T. aestivum. The durum wheat probably originated from cultivated emmer wheat
(T. turgidum var dicoccum) after several spontaneous mutations.
1
Crop Improvement –II(Rabi Crops)
Botany:
It is grown in Rabi season. It is annual plant growing up to height of 0.6 to 1.8 mts
with adventitious roots. The stem is cylindrical culm with hollow internodes. Leaves are
opposite with parallel venation. Leaf consists leaf sheath, blade, ligule and pair of auricles at
the base of leaf blade. The uppermost leaf of plant is called flag leaf. Tillers are produced
from the underground nodes by the axillary buds of the plant.
The inflorescence is of spike or ear type in which sessile spikelets are arranged
acropetally in zig-zag manner on mother axis i.e. rachis. Each spikelet consists of pair of
outer glumes which encloses 3 to 5 florets. Central one or two are sterile and remaining
florets are fertile. Each floret consists of outer glumes represented as scales, awned lemma
(flowering glume) and a palea. The lemma and palea enclose three stamens (having thread
like filaments and versatile anthers),single carpel (with bifid stigma) and two lodicules. The
lodicules help for opening of flowers. After fertilization, ovary develops into caryopsis,
which is a single seeded fruit with pericarp.
Crossing Technique:
Wheat is a self-pollinating crop. Flowering begins in the upper part of the spike and
proceeds in both the directions. Flowering on a spike is over within 2-3 days. For
emasculation roughly ¼ to 1/3 upper part of spike is clipped and a few basal, immature
florets are removed. In the remaining 5-6 pairs of florets, the central florets are also removed
and the emasculation is carried out in the remaining lateral florets of each spikelet. For this,
glumes are clipped back and anthers are removed with fine pointed forceps. The emasculated
spike is covered by a pollination bag. After 1-2 days, the stigmas are visible then the
emasculated spike is covered by pollination bag. Thereafter in 1-2 days, the stigmas are
visible and the emasculated spike is ready for pollination.
Pollination is done with the fresh pollen as pollen grains remain viable for a short
period (1-3 minutes). For preparing male spike, a spike showing a few protruding anthers is
removed from the male parent. Its glumes are cut without damaging the anthers. It is held in
vertical position/stalk inserted in ground under sunlight for a few minutes. During this
process, anthers emerge out of cut glumes. The upper portion of the pollination bag on the
emasculated spike is cut with scissors. Through this opening, the male spike shedding pollen
is inserted and shaken over the emasculated spike by twirling motion and the bag is closed
with the help of U clip. This simple and fast process of emasculation and pollination is
commonly followed at most of wheat breeding stations.
Breeding objectives:
i) High grain yield.
ii) Early maturity with short duration.
iii) Photo and thermo-insensitive varieties.
iv) Resistant to diseases like rust, loose smut, leaf blight etc. and pests like aphids,
armyworms and gujia weevil etc.
v) Responsive to high doses of fertilizers.
vi) Semi-dwarf varieties having synchronies productive tillers.
vii) Resistant to water logging and shattering.
viii) Good milling and baking quality i.e. suitable for chapatti and bread making.
ix) Amber grain colour and grain with high protein and lysine content.
x) Salt and drought tolerant varieties.
Breeding achievements:
1) Introduction: Sonora 64, Lerma rajo 64A, HI 977, Sonalika, Kalyansona Malvika
2) Pure line selection: NP 1,6,12, HB 208, K 852, Mondya 3-2, Motia, Gulab, Baxi-
288-18,
3) Pedigree selection after hybridization:
a) T. durum. N 59, MI 5749, Raj 1555, NIDW 15 (Panchwati)
b) T. aestivum: NI 747-19, Lok 1, HD 2189, HD 2278, Prgato (DWR 39),
NI-1917 (Kadwa)
4) Interspecific hybridisation
a) T. durum x T. polonicum : MACS 9 (d)
b) T. durum x T. dicoccum : Jay (d)
c) T. durum x T. dicoccum x T. aestivum : Niphad 4
5) Interspecific hybridization and back crossing
(T. durum x T. aestivum) : NP 890
6) Multiline varieties : NI 5439 Kharachiya 65
7) Mutation breeding: Pusa Lerma, Sharabti Sonora : NP 111, NP836
Assignment :
1) Dissect the floral parts of given sample and mount them on black mounting
paper. Draw the figures and label properly.
2) Practice hybridization technique in field and laboratory on given sample.
.........................
OAT
(Avena sativa)
Family : Gramineae
Genus : Avena
Species : sativa
Chromosome No. : 2n = 42
Oats are supposed to be of Asian origin and from there it spread to most of the
countries of the world like USA, Russia, Canada, Poland, Australia, France and Germany.
Cultivated species :
The Oats are classified according to their chromosome numbers and there are three
main groups of cultivated oats :
Group I :
This group of Oats contains 7 haploid chromosomes and the important of them are
Avena brevis (short oats), A. weistii (desert oat), A. strigosa (sand oats) and A. nudibrevis
(small naked seeded oats) etc.
Group II :
The oats belonging to this group contain 14 haploid chromosomes. The prominent
types of this group are Avena barbata (slender oat), A. absyssinica (Absyssinian oats).
Group III :
These types have 21 haploid chromosomes and the important members of this group
are Avena fatua (common wild oats), A. sterilis (wild red oats), A. sativa (common oats), A.
byzantina (cultivated red oats) and A. nude (hull less oat).
Inflorescence :
Inflorescence of the oat plant is panicle composed of a central loose, open rachis with
five to seven nodes, from which branches areas bearing spikelets. Each lateral branch
terminates in a single apical spikelet. Other spikelet is born on second or third-order of
branches. Each panicle may have 20 to 50 spikelets. Each spikelet consists of several florets
enclosed in two empty glumes, with the tip of one glumes extending slightly above the other.
Florets within each spikelet are arranged alternatively upon a central axis, the rachilla, and
usually the two basal florets are fertile. The flowers are perfect zygomorphic, bracteates and
hypogenous. The flower consists of a lemma and palea, two lodicules, three stamens and one
pistil.
Spikelet :
Three to four florets are present in each spikelet, but the third or fourth floret is sterile,
two glumes cover these florets.
Glumes :
The two outer bracts of the spikelet are broadly lanceolate, pointed boat shaped,
usually labours and arched. The glumes may be pale, yellow or red.
Lemma :
Lemma is a rigid structure which enclosed the rachilla at the base of the flower. Its
primary function is to protect the caryopsis. Lemma is bified and varies in colour, being
white, yellow gray or red to black. It may be awn or awnless.
Palea :
One membranous palea is present opposite to lemma. Primary function of palea is to
protect the caryopsis.
Lodicules :
Two small, smooth, pointed and shinning lodicules are present at the base inside the
floret, mature lodicules are thick at the base and pointed at the tip. The action of the lodicules
in opening the flowering glumes is not so important in self fertilized plants.
Androecium :
There are three stamens present. Stamens first appear as papillae upon the apex of the
floral axis above the flowering glues primodia. The anther consists of four locules. The
filament is attached to the central axis at its lower extremity.
Gynoecium :
There is one ovary with bifid stigma. The ovary is elliptical in cross section. Long
monocellular, epidermal hairs entirely cover the ovary and also present on the interior surface
and base of styles. The tip of style and the inner surface nearly to the base, are covered with
stigmatic branches. A single sessile anatropous ovule is located inside the ovary.
Crossing Technique :
Emasculation :
Since anthesis normally occurs in the afternoon, emasculation should be done in late
forenoon or early afternoon. Select those spikelet from a panicle in which anthesis is
expected one or two days after emasculation. Keep only one floret within a spikelet. By
applying light pressure on the dorsal glumes, separate glumes, palea and lemma and remove
all the three anthers with the forceps. When emasculation is delayed until very shortly before
the time of normal anthesis, the floral structure, being better developed will likely to be less
injured by operation than they would be if manipulated a day or more before pollination.
Cover the emasculated floret with a glassine bag to prevent contamination from foreign
pollen and tagging is to be done.
Pollination :
Researchers have reported different time interval for pollinating the emasculated
floret. Few reported optimum time between emasculation and pollination as one to three
days, while others suggested emasculating the floret in the morning and pollinating in the
evening. Anthers from desired male parent are collected. The anthers will be yellow plump.
Separate lemma and palea of emasculated floret and place the collected anthers with the help
of forceps in the inner side of the lemma. Cover the pollinated floret with the same glassine
bag used in emasculation. Approach method of pollination is also used in oats. In approach
method remove secondary floret and the anthers of the primary floret. The upper portion of
each spikelet, after emasculation is removed by clipping glumes, lemma and palea just above
the stigma. The pollens from the male parent shed directly on the stigma of erect clipped
spikelets.
Breeding objectives :
The broad objectives in breeding spring and winter varieties of oats are high grain
yield, earliness, lodging and shattering resistance, disease resistance and quality. Winter
hardiness and forage production are additional objectives in the breeding of winter oats.
Achievements :
.........................
BARLEY
(Hordeum vulgare L.)
Fertility of the lateral spikelets forms the basis of barley classification and the
cultivated barley may be classified into three main groups viz.,
i) Six rowed barley (H. vulgare L. emend, Lam)
ii) Two rowed barley (H. distichum, L. emend, Lam)
iii) Irregular barley (H. irregular, E. Aberg and Wiebe)
Botany :
The cultivated barley plant briefly consists of the following parts :
1) The roots, both seminal and permanent.
2) The stem (culm), cylindrical with hollow internodes, and from 5 to 7 solid nodes.
3) Leaves, borne alternatively on opposite sides of the stem and arising at each node.
4) The spike (head) at the top of the stem, consisting of the flowers arranged in
spikelets, three of which are attached at each node of flat zig-zag rachis.
5) The spikelet, having two glumes and the floret.
6) The floret, consisting of the lemma and the palea, which enclose the male and
female flower parts.
7) The kernel, consisting of the caryposis in naked barleys but including the lemma, the
palea and the rachilla. Which adhere to the caryopsis, in hulled barleys (Reid and Wiebe,
1979).
Each spikelet is single flowered and consists of two glumes and a floret. In six-rowed
barley, three spikelets are attached at each node of the rachis, and these triplets alternate from
side to side of the rachis. In two rowed barely, only the central spikelet of a triplet is fertile.
