PBG 301  

Principles and Methods of Plant Breeding 2+1

Theory (2017

Lec 1: Objectives and role of plant breeding - historical perspective – activities in Plant
Breeding

Plant breeding is defined as a science as well as an art of improving the genetic

makeup of plants in relation to their economic use. Plant breeding can be accomplished
through many different techniques ranging from simply selecting plants with desirable
characteristics for propagation, to more complex molecular techniques. Plant breeding has
been practiced for thousands of years, since near the beginning of human civilization. It is
now practiced worldwide by individuals such as gardeners and farmers, or by professional
plant breeders employed by organizations such as government institutions, universities, crop-
specific industry associations or research centers. Simmonds (1979) defined that plant
breeding is considered as a current phase of crop evolution.
International development agencies believe that breeding new crops is important for
ensuring food security by developing new varieties that are higher-yielding, resistant to pests
and diseases, drought-resistant or regionally adapted to different environments and growing
conditions.
The plant breeding field can be divided in to three important areas.
1. Plant genetic resources (or) germplasm
2. Breeding techniques
3. Seed production techniques
1.Germplasm: It is the total variability found in plant species. It deals with collection,
conservation, evaluation, documentation and utilization of cultivated and wild relatives of
crop plants.
2. Breeding techniques: Deals with various genetic principles and procedures of crop
improvement.
1. General breeding methods: This includes introduction, selection, and
hybridization (inter varietal hybridization)
2. Special breeding techniques: Mutation breeding, polyploidy, wide hybridization
and other special techniques like tissue culture and genetic engineering in crop
improvement.
 3. Seed production techniques: Deals with principles and methods of seed
production.
Objectives of plant breeding:
The main objective of plant breeding is to develop superior plants over the existing
ones in relation to their economic use. Thus, crop improvement with improved agronomic
characters and resistance against biotic and abiotic stresses are the main objective and
common objective of any breeding programme.
A brief account of some important objectives are given below
1. Increased yield
Majority of our breeding programmes aims at increased yield. This is achieved by
developing more efficient genotypes. The classical examples are utilization of Dee Gee Woo
Gen in rice and Norin10 in wheat. Identification and utilization of male sterile lines for
hybrid variety development.
2. Improving the quality – it differ from crop to crop
 Rice -milling, cooking quality, aroma and grain colour
 Wheat- milling and baking quality and gluten content.
 Barley – malting quality
 pulses -Protein content and improving sulphur containing amino acids
 oilseeds- PUFA (Polyunsaturated fat) content
3. Elimination of toxic substance
 HCN content in jowar plants
 Lathyrogen content in Lathyrus sativus ( β-N-oxalyl-amino-L-alanine- BOAA)
 Erucic acid in Brassicas
 Cucurbitacin in cucurbits
 Gossypol in cotton
4. Resistance against biotic and abiotic stresses
 Biotic stress: Evolving pests and diseases resistant varieties there by reducing cost of
cultivation, environmental pollution and saving beneficial insects.
 Abiotic stress: It is location specific problem. Soil factors and edaphic factors
sometimes poses severe problems. Breeding resistant varieties is the easy way to
combat abiotic stress.
5. Agronomic characteristics – modification of agronomic characters such as plant height,
tillering, branching, erect or trailing habit etc., is often desirable. In cereals dwarfness highly
associated with lodging resistant and fertilizer responsiveness. – E.g. Rice, Bajra
6. Photo and thermo- insensitivity: Photo and thermo –insensitive wheat and photo
insensitive rice has permitted their cultivation in new areas. Eg. Redgram, sorghum.
7. Synchronized maturity – one time harvest eg. Pulses
9. Non-shattering nature – Avoid wastage during harvesting due to over maturity eg.
Green gram, Brassicas
11. Determinate Growth habit –determinate growth – Pulses
12. Elimination or introduction of dormancy –Groundnut
Roll of plant breeding
Since the cultivable land is shrinking and there is no scope for increasing the area
under cultivation, the only solution to meet the food requirement is by increasing the crop
yield through genetic improvement of crop plants. There are two ways by which yield
improvement is possible.
1. Enhancing the productivity of crops
This can be done
a) By the proper management of soil and crops involving suitable agronomic practices and
harvesting physical resources.
b) By using high potential crop varieties created by appropriate genetic manipulation of crop
plants.
2. Stabilizing the productivity achieved
This is done by using crop varieties that are bred especially for wide adaptation or for
specific crop zones to offset the ill effects of unfavorable environmental conditions prevailing
in the areas.
Plant breeding, the past, present and future scopes
Indian agriculture remained stagnant particularly during early sixties. Long spells of
severe drought and serious outbreak of disease in some parts of the country led some
futurologists to state that a possible doom in India by the end of the decade. However, we
achieved breakthrough in crops such as rice, wheat, pearlmillet, jowar and maize.
1. The indica x japonica cross derivative ADT 27 is the first high yielding rice of Tamil
Nadu. The identification of Dee Gee Woo Gen and release of Wonder rice IR 8 (Peta
x DGWG) changed the scenario from poverty to problem of plenty.
2. Like wide identification of dwarfing gene in Japanese wheat variety Norin-10 by
Borlaug and breeding of Mexican dwarf wheat varieties led to the release of wheat
varieties life Kalyan sona in India.
 3. In pearl millet, breeding by male sterile line Tift 23A at Tifton, Georgia by Burton
and his coworker and later on its introduction to India led the release of hybrid bajra
HB1 to HB4, which increased bajra production many fold. In Jowar, breeding of first
male sterile line combined kafir 60A and its introduction into India led to the release
of first hybrid sorghum CSH 1 (CK 60A x IS 84) during 1970s.
4. At present we are in search of alternate source of cytoplasm in almost all crops to
breed hybrids with new source of cytoplasm to prevent the possibility of appearance
of new pest and diseases. Thus, the future of plant breeding is a challenging task. The
deployment of innovative breeding techniques will be a new tool to assist the
conventional breeding techniques.
Undesirable consequences of Plant Breeding
1. Genetic erosion: Disappearance of land races due to introduction of high yielding
varieties. Eg. Introduction of IR 20 rice led to disappearance of land races of samba rice.
2. Narrow genetic base: Genetic vulnerability to pest and diseases.
Tift 23A - Bajra - Susceptible downy mildew, T cytoplasm - Maize - susceptible to
Helminthosporium
3. Minor disease and pest become major due to intensive resistance breeding RTV (Rice
Tungro Virus) Grey mold in Bengalgram
4. Attainment of yield plateau: No more further increase in yield.
History of Plant Breeding
It started when man first chose certain plants for cultivation. There is no recorded history
when the plant breeding started.
 As early as 700 BC Babylonians and Assyrians artificially pollinated the date palm.
 In 1717 Thomas Fairchild produced the first artificial hybrid popularly known as
‘Fairchild mule’ by crossing carnation with sweet William.
 Joseph Koelreuter, German made extensive crosses in Tobacco between1760 - 1766
and emphasized hybrid vigour in F1.
 Thomas Andrew Knight (1759-1835) was the first man to produce several new fruit
varieties by using artificial hybridization.
 Le Couteur, a farmer of the Isle of Jersey published his results on selection in wheat
in the year 1843. He concluded that progenies from single plants were more
uniform than the remaining population.
 Patrick Shireff a Scotsman practiced individual plant selection in wheat and oats
and developed some valuable varieties.
 Loui de -Vilmorin (1856) proposed the Vilmorin principle of Vilmorin isolation
principle which is basic for progeny test.
 Nilsson-Ehle and his associates in Swedish Seed Association, Svalof Sweden (1890)
refined the single plant selection.
 In 1903 Johannsen the famous ‘pure line theory’ provided the genetic basis for
individual plant selection. Which states that a pure line is progeny of a single self
fertilized homozygous plant. He proposed this theory based on his studies in
Phaseolus vulgaris.
 1908 East and G.H. Shull studied the inbreeding in maize paved the way for the
development of hybrids in maize and later several other crops.
 1927 – Muller explained mutagenic action of X rays
 1928 - Stadler produced mutations in barley
 First mutation breeding programme launched in 1929 in Sweden by Ake Gustafsson
and co-workers.
 1928 Inter specific cross by Karpechenko (Radish x Potato)
 1937 doubling action of colchicine discovered independently by Blakeslee and
Nebel.
 During 1960’s Norman Borlaug and his associates developed Mexican semi dwarf
wheat varieties, which paved the way for green revolution in wheat. The dwarfing
gene was isolated from Japanese wheat variety Norin 10. Later on this Mexican dwarf
were introduced in the India by Dr. M.S.Swaminathan and a number of high yielding
wheat varieties like Kalyan Sona, Sonalika were developed.
 In rice the identification of dwarf gene Dee Geo Woo Gen from a dwarf , early
maturing variety of japonica rice from Taiwan. By using this gene Taichung Native 1
(TN1) was developed in Taiwan and IR 8 developed at IRRI Philippines were
introduced in India in 1966.
 Somatic hybridization -1972 -Carlson and co-workers – fusing the protoplasts of
Nicotiana glauca x N. langsdorffii
 Nobilisation in sugarcane by C.A. Barber and T.S.Venkataraman is another
monumental work in plant breeding.
 1996 – Genetically modified insect resistant varieties of cotton, maize and soyabean
commercially cultivated in U.S.A.
History of Plant Breeding in India
Organized agricultural research in India dates back to 1871, when the government of
India created the Department of Agriculture.

 The first scientist to be appointed in the department (in 1892) was an agricultural chemist.
In 1905, the Imperial Agricultural Research Institute was established in Pusa, now in
Bihar, this was the first agricultural research institute in the country. The buildings of this
institute were damaged by an earthquake in 1934; the institute was, therefore, shifted to
its present location in New Delhi in 1936. The name of the institute was changed to its
present one. i. e. Indian Agricultural Research Institute, in the year 1946.
 Agricultural colleges were established at Kanpur, Pune, Sabour, Llyalpur and
Coimbatore between the years 1901 and 1905.
o The main objective of these colleges was to impart agricultural education and
training. About this time, a number of Agricultural Research Farms were
established in each province. But the progress in Agricultural research was
disappointingly slow.
o In view of this, the Imperial Council of Agricultural Research was established in
1929, its name was changed to the present Indian Council of Agricultural
Research (ICAR) in 1946.
 The Indian Central Cotton Committee was established in 1921. The committee carried
out many notable researches on breeding and cultivation of cotton, E. g Development of
70 improved varieties of cotton. Encouraged by these results, central commodity
committees were set up on jute, sugarcane, tobacco, oilseed, coconut, arecanut, spices ,
cashewnut and lac. ( a total of 9 committees). Subsequently, these committees were
merged in the ICAR.
 In 1956, a project for Intensification of Regional Research on Cotton, Oilseed and
Millets (PIRRCOM) was initiated in order to intensify research on these crops. The
PIRRCOM was located at 17 different centres spread throughout the country; it focused
on cotton, castor, and groundnut, Brassica sp, Gingelly (Til), torai, taramira, jowar, and
bajara.
 The all Indian Coordinated Maize Improvement Project was started in 1957, with the
objective of exploiting heterosis. The first hybrid maize varieties developed under the
project were released in 1961. The phenomenal success of this project prompted the
ICAR to initiate coordinated project, for the improvement of other crops as well.
 The ICAR was reorganised twice in 1966 and 1973 with a view to improve its efficiency.
 The first agriculture university was established in 1960 at Pantnagar, Nainital (U.P).
Subsequently, such universities were established in most other states of the country.
 In some states, E. g U.P and Maharashtra, there are 4 universities each. The agriculture
university have the responsibility for education, research and extension in the different
areas of agriculture.
 In addition, over two dozen different Central Research Institute of ICAR are engaged in
crop improvement activities.
Activities in Plant Breeding
The desired changes in genotypes of crop species and the consequent benefits to
farmers are brought about by a series of interrelated and largely interdependent activities,
namely,
1) Creation of variation – Genetic variation is a pre request for any genetic
improvement in a crop. Hence in any plant breeding programme, this is the first step
unless genetic variation pre-exists in the breeding programme. This can be created by
domestication, germplasm collection, plant introduction, hybridization (intervarieteal,
distant, somatic), mutation, polyploidy, somaclonal variation and genetic engineering.
2) Selection - The next step consists of identification and isolation of plants having the
desirable combination of characters, and growing their progeny; this is called Selection.
Selection is ordinarily based on phenotype, but marker-assisted selection is based on
genotype of the concerned trait(s). The efficiency of selection determines the success of a
breeding program. Various breeding methods have been designed to increase the efficacy
of selection. Selection finally yields improved lines strains or populations that must be
evaluated for their performance.
3) Evaluation – Newly selected lines/strains/populations are evaluated for yield and
other traits and their performance is compared with the existing best varieties called
checks. Evaluation is a stepwise process; in India, it is ordinarily conducted at several
locations for three or more years under the concerned All India Coordinated Crop
Improvement project. IF the new line/strain performed superior than checks, it is released
as a new variety; the seeds can now be multiplied and more importantly certified for
quality.
4) Multiplication - It large scale production of quality seed of the released and
notified variety. Seed production is usually done by seed production agencies in a step-
wise manner, and the seed is certified by a seed certification agency
 5) Distribution – Certified seed is ultimately sold to the farmers who use it for
commercial crop cultivation. This activity alone makes it possible to reap the economic
benefits from the above activities in the form of (1) enhanced and (2) stable production of
(3) superior produce (4) often at lower costs.
Lec 2 : Centres of origin – contribution of Vavilov, Harlan, Zhukovosky – law of
homologous series
The cultivation of plants is one of man’s oldest occupations and probably began when
he selected some plants for his use. One of the old belief regarding to the origin of cultivated
plants was that they came to man as a gift from God. By the end of 18th century people
started questioning about the origin of cultivated plants.
Darwin (1868) considered that the cultivated plants arose by profound modifications
in the wild plant.
Alphonse de Candolle (1863) a Swiss botanist first attempted to solve the mystery
about evolution of crop plants. In his “ Origin of cultivated plants” he studied 247 plant
species of cultivated plants.
He classified the economic plants into six classes;
1. Plants cultivated 4000 years ago.
2. Plants cultivated 2000 years ago.
3. Plants cultivated less than 4000 years.
4. Plants cultivated 2000 to 4000 years.
5. Plants cultivated before the time of Columbus.
6. Plants cultivated after the time of Columbus.
It is N.I.Vavilov who proposed the concept of ‘centres of origin’. He proposed
the concept based on his studies of a vast collection of plants at Institute of Plant Industry,
Leningrad. The concept is that crop plants evolved from wild species in the area showing
great diversity and that place is termed as primary centre of origin. Later on from the
primary centre the crops moved to other places due to the activities of man.
There are certain areas where some crops exhibit maximum diversity of forms but this
may not be the centre of origin for that particular crop. Such centres are known as
Secondary centres of origin. E.g. cow pea (China)
The primary centre of origin for this crop is Africa but India exhibits maximum
diversity for this crop.
 Vavilov centers are regions where a high diversity of crop wild relatives can be found,
representing the natural relatives of domesticated crop plants. Nikolai Vavilov initially
identified 8, later in 1935 Vavilov divided the centers into 12, giving the following list:

1. Chinese center
2. Indian(Hindustan) center
3. Indo-Malayan center
4. Central Asiatic center
5. Persian center
6. Mediterranean center
7. Abyssinia center
8. South American center
9. Central American center
10.Chilean center
11.Brazilian center
12. US center

Vavilov originally proposed Eight main centres of origin. Eight main centres
of origin are
recognised by
Vavilov, they are:
1.China
2.Hindustan
3.Central Asia
4.Asia minor
5.Mediterranian
6.Abyssinya
7.Central
America
8.South America

1. The China centre: It consists of the mountainous regions of central and western China and
the neighbouring low lands. It is the largest and oldest independent centre.
The crops originated in this centre are:
I .Primary centre of origin are: ii. Secondary centre of origin are:
Soybeans Maize
Radish Cowpea
Proso millet Turnip
Opium Sesame
Brassica
Onion
2.The Hindustan Centre: This includes Burma, Assam, Malaya, Java Borneo, Sumatra and
Philippines, but excludes North West India, Punjab and North Western Frontier Provinces.
The crops originated in this centre are:
i. Primary centre of origin are: ii. Secondary centre of origin are:
Rice Cucumber
Redgram Radish
Chickpea Noble canes
Cowpea Cotton (Gossypium arboreum)
Greengram Hemp
Turmeric Coconut
3.The Central Asia Centre: It includes North West India, all of Afghanistan, the Soviet
Republics of Tadjikistan and Tian Shan. It is also known as the Afghanistan centre of origin.
The crops originated in this centre are:
i. Primary centre of origin are: ii. Secondary centre of origin are:
Wheat Rye
Pea
Broad bean
Green gram
Sesame
Safflower
Cotton(G.herbaceum)
Onion
Garlic
4.The Asia Minor Centre: This is also known as the Near East or the Persian Centre of
Origin. It includes the interior of Asia Minor, the whole of Transcaucasia, Iran and Highlands of
Turkmenistan. The crops originated in this centre are:
i. Primary centre of origin are: ii. Secondary centre of origin are:
Triticum Rape
Rye Black Mustard
Alfalfa Turnip
Cabbage
Oats
5.The Mediterranean Centre: The crops originated in this centre are:
i. Primary centre of origin are:
Many valuable cereals and legumes such as;
Durum Wheat Chikpea
Emmer Wheat Beets
Barley Peppermint
Lentil
Pea
Broad bean
6.The Abyssinian Centre: It includes Ethiopia and hill country of Eritrea. The crops originated in
this centre are:
i. Primary centre of origin are: ii. Secondary centre of origin are:
Barley Broad bean
Sorghum
Pearl millet
Lentil
Khesari
Sunflower
Castor
Coffee
Okra
7.Central American Centre: This includes South Mexico and Central America.It is also
referred to as the Mexican Centre of Origin. The crops originated in this centre are:
i. Primary centre of origin are:
Maize
Lima bean
Melons
Pumpkin
Sweet Potato
Arrowroot
Cotton (G.hirsutum)
8. The South American Centre: This centre includes the high mountainous regions of Peru,
Bolivia, Ecuador, Colombia, parts of Chile, and Brazil and whole of Peraguay The crops
originated in this centre are:
i. Primary centre of origin are:
Potato
Maize
Lima bean
Peanut
Egyptian cotton (G.barbadense)
Tobacco
Tapioca
 Later in, 1935, Vavilov divided the Hindustan Centre of Origin into two centres, viz.,
Indo Burma and Siam- Malaya-- Java Centre of Origin. The South American Centre was
divided into three centres, namely, Peru, Chile and Brazil-Peraguay Centres of Origin. At
the same time he introduced a new centre of origin, the U.S.A. Centre of origin. Two plant
species, Sunflower(Helianthus annuus) and Jerusalem Artichoke (H.tuberosus) originated in
the U.S.A. Centre of origin.
Thus the centres of origin may be more appropriately called the centres of diversity.
The centres of diversity may not be the centres of origin of the species concerned, but they
are the areas of maximum diversity of the species. Within the large centres of diversity, small
areas may exhibit much greater diversity than the centre as a whole. These areas are known
as Microcentres .
OBJECTIONS TO VAVILOV’S THEORY
According to Vavilov whenever a crop plant exhibits maximum diversity, that place
is the centre of origin for that crop. But this view is no longer valid. E.g. maize and tomato.
For maize the centre of diversity is Peru but archeological evidence shows Mexico as
centre of origin. For tomato, South America is considered to be primary centre of origin but
it is Mexico as per archeological evidence.
Secondly Vavilov stated that primary centre is marked by a high frequency of
dominant genes in the centre and recessive genes towards the periphery. But it is not so. E.g.
Wheat, maize, oil palm
Vavilov’s claim that centre of origin confined to mountainous regions only. But this
is not the case. For E.g. Maize exhibits maximum diversity in plains
Many crops have more than one centre of origin E.g. Balsam, Sorghum. In some
crops centre of domestication cannot be determined for want of suitable evidence.
`To counter the objection, Zhukovsky student of vavilov has proposed ‘mega centre’ theory.
He divided the world into 12 regions. Mega gene centres were the places where cultivated
plant species exhibit diversity and micro gene centre is the place where wild species occur.
Harlan stated that each crop may have been repeatedly domesticated at different
times in different locations or may have been brought into cultivation in several regions
simultaneously. We cannot pin point a single centre of origin. Harlan developed the idea of
‘Centre’ and ‘Non- centre’. According to him ‘centre’ means places of agricultural origin
and ‘non centre’ where agriculture has been introduced. Harlan divided the world in to
three centres and three non centres.
LAW OF HOMOLOGOUS SERIES:
 This is proposed by N.I Vavilov. According to this law “the characters found in one
species also observed in other related species”. Thus diploid, tetraploid and hexaploid wheats
show a series of identical characters. So also in case of diploid and tetraploid cotton.
Similarly genus Secale duplicates the variation found in Triticum.
Harlan's most recent theory (1992)
 Certain biomes or vegetation types may have been more conducive to domestication
than others.
 A biome  is a major regional terrestrial community with its own type of climate,
vegetation and animal life. Biomes are not sharply separated, but merge gradually
into one another over what is called an  ecotone  

Plant Genetic Diversity

Variety of genes and genotypes found in a crop species. Genetic diversity – broad
genetic base to population. N.I. Vailov (1926, 1951) realized the importance. Proposed eight
main centers of diversity and three subsidiary centers.

Vavilov not covered Africa, Australia.

Primary Centres of Diversity.

- Regions of vast genetic diversity

- Large number of dominant genes
- Mostly have wild characters
- Less crossing over
- Natural selection operates.
Secondary centers.

- lesser genetic diversity

- large number of recessive genes
- have desirable characters
- more crossing over
- both natural and artificial selections operate.
Microcenters

- Small areas – tremendous genetic diversity.

- Study of evolution of cultivated species.
 Vavilov – parallel series of variation – Law of homologous series of variation.
Particular variation in a crop another related spread
Lec 3: Plant genetic resources – importance – germplasm – types – activities – gene
erosion - gene bank – collection - conservation – types of conservation –Plant genetic
resources.

Plant genetic resources (PGR) are the basic materials that are essential for
development of improved crop varieties designed to combine high yield potential with
superior quality, resistance to diseases and pests, and also better adaptation to abiotic stress
environments. Their continued availability to plant breeders is necessary not only for
sustaining advances in crop productivity but also for stabilising production in the country.
These resources of known or potential use to man constitute a broad spectrum of diverse gene
pools representing assemblage of landraces, primitive cultivars, varieties of traditional
agriculture as well as wild and weedy relatives of crop plants. In the last two decades or so,
much attention has been drawn to indigenous locally adapted cultivars in particular because
of the useful genetic variation they contain as an invaluable resource for present and future
plant breeding, and the rapid rate at which they are disappearing through replacement by high
yielding varieties. In addition, the natural habitats of wild relatives of crop plants are
continuously getting eroded threatening survival of these populations. This diversity is not
yet adequately represented in the existing collections at national, regional and international
levels. Indian national programme on genetic conservation aims at exploring and collecting,
classifying, evaluating, conserving and documenting this natural heritage for its current and
future use. All these operations constitute a chain of activities that are now better understood
and carried out by the national and international centres mandated with such responsibilities.
The last thirty years have seen the great upsurge of this activity, with more awareness
generated by the FAO, Biodiversity International (IPGRI), and the IARCs and also by the
IUCN, UNESCO and the WWF in their concern for conservation of biodiversity with
particular reference to in-situ aspects. Equally important in this context has been the
phenomenal growth in biotechnology during the past two decades which has also created new
awareness about the value of plant genetic resources since sexual process of fertilisation and
recombination was no longer a pre-requisite to shuffling of desirable traits.

A broad outline of plant genetic resources' activities has already been presented in the
first chapter as an introductory part. In the following chapter, importance of geographical
areas of diversity of crop plants and the richness of this genetic wealth in the Indian
subcontinent has been reflected. Subsequent chapters deal largely with the methodologies and
approaches that are followed in executing PGR activities, viz. germplasm collection,
introduction, exchange and quarantine, characterization and evaluation, maintenance,
documentation, conservation and utilisation. In this chapter, the work carried out by the
National Bureau of Plant Genetic Resources (NBPGR), the national nodal organisation for
such activities, and its coordinating role in the management and monitoring of these activities
has been highlighted with a view to focussing attention on the newly emerging Indian
National Plant Genetic Resources System.
Brief history in India
Indian interest and abiding concern in the collection and utilisation of plant genetic
resources dates back to the early decades of this century (Howard and Howard, 1910), though
botanical accounts on available flora and the economic plants/products had been documented
much earlier (Hooker, 1872-97; Watt, 1889-93). However, it was late Dr. B.P. Pal who truly
focussed attention on the use of germplasm variability in crop improvement in national
context. The publication of his paper, ‘The search for new genes', in fact, paved the way for
augmenting genetic diversity for use in plant breeding (Pal, 1937; Pal and Singh, 1943).
It was primarily due to his foresight and wisdom that a nucleus Plant Exploration and
Collection Unit was established in 1946 in the Division of Botany at the Indian Agricultural
Research Institute, New Delhi. This unit became a regular wing in 1956 that was raised to the
status of a Division of Plant Introduction in 1961. The late Dr. Harbhajan Singh dedicated
his entire services to operate and boost these activities from the beginning and particularly so
during the 1960s-1970s. (Singh and Hardas, 1970; Singh, 1973). Dr. M.S. Swaminathan and
Dr. A.B. Joshi further strengthened the foundations of these activities. To serve the needs of
the ICAR crop research institutes, all India coordinated crop improvement projects and state
agricultural universities, the Indian Council of Agricultural Research created a separate
organisation named as National Bureau of Plant Genetic Resources (NBPGR) in 1976
alongwith two other Bureaus concerned with animal and fish genetic resources.
PLANT GENETIC RESOURCES

Sum total of genes in a crop species

- ‘Genetic resources’ or ‘ gene pool’, ‘genetic stock’, ‘germplasm’

- Whole library of different alleles of a species
- Basic materials for initiation of breeding programme.
Features:

- Entire genetic variability

- Land races, modern cultivars, obsolete cultivars, breeding stocks, wild forms,
species
- Cultivated & wild species
- Collected from centers of diversity, gene bank, gene sanctuaries, farmers field,
market and seed companies.
- Basic material
Classification of gene Pool

1) Area of collection
2) Domestication
3) Duration of conservation
4) Crossability in breeding program.
5)
1) Area of collection: a) Indigenous b) exotic
2) Domestication: a) Cultivated b) wild
3) Duration of conservation:

a) Base collection: Base collections include maximum number of

accessions available in a crop . Long term storage upto 50 years or more. Seed
viability not less than 95% , Seeds with 5 + 1 % moisture content stored at -18OC to –
20OC. These collections are distributed only for the purpose of regeneration.
It is also known as principle collection and refers to the whole collection.
b) Active collections
This category of germplasm is actively utilized in breeding programmes and are
conserved for medium term (8-10 years or more). These collections are stored at zero
degree cesius with moisture content around 8%. Germination test is carried out after
every 5-10 years to assess the reduction in seed viability.

c) Working collection:
These collections are frequently utilized by breeders in their crop improvement
programmes. These are stored for short term ( 3 to 5 years ). The seed is stored at
5OC – 10OC with moisture content of 8-10%.

4) Crossability in Breeding Program

1) Primary gene pool GP1: Intermating is easy – production of fertile hybrids. Same species
or closely related.

2) Secondary gene pool GP2: Partial fertility on crossing with GP1 plants related species.
3) Tertiary gene pool GP3: Sterile hybrids on crossing with primary gene pool.
Needs special techniques.
Components of Genetic Resources

Various plant materials constituting gene pool.

1) Land races: Primitive cultivars.

- evolved under sub resistance agriculture

- High level of genetic diversity – diseases, pests
- Broad genetic base
- Less uniform
- Low yielders
2) Obsolete cultivars

- Improved varieties of recent part

- Replaced by new varieties.
- Wheat varieties K68, K65, pb591 – Traditional Tall before Mexican wheat
attractive grain color and good chapatti making.
3) Modern cultivars

- Currently cultivated high yielding varieties.

- High yield potential, uniformity, parents in breeding program Narrow genetic
base
- low adaptability.
4) Advanced breeding lines

Pre-released plants developed by plant breeders. Not yet ready for release.

5) Wild forms of cultivated species.

High degree of resistance.

6) Wild relatives.