The flower is enclosed in lemma and palea. The pistil has a two branched feathery stigmas.
Three anthers are attached to the long slender filaments. The spike of barely besides being
characterized as six-row and two-row, is also described as hooded vs. awned. The hood is a
three-lobed appendage at the tip of the lemma. The hood may be either slightly elevated on a
short awn segment or sessile.
Crossing :
Flowering of barley spikelets begins in the central florets of the upper of the spike and
proceeds in both the directions with the swelling of lodicules at the base of the florets, the
flowers open and the filaments elongate. During elongation of the filament and emergence of
the anthers, the dehiscence germinated five minutes after it reached the stigma, that within 10
minutes the two male gametes had entered the pollen tube. Within 40 minutes, the male
gamete has reached the site of micropyle. Within 45 minutes, the male gamete had entered
the egg sac. The first division of the fertilized egg completes five hours after pollination.
The emasculation is done in the early stage of the spike development when the spike
is slightly visible through the covering of flag leaf. The upper 1/3 portion of the spike is
removed. The emasculated spike is covered with butter paper bag. After 2-3 days, when the
stigma is protruding. The spike is ready for pollination are carried out by twirling a spike
with ripe with ripe anthers over the emasculated spike.
Breeding Objectives :
i) Yield improvement
ii) Increased adaptability
iii) Resistance to yellow rust, aphid and nematode
iv) Improvement in nutritional quality
v) Improvement in attributes related to malt industry
Achievements of Barley :
.............................
EXERCISE NO. 2
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN CHICKPEA AND LENTIL
CHICKPEA
(Cicer arietinum)
Family : Leguminoceae
Genus : Cicer
Species : arietinum
Chromosome Number : 2n = 16
Cultivated species:
Related species : C. reticulatum
C. pinnatifidum
C. songaricum
Two main categories of Chickpea are recognized which are distinguished mainly by
their seed characteristics. They are
1) Desi types, which are relatively smaller, angular seeds with rough yellow to
brown coloured testas.
2) Kabuli types, with large, more rounded and cream coloured seeds.
Wild species :
The wild species of Cicer closely related to chickpea are :
i) C. bijugum
ii) C. echinospermum
iii) C. ecticulatum
Origin:
The chickpea is most probably originated in an area of present day south-eastern
Turkey and adjoining Syria.
Botany:
Roots are robust and long. Stems are branched, flexuous or straight, erect to prostrate
and usually ribbed. In general, height ranges from 20 to 100 cm. Three types of branching are
defined. They are primary, secondary and teritiary branches. The leaves include rachis and
leaflets. The average rachis length is 3-7 cm. Each rachis has on average 10-15 leaflets,
inserted on small pedicels. The leaf is pseudoimparipinnate i.e., the ending terminal leaflet is
not in true terminal position, but in sub terminal position (the central vein oblique to the
rachis).
The flowers are papilionaceous. They are solitary in axillary racemes. Double flowers
are rare, but are very much sought after by the breeders as possible sources of yield increase.
The calyx has five deep lancelolate teeth. Peduncle and calyx are hairy. Generally, corolla is
white. The vexillum is obovate, 8-11 mm long and 7-10 mm wide. Wings are obovate, 8-9
mm long. The keel is 6-8 mm long.
The androecium is diadelphous [(9) + 1]. The ovary is ovate, pubescent, 2-3 mm long,
and 1-1.5 mm wide. It has 1-3 ovules, rarely 4. The style is 3-4 mm long, generally glabrous.
The stigma is globose. Number of pods/plant is highly variable, generally between 30 and
150 depending on the year, location, sowing time and other factors. The seed is beaked, and
very frequently ramhead shaped and strongy wrinkled or ribbed. Pod size has been found to
be a stable charcter and based on this, two goups viz., macrocarpa and microcarpa have been
postulated in C. arietinum.
Crossing Technique :
Crossing in chickpea is usually difficult and time consuming as is the case with most
of the legumes. For emasculation, the bud is hold in left hand and gentle pressure is exerted
to open the standard and wing petals. The keels are opened with forceps and the stamens are
removed. Pollens are collected from half-open flowers and put on stigma. For better results, it
is suggested that the crossing should be attempted after formation of first pod on the plant and
as far as possible and large bud sized cultivars should be used as female parent. In Chickpea
anthesis starts between 9 and 10 am and may continue up to 3 pm. The flowers remain open
for two days, the flowering process over early on the second day. The plant is primarily self
pollinated as anther dehiscence takes place forty hours prior to opening of flowers. The
process of anther dehiscence prior to opening of flowers termed as cleistogamy has been
recorded in the species.
Breeding Objectives :
(i) Increased seed yield.
(ii) Increased biomass, tall, erect and compact cultivars
(iii) Resistance to diseases
(a) Ascochyta blight.
(b) Fusarium wilt.
(c) Root rot.
(d) Botrytis grey mould
(iv) Resistance to insect pests:
(a) Pod borer.
(v) Tolerance to stress environments:
(a) Cold
(b) Heat
(c) Drought
(d) Saline and alkaline soils.
(vi) Mechanical Harvesting
Assignment :
1) Dissect the floral parts of given sample and mount them on black mounting paper.
Draw the figures and label properly.
2) Practice hybridization technique in field and laboratory on given sample.
..............................
LENTIL
(Lens culinaris Medik)
Cultivated species :
L. culiaris Medik (L. esculenta Moench) is a characteristic component of the old
World Belt of Mediterranean Agriculture. Numerous varieties of lentil are described. The
cultivars are conventionally grouped in two inter-grading clusters – (i) small seeded lentils
(sub-sp. Microsperma Barul) with small pods and small seeds (diameter 3-6mm), (ii) large
seeded lentils (sub-sp. Macrosperma Barul), with large pods and with seeds attaining 6-9
mm.
Wild species. The four wild species are delicate, small flowered annuals distributed
over South-West Asia and Mediterranean basin.
Botany :
The flower is typical papilionaceous, small, white, pale purple or purple blue. The
corolla and specially the standard is broadly obovate and measures 4-6 mm long and 3-4 mm
wide. The wing petals develop separately, rarely growing together with the keel petals. The
keel petals enclose the pistil and the stamens. The style usually develops at a right angle to
the ovary, and usually flattended on the outer side and trichoid on the inner side. The stamens
are polyadelphous or diadelphous. The ovary is flat and non-pubescent and normally contains
one to two ovules. The flower primordium is enclosed in a whorl of elongated, nearly calyx
lobes. Pods are 1 -2 cm long, oblong, flattended, with curved beak and persistent calyx.
Crossing technique :
Lentil is strictly self-pollinated due to cleistogamy and dehiscence of anthers is before
the flower opens. Wilson and Law (1972) reported less than 0.8% occasional cross-
pollination through thrips or other small insects.
The buds (one half-three fourths the length of calyx lobes) are held between the
thumb and the foreginger with the suture of the keel facing the operator. Special care is taken
not to bend or twist the peduncle. Sharp-pointed forceps are used to carefully remove one or
two calyx lobes nearest the suture side of the keel. The wings and keels are removed
individually. The standard is either removed or folded and breaking them free from the
stamina column or by pulling them outward. Manual pollinations are made immediately after
emasculation. Pollen is selected from the vigorous flowers as soon after anther dehiscence as
feasible. The keel is opened with forceps so that the stamina column and the pistil could be
removed as a unit. This brush like unit is used to transfer pollen to the exposed stigma of
emasculated flowers. The pollen laden pistil and the anthers are brushed against the trichoid
side of the stigma. After pollen transfer the standard of the maternal flower is returned its
original position around the pistil.
Breeding objectives :
1. High seed yield.
2. Bold seed size, high protein and less cooking time.
3. Early maturity.
4. Resistance to diseases :
a. Ascochyta blight (Ascochyta lentis Bon – Mon. & Vass.)
b. Rust (Uromyces fabae (Pers.) de Bary)
c. Wilt (Fusarium oxysporum f. Sp. Lentis Vasd. & Srin. Gord).
5. Resistance to insects :
a. Pod borer (Etiella zinckenella Treit)
b. Cutworm (Agrotis ipsilon (Hfn.) Ochropleura (Agrotis) Flammatra (Schiff.)
c. Aphid (Aphs craccivora Koch., A. Gossypii Gl., Myzus persicae (Sulz.)
6. Resistance to shattering
7. Tolerance to drought
Achievements :
..........................
EXERCISE NO. 3
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN FIELD PEA, RAPESEED & MUSTARD
FIELD PEA
(Pisum sativum)
Family : Leguminoceae
Sub family : Papilionaceae
Genus : Pisum
Species : sativum
Chromosome number: 2n=14
Cultivated species :
Linnaeus distinguished two species within the genus Pisum. Pisum arvense, the
coloured flower field-pea and Pisum sativum, the white flowered garden pea. Since then,
following species have been designated.
1. Pisum abyssinicum
2. P. aucheri
3. P. elatius
4. P. formosum
5. P. fulvum
6. P. humile
7. P. jomardi
8. P. transcaucasicum
All forms of peas previously accorded species status (except P. aucheri and P.
formosum which are tuber forming perennials and have been placed in genus Alophotropsis
have a diploid chromosome number of 14 and cross among themselves readily. Therefore,
they are now classified as ecotypes under P. Arense with white flowered garden pea
considered as ecotype sativum. However, the widespread use of P. sativum to designate the
garden pea would make it difficult to change this designation (Grittion, 1986).
Botany
The inflorescence is raceme arising from the axil of a leaf. The lowest node at which
flower imitation occurs is quite constant under a given set of condition and is used in
classifying the varieties with respect to flowering and fruiting duration. Most early cultivars
produce the first flower from nodes 5 to 11 and the late cultivars start flowering at about
nodes 13 to 15 (Gritton 1986).
The flowers are typical papilionaceous with green calyx comprising of five united
sepals. Five petals (one standard, two wings and two keels). The stamens are in diadelphous
(9+1) condition. Nine filaments are fused to form a staminal tube while the tenth is free
throughout its length. The gynoecium is monocarpellary, with ovules (upto 13) alternately
attached to the two placentas. Style normally bends at right angle to the ovary. Stigma is
elliptical and sticky. Early cultivars are often single flowered or bear some single and some
double flowers. Late cultivars are mostly double or triple flowered.
Crossing Technique :
Pea is strictly self-pollinated in nature. Stigma is receptive to pollen from several days
prior to anthesis until 1 day or more after the flower wilts. Pollen is viable from the time
anthers dehisce until several days thereafter.
For emasculation the plants to be selected should be vigorous and just beginning to
flower. The flower bud chosen should have developed to the stage just before anther
dehiscence, indicated by extension of petals beyond sepals. Flowers can be emasculated at
any time. The first step in emasculation is to tear away with the flower and thumb in front
and a light pressure is applied. This spreads the standard and wings to expose the keel. The
exposed keel is slit-open by tips of forceps. Pressure can be applied by the thumb and finger
on keel for increased exposure of the pistil and stamens. The 10 stamens are polled out.
Pollen can be obtained throughout the day, preferable from a freshly opened flower.
For pollen collection, it is more convenient to pick the male flowers, remove the standard and
wings, pull back the keel so that the style protrudes and use the pollen covered stylar brush as
an applicator to transfer the pollen to the stigma of the emasculated bud. Older flowers and
other flower buds not used in crossing are removed from the penduncle to increase the pod
set after crossing (Gritton, 1986).
Breeding objectives :
Garden pea
1. High green pod yield
2. Long, attractive green pods with 9 – 11 seeds / pod
3. Sweetness
4. High shelling percentage
5. Specific maturity (early and medium)
Field pea
1. High grain yield
2. Bold, attractive seeds
3. Early maturity
Resistant to diseases
1. Downy mildew (Peronospora viciae (Berk) de Bary)
2. Powdery mildew (Erysiphe polygoni DC)
3. Rust (Uromyces viciae – fabae (Pers.) Schroet and U. Pisi (pers.) Wint.)
4. Wilt (Fusarium oxyporum Schl. F. Sp. Pisi (van Hall) Snyd. & Hans).
Resistance to insect
1. Leaf miner (Phytomyza horticola Gour (= atricornis)
2. Semi-looper (Plusia ortichalea Fb.)
3. Aphids (Aphis cracivara Koch., A. Gossypii Gl.)
4. Pod-borer (Etiella zinckenella Trcit)
5. Pea stem fly (Ophiomyia phaseoli Tryon).
Important Achievements
Field pea varieties recommended for various states
State Varieties
Uttar Pradesh Adarsh, Vikas, Prakash, Rachana, KPMR400, Matar3, Pant Pea 42
Bihar Rachna, HUDP15, VL42
Maharashtra Adarsh, Vikas, Prakash, Rachana, Ambika, KPMR400
Rajasthan Rachna, Hariyal, DDR27
Madhya Pradesh Adarsh, Vikas, Prakash, Rachana, Ambika, KPMR400
Assignment :
1) Dissect the floral parts of given sample and mount them on black mounting
paper. Draw the figures and label properly.
2) Practice hybridization technique in field and laboratory on given sample.
.......................
Botany :
Brassicas have taproot system. Stem is succulent, straight and cylindrical. The leaves
are pinnati divided. Whenever they exist, trichomes are always simple. Their presence or
absence may be a good taxonomic character. A simple and well known example may be
that of B. oleracea, B. nigra and B. campestris where the first is completed glabrous and
the two others hairy. The amphidiploids where one of the parents is B. oleracea (i.e. B.
carinata and B. napus) are only very slightly hairy (Gomez Campo, 1980). The flower has
typical cruciferae formula (K2 + 2, C4, A2 + 4, G (2)). The inflorescence is racemose and
flowering is indeterminate beginning at the lowest bud on the main raceme. The
syncarpous ovary develops into a pod (silique) with two carpels separated by a false
septum.
Crossing technique :
The flowering is indeterminate and may last for two-three weeks. Stigma is receptive
for about six days (three days prior to three days after the opening of the flower). The
amphidiploids species (B. Carinata, B. napus, B. juncea) are self-compatible and self-
pollinated in nature but about 30 % cross-pollination may occur by wind and insect under
field conditions. The diploid species viz., B. nigra, B. aleracea and B. campestris are self
incompatible and consequently cross pollinated.
Selfing usually carried out by enclosing a flowering branch whose open flowers have
been removed, in a muslin cloth bag in case of amphidiploids species – which are self
compatible. In case of self – incompatible diploid species, selfing is done mostly by bud-
pollination. In bud pollination, a flowering branch whose open flowers / young pods have
been removed, is bagged by muslin cloth bag. After a few days, the bag is removed
temporarily and pollen from freshly opened flowers is applied on the stigma of young buds
which are preferably emasculated. The self incompatibility in Brassica is of sporophytic-
homomorphic type under monofactorial polyallelic series where pollen is inhabited on the
stigma. Various techniques available for obtaining a temporary break down of the self-
incompatibility character are as follows (De Nettancourt, 1972).
1. Bud pollination
2. Delayed self pollination
3. Grafting
4. Heat shocks
5. Application of carbon dioxide
6. Hormones and protein inhibitors
7. Chronic irradiation
8. Acute irradiation of styles
9. Acute irradiation of pollen mother cells
10. Tetraploidization – haploidization
11. Haploidization – diploidization.
Permanent self –compatibility can be induced by
1. Mutation of S locus
2. Modification of the genetic background
3. Tetraploidization
Breeding objectives
1. High yield
2. Early maturity
3. High oil
4. Low erucic acid and glucosinolates
5. Resistance to diseases
a. Alternaria blight (Alternaria brassicae)
b. White rust (Albugo candida)
c. Downy mildew (Pernospora parasitica)
d. Sclerotinia rot (Sclerotinia sclerotiorum)
6. Resistance to insects
a. Aphids (Lipaphis erysimi)
...........................
EXERCISE NO. 4
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN SUNFLOWER
SUNFLOWER
(Helianthus annus)
Wild relatives :
H. decapitulus
H. rigids
H. annus sub spp. annus
H. annus sub spp. lenticularis
H. annus sub spp. jaegeri
Crossing technique :
Sunflower is highly cross pollinated crop mainly through insects and to a limited
extent by wind. The flower opening starts from outer side of the head and proceeds towards
centre of the head, the heads bloom within 5-10 days depending upon size and season.
Anthesis occurs between 5 to 8 AM. The pollen grain viability lasts for 12 hours. The stigma
Practical Manual GPB-366 20
Crop Improvement –II(Rabi Crops)
remains receptive for two-three days. Selfing is done by bagging of the head. The bagging
material could be cotton cloth, tiffany bags or paper bags or cheese cloth bags or plastic
netting. Emasculation is done as follows :
Hand emasculation :
Emasculation is done by removing the anther tubes with forceps early on the morning
that the flowers open. Unemasculated flowers are removed.
Without emasculation :
Considering hand emasculation tedious, sometimes crosses are made without
emasculation. Hybrid plants are distinguished from selfed ones on the basis of vigour or the
presence of marker genes.
Pollination :
Pollination is carried out by collecting pollen from heads which are already bagged
prior to flowering. Pollen can be collected from flowering heads into paper bags by a light
tap of the hand on the back of the head. Pollination is usually done in the same morning after
emasculation. Pollen can be applied by a small piece of cotton, a camel hair brush, the corner
of the cloth bag isolator, a small section of leaf, paper or other suitable material that is dipped
in the pollen and gently drawn over the receptive surface of the stigmas. Freshly collected
pollens are more effective in pollination. Pollen can be stored without serious loss of
viability for 1-2 weeks in cork-stoppered vials at ordinary room temperature. After each
cross, care must be taken to avoid contamination by wiping the hands with alcohol and
cleaning or discarding the pollen applicator (Fick, 1978).
Breeding objectives :
i) High seed yield
ii) Early maturity
iii) Lodging resistant dwarf plant type
iv) Uniformity of plant type
v) High oil percentage
vi) Tolerance to stress conditions
vii) Resistance to bird damage
viii) Resistance to diseases : Flowing diseases are serious in India.
a) Leaf spots (Alternaria helianthi, Cladosporium cladosporoides )
b) Rust (Puccinia helianthi)
c) Root rot and damping of (Sclerotium rolfsii, Rhzoctonia bataticola, Syn.
Macrophomina phaseolina)
d) Stem rot (Sclerotinia sclerotiorum)
e) Head rot (Rhizopus spp.)
f) Powdery milder (Erysiphe cichoracearum)
Achievements :
Open-Pollinated varieties and hybrids of sunflower evolved / released
Seed yield
States for which
Variety / hybrid (Kg/ha) in rainfed Oil content (%)
recommended
areas
Variety
Morden All states 600-800 36-38
EC 68414 All states 800-1,000 40-42
EC 68415 Karnataka 800-1,000 40-42
Surya Maharashtra 800-1,000 32-35
CO 1 Tamil Nadu 500-700 38-39
CO 2 Tamil Nadu 800-1,000 38-40
TNAU-SUF 7 All States 800-1,200 38-42
GAU-SUF 15 Gujarat 800-1,200 38-42
SS 56 Maharashtra 700-900 36-38
Hybrid
BSH All states 1,000-1,500 40-42
KBSH 1 All states 1,200-1,500 42-44
KBSH 11 All states 1,000-1,500 40-42
LSH 1 Maharashtra 900-1,200 37-39
LSH 3 Maharashtra 1,000-1,500 38-40
PSFH 67 Punjab 1,000-1,500 40-42
PKVSH 27 Maharashtra 900-1,100 38-40
EXERCISE NO. 5
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN POTATO AND BERSEEM
POTATO
(Solanum tuberosum)
Cultivated species :
Potato is the most useful and important member of the family Solanaceae and it
belongs to genus Solanum. Genus Solanum consists of seven cultivated and about 154 wild
species but the commercially viable potato has only two species :
1) Solanum andigenum :
The plants of this species are characterised with thin and long stems, small and
narrow leaflets having profuse flowering and long stolons. The tubers are mostly covered
with deep sunken eyes on them. The yielding potential is very low and therefore, it is not
very common type.
2) Solanum tuberosum :
The potato cultivated the world over is an autotetraploid species, S. tuberosum. This
species is divided into two sub species viz. ssp. tuberosum (cultivated throughout the world)
and spp. andigena (confined to the hills of South America). In addition, diploid (2n=24),
triploid (2n=36) and pentaploid (2n=60) forms are also cultivated in Peru and Bolivia.
3) Solanum demissum and Solanum stenotonum are two more species which are somewhat
important as they are resistant to some types of virus and disease but they are also not in
cultivation commercially.