1) Hybrid sterility problems in crossing

2) Hybrid invariability
3) Undesirable genes with desirable alleles.
7) Mutants

Mutant gene pool Dee-Geo-Woo-Gen in rice and Norin 10 in wheat. Valuable genetic
resources. In seed propagated crops, 410 varieties have been released.
GERMPLASM
The germplasm collection is a collection of large number of genotypes of a crop
species and its wild relatives. In other words it is the sum total of hereditary or genes present
in a species. Therefore, germplasm consists of the following five types of materials: (1)
land races, (2) obsolete varieties, (3) varieties in cultivation, (4) breeding lines, and (5) wild
forms and wild relatives.
Germplasm collections are also known as gene banks or gene pool or world
collection. The term working germplasm refers to the smaller number of collections kept by
a breeder for hybridization programme.
Need for Germplasm Bank :
a) The modernisation of agriculture and evolution of high yielding varieties and hybrids led
to the replacement of the land races. For examples after the introduction of IR 20 rice for
samba season all the local varieties like karthigai samba, Toppi sampa, Rubber samba,
Thiruchengodu samba. Athur samba went out of cultivation. Along with them the
beneficial genes also vanished. This is known as genetic erosion or in other words
narrowing down of variability. So, to prevent the loss of variability in cultivated forms
and their wild relatives (Genetic erosion) it is necessary to maintain germplasm.
b) Nature has provided enormous variability for the use of mankind. We should not destroy
them and preserve them for the use of future mankind.
Germplasm conservation :
Conservation is the management, preservation and use of known genetic resources so
that they may yield the greatest sustainable benefit to the present generation, which
maintaining their potential to meet the needs and aspirations of generations to come (IUCN,
1980)
There are two methods of conserving germplasm: in situ and ex situ (Frankel and
Soule, 1981).
Conservation in situ involves the setting aside of natural reserves to conserve species
in natural habitats. This type is also classified as dynamic evolutionary conservation. Plants
and animals are conserved in entire biomes free to evolve through natural selection.
Extinction of species is deterred but this method has little impact on useful plants.
Gene Sanctuaries or Insitu conservation:
The areas of diversity are protected from trespass of human beings by fencing the area
so that the plant species are preserved under natural conditions. This is known as insitu
conservation.
E.g. Meghalaya for citrus, North Eastern Region for Musa, Oryza, Saccharum.
Exsitu conservation:
Conservation ex situ is the conservation of species out of their natural habitat (Hoyt,
1988). There are three main methods of ex situ conservation: seed banks, field genebanks and
tissue culture. Collections of germplasm using any of these methods are often called
genebanks. With the advent of biotechnology a genebank may also include a. collection of
cloned DNA fragments from a single genome and, ideally, representing the whole of the
genome.
There are two types of conservations are
Short term conservation:
Based on the viability of the seeds the gene pool is to grown once in two years or
more than two years. Each line is to be grown with proper spacing and care must be taken to
ensure self pollination, so that the genetic architecture is not altered. For example in sorghum
covering the panicle in boot leaf stage it ensures selfing.
This short term conservation is a costly affair which requires much time, labour, land
and cost. Further there is every chance for mixing up of genotypes while large number is
handled annually.
Long term conservation :
To overcome this difficulty long term preservation in the cold storage the germplasm
can be preserved. Using liquid nitrogen the germplasm can be stored for more than ten years.
Complete information about the genotypes can be computerized and this is known as
cataloguing and information retrieval system.
Exploratory Surveys :
NBPGR will arrange for survey and collection of germplasm. Explorations generally
cover those that are likely to show great diversity of forms. Tribal areas will have more
forms of diversity. Along with ICRISAT, NBPGR the TNAU has conducted exploratory
surveys for collection of small millets, sorghum and pearl millet.
The palamalai hills of Coimbatore is a rich source of diversity for sorghum.
Sorghum halapense both 2n = 20 and 2n = 40 forms are available there. The Kodaikanal hills
are having S.nitidum under natural conditions. In southern districts S.stafii is available.
Anaikatti hills are rich source of diversity for small millets. Normally during surveys the
samples collected will be of three kinds.
a) Field Sample : Seeds collected from field or farm areas where it is available.
b) Market sample : Types available in local shandies or market will be collected.
c) Storage sample : By visiting the houses of farmers the seeds stored for sowing will
be collected.
Centres maintaining germplasm
1. Institute or plant Industry, Leningrad.
2. Royal Botanic Gardens, Kew, England.
3. USDA, Beltsville.
4. ICRISAT. Sorghum, Red gram, Ground nut, Pearl millet and Bengal gram
5. IRRI - Rice
6. World vegetable centre - Taiwan - Soybean
7. Biodiversity International (IPGRI ) - International Germplasm Repository.
8. NBPGR - National Germplasm Repository.
Genetic erosion :
The loss of genetic material (genes, genotypes) from individuals or populations is
termed genetic erosion (IBPGR, 1991).
Reason for genetic erosion
1. Changing patterns of land use such as clearing of forests, housing and industrial
developments contribute to genetic erosion.
2. So does changing cultural practices particularly the widespread use of a limited number
of standard varieties in lieu of the genetically rich old and traditional populations of
cultivated species.
3. The threat of genetic erosion is real.
There are several recorded epidemics due to diminished genetic diversity resulting
into increased genetic vulnerability in major crops (NAS, 1972).
a. 1840 famine in Ireland due to potato late blight (Phytophthora infestans)
b. 1917 wheat less days in USA due to stern rust epidemics (Puccinia gram in is)
c. 1943 famine in. Bengal, India due to brown spot disease of rice (Cochliobolus
miyabeanus) and
d. a typhoon Mid 1940s complete elimination of all oats derived from the variety
Victoria in the U.S. due to the Victoria blight disease (Helminthosporium
victoriae)
e. 1970-71 southern corn leaf blight (Helminthosporiuni maydis) epidemic on all
U.S. corn hybrids carrying the T-type cytoplasmic male sterility
 f. In rice, recent epidemics associated with the widely grown and muliple-cropped
semidwarfs have been pointed out (Chang, 1979, 1984).
Lec 4 Germplasm: evaluation – use of descriptors, documentation, utilization; Agencies
– national and international; germplasm exchange – quarantine.
Germplasm activities.

1) Exploration and collection.

- Collection trips tapping genetic diversity from various sources.

- assembling at one place.
Reduction in genetic variability

a) Replacement of land races with improved cultivars

b) Modernization of agriculture – eliminates wild & weedy former.
c) Extension of farming into wild habits.
d) Grazing into wild habitats.
e) Growth of cities.
ii) Extraction: Permanent loss of a crop species.
Process
1) Sources of collection: Centres of diversity
gene bank
gene sanctuaries
seed companies
farmers fields
2) Priority of collection: Endangered areas, endangered species
3) Agencies of collection: SAU
ICAR
(Biodiversity International) IPGRI
4) Method of collection: Expeditions, personal visit to gene bank, correspondence,
exchange of material.
5) Method of sampling: Random sampling – biotic tolerant stresses
Biosed sampling – distinct morphological characters.
6) Sample rice: 95% of total diversity should be caps there. 50 –100
individuals, 50 seeds/ plant.
Merits
1) Tapping crop genetic diversity
2) New material, prevents extraction
Demerits
entry of new disease, remote areas.

2. Conservation
Protection of genetic diversity of plants from genetic erosion.
In situ Conservation: under natural conduction.
Establishment of biosphere reserves.
- Costly method, several areas have to be conserved.
Ex situ conservation: Preservation of germplasm in gene banks.

Cheaper, easy, entire genetic diversity conserved.

Seed Meristem

easy
Long term (50-100 years)
Medium term (10-15 years)
Short term (3-5 years)
Robert (1973) – Orthodox – dried to low moisture content, no loss in validity.
Eg: wheat, papaya, various beans. - recalcitrant
Drastic loss in viability with a decrease in meristem below 12-13OC
Eg: Cocoa, margo, tea, coffee, jackfruit, ruble.
Meristem
Merits – free from virus

- Vegetatively propagated crop.

- Perennial plants
- Regeneration easy
3. Evaluation:

1) to identify gene sources

2) classification of germplasm.
- by simple measures of dispersion (Range, standard deviation, SE, CV)
- by metroglyph analysis of Anderson (1957)
- D2 statistic of Maharlanobis (1936)
4. Documentation

Compilation, analysis, classification, storage, dissemination of information.

Information system.

- provides information about various activities of plant genetic resistance.

 - 7.3 million germplasm accession – 200 crop species.
Biodiversity International - (IPGRI) – descriptor

5. Distribution:

Specific germplasm – supplied on demand.

6) Utilization use of germplasm in crop improvement programme

cultivated germplasm - as a variety

- as parent

- as variant in gene pool.

Wild – Transfer of resistance.

Biodiversity International (IPGRI) – Supervised by consultative group on international

Agricultural Research (CGIAR)

CGIAR – 1972 by FAO, world bank.

UNDP to establish international research Institutes.

Biodiversity International (IPGRI) established by CGIAR in 1994.

Conducting research and to promote an International Net work of plant Genetic

Resources.

IBPGR Till 1993 – IBPGR 1974.

NBPGR

NBPGR – by ICAR – 1976 – New Delhi

1946 – plant introduction started at IARI, New Delhi.

1961 – separate division of Plant Introduction – Dr. H.B. Singh

1976 – NBPGR

Quarantine: 1914 – Destructive Insects and Pest Act. Photosanitary certificate.

NBPGR, FRI – Dehradun and Botanical survey of India. Calcutta

- Directorate of plant protection, Quarantine and storage - faredabad

- good grains & produces imported for human consumption.
Germplasm exchange
The NBPGR, New Delhi has brought out a brochure 'Guidelines for the exchange of
seed/planting material' (National Bureau of Plant Genetic Resources, 1986) and this had
been widely circulated among scientists in India. The guidelines for import of
seed/planting material for research purposes have been revised (National Bureau of Plant
Genetic Resources, 1989), in view of the enactment of New Seed Development Policy by
the Government of India which has also been circulated amongst various
scientists/institutes involved. The issuance of 'Import Permit' has been made mandatory
by the NBPGR for import of all seed/planting material for research purposes. The salient
features of the procedure are described below.
Import
All requests for indenting germplasm from abroad are to be made to the NBPGR
giving specific details of the required material, stating the source/country as well as address
of organisation/scientist through a prescribed application so that the Import Permit is issued
and sent to concerned scientist(s) for sending the same to exporting organisation or the
Bureau be requested to persue the import of desired material(s) from abroad.

When requests are sent directly by an individual scientist to any foreign source
without an 'Import Permit', the NBPGR needs to be kept informed of such requests for the
issuance of 'Import Permit'. The concerned scientist/organisation abroad is advised to take
into consideration the following requirements for mailing the material to India.

1. Only healthy, viable and clean seed material (free from soil, pests, pathogens and weeds)
are to be forwarded without any seed treatment so as to facilitate proper quarantine
examination. It may, however, be fumigated, if considered necessary.

2. The material is required to be accompanied with an 'Import Permit' and 'Phytosanitary

Certificate' with additional declaration, if any, based on crop inspection certifying that the
material is free from particular pathogen(s)/insect(s). Preferably, two copies of the
phytosanitary certificate, one inside the package and the other affixed outside for use by the
quarantine authorities are required. It should further be ensured that the package of
seed/planting material must be addressed to the Director, NBPGR, New Delhi, who has to
take delivery of the seed/planting material and conduct quarantine examination.

3. The seed/planting material should not be sent to any scientist by name directly, since one
point entry is necessary to have proper monitoring for quarantine requirements, for
documentation of passport data and national accessioning (E.C. Number assignment referring
to Exotic Collection).

4. The seed may preferably be sent by first class air mail addressed to the Director, National
Bureau of Plant Genetic Resources, Pusa Campus, New Delhi - 110012.

5. The perishable plant propagules (scion/woods, budwoods, plant rhizomes, suckers, etc.)
may preferably be sent by airfreight through any commercial airlines operating between
source country and the Indira Gandhi International Airport, New Delhi so as to avoid delay in
receipt and clearance. If unavoidable, the material can be sent on charge collect basis. An
advance intimation of the despatch of such perishable materials to the NBPGR will help in
prompt receipt and quick clearance of the material soon after its arrival. This will also avoid
payment of demurrage charges. However, in case it is not possible to send by air freight, the
same could be sent by courier service.

6. The sender should be requested to give full particulars of the seed/planting material along
with the address of the concerned scientist to whom the material is to be made available after
quarantine clearance by the Bureau.

7. Germplasm should be obtained in small quantities (3000 to 4000 seeds only) and, in case
of vegetative materials, it should not exceed six scion woods, rhizomes, etc., while in case of
rooted plants, the number should be kept to the minimum (1-2 plants each). Efforts should be
made to avoid repeat introductions. When requests are routed through the NBPGR, this
aspect is taken care of since supply of the required seed/planting material may possibly be
arranged from sources within India, depending on its availability.

Export

Exchange of germplasm involves not only introductions but also the supply of seed and other
materials to collaborating scientists/organisations abroad. While responding to such requests,
the following guidelines are to be followed:

1. Requests for seed/planting material received from concerned organisations/agencies

abroad have to be forwarded to the NBPGR with relevant information so that prompt
decision on the supply of desired materials could be taken.
2. Despatch of the seed/planting materials is also to be channelised through the Bureau so
that prompt inspection of the material could be done from quarantine angle and
phytosanitary certificate be issued.

3. Only the healthy seed material (free from diseases, pests, weeds, soil clods, plant debris,
etc.) should be sent to the Bureau in small quantity along with full details of the
material(s) and the name and address of the recipient. Quarantine clearance and despatch
normally takes about 7 to 10 days.

4. No seed dressing with insecticides or fungicides be given while despatching the seed to
the Bureau.

Quarantine: It is a strategy of control to prevent the spread of pests and diseases. It covers
all regulatory actions taken to exclude animal or plant pests or pathogens from a site, area,
country, or group of countries. For example, when animal or plant genetic resources are
imported from another country or region, there is a risk that they may contain or carry pests
or pathogens that could be damaging to agriculture. For this reason, countries use quarantine
practices to protect their agriculture and living natural resources from potential damage or
destruction
Quarantine is usually a government responsibility, and the manner in which quarantine
is executed differs among nations. National agencies responsible for plant quarantine may
have other responsibilities, such as domestic pest control; research; pesticide registration,
safety, and residue monitoring; or seed quality and labeling.
-
Lec 5: Modes of reproduction – sexual – asexual - self and cross fertilization –
significance of pollination
Modes of Reproduction
Knowledge of the mode of reproduction and pollination is essential for a plant
breeder, because these aspects help in deciding the breeding procedures to be used for the
genetic improvement of a crop species. Choice of breeding procedure depends on the mode
of reproduction and pollination of a crop species.
Reproduction refers to the process by which living organisms give rise to the offspring of
similar kind (species). In crop plants, the mode of reproduction is of two types: viz.
1) sexual reproduction and
2) asexual reproduction
I. Sexual reproduction
Multiplication of plants through embryos which have developed by fusion of male and
female gametes is known as sexual reproduction. All the seed propagating species belong to
this group.
Sporogenesis
Production of microspores and megaspores is known as sporogenesis. In anthers,
microspores are formed through microsporogensis and in ovules, the megaspores are formed
through megasporogenesis.
Microsporogenesis
The sporophytic cells in the pollen sacs of anther which undergo meiotic division to
form haploid i.e., microspores are called microspore (MMC) or pollen mother cell (PMC) and
the process is called microsporogenesis. Each PMC produce four microspores and each
microspore after thickening of the wall transforms into pollen grain.
Megasporogenesis
A single sporophytic cell inside the ovule, which undergo meiotic division to form
haploid megaspore, is called megaspore mother cell (MMC) and the process is called
megasporogenesis. Each MMC produces four megaspores out of which three degenerate
resulting in a single functional megaspore.
Gametogenesis
The production of male and female gametes in the microspores and megaspores is
known as gametogenesis.
Microgametogenesis
This is nothing but the production of male gametes or sperm. On maturation of the
pollen, the microspore nucleus divides mitotically to produce a generative and a vegetative or
tube nucleus. The pollen is generally released in this binucleate stage. The reach of pollen
over the stigma is called pollination. After the pollination, the pollen germinates. The pollen
tube enters the stigma and travels down the style. The generative nucleus at this phase
undergoes another mitotic division to produce two male gametes or sperm nuclei. The pollen
along with the pollen tube possessing a pair of sperm nuclei is called microgametophyte. The
pollen tube enters the embryo sac through micropyle and discharges the two sperm nuclei.
Megagametogenesis
The nucleus of the functional megaspore undergoes three mitotic divisions to produce
eight or more nuclei. The exact number of nuclei and their arrangement varies from one
species to another. The megaspore nucleus divides thrice to produce eight nuclei. Three of
these nuclei move to one pole and produce a central egg cell and two synergid cells on either
side. Another three nuclei migrate to the opposite pole to develop into three antipodal cells.
The two nuclei remaining in the center, the polar nuclei, fuse to form the secondary nucleus.
The megaspore thus develops into a mature female gametophyte called megagametophyte or
embryo sac. The development of embryo sac from a megaspore is known as
megagametogeneis. The embryo sac generally contains one egg cell, two synergids with the
apparent function of guiding the sperm nucleus towards the egg cell and three antipodals
which forms the prothalamus cells and one diploid secondary nucleus.
Fertilization
The fusion of one of the two sperms with the egg cell producing a diploid zygote is
known as fertilization. The fusion of the remaining sperm with the secondary nucleus leading
to the formation of a triploid primary endosperm nucleus is termed as triple fusion. The
primary endosperm nucleus after several mitotic divisions develops into mature endosperm,
which nourishes the developing embryo.
II. Asexual reproduction
Multiplication of plants without the fusion of male and female gametes is known as
asexual reproduction. Asexual reproduction can occur either by vegetative plant parts or by
vegetative embryos which develop without sexual fusion (apomixis). Thus asexual
reproduction is of two types: viz. a) vegetative reproduction and b) apomixis.
Vegetative reproduction refers to multiplication of plants by means of various
vegetative plant parts. Vegetative reproduction is again of two types: viz.
 i) Natural vegetative reproduction and
ii) Artificial vegetative reproduction.
Natural vegetative reproduction
In nature, multiplication of certain plants occurs by underground stems, sub aerial
stems, roots and bulbils. In some crop species, underground stems (a modified group of
stems) give rise to new plants. Underground stems are of four types: viz. rhizome, tuber,
corm and bulb. The examples of plants which reproduce by means of underground stems are
given below:
Rhizome: Turmeric (Curcuma domestica), Ginger (Zingiber officinale)
Tuber: Potato (Solanum tuberosum)
Corm: Arvi (Colocasia esculenta), Bunda (C. antiquorum)
Bulb: Garlic (Allium sativum), onion (A. cepa)
Rhizome: Turmeric Tuber: Potato Bulb: Onion
Sub aerial stems include runner, sucker, stolon, etc. These stems lead to vegetative
reproduction in mint (Mentha sp) rose, strawberry, banana, etc. Bulbils are modified forms of
flower. They develop into plants when fall on the ground. Bulbils are founding garlic.
Artificial vegetative reproduction
Multiplication of plants by vegetative parts through artificial method is known as
artificial vegetative reproduction. Such reproduction occurs by cuttings of stem and roots, and
by layering and grafting. Examples of such reproduction are given below:
Stem cuttings: Sugarcane (Saccharum sp.) grapes (Vitis vinifera), roses, etc.
Root cuttings: Sweet potato, citrus, lemon, etc. Layering and grafting are used in fruit and
ornamental crops.
Apomixis
Apomixis refers to the development of seed without sexual fusion (fertilization). In
apomixis embryo develops without fertilization. Thus apomixis is an asexual means of
reproduction. Apomixis is found in many crop species. Reproduction in some species occurs
only by apomixis. This apomixis is termed as obligate apomixis. But in some species sexual
reproduction also occurs in addition to apomixis. Such apomixis is known as facultative
apomixis.
There are four types of apomixis: viz.
1) parthenogenesis, 2) apogamy, 3) apospory and 4) adventive embryony.
1. Parthenogenesis. Parthenogenesis refers to development of embryo from the egg cell
without fertilization.
2. Apogamy. The origin of embryo from either synergids or antipodal cells of the embryosac
is called as apogamy.
3. Apospory. In apospory, first diploid cell of ovule lying outside the embryosac develops
into another embryosac without reduction. The embryo then develops directly from the
diploid egg cell without fertilization.
4. Adventive embryony. The development of embryo directly from the diploid cells of ovule
lying outside the embryosac belonging to either nucellus or integuments is referred to as
adventive embryony.
Modes of Pollination
The process by which pollen grains are transferred from anthers to stigma is referred
as pollination. Pollination is of two types: viz.
1) Autogamy or self pollination and
2) Allogamy or cross pollination.
I. Autogamy
Transfer of pollen grains from the anther to the stigma of same flower is known as
autogamy or self pollination. Autogamy is the closest form of inbreeding. Autogamy leads to
homozygosity. Such species develop homozygous balance and do not exhibit significant
inbreeding depression.
Mechanism promoting self-pollination
1. Bisexuality
Presence of male and female organs in the same flower is known as bisexuality. The
presence of bisexual flowers is a must for self pollination. All the self pollinated plants have
hermaphrodite flowers.
2. Homogamy
Maturation of anthers and stigma of a flower at the same time is called homogamy. As
a rule, homogamy is essential for self-pollination.
3. Cleistogamy
When pollination and fertilization occur in unopened flower bud, it is known as
cleistogamy. It ensures self pollination and prevents cross pollination. Cleistogamy has been
reported in some varieties of wheat, barley, oats and several other grass species.
4. Chasmogamy
Opening of flowers only after the completion of pollination is known as chasmogamy.
This also promotes self pollination and is found in crops like wheat, barley, rice and oats.
5. Position of Anthers
In some species, stigmas are surrounded by anthers in such a way that self pollination
is ensured. Such situation is found in tomato and brinjal. In some legumes, the stamens and
stigma are enclosed by the petals in such a way that self pollination is ensured. Examples are
greengram, blackgram, soybean, chickpea and pea.

II. Allogamy
Transfer of pollen grains from the anther of one plant to the stigma of another plant is
called allogamy or cross pollination. This is the common form of outbreeding.
Allogamy leads to heterozygosity. Such species develop heterozygous balance and
exhibit significant inbreeding depression on selfing.
Mechanism promoting cross-pollination
1. Dicliny
It refers to unisexual flowers. This is of two types: viz. i) monoecy and ii) dioecy.
When male and female flowers are separate but present in the same plants, it is known as
monoecy. In some crops, the male and female flowers are present in the same inflorescence
such as in mango, castor and banana. In some cases, they are on separate inflorescence as in
maize.
Other examples are cucurbits, grapes, strawberry, cassava and rubber. When
staminate and pistillate flowers are present on different plants, it is called dioecy. It includes
papaya, date palm, spinach, hemp and asparagus.
2. Dichogamy (from the Greek dikho-apart and gamous-marriage)
It refers to maturation of anthers and stigma of the same flowers at different times.
Dichogamy promotes cross pollination even in the hermaphrodite species. Dichogamy is of
two types: viz. i) protogyny and ii) protandry. When pistil matures before anthers, it is called
protogyny such as in pearl millet. When anthers mature before pistil, it is known as
protandry. It is found in maize, sugarbeet and several other species.
3. Heterostyly
When styles and filaments in a flower are of different lengths, it is called heterostyly.
It promotes cross pollination, such as linseed.
4. Herkogamy
Hinderance to self-pollination due to some physical barriers such as presence of
hyline membrane around the anther is known as herkogamy. Such membrane does not allow
the dehiscence of pollen and prevents self-pollination such as in alfalfa.
5. Self incompatibility
The inability of fertile pollens to fertilize the same flower is referred to as self
incompatibility. It prevents self-pollination and promotes cross pollination. Self
incompatibility is found in several crop species like Brassica, Radish, Nicotiana, and many
grass species. It is of two types sporophytic and gametophytic.
6. Male sterility
In some species, the pollen grains are non functional. Such condition is known as
male sterility. It prevents self-pollination and promotes cross pollination. It is of three types:
viz. genetic, cytoplasmic and cytoplasmic genetic. It is a useful tool in hybrid seed
production. Study of floral biology and aforesaid mechanisms is essential for determining the
mode of pollination of various crop species. Moreover, if selfing has adverse effects on seed
setting and general vigour, it indicates that the species is cross pollinated. If selfing does not
have any adverse effect on these characters, it suggests that the species is self-pollinated.
The percentage of cross pollination can be determined by growing a seed mixture of
two different varieties together. The two varieties should have marker characters say green
and pigmented plants. The seeds are harvested from the recessive (green) variety and grown
next year in separate field. The proportion of pigmented plants in green variety will indicate
the percentage of out crossing or cross pollination.
Significance of pollination
The mode of pollination plays an important role in plant breeding. It has impact on
five important aspects: viz. 1) gene action, 2) genetic constitution, 3) adaptability, 4) genetic
purity and 5) transfer of genes.
Classification of crop plants based on mode of pollination and mode of reproduction
Mode of pollination and reproduction
Examples of crop plants
A. Autogamous Species
1. Seed Propagated Rice, Wheat, Barley, Oats, Chickpea, Pea, Cowpea, Lentil, Green gram,
Black gram, Soybean, Common bean, Moth bean, Linseed, Sesame, Khesari, Sunhemp,
Chillies, Brinjal, Tomato, Okra, Peanut, etc.
2. Vegetatively Propagated Potato
B. Allogamous Species
1. Seed Propagated Corn, Pearlmillet, Rye, Alfalfa, Radish, Cabbage, Sunflower, Sugarbeet,
Castor, Red clover, White clover, Safflower, Spinach, Onion, Garlic, Turnip, Squash,
Muskmelon, Watermelon, Cucumber, Pumpkin, Kenaf, Oilpalm, Carrot, Coconut, Papaya,
etc.
2. Vegetatively propagated Sugarcane, Coffee, Cocoa, Tea, Apple, Pears, Peaches, Cherries,
grapes, Almond Strawberries, Pine apple, Banana, Cashew, Irish, Cassava, Taro, Rubber, etc.
C. Often Allogamous Species Sorghum, Cotton, Triticale, Pigeonpea, Tobacco.
Genetic consequences of self and cross-pollination
S.No. Self-Pollination Cross-Pollination

1. Self pollination leads to a very rapid increase Cross pollination preserves and
in homozygosity. Therefore, populations of promotes heterozygosity in a
self – pollinated species are highly population. Cross pollinated species
homozygous. are highly heterozygous and show
mild to severe inbreeding depression
and a considerable amount heterosis
2. Self pollinated species do not show The breeding methods in such
inbreeding depression, but may exhibit species aim at improving the crop
considerable heterosis. species without reducing
heterozygosity to an appreciable
degree
3. The aim of breeding methods generally is to Usually hybrid or synthetic varities
develop homozygous varieties. The are the aim of breeder wherever the
inbreeding mechanisams are generally under seed production of such varieties is
precise genetic control, but can be influenced economically feasible.
by both the genetic background as well as the
environment.

Lec 6. Self incompatibility – classifications – mechanisms – application – measures to

overcome and limitations
SELF INCOMPATABILITY
Self incompatibility and sterility are the two mechanisms which encourage cross
pollination. More than 300 species belonging to 20 families of angiosperms show self
incompatibility

Definition
 In self incompatible plants, the flowers will produce functional or viable pollen
grains which fail to fertilize the same flower or any other flower of the same plant.
a) Self incompatible pollen grain may fail to germinate on the stigmatic surface.
b) Some may germinate but fails to penetrate the stigmatic surface
c) Some pollen grains may produce pollen tube which enters through stigmatic surface
but its growth will be too slow. By the time the pollen tube enters the ovule the
flower will drop.
d) Some time fertilization is effected but embryo degenerates early.
Reason
Self incompatibility is appeared to be due to biochemical reaction, but precise nature
of these reactions is not clearly understood.
Classification of self incompatibility
According to Lewis (1954) the self incompatibility is classified as follows.

Self incompatibility

Heteromorphic system Homomorphic system

Distyly Tristyly Gametophytic System Sporophytic System

Heteromorphyic system
In this case there will be difference in the morphology of the flowers. For example in
primula there are two types of flowers namely PIN and THRUM.
PIN flowers have long style and short stamens while THRUM flowers have short
style and long stamens. This type of difference is known as Distyly
TRISTYLY is known in some plants like Lythrum. In this case the style of the flower
may be either short, long or medium length.
In case of distyly the only compatible mating is between PIN and THRUM. The relative
position of anthers is determined by single gene S/s. The recessive ss produces PIN and
heterozygotes Ss produces THRUM. Homozygous dominant SS is lethal and do not exist. The
incompatibility reaction of pollen is determined by the genotype of the plant producing them. Allele
S is dominant over s. This system is also known as heteromorphic - sporophytic system. Pollen
grains produced by PIN flowers will be all s in genotype as well as in incompatibility reaction. Where
as THRUM flowers will produce two types of gametes S and s but all of them would be S
phenotypically. The mating between PIN and THRUM would produce Ss and ss progeny in equal
frequencies. This system is of little importance in crop plants. It occurs in sweet potato and buck
wheat.
Mating Progeny
Phenotype Genotype Genotype Phenotype
Pin x Pin ss x ss Incom. mating -
Pin x Thrum ss x Ss 1 ss : Ss 1 Thrum
1 Pin
Thrum x Pin Ss x ss 1 Ss : ss 1 Thrum
1 Pin
Thrum x Thrum Ss x Ss Incom. mating -

Homomorphic System
Here the incompatibility is not associated with morphological difference among
flower. The incompatibility reaction of pollen may be controlled by the genotype of the plant
on which it is produced – (Sporphytic control) or by its own genotype – (Gametophytic
control).
Gametophytic System
First discovered by East and Mangelsdorf in 1925 in Nicotiana sanderae. Here the
incompatible reaction of pollen is determined by its own genotype and not by the genotype of
the plant on which pollen is produced. Generally the incompatibility reaction is determined
by a single gene having multiple allele. E.g.Trifolium Nicotiana, Lycopersicon, Solanum,
Petunia. Here Codominance is assumed
Genotype of Plant S1 S2 S3 S4
(Sporophyte)
Genotype of gametes S1 S2 S3 S4