Flowering in potato :
The potato requires long day lengths (around 16 h), abundant rainfall, and cool
temperature to flower. Under most normal growing conditions, the day length in the early
part of the season will favour constriction of the stem, and grafting of young potato shoots
onto tomato or other compatible Salanaceous plants. Among the cultivars currently used for
breeding, selecting for increased flowering and seed set does not cause reduced tuber yield.
Flowering and tuber yield are uncorrelated as are berry or seed set and tuber yield.
Crossing Techniques :
Flower buds that are mature are selected for emasculation just prior to crossing. It is
particularly important to emasculate just prior to crossing if pollinations are done in the fields
the wind can break off the stigmas before pollination occurs if they are emasculated too far
ahead of pollination. Mature buds are plump, with the petals ready to separate. The remaining
buds and opened flowers in the bunch are removed to facilitate emasculation of the selected
buds and prevent contamination of the emasculated flowers by the open flowers. There is a
limit to the number of flowers from an inflorescence that will set fruit / seed, so removing the
extra flowers increases the chances that the pollination will be successful. The petals of the
selected flowers are gently pushed apart along the sutures and the five stamens removed with
fine-pointed forceps without breaking the style. The emasculated flowers are then bagged.
Inserting a branch with one or two leaves into the bag helps in maintaining a humid climate
inside the bag. In fully self-sterile parents, emasculation is unnecessary.
Pollination can be done at any time of the day as long as the temperature is not too
high. Open flowers are collected from the plant to be used as a male. The flowers are laid out
to dry overnight. The following morning the pollen is collected from them by shaking into
gelatine capsules such as those used in the pharmaceutical industry (other small tubes can
also be used). For large quantities of flowers, the pollen is shaken out by placing the flowers
in the top section of a sieve, and the sieve is then shaken at high speed. The pollen falls
through and is collected in the smaller capsules or tubes for storage. For smaller quantities of
flowers, a modified toothbrush or doorbell buzzer is used to vibrate the pollen free. The
flowers are inverted over glassine paper and the vibrating portion of the toothbrush or buzzer
is touched to the anthers. The pollen falls onto the slick paper and is then transferred into the
capsules or tubes. Pollen can be stored desiccated in the refrigerator for 1 – 2 weeks and in
the freezer for 6 months to a year. To make the pollination, the stigma is dipped in the pollen
in the capsule or tube, and then the pollination tag is attached and the bag is placed over the
flower and left on until the fruit is harvested. Setting of seed may be observed in about 7 – 10
days. Average seed set per berry varies with the cultivar, but levels of 50 – 200 seed per fruit
maybe obtained.
The objective of TPS is to have completed homogeneous progeny. This can best be
accomplished by the use of 4x families from 4x x 2n crosses, where the 2x parent produces
2n gametes. It is important that both parents be adapted to the area where the homogenous
progeny are going to be grown. Studies have shown that higher seeding vigour and tuber
yields resulted from this approach compared to progeny produced from 4 x x 4x crosses or
progeny obtained from open pollinated seed.
Year of
Variety Salient features and adaptability
release
Kufri 1997 Medium-maturing, resistant to late blight and excellent for
Chipsona 1 processing, North India plains
Kufri 1997 Medium-maturing, resistant to late blight and excellent for
Chipsona 2 processing, Uttar Pradesh and Bihar
Kufri 1997 Medium to late maturing and resistant to late blight, North
Giriraj western hills
HT/92-61 2003 Hybrid, heat tolerant, resistant to leaf hopper and mites, high dry
matter content, suitable for making French-fries
JW160 2003 White hybrid, having field resistance to late blight, excellent
keeping quality
MS/92-2105 2003 Red skinned, high-yielding hybrid, oval attractive tubers, field
resistance to late blight
SM/87-185 2003 Late-blight resistant, white tuber hybrid with high dry matter
content and better keeping quality
BERSEEM
(Trifolium alexandrium)
Cultivated species :
Berseem, known as king of fodder crops, is popular among livestock farmers of the
world. It belongs to the clover group and internationally famous as Egyptian Clover.
Botanically it is known as Trifolium alexandrinum L. Berseem is one of the oldest cultivated
clovers, domesticated in Egypt and later introduced into many other parts of the world.
Berseem belongs to the family leguminosae and genus Trifolium which consists of nearly 290
species as most important forage legumes. The most important species for forage and pasture
are berseem (Trifolium alexandium), Shaftal (T. resupinatum), White clover (T. repens), Red
clover (T. pratense), Crimson clover (T. incarnatum), Alsike clover (T. hybridum) and
Subterraneum clover (T. subterraneum) etc.
Origin :
Berseem is believed to be originated in Asia minor and from there it was introduced
to Egypt. Because of its introduction from Egypt it is famous as Egyptian clover and it is
gaining its increasingly importance as rabi crop.
Berseem doesn't have original wild forms.
Botany :
It is a fast growing annual crop with 30-60 cm plant height. The stem is hollow and
succulent. Both basal and /or stem branching is observed. Roots do not extend beyond two
feet in general and contains nodules. It is sparingly hairy and commonly possess trifoliate,
petiolate leaves. Leaves are membranous, oblong-elliptical to oblong-lanceolate and are
arranged alternately except the uppermost leaf. Leaflets are mucronate at the apex and
denticulate in. Inflorescence is head, terminally or auxiliary located and pedunculate with
conical to ovoid in shape. There is a small involucre at the base of the head. Calyx tube
displays ten prominent nerves while the corolla is almost double the height of the calyx. Each
inflorescence contains around 100 papilionaceous flowers, white in colour with around 1cm
length. At maturity each floret contains one single seed. Seeds are solitary and small in size.
Seed is egg shaped, yellowish in colour and is of around 2mm in length. In berseem white
coloured flowers are produced in cluster which are hermaphrodite in nature with five fused
sepals and five free petals. The upper large petal which covers the rest of the petals in bud
stage is called standard petal, while two bottom petals are fused together and formed a boat-
like structure called the keel. The stamens are always ten in number and their filaments are
fused in a group of 9+1. Berseem is a cross pollinated plant and is entomophilous in nature.
Anthesis occurs in the morning hours which coincides with maximum pollinator activity,
leads to seed setting.
Based on regeneration capacity and branching pattern three different ecotypes viz.,
Mescavi, Fahli and Saidi are reported in berseem. The mescavi type has very good
regeneration potential and is capable of 5-6 cuts with basal or crown branching pattern and is
the most popular type with large number of varieties in India belong to this group. Fahli is a
stem branching type with low regeneration potential and is suitable for single cut only. Saidi
is having moderate regeneration capacity allowing 2-3 cuts and possess both basal and stem
branching.
Achievements:
Diploid varieties like Meskavi, Fahali, Sauidi, Zaidi, BL-1, BL-2, BL-10, BL-22, BL-30, BL-
92, JB-3, JB-4, IGFRI-S-99-1, UPB-101, UPB-103, UPB-104, UPB-1905, and Khadrabi are
very popular but newly evolved high yielding tetraploid varieties like Pusa Giant, T-526, T-
724, T-780, T-529, T-560, T-561, T-674, T-678, T-730 etc. are very promising and give
about 50 per cent higher fodder yield.
......................
EXERCISE NO. 6
EMASCULATION AND HYBRIDIZATION TECHNIQUES
IN SUGARCANE AND COWPEA
SUGARCANE
(Saccharum spp.)
`
Family: Gramineae
Genus: Saccharum
Cultivated species :
There are three cultivated and two wild species of sugarcane. Their brief description is
a follows (Rao et. al. 1983; Purseglove, 1988).
Wild species :
1. Saccharum spontaneum (2n = 40 to 128).
2. Saccharum robustum (2n = 60 to 194).
Botany :
Induction of flowering. Lack of flower induction and synchronization are barriers in
sugarcane crossing programme. Flowering of sugarcane is rare in subtropics. Experiments of
photoperiod requirements have indicated that sugarcane can be classified as intermediate day
length plant (IDP) on the basis of initial induction and as short day plant (SDP) based on the
development of the panicle and flowers. A dark period around 12.30 hr in general, has been
found necessary for induction of flowering. Sites have been identified in India, Brazil,
Barbados, Hawaii, Fiji, Indonesia and Philippines where most clones of sugarcane flower. In
India Coimbatore has been chosen as the ideal place for natural profuse flowering and good
seed setting in sugarcane clones. In varieties difficult to flower, exposing the plants to 4 hr
extra- darkness in continuation of normal night for 6 – 8 weeks at the transformation stages
has been found effective (Rao et. al. 1983). Synchronization of flowering between early and
late flowering varieties is possible by manipulation of 4 hr extra darkness and 4 hr extra light.
Floral biology :
The inflorescence of sugarcane is an open, branched panicle and is called as an arrow
due to its shape which is like an arrow. Flowering is seasonal and takes place when the day
length decreases. In the northern hemisphere the flowering coincides with the onset of winter
(Oct.-Nov.) and in the southern-hemisphere in May-June.
The spikelets open about sunrise, beginning at the top of the panicle and proceeding
downwards and from the tips of the branches inwards, over a period of 5 – 15 days.
Approximately1/6 to 1/10th of the panicle opens each day. The swelling of the lodicules by
water uptake causes the glumes to be pushed apart and the stigmas come out. The anthers
dehisce about three hours after the elongation of the filaments. High humidity delays an
thesis. Natural pollination is by wind.
Crossing techniques : Following techniques are used to facilitate convenient handing of the
parents.
Marcotting :
It was first used in India and now is used in many countries. Generally, a plastic
sleeve containing a growth medium is secured about five to ten nodes above the bas of the
stalk to induce rooting.
Potted plants :
For clones and species not responding to the sulphurs acid technique or marcotting,
the sugarcane clones are grown in small containers.
Crossing in field :
This system is common in India. Pollen proof enclosures made of cloth (cloth
lanterns) are used to cover the arrow of female and male parent before anthesis. Male arrow
(which is also protected) is introduced into this lantern and it is shaken for 5 – 6 days once in
a day.
The crossing may be done either with the arrows attached to the parent plants or with
the arrows severed and transported to a central crossing area and maintained in living
condition by means indicated above. Female and male arrows can be enclosed in a common
lantern if they are planted close to each other.