Incompatible
reaction of pollen S1 S2 S3 S4

Incompatible reaction
Of style S1 S2 S3 S4

Mating S1S2 x S1 S2 - Fullly Incompatible

S1S2 x S1 S3 - Partially compatible
S1S2 x S3 S4 - Fully compatible.
Sporophytic System
Here also the self incompatibility is governed by a single gene S with multiple alleles.
More than 30 alleles are known in Brassica oleracea. Here dominance is assumed.
The incompatibility reaction is determined by the genotype of the plant on which
pollen grain is produced and not by the genotype of the pollen. This system is more
complicated. The allele may exhibit dominance, co-dominance or competition. This system
was first reported by Hugues and Babcock in 1950 in Crepis foetida and by Gerstal in
Parthenium argentatum. The sporophytic system is found in radish, brassicas and spinach.
Lewis has summarized the characteristics of sporophytic system as follows.
1. There are frequent reciprocal differences.
2. Incompatibility can occur with female parent
3. A family can consist of three incompatibility groups.
4. Homozygotes are a normal part of the system
5. An incompatibility group may contain two genotypes.
MACHANISM OF SELF INCOMPATABILITY
This is quite complex and is poorly understood. The various phenomena observed in
Self Incompatibility is grouped in to three categories.
a) Pollen - Stigma interaction
b) Pollen tube - Style interaction.
c) Pollen tube - Ovule interaction.
a) Pollen - Stigma interaction :
This occurs just after the pollen grains reach the stigma and generally prevents pollen
from germination. Prviously it was thought that binucleate condition of pollen in
gamatophytic system and trinucleate condition in sporophytic system was the reason for self
incompatability. But later on it was observed that they are not the reason for S1. Under
homomorphic system of incompatability there are differences in the stigmatic surface which
prevents pollen germination. In gametophytic system the stigma surface is plumose having
elongated receptive cells which is commonly known as wet stigma. The pollen grain
germinates on reaching the stigma and incompatability reaction occurs at a later stage.
In the sporophytic system the stigma is papillate and dry and covered with hydrated
layer of protein known as pellicle. This pellicle is involved in incompatability reaction. With
in few minutes of reaching the stigmatic surface the pollen releases an exince excudate which
is either protein or glycero protein. This reacts with pellicel and induces callose formation
which further prevents the growth of pollen tube.
Pollen - Stigma interaction
Gametophytic Sporophytic
System System
Stigma surface Plumose Commonly Stigma surface Papillate and dry. Covered
known as wet Stigma with hydrated layere of protein known as
pellicle which involves in incompatibility
reaction.
Pollen grain germinates and Pollen grain releases exine exudate which is
incompatibility reaction occurs at a protein or Glycero-protein.
later stage.
This protein reaction with pellicle and induces
callose formation and arrests growth of pollen
type.
b) Pollen Tube - Style interaction :
Pollen grains germinate and Pollen tube penetrates the stigmatic surface. But in
incompatible combinations the growth of pollen tube is retarded with in the style as in
Petunia, Lycopersicon, Lilium. The protein and poly saccharine synthesis in the pollen tube
stops resulting in bursting up of pollen tube and leading to death of nuclei.
c) Pollen tube - Ovule interaction :
In Theobroma cacao pollen tube reaches the ovule and fertilisation occurs but the
embryo degenerates later due to some biochemical reaction.
Relevance of Self incompatibility in Plant Breeding :
Self incompatibility may be used for Hybrid seed production.
a) By planting two self incompatible but cross compatible varieties alternatively seeds
obtained from both the lines are hybrids.
b) Alternatively by planting a self incompatible variety along with self compatible variety,
the seeds obtained from self incompatible line will be a hybrid.
Hybrid seed production was made in brassicas, clover, Trifolium Solanaceous and
Asteraceae crops. But there are certain difficulties in this.
a) Production and maintenance of inbred line by hand pollination is tedious and costly.
b) Continuos selfing leads to break down of self incompatibility and self fertile lines will
appear.
c) Environmental factors such as high temperature and high humidity reduce self
incompatibility.
d) Bees often prefers to stay with in particular parental line which in turn increases the
proportion of selfed seed.
Elimination of Self incompatibility :
1) In a single gene gametophytic system by doubling the chromosome number we can
elimate self incompatibility.
2) By induced mutagenesis to produce self fertile lines.
3) By transferring self compatible alleles from related species thro’ back cross breeding.
Overcoming Self incompatibility
1. By bud pollination : Application of matured pollen to immature stigma.
2. By surgical technique : Removal of the stigmatic surface E.g. Brassicas or removal of
style E.g. Petunias.
3. End of season pollination : In some cases self incompatibility is reduced towards the end
of flowering period. Pollination at that time may be successful.
4. Use of high temperature : Exposure of pistil to 600C will induce pseudo fertility.
5. Irradiation :Grafting : Grafing of a branch to another branch.Double pollination :
Application of a mixture of incompatible and compatible pollen grains.
Lec 7 -Sterility – male sterility – introduction – classification – CMS,GMS,CGMS
-inheritance and applications
MALE STERILITY
Male sterility is characterized by nonfunctional pollen grains, while female gametes
function normally. It occurs in nature sporadically.

Morphological features of male sterility:

The male sterility may be due to mutation, chromosomal aberrations, cytoplasmic

factors or interaction of cytoplasmic and genetic factors. Because of any of the above reasons
the following morphological changes may occur in male sterile plants.

1. Viable pollen grains are not formed. The sterile pollen grains will be transparent and
rarely takes up stain faintly.
2. Non dehiscence of anthers, even though viable pollens are enclosed within. This may
be due to hard outer layer which restrict the release of pollen grains.
3. Androecium may abort before the pollen grains are formed.
4. Androecium may be malformed, thus there is no possibility of pollen grain formation.
Kinds of male sterility, maintenance and uses:
Male sterility may be conditioned due to cytoplasmic or genetic factors or due to
interaction of both. Environment also induces male sterility. Depending on these factors
male sterility can be classified in to
a) Cytoplasmic male sterility (CMS)
b) Genetic male sterility (GMS)
i) Environmental induced male sterility which is again sub divided in to
i) TGMS (Theromosensitive)
Two line breeding
ii) PGMS (Photo sensitive)
ii) Transgenic male sterility
c) Cytoplasmic-genetic male sterility (CGMS) (Three line breeding, A , B and R line)
A line or MS line: This term represents a male sterile line belonging to any one of the above
categories. The A line is always used as a female parent in hybrid seed production.
B line or maintainer line: This line is used to maintain the sterility of A line. The B line is
isogenic line which is identical for all traits except for fertility status.
R line and restoration of fertility: It is otherwise known as Restorer line which restores
fertility in the A line. The crossing between A x R lines results in F 1 fertile hybrid seeds
which is of commercial value.
1. Cytoplasmic Male Sterility (CMS):
It occurs due to the mutation of mitochondria or some other cytoplasmic factors
outside the nucleus. Nuclear genes are not involved here. There is considerable evidence
that gene or genes conditioning cytoplasmic male sterility. Particularly in maize DNA reside
in mitochondria and may be located in a plasmid like element.
Genetic structure

Sterile
Maintenance
x

f F

A line sterile B line fertile

f
sterile
Since mother contributes the cytoplasm to the offspring, the sterility is transferred to the F1
Uses:
Since there are no R lines available, this type of sterility is useful only in crops where
seed is not the end product. For example in onion and many ornamental plants the hybrids
developed exhibit maximum hybrid vigour with respect to longer vegetative duration and
larger flower size and larger bulb size. In certain ornamental species, or in species where a
vegetative part is of economic value. Cytoplasmic male sterility has successfully been
exploited in maize for producing double cross hybrids.

GENETIC MALE STERILITY : (GMS)

Genetic male sterility is normally governed by nuclear recessive genes ms ms.
Exception to this is safflower where male sterility is governed by dominant gene Ms Ms.
This type of male sterility is used in Redgram and Castor for production of hybrids.

Genetic structure :
A line

ms
ms

In Redgram there are number of GMS lines are available. E.g. Ms Co5, Ms T21
Maintenance :

In genetic male sterility, the sterile lien will be maintained from heterozygous
condition. The genetic structure of heterozygous line will be.

Ms
ms

When this heterozygous line is grown in the field it will segregate in the ratio of 1
Fertile : 1 sterile.
Sterile Fertile

ms Ms
ms ms
1 : 1

The pollen from the Fertile line will pollinate the sterile line and as a result seed set
will be there in the sterile line. These seeds are to be harvested and used for hybrid seed
production.
For hybrid seed production, the seeds collected from sterile plants will be grown using
double the seed rate since it will segregate in the ratio of 1 fertile : 1 sterile line. At the time
of flowering, the fertile line will be identified by yellow plumpy anthers and removed from
the field. Only the sterile line will remain in field. These will be pollinated by the R line and
the F1 obtained will be hybrid redgram
.
Utilisation: Hybrid Development. Eg: Redgram
Ms T21 x ICPL 87109
A line R line

ms Ms
F ms F Ms

Ms
F ms
 Hybrid
CoRH 1
Utilization in Plant Breeding
Genetic male sterility may be used in hybrid seed production. The progeny from ms
ms x Ms ms crosses are used as female, and are inter planted with a homozygous male fertile
(Ms Ms) pollinator. The genotypes of ms ms and Ms ms lines are identical except for the ms
locus, i.e., they are isogenic ; they are known as male sterile (A) and maintainer (B) lines,
respectively. The female line would, therefore, contain both male sterile and male fertile
plants ; the latter must be identified and removed before pollen shedding. This is done by
identifying the male fertile plants in seedling stage either due to the pleiotropic effect of the
ms gene or due to the phenotypic effect of a closely-linked gene. Pollen dispersal from the
male (pollintor) line should be good for a satisfactory seed set in the female line. however,
generally pollen dispersal is poor and good, closely-linked markers are rare. Rouging of male
fertile plants from the female lines is costly as a result of which the cost of hybrid seed is
higher. Due to these difficulties, genetic male sterility has been exploited commercially only
in a few countries. In USA, it is being successfully used in Castor. In India, it is being used
for hybrid seed production of arhar by some private seed companies, e.g., Maharashtra
Hybrid Seed Co. Ltd., India, produced and sold 50 Q seed of a hybrid variety of arhar,
Suggestions have been made for its use in several other crops, e.g., Cotton, barley, tomato,
sunflower, cucurbits etc., but it is not yet practically feasible.
DIFFICULTIES IN USE OF GMS
1. Maintenance of GMS requires skilled labour to identify fertile and sterile line.
Labelling is time consuming and costly

2. In hybrid seed production plot identification of fertile line and removing them is costly.
3. Use of double the seed rate of GMS line is costly.
4. In crops like castor high temperature leads to break down of male sterility.
CYTOPLASMIC – GENEIC MALE STERILITY
This is a case of cytoplasmic male sterility where dominant nuclear gene restores
fertility. This system is utilised for the production of hybrids in bajra, jowar, maize, rice,
wheat and many other crops.
Genetic Structure
A line

ms
ms

Male sterile.
Maintenance
A line B line
ms x ms
S ms F ms
sterile Fertile

ms
S ms

Male sterile line

The A line which is male sterile is maintained by crossing it with isogenic B line
which is also known as maintainer line. The B line is similar to that of A line in all
characters (isogenic) except fertile cytoplasm.
UTILISATION :
The male sterile. A line is crossed with R line ( Restorer) Which restores fertility in
F1.
A line R line

ms Ms
S ms Ms
F
Sterile Fertile
Ms
S ms

Hybrid
Fertile
DIFFERENT TYPES OF MATING IN CGMS LINE

ms ms ms
S ms x F ms _____ S ms

Sterile Fertile Sterile

ms Ms Ms
S ms x S/F Ms ______ S ms

Sterile Fertile Fertile

ms Ms
S ms x S/F ms

Ms ms
S ms and S ms

Fertile Sterile.
Transfer of Male Sterility from Exotic lines to Nature lines:
Most of the times the MS lines obtained from other countries may not be suitable to our
condition. Examples are:
Crop Source of cytoplasm Drawbacks
Maize Texas Cytoplasm Susceptible to Helminthosporium leaf blight
Sorghum Combined kafir Black glumes and chalky endosperm
Pearlmillet Tift 23 A (Tifton) Susceptible to Green ear & downy mildew
Rice Wild abortive incomplete panicle exertion
Sunflower H petiolaris, H gigantis
Tobacco Microcephalan Reduced vigour in F1 hybrids
Wheat Aegilops caudata Susceptible to pistiloidy
Due to these drawbacks, the well adapted local lines should be converted into male
sterile lines. This can be done by repeated back crossing of the local lines to the exotic MS
lines.
Transfer of Male Sterility to a New Strain – back cross breeding
Maintenance of Male Sterile Line or A line: Since A line does not produce pollen, seed is
not formed for maintaining A line. It has to be crossed with its fertile counterpart having
similar nuclear genes with fertile cytoplasm which is known as B-line.
Production of Hybrid seed: For production of hybrid seed, A-line has to be kept as female
parent and the pollen parent should posses the restorer genes in order to induce fertility and
seed development in the next generation. Such line is known as restorer line and denoted as
‘R'line. The A line & R line should be of different genetic constitution and should be able to
give maximum heterosis
Limitations of CGMS lines.
1. Fertility restoration is a problem. E.g. Rice.
2. Seed set will be low in crops like Rice where special techniques are to be adopted to
increase seed set.
3. Break down of male sterility at higher temperature.
4. In crops like wheat having a polyploidy series it is difficult to develop effective R line.
5. Undesirable effect of cytoplasm. E.g. Texas cytoplasm in maize became susceptible to
Helminthasporium. In bajra Tift 23 A cytoplasm became susceptible to downy mildew.
6. Modifier genes may reduce effectiveness of cytoplasmic male sterility.
LINE BREEDING
The process of using different lines (genotype) and producing hybrid is known as line
breeding. This terminology is used in production of rice hybrids.
The different kinds of line breeding are.
a) One line method
b) Two line method
c) Three line method
a) One line method of rice breeding :
Rice hybrids can be developed and propagated through the following concepts.
- Vegetative propagation. This can be done by ratooning followed by stubble
planting.
- Micropropagation employing tissue culture technique.
- Anther culture hybrids. The anthers of F 1 hybrid can be cultured and plant lets
developed.
- Apomictic lines.
b) Two line method of rice breeding.
Two line hybrids can be evolved through application of gametocides and use of
environmentally induced genic male sterility.
To the selected female parent pollen suppressors can be sprayed at the time of
flowering so that it will arrest the production of pollen and thus temporary male sterility is
induced. The best combiner is used as a male parent and hybrid is produced.
The EGMS system is used successfully in china. Both TGMS and PGMS lines were
identified. In this system male sterility is mainly controlled by one or two pairs of recessive
nuclear genes and has no relation to cytoplasm. In this system only two lines viz. male sterile
and Restorer lines are used. Maintainer line is not needed because by growing the male
sterile line in suitable atmosphere the sterility is maintained. In this method there is no
negative effect due to sterile cytoplasm.
Three line method or CGMS System
This system nowadays known as CGMS system involving three lines viz.
a) Cytoplasmic genic male sterile line. b) Maintainer or B line and c) Restorer line.
TRANSFER OF MALE STERILITY OF A NEW STRAIN
Lec 8: TGMS, PGMS, Gametocides, Transgenic male sterility and
applicationsTemperature Sensitive Genetic Male Sterility (TGMS):
Plants are sterile when temperature exceeds 32˚C/24˚C (day /night) and becomes
fertile when the temperature is below 24˚C/18˚C (day /night). However, in few cases,
sterility is observed at lower temperature and fertility is observed at higher temperatures.
Such type of male sterility is referred to as “Reverse TGMS type”. This can be utilized in
tropical and subtropical countries, where there are large temperature differences across
locations, regions and seasons and at different attitudes. It is used to development of two –
line hybrids.
Photoperiod Sensitive Genetic Male Sterility (PGMS):
The line is sterile when the photoperiod (day light) exceeds 14 hrs and same line becomes
fertile when subjected to photoperiod of < 13 hrs. PGMS is useful and can be deployed in
termperate countries where the day length differs considerably during different seasons.
TGMS and PGMS are used for development of hybrid rice in China during eighties.
Trangenic Genetic male sterility (TrGMS)
Transfer of gene into an genome of aan organism by recombinant DNA technology is
called transgene.
Barnase/Barstar system is good example of transgenic male sterility.
Barnase gene of Bacillus amyloliquefaciens encodes an RNase.
Barnase gene is driven by TA29 promoter expressed only in tapetum cells causing
their degeneration.
Transgenic tobacco and Brassica napus plants with Barnase genes where complete
male sterile. Another gener Barstar from the same bacterium encodes a protein which is
inhibitor of Barnase Rnase. Hence the transgenic plants expressing both Barstar and Barnase
are fully male fertile.
Barnase gene has been linked with bar gene, which is resistant to specific herbicide
phosphinothricin. Male sterile line can be maintained by crossing it with any male fertile line.
Resultant progenies are 1:1 sterile: fertile. Fertile can be easily eliminated by
herbicide spray.
Gametocides/ Chemically Hybridizing Agents

Chemical induction of sterility in plants has been of interest since 1950 when the
potential for selective male sterility was first demonstrated. Various terms have been used
since 1950 to describe chemicals that induce male. These criteria are essential to sterility in
plants. The most commonly used maximize the efficiency of hybrid seed term is gametocide
(or) selective gametocide. This terminology was introduced by Eaton (1957) who
demonstrated the potential of producing Gossypium hirsutum hybrids through the use of
sodium-a, ~-dichloro- isobutyrate (FW-450). Over years many investigator have used such
terms as male sterilant, selective male sterilant, pollen suppressant, pollenicide and
androcide. Later, the term Chemical Hybridizing Agents (CHAs) is used after the entire
primary objective is to produce a hybrid.
The first reports of chemically induced male sterility were those by Moore (in 1950)
and Nylor induced male sterility in maize using Maleic hydrazide (MH). Laibach and
Kribban reported that αNAA and ᵝ- IAA increased the proportion of staminate flowers in
cucumber (Cucumis sativus).
Important CHAs
1. Zinc methyl arsenate and sodium methylarsenate are commercially used. Sodium
methyl arsenate has been popular in rice hybrid production (usually at 150 mg/l) as foliar
spary 15 days before heading. Due to toxic effect 5 day before heading is more effective.
2. Ethephon (Ethrel) : It is used in barley, mustard, oats etc., 750+4000mg/l and 0.2 to
12kg/ha depending on crop. Ethrel produces some adverse effects like delayed vegetative
growth and low female fertility. Etheral spray before meiosis limits its adverse effect on
female fertility.
3. Gibberellic Acid (GA3) : It is commonly applicable maize, barley, wheat, rice and
sunflower. It should be sprayed before meiosis initiation after meiosis it is in effective.
4. LY195259: It is very effective but negative effects on seed set and seed quality
5. RH0007 (Hybrex) Used commercially in wheat. 0.2 kg/ha
Advantages:
1. Any line can be used as female parent. The lengthy and cumbersome production of
CMS, GMS and CGMS lines for hybrid seed production becomes unnecessary
2. Any line can be used as female parent and any line can be used as male parent,
restorer gene not required
3. Hybrid seed production only based on two lines
4. Maintenance of parental line achieved through selfing.
5. In CHAs F2s are fully fertile. This would allow for commercial cultivation.
Limitations:
1. The expression and duration of CHA- induced male sterility is stage specific
2. Vulnerable to prevailing environmental condition
3. Incomplete male sterility might lead to the production of selfed seed on the female
parent
4. Many CHAs are toxic to plants and animals
5. Some CHAs eg arsenicals and WL 84811 may produce carryover residual effect in
F1 seeds
6. Sme CHAS stimulate neoplamic growth affect human growth
7. CHAs are generally genotype- dose and application stage specific.

Lec 9 - Apomixis – introduction - classification-applications; Parthenocarpy and its

types.
INTRODUCTION:
 Apomixis, derived from two greek word “APO” (away from) and “mixis” (act of
mixing or mingling). It refers to the occurrence of an sexual reproductive process in the
place of normal sexual processes involving reduction division and fertilization. In other
words apomixis is a type of reproduction in which sexual organs of related structures take
part but seeds are formed without union of gametes. Seeds formed in this way are vegetative
in origin. When apomixis is the only method of reproduction in a plant species, it is known as
obligate apomixis. On the other hand, if gametic and apomictic reproduction occur in the
same plant, it is known as facultative apomixis. The first discovery of this phenomenon is
credited to Leuwenhock as early as 1719 in Citrus seeds.
Apomixis is widely distributed among higher plants. More than 300 species
belonging to 35 families are apomictic. It is most common in Gramineae, Compositae,
Rosaceae and Rutaceae. Among the major cereals maize, wheat and pearl millet have
apomictic relatives.

.
Types of apomixis :
Mainly three types of apomixis phenomenon are suggested by Maheshwari (1954) :
1. Recurrent Apomixis :
An embryo sac develops from the MMC or megaspore mother cell
(archesporial cell) where meiosis is disturbed (sporogenesis failed) or from some adjoining
cell (in that case MMC disintegrates). Consequently, the egg-cell is diploid. The embryo
subsequently develops directly from the diploid egg-cell without fertilization. Somatic
apospory, diploid parthenogenesis and diploid apogamy are recurrent apomixis. However,
diploid parthenogenesis / apogamy occur only in aposporic (somatic) embryo-sacs.
Therefore, it is the somatic or diploid aposory that constitutes the recurrent apomixis. Such
apomixis occurs in some species of Crepis, Taraxacum, Poa (blue grass), and Allium (onion)
without the stimulus of pollination. Malus (apple), and Rudbeckia where pollination appears
to be necessary, either to stimulate embryo development or to produce a viable endosperm.
2. Non -recurrent Apomixis :
An embryo arises directly from normal egg-cell (n) without fertilization. Since an
egg-cell is haploid, the resulting embryo will also be haploid. Haploid parthenogenesis and
haploid apogamy, and androgamy fall in this category. Such types of apomixis are of rare
occurrence. They do not perpetuate and are primarily of genetic interest as in corn.
3. Adventive Embryony :
 Embryos arise from a cell or a group of cells either in the nucellus or in the
integuments, e.g. in oranges and roses. Since it takes place outside the embryo sac, it is not
grouped with recurrent apomixis, though this is regenerated with the accuracy. In addition
to such embryos, regular embryo within the embryo sac may also develop simultaneously,
thus giving rise to poly-embryony condition, as in Citrus, Opuntia.
Now, different apomictic phenomena in each of the recurrent and non-recurrent
apomicts are considered in relation to the development of the embryo sac or embryo.

Development of apomictic embryo sac

1. Apospory :
It involves the development of embryo sac either from the archesporial cell or from
the nucellus, or from other cell. It is of two types :
(i) Generative or haploid apospory: If the embryo sac develops from one of the megaspores
(n), the process is called generative or haploid apospory. Since it cannot regenerate, as it is
haploid and fertilization fails, the process gives rise to non-recurrent apomicts.
(ii) Somatic or diploid apospory: When diploid embryo sac is formed from nucellus or other
cells, the process is termed as somatic or diploid asopory. Since it regenerates without
fertilization, it is recurrent.
Development of apomictic embryo
1. Parthenogenesis :
This refers to the development of embryo from egg-cell without fertilization, e.g. in
some cases in corn, wheat, tobacco. This is also of two kinds :
(i) Haploid parthenogenesis: The embryo develops from egg-cell without fertilization in a
haploid embryo-sac produced by generative apospory. It is non-recurrent in nature.
(ii) Diploid parthenogenesis: The embryo develops from egg-cell without fertilization in a
diploid embryo-sac arising from somatic apospory. It is recurrent type.
2. Apogamy :
This is related to the development of embryo not from the egg-cell but from any one
of the synergid or antipodal cells within the embryo sac, without fertilization. This is haploid
or diploid. In the haploid apogamy, the embryo arises from any cell other than the egg-cell
without fertilization in haploid embryo -sac formed by generative apospory. By virtue of its
haploid nature, it is also non-recurrent apomixis. Whereas in case of diploid apogamy,
embryo develops from any cell other than the egg-cell without fertilization in a diploid
embryo-sac developed by somatic apospory. It is recurrent type.
3. Androgamy :
It is the development of embryo neither from egg cell nor from synergids or
antipodals, but from one of the male gametes itself, inside or outside the embryo-sac. Since it
is haploid, it is non-recurrent apomixis.
In another phenomenon, i.e. parthenocarpy, seedless fruits are formed from ovary
without fertilization. Normally, fertilization stimulates the ovary to become enlarged and
form fruit. But in case of parthenocarpy, such stimulation may be received even from
incompatible pollination.
Genetics of apomixis : Crosses between amphimicts and apomicts belonging to the same
species, segregate for the two types of individuals in advanced generations. This suggests
that apomixis is a genetically controlled phenomenon in plants. Stebbins (1958) states that,
as a rule, the apomictic condition is recessive to sexuality, although polyploid apomicts show
tendency towards dominance. However, this recessiveness is not usually due to a monogenic
difference. Since there is frequent reversion of apomicts to normal sexuality or sterility or
some abnormal genetic behaviour in crosses involving in apomict and an amphimict or
involving two apomicts of diverse origins, it appears that a successful apomictic cycle is the
result of an interaction of many genes which tend to break on hybridization. It is only in the
relatively simple types of apomixis, like adventive embryony and vegetative reproduction
that simple genetic beheaviour can be expected. Recently, Vardy et al. (1989) recorded three
recessive genes with additive effects responsible for parthenocarpy.
Advantages of apomixis in plant breeding : The two sexualprocesses, self-and cross-
fertilization, followed by segregation, tend to alter the genetic composition of plants
reproduced through amphimixis. Inbreeding and uncontrolled out breeding also tend to break
heterozygote superiority in such plants. On the contrary, apmicts tend to conserve the genetic
structure of their carriers. They are also capable of maintaining heterozygote advantages
generation after generation. Therefore, such a mechanism might offer a great advantage in
plant breeding where genetic uniformity maintained over generation for both homozygosity
(in varieties of selfers), and heterozygosity (in hybrids of both selfers and outbreeders) is the
choicest goal. Additionally, apomixis may also affect an efficient exploitation of maternal
influence, if any, reflecting in the resultant progenies, early or delayed because it causes the
perpetuation of only maternal individuals and maternal properties due to prohibition of
fertilization. Maternal effects are most common in horticultural crops, particularly fruit trees
and ornamental plants.
 Thus, in short the benefits of apomixis, insofar as their utility in plant breeding is
concerned, are :
1. Rapid multiplication of genetically uniform individuals can be achieved without risk of
segregation.
2. Heterosis or hybrid vigour can permanently be fixed in crop plants, thus no problem for
recurring seed production of F1 hybrids.
3. Efficient exploitation of maternal effect, if present, is possible from generation to
generation.
4. Homozygous inbred lines, as in corn, can be rapidly developed as they produce sectors of
diploid tissues and occasional fertile gametes and seeds.

Exploitation of apomixis in crop improvement :

The use of apomixis in crops in a follow-up process, after a variety or hybrid is
evolved, as reflected by the benefits it renders. Therefore, our aim in this section is to deal
with only apomixis as a tool to plant breeding.
With a view to exploit apomixis in sexual crops, it needs to detect and identify an
apomictic phenomenon, occurring spontaneously in any plant, or, to incorporate it artificially,
perhaps through hybridization between apomicts and amphimicts.

Detection of apomixis : Positive evidence for the presence or absence of apomixis can be
obtained only from an intensive screening of a large number of plants in a variety/hybrid.
The screening involves a careful and systematic tracing of steps for the development of
embryo-sac and embryo, through microtomy of ovule, right from megaspores to embryonic
development. as such, therefore, it is a most tedious job requiring a lot of patience and
persistence indeed.
It should however be noted that it is only recurrent apomixis, namely diploid forms of
apospory / parthenogenesis / apogamy / adventive embryony and vegetative propagation
which are beneficial for plant breeding purposes. The simple reason being that it is these
which produce viable diploid embryos without fertilization and thus can continue to
perpetuate over generations. Nonrecurrent apomixis are of academic use.
Maintenance and transfer of apomixis : Once an apomict plant is detected its inheritance
should promptly be studied by crossing a half or few flowers with the pollen obtained from
normal plants and going through the segregation pattern in F 2 and onward generations. The
remaining flowers may thoroughly be checked and seeds collected on maturity. The true
nature of such plants would become distinct only after progeny tests. A true apomictic plant
will automatically produce mother apomictic progenies which can be maintained without
difficulty.
With regard to transfer of apomixis, substantial evidence is available for the hybrid
origin of many of the apomicts. Nevertheless, there is no evidence at all the hybridization by
itself can induce apomixis (Stebbins, 1950). Situation is further aggravated by the unstable
nature of apomicts since there is every likelihood of the breaking down of interacting gene
complexes conditioning apomixis, as stated earlier. Therefore, possibilities of introducing
apomixis in non-apomicts are the least but not totally absent.