Breeding objectives :
1. High cane yield.
2. Moderate high sucrose content
3. Early to full season maturity
4. Resistance to diseases.
a. Red rot (Physalospora tucumanesis)
b. Smut (Ustilago scitaminea Sydow).
c. Wilt (Cephalsporium sacchari Butler)
d. Mosaic (a viral disease)
e. Ratoon-stunting disease (caused by a bacteria)
f. Grassy – shoot disease
5. Resistance / tolerance to insect pests
a. Shoot borer
b. Cane borer
c. Pyrilla
d. Mealy bugs
e. Whiteflies
f. Termites
g. White grub
6. Tolerance to Aboitic stresses
a. Drought
b. Salinity
c. Flooding
d. High temperature
Achievement :
i) Sugarcane breeding institute has been the source of germplasm and genetic
variability for selection of varieties suited to different agro-climatic zones of the
country. The spread of Co canes to foreign countries began when Co 285 was taken
to Cuba and USA (Florida) for cultivation. Varieties bred at Coimbatore are / were
being used in 28 other countries either for commercial cultivation or as parents. Co
419 released in 1933 became the most popular variety in tropical India and was
rightly hailed as the wonder cane the world over.
ii) Two outstanding varieties viz., Co 658 for Tamil Nadu and Co 740 for Maharashtra
were released in 1940s. Co 740 continues to be cultivated in Maharashtra even now.
iii) Co 997 and Co 1148, released during 1950s, became ruling varieties in Andhara
Pradesh and North India respectively. Co 1148 remained the most predominate
variety in sub-tropical region for over four decades.
iv) Co 6304, a high yielder, became the most important variety in Tamil Nadu replacing
Co 419.
v) Varietal evaluation for juice quality conduced across seasons helped in the
indemnification of high sucrose varieties viz. Co 7204, Co 7704, CoA 7601, CoC
671, Co 8336, Co 8338 etc.
vi) Co 86249, an elite variety with resistance to red rot and high reasonability has been
evolved by the Institute and notified for release in the East Coast zone, It is also
serving as a source of resistance to red rot in the breeding programmes.
vii) Co 86032 Combining high yield and quality evolved by the institute and identified
by the AICRP (S) has been notified by the Central Sub-Committee on Crop
Standards, Notification and Release of Varieties of Agricultural Crops and is
occupying a major area in Tamil Nadu (90%), Karnataka, Maharashtra and Gujarat.
COWPEA
Botanical name : Vigna unguiculata
Family : Leguminaseae
Genus : Vigna
Species : unguiculata (sinense)
Chromosome No. : 2n = 2x = 22
Cultivated species :
Cowpeas belong to the botanical species Vigna unguiculata (L.) Walp. There are
more than 20 synonyms for V. Unguiculata
Verdcourt (1970) subdivided V. unguiculata into five sub-species as.
Unguiculata
Cylindrica
Cultivated species
Sesquipedails
Dekindtiana
Wild species
Mensensis
Marechal and Colleagues (1978) do not consider the three cultivated subspecies as distinct
and grouped under one subspecies V. unguiculata subsp. Unguiculata and differentiate them
by the intraspecific category ‘cultigroup’.
Crossing technique :
Cowpea flowers are large and showy. Mostly flowers open between 7 and 9 am. On
cloudy days the flowers may open in the afternoon. Though the flowers open late in the
morning, the dehiscence of the anthers is much earlier. It may vary from 10 pm to 0.45 am.
The dehiscence is influenced by environmental factors like presence of moonlight, a clear sky
and a dry warm atmosphere. During dark nights the dehiscence tends to be delayed. Due to
dehiscence taking place before the opening of flowers, the cowpea is self-pollinated in nature.
Since the dehiscence of anthers is much in a advance of the blooming, the
emasculation needs to be carried out in mature flower buds in the preceding evening. The
flower buds likely to bloom the next day (recognized by large size, the yellowish colour of
the back of the standard petal) is selected for emasculation. The bud is held between the
thumb and the forefinger with the keel side upper most. A needle is run along the ridge where
the two edges of the standard unite. One side of the standards is brought down and secured in
position with thumb. Same thing is done with one of the wings. After this the exposed keel is
slit on the exposed side, about 1 / 16 inch from the stigma. A section of keel is also brought
down and secured in position under the end of thumb. Now 10 stamens are seen. They are
removed with pointed forceps. Afterwards, the disturbed parts of standard, wing and keel are
brought in original position as far as possible. To prevent drying out of the emasculated bud,
a leaflet may be folded and pinned around the bud. A tissue paper can be used to cover and
protect the bud.
Pollination is down next morning from a freshly opened flower. The standard and
wings of male flower are removed. By slight depression of the keel, stigma covered with
pollen grains protrudes out. This itself can be used as a brush for pollination. Cowpea flowers
are highly sensitive and drop off easily with slight mechanical disturbance or injury.
Therefore, much labour and time should be devoted to get enough crossed seed
(Krishnaswamy, 1970).
Under improved techniques of Rachie et. al. (1975), the time taken for both
emasculation and pollination has been reduced substantially and the pod set has increased
from 18.6 to 26.1 per cent. Mishra et al. (1985) noted parental selectivity in hybridization
indicating that certain lines produce more pods and seeds/ pod when used as female parents.
Breeding objectives :
1. High green pod yield (vegetable type varieties)
2. High seed yield (dry-seed type varieties)
3. High fodder yield (fodder type varieties)
4. Dual purpose (seed and vegetable type and seed and fodder)
5. Earliness
6. Appropriate plant type (erect, determinate for vegetable and seed type cultivars and
spreading type for fodder type cultivars).
7. Resistance to diseases.
a. Anthracnose (Colletotrichum lindemuthianum)
b. Cercospora leaf spot (Cersospora cruenta)
c. Powdery mildew (Erysiphe polygoni)
d. Fusarium wilt (Fusarium oxysporum)
e. Ascochyta blight (Ascohyta phaseolorum)
f. Bacterial blight (Xanthomonas campestris)
g. Bacterial pustules (Xanthomonas phaseoli)
h. Cowpea yellow mosaic virus.
8. Resistance to insects
a. Hairy caterpillar
b. Leaf hoppers
c. Aphids
d. Thrips
e. Bruchids
f. Pod borer
g. Pod sucking insects
9. Better seed quality (acceptable to consumers)
Medium to large seed size, uniformly white / creamy / light red without black / brown
scare around hilum.
10. Development of elite, high yielding ‘Plant type’ as a composite of following (Rachie
and Rawal, 1976).
- Tall vigorous plant
- Deeply penetrating tap root
- Lodging and shattering resistance
- Low branching and or short branching
- Narrow leaves
- Short peduncles
- Profusion of peduncles at nodes
- Multiple podding of racemes
- Long pods with many seeds
- Weathering resistance pods and seeds
- Medium or medium small seeds of good quality.
Achievement :
Grain purpose varieties : Pusa-152, Pusa Sawani (T-5269), CO.1, CO.2, CO.3,CO.4, K.11,
K.14, RC.19
Vegetable purpose : Pusa Phalguni, Pusa Barsati, Pusa Do Fasli, Pusa Komal.
Fodder purpose varieties : Rassian Giant, T2, EC4216, K-391 and Cowpea-4.
Fodder purpose varieties at IGFRI Jhansi : Bundela Lobia-1, Bundela Lobia-2
.....................
EXERCISE NO. 7
EMASCULATION AND HYBRIDIZATION TECHNIQUE IN
SAFFLOWER
SAFFLOWER
(Carthamum tinctorius)
Family : Compositae
Genus : Carthamum
Species : Tinctorius
Chromosome Number : 2n = 2x = 24
Cultivated species:
Carthamum tinctorius L (2n = 2X = 24)
Wild Species
C. palaestinus
C. oxycantha
C. lanatus
C. flavenscens
Origin:
Safflower has been grown for many centuries from Egypt in north Africa eastward to
India. Safflower is believed to have two centers of origin, Ethiopia & Afghanistan.
Botany:
It is annual, erect herb having spreading type of branching. Stem is cylindrical,
slightly ribbed with spiny leaves. Leaves are simple, oblong to lanceolate. Their margin is
spiny and incised. Main stem terminates into flowering head called capitulum. Inflorescence
is of racemose type called head or capitulum without ray florets. The whole capitulum is
surrounded by number of overlapping, green, leaf like serrated (Mostly spiny) structures
called bracts which collectively known as involucre. Single capitulum may contain 20 to 200
disc florets.
Disc florets are sessile, tubular and mostly bisexual. The calyx is rudimentary. Corolla
is epigynous with 5 petals united (Gamopetalous) to form a tube. Upper expanded portion of
corolla is called limb, which is having different colours like yellow, orange, red or white
depending upon variety. It changes its colour after fertilization. Stamens are five, in
syngenious condition having hairy filaments and are epipetalous. Pistil is bicarpellary and
syncarpus with inferior ovary (Unilocular) having long bifid hairy stigma. After fertilization,
ovule is converted into dry indehiscent, cypsela type fruit.
Flowering:
It is often cross-pollinated crop. Marginal florets open first followed by florets in
central (centripetal order). It is completed within 1 to 5 days. The opening of florets takes
place in the morning hours between 9 to 10 a.m. The style elongates and stigma emerges
from corolla tube. At the same time, corolla opens and anthesis takes place. However, hairy
portion of style is still within tube.
Breeding objectives:
1) High seed yield of oil contents
2) Wide adaptability
3) Development of early and non-spiny varieties
4) Tolerance / Resistance to
A) Diseases
i) Wilt ii) Leaf spot (particularly alternaria)
iii) Rust iv) Powdery mildew
B) Pest
i) Gujea weevil ii) Aphids
iii) Heliothis armigera iv) Hairy caterpillar
v) Capsule vi) Army worm
5) Tolerance to abiotic stresses:
i) Moisture stress (drought)
ii) Thermo-insensitiveness i.e. for extreme temperatures
6) Development of appraisal type genotypes (to accommodate more plant population)
7) Development of stable GMS lines
8) Improvement in oil quality
Crossing Technique :
Emasculation:
Emasculation is done in previous day of anthesis in evening. The selected parents are
raised in crossing block. The selected capitulum is labeled and bagged. At the time of
emasculation, first involucore bracts are clipped off and disc florets are exposed. Generally
one marginal whorl of florets, which is likely to open on same day, is kept on disc and other
florets are nipped off. The anther tube surrounding the stigma and style is punctured
carefully at the base and slit is opened upward. This helps to remove entire column along
with surrounding corolla. During emasculation, some pollens may shed in the emasculated
flower which are removed by rinsing the florets with jet of water or 57 per cent alchohol and
then rinsed by water. The required numbers of florets are emasculated and head is properly
bagged and labeled.