Lec 10 – Polygenic variation – components of variance-phenotypic, genotypic and

variance -heritability and genetic advance

The phenotype may be described according to a mathematical model to facilitate

statistical analysis and interpretation. The phenotypic mean i.e. X of a given genotype from
the trial may be expressed as m,
x = μ + g +e +ge where,
x = phenotypic mean
μ = general population mean
g = effect of genotype
e = effect of environment
ge = interaction between genotype and environment

Analysis of variance for genotypes grown in a replicated trial according to rbd

Source of Variation d.f. Expectation of MS
Replications r-1 σ + gσr2
e
2

Genotypes g-1 σe2 + rσg2

Error (r-1) (g-1)
σe2
Total (rg)-1

g and r are the number of genotypes and replications respectively; and σe2, σr2 and σg2
denote the variances due to error, replications and genotypes respectively.
 Genotypic variance (σg2)=(MS due to genotypes – MS due to error)/R

Phenotypic variance (σp2) = σg2 + σe2

The environment and quantitative variation

All genes are expressed in an environment (phenotype = genotype + environmental
effect). However, quantitative traits tend to be influenced to a greater degree than qualitative
traits. It should be pointed out that, under significantly large environmental effects,
qualitative traits (controlled by one or a few major genes) can exhibit a quantitative trait
inheritance pattern. A strong environmental influence causes the otherwise distinct classes to
overlap.
Variance components of a quantitative trait
The genetic properties of a population are determined by the relative magnitudes of the
components of variance. In addition, by knowing the components of variance, one may
estimate the relative importance of the various determinants of phenotype. K. Mather
expressed the phenotypic value of quantitative traits in this commonly used expression:
P (phenotype) = G (genotype) + E (environment)
Individuals differ in phenotypic value. When the phenotypes of a quantitative trait are
measured, the observed value represents the phenotypic value of the individual. The
phenotypic value is variable because it depends on genetic differences among individuals, as
well as environmental factors and the interaction between genotypes and the environment
(called G × E interaction).
Total variance of a quantitative trait may be mathematically expressed as follows:
VP = VG + VE + VGE
where VP = total phenotypic variance of the segregating population, VG = genetic variance,
VE = environmental variance, and VGE = variance associated with the genetic and
environmental interaction. The genetic component of variance may be further partitioned into
three components as follows:
VG = VA + VD + VI
where VA = additive variance (variance from additive gene effects), VD = dominance
variance (variance from dominance gene action), and VI = interaction (variance from
interaction between genes). Additive genetic variance (or simply additive variance) is the
variance of breeding values and is the primary cause of resemblance between relatives. Hence
VA is the primary determinant of the observable genetic properties of the population, and of
the response of the population to selection. Further, VA is the only component that the
researcher can most readily estimate from observations made on the population.
The total phenotypic variance may then be rewritten as:
VP = VA + VD + VI + VE + VGE
Concept of heritability
A phenotype observed is an interaction between the genes that encode it and the environment
in which the genes are being expressed. Plant breeders typically select plants based on the
phenotype of the desired trait, according to the breeding objective. Sometimes, a genetically
inferior plant may appear superior to other plants only because it is located in a more
favourable region of the soil. This may mislead the breeder. In other words, the selected
phenotype will not give rise to the same progeny. If the genetic variance is high and the
environmental variance is low, the progeny will be like the selected phenotype. The converse
is also true. If such a plant is selected for advancing the breeding program, the expected
genetic gain will not materialize. Quantitative traits are more difficult to select in a breeding
program because they are influenced to a greater degree by the environment than are
qualitative traits. If two plants are selected randomly from a mixed population, the observed
difference in a specific trait may be due to the average effects of genes (hereditary
differences), or differences in the environments in which the plants grew up, or both.
Definition
Heritability is defined as a fraction: it is the ratio of genetically caused variation to total
variation (including both environmental and genetic variation).
Types of heritability
There are two different estimates of heritability.
1. Broad sense heritability.
Heritability estimated using the total genetic variance (VG) is called broad sense
heritability. It is expressed mathematically as:
H = VG/VP
It tends to yield a high value. Some use the symbol H2 instead of H.

2. Narrow sense heritability.

Because the additive component of genetic variance determines the response to
selection, the narrow sense heritability estimate is more useful to plant breeders than
the broad sense estimate. It is estimated as:
h2 = VA/VP
However, when breeding clonally propagated species (e.g., sugarcane, banana), in which both
additive and non-additive gene actions are fixed and transferred from parent to offspring,
broad sense heritability is also useful. The magnitude of narrow sense heritability cannot
exceed, and is usually less than, the corresponding broad sense heritability estimate. The
estimates are expressed as a fraction, but may also be reported as a percentage by multiplying
by 100. A heritability estimate may be unity (1) or less.
Factors affecting heritability estimates
1. Genetic population: The amount of genotypic variance present for a trait in a
population influences estimates of heritability. Parents are responsible for the genetic
structure of the populations they produce. More divergent parents yield a population
that is more genetically variable.

2. Sample size: Because it is impractical to measure all individuals in a large population,

heritabilities are estimated from sample data. To obtain the true genetic variance for a
valid estimate of the true heritability of the trait, the sampling should be random. A
weakness in heritability estimates stems from bias and lack of statistical precision.

3. Method of computation
Heritabilities are estimated by several methods that use different genetic populations
and produce estimates that may vary.
Calculations:

Genotypic Co-efficient of variation

σg2
GCV:= x 100
μ
Phenotypic Co-efficient of variation

σp2
PCV= x 100
μ

Heritability
σg2
h2 = x 100
σp2
Genetic Advance
σg2
GA = x K where,
√σp2
K = Selection differential which is constant for the known selection intensity
(k at 5% selection intensity = 2.06).
Lec 11- Plant introduction as a breeding method – types of introduction – objectives –
quarantine - acclimatization – achievements - merits and demerits.

PLANT INTRODUCTION

Definition
Taking a genotype or a group of genotypes in to a new place or environment where
they were not grown previously.
Thus introduction may involve new varieties of a crop already grown in that area, a
wild relative of the crop species or totally a new crop species for that area.
E.g. a) Introduction of IRRI rice varieties.
b) Introduction of sunflower wild species from Russia
c) Introduction of oilpalm in to Tamil Nadu.
Primary Introduction
When the introduced crop or variety is well suited to the new environment, it is
directly grown or cultivated with out any alteration in the original genotype. This is known
as primary introduction. E.g. IR. 8, IR 20, IR 34, IR 50 Rice varieties. Oil palm varieties
introduced from Malaysia, Mashuri rice from Malaysia.
Secondary Introduction
The introduced variety may be subjected to selection to isolate a superior variety or it
may be used in hybridization programme to transfer some useful traits. This is known as
secondary Introduction.
E.g. In soybean EC 39821 introduced from Taiwan is subjected to selection and
variety Co1 was developed.
In rice ASD 4 is crossed with IR 20 to get Co 44 which is suited for late planting.
Objectives of Plant Introduction
1. To introduce new plant species there by creating ways to build up new industries.
E.g. Oil palm
2. To introduce high yielding varieties to increase food production. E.g. Rice and wheat.
3. To enrich the germplasm collection. E.g. Sorghum, Groundnut.
4. To get new sources of resistance against both biotic and abiotic stresses. E.g. NCAC
accessions to have rust resistance in groundnut. Dasal rice variety for saline resistance
5. Aesthetic value - ornamentals.
Plant Introduction Agencies
Most of the introductions occurred very early in the history. In earlier days the
agencies were invaders travelers, traders, explorers, piligrims and naturalists Muslim invaders
introduced in India cherries and grapes. Portuguese introduced maize, ground nut, chillies,
potato, sweet potato, guava, pine apple, papaya and cashew nut. East India Company brought
tea. Later Botanic gardens played a major role in plant Introduction
A centralised plant introduction agency was initiated in 1946 at IARI, New Delhi.
During 1976 National Bureau of Plant Genetic Resources (NBPGR) was started. The bureau
is responsible for introduction and maintenance of germplasm of agricultural and
horticultural plants. Similarly Forest Research Institute, Dehradun has a plant introduction
organisation which looks after introduction, maintenance and testing of germplasm of forest
trees. Besides NBPGR the Central Research Institutes of various crops also maintain
working germplasm. All the introductions in India must be routed through NBPGR, New
Delhi. The bureau functions as the central agency for export and introduction of germplasm.
At International level Biodiversity International (old name IBPGR) with head
quarters at Rome, Italy is responsible for plant introduction between countries.
Procedure for plant Introduction
The scientist / University will submit the requirement to NBPGR. If the introduction
is to be from other countries, NBPGR will address IBPGR for effecting supply. The IBPGR
will assign collect the material from the source and quarantine them, pack them issue
phytosanitary certificate suitably based on the material and send it to NBPGR. The NBPGR
will assign number for the material, keep part of the seed for germplasm and send the rest to
the scientist.
There are certain restrictions in plant introduction. Nendran banana from Tamil Nadu
should be not be sent out of state because of bunchy top disease. Similarly we cannot import
Cocoa from Africa, Ceylon, West Indies, Sugarcane from Australia, Sunflower from
Argentina.
Functions of NBPGR
1. Introduction maintenance and distribution of germplasm
2. Provide information about the germplasm through regular publications.
3. Conduct training courses to the scientist with regard to introduction and maintenance of
germplasm.
4. Conduct exploratory surveys for the collection of germplasm.
5. To set up Natural gene sanctuaries.
Merits and demerits of plant introduction.
Merits.
1. It provides new crop varieties which are high yielding and can be used directly
2. It provides new plant species.
3. Provides parent materials for genetic improvement of economic crops.
4. Enriching the existing germplasm and increasing the variability.
5. Introduction may protect certain plant species in to newer area will save them from
diseases. E.g. Coffee and Rubber.
Demerits
1. Introduction of new weed unknowingly.
E.g. Argemone mexicana, Eichornia Parthenium
2. Introduction of new diseases :
Late blight of potato from Europe.
Bunchy top of banana from Sri Lanka
3. New pests : Potato tuber moth came from Italy
4. Ornamentals becoming weeds : Lantana camara
5. Introduction may cause ecological imbalance E.g. Eucalyptus.
ACCLIMATIZATION
When superior cultivars from neighbouring or distant regions are introduced in a new
area, they generally fail initially to produce a phenotypic expression similar to that in their
place of origin. But later on they pickup and give optimal phenotypic performance, in other
words they become acclimatized to the new ecological sphere. Thus acclimatization is the
ability of crop variety to become adapted to new climatic and edaphic conditions.
The process of acclimatization follows an increase in the frequency of those
genotypes that are better adapted to the new environment.
The success of acclimatization depends upon two factors
i) Place effect
ii) Selection of new genotypes.
Lec 12 - Genetic basis of self pollinated crops – Vilmorin principle of progeny selection
- Johannsen’s pure line theory

METHODS OF BREEDING AUTOGAMOUS CROPS- Self pollinated crops

The following are the methods of breeding autogamous plants.

1. Introduction

2. Selection with out hybridization

a) Pure line selection
b) Mass selection

3. Hybridization and selection

 i) Inter varietal
a) Pedigree Method
b) Bulk Method.
c) Single Seed Descent Method.
d) Modified Bulk Method
e) Mass - Pedigree Method.
ii) Interspecific hybridization
4. Back cross method
5. Multiline Varieties
6. Population approach
7. Hybrids.
8. Mutation breeding
9. Polyploidy breeding
10. Innovative techniques
SELECTION IN SELF POLLINATED CROPS
To get successful results by selection there are two pre-requisites.
a) Variation must be present in the population.
b) The variation must be heritable.
HISTORY OF SELECTION :
Selection was practiced by farmers from ancient times. During 16th century Van
Mons in Belgium, Andrew knight in England and Cooper in USA practiced selection in crop
plants and released many varieties.
Le coutier, a farmer of island of New Jersey published his results on selection in
wheat in the year 1843. He concluded that progenies from single plants were more uniform.
During the same period Patrick shireff, a scotsman practiced selection in wheat and oats and
developed some valuable varieties.
During 1857 Hallet in England practiced single plant selection in wheat, oats and
barley and developed several commercial varieties.
About this time Vilmorin proposed individual plant selection based on progeny
testing. This method successfully improved the sugar content in sugar beet. His method
was called as Vilmorin isolation principle. He emphasised that the real value of a plant can
be known only by studying the progeny produced by it. This method was successful in sugar
beet but not in wheat. This shows the in-effectiveness of selection in cross pollinated crops.
Today progeny test is the basic step in every breeding method.
PURE LINE THEORY

A pure line is the progeny of a single self fertilized homozygous plant.

The concept of pureline was proposed by Johannsen on the basis of his studies with
beans (Phaseolus vulgaris) variety called Princess. He obtained the seeds from the market
and observed that the lot consisted of a mixture of larger as well as smaller size seeds. Thus
there was variation in seed size. Johannsen selected seeds of different sizes and grown them
individually. Progenies of larger seeds produced larger seeds and progenies from smaller
seeds produced small seeds only.
This clearly showed that there is variation in seed size in the commercial lot and it has
a genetic basis. He studied nineteen lines al together. He concluded that the market lot of the
beans is a mixture of pure lines.
He also concluded whatever variation observed with in a line is due to environment
only. Confirmatory evidence was obtained in three ways.

In line 13 which is having 450 mg seed wt he divided the seeds on weight basis. He
divided the line into seeds having 200, 300, 400 and 500 mg weights and studied the
progenies. Ultimately he got lines having weight ranging from 458 to 475. Thus the
variation observed is purely due to environment.
The second evidence was that selection with in a pure line is ineffective. From a pure
line having 840 mg selection was made for large as well as small seeds. After six
generations of selection the line for large seed as well as for small seed gave progenies
having 680-690 mg. Thus it was proved that selection within a pure line is ineffective.
 In third evidence when parent - offspring regression was worked in line thirteen. It
worked to zero indicating that variation observed is non heritable and it is due to environment
only.

ORIGIN OF VARIATION IN PURELINES

1. Mechanical mixtures.
2. Natural hybridization.
3. Chromosomal aberrations.
4. Natural mutation.
5. Environmental factors.
EFFECT OF SELF-POLLINATION ON GENOTYPE
Self-pollination increases homozygosity with a corresponding decrease in
heterozygosity. For example an individual heterozygous for a single gene Aa is self
pollinated in successive generations, every generation of selfing will reduce the frequency of
heterozygote Aa to 50 percent of that in the previous generation. There is a corresponding
increase in homozygotes AA and aa. As a result, after 10 generations of selfing virtually all
the plant in the population will be homozygous AA and aa.
No. of Frequency(%) Frequency (%)
generations
of selfing AA Aa aa Homozygote Heterozygote
0 0 100 0 0 100
1 25 50 25 50 50
2 25 + 12.5 25 25 + 12.5 75 25
This can be calculated by the formulae
[2m - 1) / 2m ]n
where m = No. of generations of self-pollination.
 n = No. of genes segregating.
When number of genes are segregating together, each gene would become
homozygous at the same rate as Aa. Thus the number of genes segregating does not affect
the percentage of homozygosity. Similarly linkage between genes does not affect the
percentage of homozygosity in the population.
Genetic advance under selection
Normally selection is practiced based on the phenotype of the individual plant. The
phenotype in turn is the result of joint action of genotype and environment i.e.,
VP = VG + VE
Where P= Phenotype
G = Genotype
E = Environment
Genetic advance:
It is the improvement in the mean genotypic value of the selected families over the
base population is known as genetic advance under selection.

Genetic advance under selection depends upon

a) Genetic variability among different plants or families in the base population.
b) The herbitability of the character under selection.
c) The intensity of selection i.e., the proportion of plants or families selected.
Genetic advance under selection may calculated as follows.
GS = (K) (P) (H)
Where GS = Genetic advance under selection
k = Selection differential
P = Phenotypic standard deviation of the base population
H = Heritability of the character under selection.
Lec 13 - Breeding methods for self pollinated crops without involving artificial
hybridization: Pure line selection – procedure – merits and demerits – achievements;
Mass selection in self pollinated crops – procedure - types – comparison of mass and
pureline selection – achievements.
PURELINE SELECTION
A large number of plants are selected from a self pollinated crop. The selected plants
are harvested individually. The selected individual plants are grown in individual rows and
evaluated and best progeny is selected, yield tested and released as a variety.
CHARACTERISTICS OF PURELINES
1. All plants within a pure line have the same genotype.
2. The variation with in a pureline is environmental and nonheritable.
3. Purelines become genetically variable with time due to natural hybridization, mutation
and mechanical mixtures.