Mass emasculation:
Hand emasculation is time consuming and causes injury to florets as a result of which
seed setting is poor. For mass emasculation, cut off free laminae of involucores bracts and
cover capitulum with plastic bag and tie it till opening of flowers (in case of open capitulum,
the anthers dehiscence coincides with the elongation of style so that stigmas are covered with
pollen of same floret during anthesis). In this method, plastic bag prevents anthesis of flower
due to increased humidity inside bag. Receptive stigma of such flowers is immediately
pollinated by desired pollens just after removing polythene bag. Continue this procedure for 2
to 3 consecutive days for complete pollination. Then remove plastic bags within a week.
Pollination:
Pollen grains from desired selfed male parents are collected in petridish and dusted
over the stigma of the emasculated flowers on next day morning i.e. 9 to 10 a.m. Sometimes,
male capitulums (shedding pollens) are also used for pollination by hand repeat pollination
for 1 to 3 times for effective crossing. Then the head is bagged and properly labelled after
every pollination.
Achievements:
1) Pure line selection: N7, N 62-8, Bhima (81), Manjira
2) Pedigree selection after hybridization: Tarea Annegiri 1, Girna
3) Development of Commercial hybrids by using GMS: DSH 129
Assignment :
1) Dissect the floral parts of given sample and mount them on black mounting paper.
Draw the figures and label properly.
2) Practice hybridization technique in field and laboratory on given sample.
...................
EXERCISE NO. 8
HANDLING OF GERMPLASM AND SEGREGATING POPULATIONS
BY DIFFERENT METHODS LIKE
PEDIGREE, BULK AND SINGLE SEED DECENT METHODS
1) PEDIGREE METHOD :
The pedigree may be defined as a description of the ancestors of an individual and it
generally goes back to some distant ancestor or ancestors in the past.
In Pedigree method, a detailed record of the relationship between the selected plants
and their progenies is maintained. Individual plants are selected from F2 and subsequent
generations and their progenies are tested. During the entire operation a record of all the
parent offspring relationship is kept. This is known as a Pedigree record. Individual plant
selection is continued till the progenies do not show segregation.
F1 generation:
F1 seeds are space planted so that each F1 plant produces the maximum F2 seed.
Generally 15-30 F1 plants are raised.
F2 generation:
In F2 2000-10000 plants are space planted to facilitate selection. About 100-500
plants are selected and their seeds are harvested separately.
F3 generation:
Individual plant progenies are space planted. Each progeny should have about 30 or
more plants. Individual plants are selected with superior characteristics. The number of plants
selected in F3 should be preferably less than the number of F2 selection. If number of
superior progenies is small the whole cross may be rejected.
F4 generation:
Individual plant progenies are space planted. Desirable plants are selected mainly
from superior progenies. The number of plants selected in F1 is generally much lower than
the number of F4 progenies. Progenies with defects and undesirable characteristics are
rejected. Emphasis is given on selection of desirable plants from superior progenies.
F5 generation:
Individual plant progenies are generally planted according to the recommended
spacing. Three or more rows are grown for each progeny to facilitate comparison among
progenies. Many families may have become reasonably homozygous and may be harvested in
bulk. The number of progenies must be reduced to a size manageable in preliminary yield
trials, which is usually of 25-100 progenies.
F6 generation:
Individual plant progenies are planted in multi row plants and evaluated visually.
Progenies are harvested in bulk, since they would have become almost homozygous.
Progenies showing segregation may be eliminated. Preliminary yield trials may be planted for
these reasonably homozygous progenies which are and have enough seed. Inferior progenies
are eliminated based on yield data from preliminary yield trial or visual evaluation.
F7 generation:
Preliminary yield trial with three or more replications is conducted to identify few
superior lines. Standard commercial variety must be included as a check for comparison.
F8 to F10 generation:
Superior lines are tested in replicated yield trials at several locations for two to five
years. The line superior to best check in yield and other characteristics would be
recommended for release of new variety.
F11 generation:
The breeder usually multiplies its seed during its last year of trial when a strain is
likely to be released as a variety. Thus in F11 and F12 seed is multiplied for distribution to the
farmers.
Assignment :
Draw schematic representation of the pedigree method of handling the segregating
generation
Bulk Method:
In the bulk method F2 and subsequent generations are harvested in mass or as bulks to
raise the next generations. At the end of bulking period individual plants are selected and
evaluated in similar manner as in the pedigree method. In this method artificial selection is
not practiced.
Procedure:
Hybridization:
Parents are selected according to the objectives of the breeding programme. Simple or
complex crosses are made depending on the number of parents involved.
F1 generation:
F1 is space planted and harvested in bulk. The number of F1 plants should be as large
as possible usually more than 20 plants should be grown.
F2 to F6 generation:
F2 to F6 are planted at commercial seed rates and spacing. These generations are
harvested in bulk. Artificial selection is not done. The population size should be as larges as
possible in each generation, i.e. 30 to 50 thousand plants.
F7 generation:
About 30 to 50 thousand plants are space planted and about 1000 to 5000 plants are
selected with superior phenotype and their seed is harvested separately. Selection is based on
phenotype of plants, grain characteristics, disease resistance etc.
F8 generation:
Individual plant progenies are grown in single or multi row plots. Most of the
progenies would be reasonably homozygous and are harvested in bulk. Weak and inferior
progenies are rejected on the basis of visual evaluation. Only 100 to 300 plant progenies are
selected with desirable characteristics.
Preliminary yield trial is conducted. Standard commercial varieties are used as
checks. The yield is used as basis of selection of superior progenies. Quality test may be
conducted.
F10 to F12 generation:
Replicated yield trials are conducted over several locations using standard commercial
varieties as checks. The lines are evaluated for important characteristics in addition to yield
i.e. disease resistance, quality etc. If a line is superior to the standard varieties in yield trials,
it would be released as a new variety.
F13 generation:
The seed of the released variety is increased for distribution to the cultivators.
In this method a single seed from each of the one to thousand F2 Plants is bulked to
raise the F3 generation. Similarly, in F3 and subsequent generations one random seed is taken
from every plant present in the population and planted in bulk to raise the next generation.
This procedure is followed till F5 or F6 when the plants would have become nearly
homozygous. In F5 or F6 a large number of individual plants are selected and individual plant
progenies and the number of progenies is sufficiently reduced to permit replicated trial in the
next generation. Individual plants may be selected only from outstanding families showing
segregation. Thus, preliminary yield trials and quality tests being conducted in F7 or F8 and
co-ordinated yield trials in F8 or F 9 generations.
The objective of the single seed descent method is to advance the generation of
crosses rapidly. At the end of the scheme a random sample of homozygous or merely
homozygous genotypes lines is obtained. F2 and subsequent generations are grown at very
high plant densities as vigour of plant is not important. In each year 2-3 generations may be
raised using off season nurseries and green house facilities.
Assignment :
Draw the diagrams of scheme for Bulk method of Breeding & single seed descent
method
...................
EXERCISE NO. 9
STUDY OF FIELD TECHNIQUES FOR SEED PRODUCTION AND
HYBID SEED PRODUCTION IN RABI CROPS
When space isolation is not possible, time isolation can be used. In case of time
isolation, different sowing is done so that an thesis in a seed field does not coincide with that
in the other fields. Obviously, the flexibility in time isolation is determined by the length of
the crop season. In almost all crops, late sowings are not admissible due to yield reductions
and greater chance of occurrence of biotic and abiotic stresses.
When both time and space isolations are not possible, mechanical barriers may be
used. Mechanical barrier is generally achieved by growing thick stands of fast growing crops
having greater height e.g., Sesbania in the case of hybrid rice. Occasionally, use of walls or
artificial barriers like cloth or plastic sheets may also be used.
Before the emergence of seed technology, what was considered to be adequate
isolation was solely based on practical experience. With expansion in knowledge,
experimental evidence was accepted as the basis for such decisions. For example, in the case
of wheat, the earlier recommendation for isolation distance was three meters. But
experiments suggested that in case of loose smut infestation, the appropriate isolation
distance should be 150 meters. In general, the greater the isolation distance, the smaller is the
possibility of contamination. However, it is difficult to achieve complete isolation for seed
plots of a given variety in the crop production regions of a crop. Therefore, seed production
plots are so isolated as to keep level of contamination below the prescribed minimum for the
concerned seed class.
Commonly used male : female ratio in the hybrid seed production of some crops.
Field inspection
Field inspection refers to the scrutiny of seed production plots by a team of qualified
persons. The primary objective of field inspection is to ensure that the seed production
pertains to the designated variety and that it has not been contaminated genetically physically
beyond certain specified maximum limits. Field inspections also ensure that the steps
necessary to minimize genetic and physical contamination have been taken properly and in
time to make them effective. The aims of field inspections are fulfilled by verifying the
following about the seed crop.
1. It has been raised from such seed whose source is approved.
2. It has been grown in a field area, which satisfies the prescribed land requirements.
3. In case of hybrids, the planting is as per the prescribed male : female ratio.
4. The prescribed isolation or border rows (in case of hybrids) have been provided.
5. Seed crop has been properly rouged to remove off-types, objectionable weeds and
inseparable other crop plants so as to conform to the maximum limits of standards
prescribed for these factors.
6. The crop is true to the varietal characteristics descriptive of the concerned variety.
7. The seed crop is harvested properly to avoid mechanical admixture.
8. The disease incidence, particularly of specified disease, is below the maximum
permissible limits.
9. The crop complies with such other special requirements that may be specified for the
crop concerned.
.........................
EXERCISE NO. 10
ESTIMATION OF HETEROSIS, INBREEDING
DEPRESSION AND HERITABILITY
Introduction :
Heterosis refers to the superiority of F1 hybrid in one or more characters over its
parents. In other words, heterosis refers to increase in fitness and yield over parental values.
Heterosis is also called as hybrid vigour. There are three possible genetic causes of heterosis,
viz. dominance, over dominance & epistasis. Out of these causes, dominance is widely
accepted. In crop plants, there are three main ways of fixing heterosis, viz. asexual
reproduction, apomixis and polyploidy.
Heterosis is estimated in four ways, viz. (1) over mid parent, (2) over better parent,
(3) over commecial cultivar, and (4) over commercial hybrid(check). These are called as
average heterosis, heterobeltiosis, useful heterosis (economic heterosis) and standard
heterosis, respectively; Standard heterosis is estimated in those crops where hybrids are
already available for comparison. Various types of heterosis are estimated as follows:
F1 - MP
1. Mid-parent heterosis = -------------------- X 100
MP
F1 - BP
2. Heterobeltiosis = -------------------- X 100
BP
F1 - CC
3. Useful or economic heterosis = -------------------- X 100
CC
F1 - SH
4. Standard heterosis = -------------------- X 100
SH
In plant breeding, useful and standard heterosis has direct practical significance.
Positive heterosis denotes the excelled performance of hybrid suggesting increased value of
particular character eg. + ve heterosis for grain yield, fruits/tree, branches /tree etc.indicates
high yielding ability of hybrid over parents. Where as, Negative heterosis shows reduced
performance of hybrid as compared to parents. Such type of – ve heterosis is useful where
we want the hybrid should have reduced performance for particular character viz. dwarfness,
earliness in flowering /maturity etc.
F1 – F2
Inbreeding depression = ------------------- X 100
F1
Where F1 = Mean value of F1 Cross
F2 = Mean value of above F1 cross in F2 generation
Solved Problems
Problem –1
In rice, grain yields (q/ha) of parents (P1 and P2), their F1 and F2 progenies are given
below:
Parent 1 Parent 2 F1 Hybrid F2 Progeny
18.94 22.69 29.38 15.18
18.94 + 22.69
Mean of Parents (MP) = ---------------------------
2
= 20.81
29.38 – 20.81
= ------------------- X 100
20.81
Heterosis = 41.12%
F1 - BP
Heterobeltiosis = ----------------- X 100
BP
Better Parent Value = 22.69
29.38 – 22.69
= ------------------------ X 100
22.69
Heterobeltiosis = 29.48 %
F1 - F2
Inbreeding depression = -------------- X 100
F1
29.38 – 15.18
= ---------------------
29.38
= 48.33 %
Answer :
Average heterosis = 41.12%
Heterobeltiosis = 29.48%
Inbreeding depression = 48.33%
Problem -2
Problem 3 :
What will be the yield of F2, when Inbreeding Depression is 39.77 per cent and the
grain yield of F1 is 24.68 g/plant?
Problem 4:
Work out the yield of F1, when heterobeltiosis is 67.80 per cent and grain yield of
better parent is 52.45 g/plant
Problem 5:
Explain the genetic reason for reduced yield in F2 generation as compared to F1.
ESTIMATION OF HERITABILITY
Introduction:
Heritability is an index of the transmission of characters from parents to their
offspring. Heritability is of two types, viz. broad sense heritability and narrow sense
heritability. Broad sense heritability is the percentage ratio of genotypic variance to the
phenotypic variance, whereas narrow sense heritability is the ratio of additive variance to the
phenotypic variance.
VG
Heritability (bs) = --------------- x 100
Vp
From generation mean analysis, the heritability is worked out with the help of
following formula
VF2 – VF1
Heritability (bs) = --------------- x 100
VF2
1 D + 1 H 1 – 1 H2 – 1 F
2 2 2 2
Heritability (ns) = -------------------------------------- x 100
1 D + 1 H 1 – 1 H2 – 1 F + E
2 2 4 2
2V gca
Heritability (ns) = ---------------------------- x 100
2V gca + V sca + V E
V gca
Heritability (ns) = ---------------------------- x 100
V gca + V sca + V E
D
Heritability (ns)as per Mather (1949) = ---------------------------- x 100
D + H + E
Solved Problems
Problem –1
In Pigeonpea, 33 genotype were evaluated for grain yield in RBD with three
replication and following mean square values were obtained for genotypes and error:
MSS treatments (genotypes) = 16.47, MSS error = 2.83, X = 11.68
Find out the value of heritability
Solution :
First we have to calculate genotypic and phenotypic variances as follows:
MSS tr – MSS e
Genotypic Variance (VG) = ------------------------
Replication
16.47 – 2.83
= -----------------
3
= 4.547
VG
Heritability = --------- x 100
VP
4.547
Heritability = --------- x 100
7.377
= 61.64%
Heritability (bs) = 61.64%
Problem – 2
In a 8 x 8 diallel cross of cotton, following parameters were obtained for fibre length
D H1 H2 F E
6.47 3.39 2.86 2.00 0.61
Calculate heritability in narrow sense.
Solution :
From diallel analysis, heritability is estimated with the help of following formula:
1 D + 1 H 1 – 1 H2 – 1 F
2 2 2 2
Heritability (ns) = ----------------------------------- x 100
1 D + 1 H 1 – 1 H2 – 1 F + E
2 2 4 2
6.47 + 3.39 - 2.86 - 2.00
2 2 2 2
= --------------------------------------------------- x 100
6.47 + 3.39 - 2.86 - 2.00 + 0.61
2 2 4 2
4.93 – 2.43
= -----------------------------------
4.93 – 1.715 + 0.61
2.500
= ---------------- = 65.36%
3.825
Problem –3
Genotypic and phenotypic variances and covariances of two characters (x and y) are
given below:
Solution:
GVX
Heritability of X = --------- x 100
PVX
3.252
= --------- x 100
5.044
= 64.47%
GVy
Heritability of Y = --------- x 100
PVy
4.720
= --------- x 100
5.520
= 85.51%
Problem – 4
Following estimates were obtained from generation mean analysis
Solution:
VF2 – VF1
Heritability (bs) = --------- ----- x 100
VF2
0.218 – 0.051
= --------- --------- x 100
0.218
0.167
= --------- ---- x 100
0.218
= 76.60%
1D
2
Heritability (ns) = --------- ----- x 100
VF2
0.084
2
= --------- --------- x 100
0.218
0.042
= --------- ---- x 100
0.218
= 19.26%
Problem – 5
Calculate broad sense heritability from the following estimates obtained from
generation mean analysis .
D = 0.842, H = 1.465, E = 0.072
Solution:
D
Heritability = --------- ----- x 100
D+H+E
0.842
= --------- ---------------------- x 100
0.842 + 1.465 + 0.072
0.842
= ----------
2.379
= 35.39%
Problem – 6
The following estimates were obtained from a diallel analysis
The following formula is used for calculation of heritability (ns) from above estimates
obtained either from diallel cross or partial diallel cross.
2 V gca
Heritability = --------- ------------------------ x 100
2 V gca + V sca + V e
2 (381.26)
= --------- ------------------------ x 100
2 (381.26) + 147.43 + 3.65
762.52
= --------- ------------------------ x 100
762.52 + 147.43 + 3.65
762.52
= --------- ---------- x 100
913.6
= 83.46%
Problem – 7
In Rice, following estimates were obtained for Line X Tester analysis.
V gca = 34.15, V sca = 12.27, V e = 0.32
Calculate heritability in narrow sense.
Solution:
The following formula is used for estimation of heritability from above estimates
obtained from line X tester analysis.
V gca
Heritability = --------- ------------------------ x 100
V gca + V sca + V e
34.15
= --------- ------------------------ x 100
34.15 + 12.27 + 0.32
34.15
= --------- -------
46.74
= 73.06%
Problem –8
In a study, following estimates of genotypic, environmental, dominance and epistatic
variances were obtained for a character.
Variance VG VE VD VI VP
Estimates 160 240 40 15 400
Calculate heritability in narrow sense.
Solution
The following relationship is observed in variances
VP = VG + VE
VG = VA + VD + VI
VG = 160, VE = 240, VD = 40, VI = 15
VP = 160 + 240 + 400
160 = VA + 40 + 15
or VA = 160 –55
= 105
VA
In narrow sense heritability = ---------- X 100
VP
105
= --------- x 100
400
= 26.25
Ans. Heritability (ns) = 26.25%
Variance VG VE VD VI
Estimates 210 190 60 25
..........................
EXERCISE NO. 11
LAYOUT OF FIELD EXPERIMENTS
Techniques in conducting field trials:
The proper conduct of field trials is of major interest to the plant breeder. In his search
for a new variety the breeder usually finds it necessary to grow a very large assortment of
experimental strains. Most of the strains will be inferior in some respects. If their undesirable
features can be recognized, they may immediately be eliminated from further consideration.
In ordinary practice, the procedure is first to grow large numbers of new strains, which have a
limited seed supply, in small observation plots where the breeder evaluates their maturity,
height, lodging, disease resistance, and other characteristics including over-all vigour. From
these visual observations the breeder selects what appears to him to be the superior strains.
The superior strains are then grown in replicated field trials to determine more accurately
their potential performance, including yield, in comparison with standard commercial
varieties. Since replicated field trials are more expensive to conduct, fewer strains are tested
in them in comparison with the very large number of strains that may be grown in the
preliminary observation nurseries. Even when outstanding experimental strains are
encountered, their yield superiority over the best commercial varieties will generally be
small. This need to measure small yield differences accurately is most important in advanced
trials in which only elite strains are being tested. By this time the breeder might have already
eliminated the strains those were found grossly inferior by the observation in nurseries and in
preliminary yield trials.
Nursery vs. Field Plots.
Nursery plots are small. Single or multiple row plots in which varieties of field crops
are grown for observation or yield trials. The size of the plots will vary with the crop, the
amount of seed available, and the nature of the observations which the breeder expects to
make. The nursery plot is used when (a) the seed supply of the strain is limited, and when (b)
a large number of strains are to be tested.
Field plots are of such size and shape that they may be planted and cultivated with
standard farm implements. Usually, field plots vary from 1/10 to 1/100 acre in size. Field
plots more closely related actual field conditions than do nursery plots. They are valuable as
observation plots, because their size makes it easy for the breeder to make visual observations
of the performance of a variety. They are useful for making preliminary seed increase. Field
plots require more seed and are more expensive for testing a given number of varieties than
are nursery plots. In general, field plots are used only for testing a few elite experimental
strains and standard varieties, after the superiority of a strain or variety has been
demonstrated in nursery plots.
Principles in Plot Technique:
The purpose of conducting variety performance trials is to measure comparative
yields, maturity, height, lodging, disease resistance, and other characteristics of varieties and
experimental strains of particular crop.
In order to have accurate results, the experimenter must follow careful and proven
procedures that are uniformly carried out with all the strains included in the test and he must
eliminate personal bias in recording notes and in interpreting the data.
Soil variability:
The variation in the soil is one of the important sources of error in field plot trials.
Even in small plots the soil may differ in fertility, drainage and texture and plants of similar
variety growing within a few feet of each other may perform differently. Previous soil
treatments often leave residual effects that affect the growth of the succeeding crop.
Therefore, experimental site used for performance trials should be selected carefully
considering topography, drainage, fertility, previous treatments and uniformity. It is often
helpful to observe the uniformity of the preceding crop before selecting the exact area to be
used for a performance trial. Generally, plots that are long and narrow will most effectively
sample the soil variations, if the long dimension of the plot is in the direction of the gradient
in soil fertility.
B) Competition and Border Effect:
Plants of different varieties in adjacent rows compete for the soil moisture and plant
nutrients. A vigorously growing variety may adversely affect the performance of a variety in
an adjacent row, especially if moisture or nutrients are limited. Tall growing varieties may
shade shorter varieties in adjacent rows. The performance of varieties growing in adjacent
rows may also be affected by differences in maturity, lodging or type of growth. To reduce
the error resulting from competition between varieties, it is a common practice to plant
nursery yield tests in three- row plots and harvest only the center row, or to plant four-row
plots and harvest two center rows. To eliminate border effect, it is common practice to plant
along sides of the plots several rows of standard variety which are discarded before harvest.
Ends of the plots are also discarded before harvest.
Replication:
The recorded yield of a plot is always subject to some error in the conduct of yield
trials. Depending upon the extent and the direction of the error, true yield of an individual
plot will be either larger or smaller than the recorded yield. If the error is due to chance, it
may be expected that the yield of different individual plots of the variety will fluctuate
around the true yield. If the yields of several plots of the variety are averaged, the chance
fluctuations will be less. For this reason, the mean yield of several plots of a variety is a
better estimate of the true yielding ability of a variety than the yield of a single plot. The
number of times a treatment (variety) is repeated in an experiment is commonly referred to as
the number of replications. This may range from design of the experiment, the accuracy
desired in the yield data, and the amount of land and seed available. In most standard yield
trials, either four or five replications are planted. Replication is necessary to sample
effectively the variations in soil fertility. Replication provides the means for experiment.
Adequate number of replications are used for performance trials that are harvested for yields.
.Assignment :
Give Diagrammatic representation of field plot designs.
1. Completeness: The breeder should be able to identify from his record the
parentage of particular strains as well as their current performance. The notes recorded on
performance will vary with the crop, but generally they should include such observations as
height, lodging, relative earliness of maturity, reaction to prevailing diseases or pests, and
over-all vigour. If a yield test, they will also include yield and quality evaluation of the grain,
fibre, or forage. Special identifying characteristics may be desirable to note, even though they
have little or no relation to performance.
.................
EXERCISE NO. 12
STUDY OF QUALITY CHARATERS, STUDY OF DONAR PARENTS
FOR DIFFERENT CHARACTERS
1. Market quality :
The market quality refers to fitness of a product for marketing. It includes uniformity
in shape, size, colour and texture in food and vegetable crops.
2. Industrial quality :
The industrial quality includes suitability for baking in wheat, malting in barley,
crushing in sugarcane, canning in fruit crops, etc.
3. Nutritional quality :
The nutritional quality refers to the suitability or fitness of a plant product for human
and animal consumption.
1) Wheat : In wheat, white or amber grain colour medium to bold size, and lustrous
appearance are important features for good market quality. High lysine content and good
baking quality are essential for use in biscuit and bread manufacturing.
2) Barley : In malting barley, low protein content and high extract of soluble
oligosaccharides after malting are desirable characters. Low protein produces less haze in
beer and high oligosaccharides are suitable for fermentation.
3) Pulses : In pulse crops attractive shape, size and colour of grains, high protein contents;
high methionine and tryptophan, and less flatulence are desirable characters.
4) Oilseeds : In oilseed crops, attractive shape, size and colour of seeds, high oil content
free from antinutritional factors and more proportion of unsaturated fatty acids are the
important quality characters.
5) Sugarcane : Moderate hardness, long internodes, optimum (low) fibre for milling, sucrose
ratio, high sucrose content and good quality of juice are desirable traits in sugarcane.
6) Forage crops : Greater nutritive value, more palatability and freedom from toxic
substances are the desirable characters in forage crops.
Thus, quality differs according to economic use of plant product. There are four
major goals of breeding for improved nutritional quality. These are breeding for (1) high
content and quality of protein, (2) high content and quality of oil, (3) high vitamin contents
and (4) low toxic substances which are harmful for human health.
Breeding methods :
Breeding methods used for improvement of quality do not differ from methods used
for any other character. Breeding methods used for improvement of quality traits include
backcross, pedigree method, single seed descent, recurrent selection, progeny selection and
mutation breeding.
Quality Assessment:
The system for quality assessment in plant breeding programs should be quick, cheap
and economical of material since plant breeder thousands of stocks each year, having limited
time and limited material for testing.
1) Organoleptic Characters:
Effected to characterize flavours chemically have failed. So the breeder, aided by a
taste panel looks at, smells and taste his fruits and vegetables and pulses and takes decisions.
If material to be examined after processing, he will take care to standardize the preparation
procedure as closely as possible.
B) Wild species:
i) H. argophyllus Drought tolerance
ii) H. praecox Resistance to alternaria rust &
downy mildew.
iii) H. giganteus Resistance to Sclerotia wilt.
iv) H. petiolaris Source for high oil content & for
alteration of fatty acid composition,
Cytoplasmic male sterility
v) H. debilis Source for salt tolerance
vi) H. tuberosus Source of resistance to leaf spot /
blight, Downy mildew.
5) SAFFLOWER A) Cultivated species
i) Carthamus tinctorius L.
B) Wild species
i) C. oxyacantha For resistance to leaf spot disease.
ii) C. palaestinus Eig For seed dormancy & earliness
(drought avoidance)
iii) C. paleestinus For resistance to stem fly
C. flavescens
iv) C. flavenscens For cold tolerance
v) C. lanatus For resistance to rust.
Chickpea :
Donors for different characters
Lines Characters
ILC 72., 196, 201, 202, 3279, 3346, ICC 8920, Tall upright growth
8922, G 130, Caina, NEC 249, P 336, 6099,
6308.
ICC 364, 552, 4945, 4951, 8284, P 271, 311, Double pods
1482, JG 62.
ICC 11520, 11521, 12206, 12208, 12212, 12213, Multiseeds
NEC 989, P 99, 431, 1198-1, ILC 194, 306,
2484, 2552, 2647
ILC 95, 96, 97, 99, 100, 101, 148, 149, T 3, K Bold seed
850, Rabat, L 144.
Multiple disease resistance
ICC 12237, 11269 Fusarium wilt, dry root rot, black root rot
ICC 1069 Fusarium witl, Ascochyta blight, Botrytis
gray mold
ICC 10466 Fusarium wilt, dry root rot, stunt
ICC 858, 959, 4918, 8933, 9001 Fusarium wilt, Sclerotinia stem rot
Insect resistance
ICC 506, 1381, 4856, 5264, 6663, 7510, 7559, Pod borer
7966, 10667, 10761, 10870
ILC 726, 1776, 2319, 2618 Leaf miner
G 109-1 Bruchids
P 636, H 208, PGM 442, BG 305, L 550, BG Root rot nematode
405
Environmental stresses
ICC 4973, 5003, 11514, BG 2, 209, 390, Ujjain Salt
24, NP 57
ICC 4958, 10448, C 214, H 208, G 24 Drought
ILC 666, 668, 1071, 2487, 2505, 3081, 3287 Cold
Annigeri, 850-3/27, H 208 Heat
C 214 Frost
Lentil :
Important donors for A lentil Breeding Programme
...........................
EXERCISE NO. 13
VISIT TO SEED PRODUCTION PLOTS
Certification Plot
EXERCISE NO. 14
VISIT TO AICRP PLOTS OF SAFFLOWER AND CHICKPEA
A) SAFFLOWER
B) CHICKPEA
EXERCISE NO. 15
VISIT TO AICRP PLOTS OF SUNFLOWER AND RABI SORGHUM
A) SUNFLOWER
B) RABI SORGHUM
SYLLABUS
Practical
Floral biology, emasculation and hybridization techniques in different crop species
namely Wheat, Oat, Barley, Chickpea, Lentil, Field pea, Rajma, Horse gram, Rapeseed
Mustard, Sunflower, Safflower, Potato, Berseem. Sugarcane, Tomato, Chilli, Onion;
Handling of germplasm and segregating populations by different methods like pedigree, bulk
and single seed decent methods; Study of field techniques for seed production and hybrid
seeds production in Rabi crops; Estimation of heterosis, inbreeding depression and
heritability; Layout of field experiments; Study of quality characters, study of donor parents
for different characters; Visit to seed production plots; Visit to AICRP plots of different field
crops
Teaching Schedule
a) Theory
Lecture Topic Weightage (%)
b) Practical
Experiment Exercise
1 Emasculation and hybridization techniques in wheat, oat & barley
2 Emasculation and hybridization techniques in chickpea & lentil
3 Emasculation and hybridization techniques in field pea, rapeseed & mustard
4 Emasculation and hybridization techniques in sunflower
5 Emasculation and hybridization techniques in potato &berseem
6 Emasculation and hybridization techniques in sugarcane & cowpea
7 Emasculation and hybridization techniques in safflower
Handling of germplasm and segregating populations by different methods like
8
pedigree, bulk and single seed decent methods
Study of field techniques for seed production and hybrid seeds production in
9
Rabi crops
10 Estimation of heterosis, inbreeding depression and heritability
11 Layout of field experiments
12 Study of quality characters, study of donor parents for different characters
13 Visit to seed production plots
14 Visit to AICRP plots of Safflower & Chickpea
15 Visit to AICRP plots of Sunflower & Rabi sorghum
Suggested Readings:
Sr. Title of Book Author/Authors Publisher
No
1. Crop Breeding and HariHar Ram KalyaniPublication New
Biotechnology Delhi.
Place :