General steps for making a pureline selection

First Season : From the base population select best looking plants having the desirable
characters. Harvest them on single plant basis.
Second Season : The selected single plants are grown in progeny rows and estimate the
performance. Reject unwanted progenies.
Third Season : Repeat the process of second season.
Fourth Season : Grow the selected single plants in replicated preliminary yield trial along
with suitable check or control variety.
Fifth Season : Conduct regular comparative yield trial along with check variety and select
the best culture.
Sixth Season : Conduct multilocation trial in different research stations along with local
check.
Seventh Season : Conduct Adaptive Research Trial in farmer’s field. Fix the best yielder
and release it as a variety thro’ Variety Release committee.

Advantage of Pureline Selection.

1. Achieves maximum possible improvement over the original variety.
2. Extremely uniform in appearance.
3. Because of the uniformity, a variety is easily identified and seed certification is easy.
Disadvantages :
1. It does not have wide adaptability because improvement is made only in the local variety.
2. Time required for developing a variety is more when compared to mass selection.
3. Depending on the genetic variability present in the base population only the improvement
is made. If there is no genetic variability improvement cannot be made.
4. Breeder has to spend more time compared to mass selection.
b) MASS SELECTION
Here a large number of plants having similar phenotype are selected and their seeds
are mixed together to constitute a new variety. Thus the population obtained from selected
plants will be more uniform than the original population. However they are genotypically
different.
Steps :
First season :
From the base population select phenotypically similar plants which may be 200 -
2000. Harvest the selected plants as a bulk.
Second season :
The bulk seed is divided into smaller lots and grown in preliminary yield trial along
with control variety. Dissimilar phenotypes are rejected. Higher yielding plots are selected.
Third to Sixth Season :
With the selected lots conduct yield trials along with appropriate check or control.
Select the best one and release it as a variety.
Types of mass selection
Positive mass selection:
1) Desirable plants are selected from a mixed population
2) Base materials is old varieties or land races
Negative mass selection
1) Undesirable off types are removed from a mixed population
2) Used for varietal purification and certification programme
Merits of Mass Selection :
1. Varieties developed will be having more adaptability since each plant is genotypicaly not
similar. They have buffering action against abnormal environment.
2. Time taken for release of a variety is less.
3. The genetic variability present in the original population is maintained.
Demerits :
1. Compared to pure line variety they may not be uniform.
2. In the absence of progeny test we are not sure whether the superiority of selected plant is
due to environment or genotype.
3. May not be as uniform as that of a pureline variety and certification is difficult.

COMPARISON BETWEEN PURELINE AND MASS SELECTIONS

Pureline selection Mass selection
1. The new variety is a pureline The new variety is a mixture of
purelines.
2. The new variety is highly uniform. In The variety has genetic variation of
fact, the variation within a pureline quantitative characters, although it is
variety is purely environmental. relatively uniform in general
appearance.
3. The selected plants are subjected to Progeny test is generally not carried out.
progeny test.
4. The variety is generally the best The variety is inferior to the best
pureline present in the original pureline because most of the purelines
population. The pureline selection included in it will be inferior to the best
brings about the greatest improvement pureline.
over the original variety.
5. Generally, a pureline variety is Usually the variety has a wider
expected to have narrower adaptation adaptation and greater stability than a
and lower stability in performance pureline variety.
than a mixture of purelines.
6. The plants are selected for the The selected plants have to be similar in
desirability. It is not necessary they phenotype since their seeds are mixed to
should have a similar phenotype. make up the new variety.

7. It is more demanding because careful If a large number of plants are selected,

progeny tests and yield trials have to expensive yield trials are not necessary.
be conducted. Thus it is less demanding on the breeder.
Lec 14 - Breeding methods of self pollinated crops involving artificial hybridization:
Creating variability in self pollinated crops - Hybridization and selection – objectives
types – choice of parents – combining ability - combination breeding and transgressive
breeding – steps in hybridization - kinds of emasculation.

HYBRIDIZATION AND SELECTION

Objective of hybridization
The chief objective of hybridization is to create variation. When two genotypically
different plants are crossed, the genes from both the parents are brought together in F 1.
Segregation and recombination produce many new gene combinations in F2 and subsequent
generations.
The degree of variation produced depends on the number of heterozygous genes in F1.
The number of heterozygous genes in F1 in turn depends on number of genes for which the
two parents differ. If the parents are not related they may differ for several genes.
Combination breeding
The main aim of combination breeding is the transfer of one or more characters into a
single variety from other varieties. These characters may be governed by oligogenes or
polygenes. In this approach, increase in yield is obtained by correcting the weaknesses in
the yield contributing traits like tiller number, grains per panicle, seed weight of the
concerned variety. Example for combination breeding is disease resistance achieved by
backcross breeding. Pedigree method is also another example.
Transgressive breeding
Transgressive segregation is the production of plants in F2 generation that are superior
to both the parents for one or more characters. Such plants are produced by the accumulation
of favourable genes from both the parents as a consequence of recombination. In this case
the parents involved in hybridization must combine well with each other and preferably be
genetically diverse. This way, each parent expected to contribute different plus genes which
when brought together by recombination gives rise to transgresive segregation. The pedigree
method as well as population approach are designed to produce transgresive segregants.

Simple cross = crossing between two parents

Convergent or complex cross – crossing involing more than two parents
A/B – Simple corss
A/B//C/D – Double corss
A/B//C = three way corss
A*3/B – A is back crossed three times as recurrent parent
A/2*b – B is back crossed two times as recurrent parent
PROCEDURE OF HYBRIDIZATION
1. Set up your objective.
2. Selection of parents.
3. Evaluation of parents.
4. Sowing plan.
5. Emasculation and dusting.
6. Labelling and bagging.
7. Harvesting and storage of seeds.

1. Objective
Based on the requirement, set your objective. Because based on the objective only the
selection of parents is done. If it is resistance breeding one of the parents must be a donor.
2. Selection of parents
Normal practice is, the female parent will be a locally adapted one in which we can
bring in the plus genes. In case of intervarietal hybridization geographically diverse parents
will be selected so as to get superior segregants.
3. Evaluation of parents
In case of parents which are new to the region they must be evaluated for their
adaptability. Further to ensure homozygosity, they must be evaluated.
4. Sowing plan
If the flowering duration is same, simultaneous sowing of both the parents can be
done. Otherwise staggered sowing is to be followed. The normal practice is to raise the
ovule parent in the centre of the plot in rows and on the border pollen parent for each
combination.
5. Emasculation and dusting
Emasculation is the removal of immature anthers from a bisexual flower. Depending
on the crop the emasculation practice differs. Normal practice of hand emasculation and
dusting of pollen is done. Depending on the time of anthesis the time of emasculation differs.
For E.g. in rice the anthesis at Coimbatore takes place between 7.00 to 10.00 A.M. So the
emasculation is done at around 6.30 A.M. and dusting of pollen is done immediately.
An efficient emasculation technique should prevent self pollination and result ina high
percentage of seed set on pollination.
 Different emasculation techniques are
i. Hand emasculation – It can be done for relatively large flowers. Anthers and stamens
are removed with the help of forceps. Emasculation is done before the anthers are
mature and the stigma has become receptive. This minimize accidental self pollination.
Generally emasculation is done in the evening between 4 and 6 P.M one day before
anthers of the flower are expected to dehisce or mature and stigma is likely to become
fully receptive. Care must be taken to remove all the anthers from the flowers without
breaking them and most important gynoecium must not be injured.
ii. Suction method: Useful in species of small flower. Emasculation is done in the
morning just befor or immediately after the flower open, Petals are removed with
forceps and exposed anther and stigma. A thin rubber or glass tube attached to a suction
hose is used to suck the anthers th tube is also passed over the stimas to suck any pollen
grains present on their surface. Th suction may be produced by an aspirator attached to
a water tap or by a small suction pump. With suction method, considerable amount of
self pollination (up to 15 %) is likely to occur.
iii. Hot water method: Pollen grains are more sensitive than the female reproductive
organs to both genetic and environmental disturbances. Kill pollen grains with hot
water without damaging female reproductive organs. In this method temperature may
vary crop to crop. Eg. Sorghum – 42- 48°C for 10 min ; Rice 40-44°C for 10 min .
:Hot water treatment is given before anthers dehise and prior to opening of the flower
iv. Alcohol treatment: Rarely used method consists of immersing the flower or the
inflorescence in alcohol of a suitable concentration for a brief period followed by
rinsing it with water. Eg. Sweer clover immersion of the inflorescence in 57 % alcohol
for 10 sec was highly effective.
v. Cold treatment: Like hot water treatment, kill pollen grains without damaging
gynoceium with cold water treatment. Eg rice 0-6 °C; Wheat 0-2°C for 15-24 hrs. it is
less effective than hot water treatment.
vi. Genetic emasculation: CMS, GMS and CGMS may be used to eliminate the necessisty
of emasculation. SI, protogyny facility also help crossing without emasculation.
vii. Chemical emasculation: many chemicals eg Sodium methl arsenate, ethephon, GA3,
Hybrex etc. These chemicals are called chemical hybridizing agents.
6. Labelling and bagging
Immediately after hybridization put a label indicating the parents and date of
crossing. Put appropriate cover to prevent foreign pollen, contamination.
7. Harvesting and storage of seeds
Normally 15-20 days after crossing the seeds will be set. In the case of pulses the
crossed pods can be easily identified by the shrunken nature of pod and seed set will be
reduced. Harvest of crossed seeds must be done on individual plant basis. Seeds collected
from individual plants are to be stored in appropriate containers with proper label and stored.

Lec 15 - Pedigree breeding – procedure – mass pedigree – merits – demerits –

achievements; Bulk breeding – procedure – merits – demerits – achievements.
Lec 16 - Comparison of pedigree and bulk breeding methods.  Single Seed Descent
(SSD) method – procedure – application – merits and demerits.
a) PEDIGREE METHOD :
In this method, individual plants are selected from F2 and subsequent generations and
their progenies are tested. During this process details about the plants selected in each
generation is recorded in Pedigree Record. By looking into Pedigree record we can know
about the ancestry of the selected plants.
For maintenance of pedigree record the basic thing required is Crossing Ledger. This
Ledger gives the details about parentage, Season in which the cross is made.
Sl.No. Cross Number Parentage
1 X S 9801 Co2 x MS 9804
2 X S 9802 Co4 x C152
3 X S 9803 Co1 x Co4
X = Cross
S = Summer Season.
98 = Year
1 = Serial No.of Cross.
There are several ways to maintain the pedigree Record. The selection of plants starts
from F2 onwards. The details about selected plants can be recorded as follows. E.g. F 2 X S
9801 - 7. Here the 7 denotes seventh plant selected.
In F3 if selection is made from the 7th plant of cross X S 9801 it can be
recorded as F3 X S 9801 - 7 - 4. The number four indicates that fourth plant of 7th plant of
F2 is selected. This can be followed till F4 or F5 generations. After F4 or F5 the selected
plants are bulked to form a family.
In the pedigree record all the biometerical data like plant height, number of branches,
No. of pods / plant, pod length, seeds / pod, pod weight, seed weight are recorded.
Merits of Pedigree Method :
 1. Gives maximum opportunity to the breeder to use his skill and judgement for the
selection of plants.
2. Well suited for characters which are simply inherited
3. Transgressive segregants can be easily identified thro’ records.
4. Information about inheritance is precisely obtained.
Demerits :
1. Maintenance of pedigree record is time consuming and limits handling of larger
population.
2. The success in this method is largely dependent on skill of the breeder. There is no
opportunity for natural selection.
3. Selection for yield in F2 and F3 is ineffective. If care is not taken to maintain larger
population, valuable materials may be lost.
PEDIGREE METHOD PROCEDURE

F1 Generation : The F1 seeds are space planted so that full expression of F1 can be had. It
is advisable to raise the parents involved in the cross to raise as border rows so that
dominance and other characters can be studied. The F1s are harvested as single plants.
F2 generation : In F2, 2000 to 10,000 plants per cross are planted. About 100 - 500 plants
are selected and harvested on single plant basis. The selection in F 2 depends upon the skill of
the breeder. The selection intensity may be 5 to 10%.
F3 generation : Individual plant progenies are space planted. Again desirable plants are
selected. From F3 onwards the term family is introduced. The line selected from each cross
is termed as family.
F4 generation : Similar to F3.
F5 generation : Many families would have attained homozygosity and may be harvested as
row bulk.
F6 generation : The row bulk may be assessed in multi row trial. The families exhibiting
segregation may be isolated and studied separately.
F7 generation - RRYT
F8 generation - PYT, CYT 3 seasons.

Basis of selection :
Depending upon the objective, selection is to be made in segregating generation.
For insect and disease resistance part of the seeds may be reserved in segregating generation
and the rest may be subjected to epiphytotic conditions. The families exhibiting resistance
may be identified and the reserve seeds may be used for further selection and testing.
Early generation testing:
If superior families are identified in F3 or F4, they can be tested for desirable
characters and this is known as early generation testing.
Shuttle breeding :
This is followed especially in disease or insect resistance breeding. For e.g. at
Coimbatore YMV in blackgram is in epidemic form during summer season only. Whereas at
Vamban (Pudukkottai) the YMV is epidemic during kharif season. So instead of waiting for
next summer at Coimbatore the materials can be tested at Vamban during kharif and thus one
season is saved.
Off season nursery :
Some crops may be season bound. But it may be non - season bound in certain agro -
climatic zone. For e.g. Thalai virichan cholam. (S.roxburghii) is season bound at
Coimbatore. It has to be sown during July - August and harvested during December -
January. But this S.roxbughii is non - season bound in Yercaud. So to save one season, the
segregating material can be raised during Rabi summer at yercaud. This method is otherwise
known as rapid generation advancement (RGA).
b) BULK METHOD
In this method F2 and subsequent generations are harvested as bulk to grow the next
generation. The duration of bulking may be 6 - 7 generations. Selection can be made in each
generation but harvest is done as bulk. This is similar to mass selection . At the end of
bulking period single plant selection is made and tested for yielding ability.

If bulking period is long say 20 - 30 seasons, then natural selection acts on the
homozygous lines.
In this method the breeder uses his skill for selecting the plants and at the same time
there is no pedigree record. This saves much time and labour.
Merits of bulk method :
1. Simple, convenient and inexpensive
2. By inducing artificial epiphytotic conditions undesirable or weaker genotypes can be
eliminated.
3. If bulking period is longer natural selection operates and desirable genotypes are
selected.
4. No pedigree record is maintained.
5. Since large population is grown there is chance for appearance of transgressive segregants
which will be superior than parents or F2
Demerits :
1. Takes much longer time to develop a new variety.
2. In short term bulk there is no chance for natural selection.
3. A large number of progenies are to be selected in each generation which requires much
labour, time and space.
4. We cannot get information on inheritance.
c) SINGLE SEED - DESCENT METHOD
It is the modification of the bulk method. In this method a single seed from each of
the F2 plants is collected and bulked to raise F3 generation. Similarly single seed from each
F3 plant is collected and carried forward to F 4. This procedure is followed till F 6 or F7. After
wards single plant selection is
made and studied in progeny
rows.

In this Scheme the main features

are:
1. Lack of selection till F6 or F7
when the population becomes
homozygous.
2. Each F2 plant is represented till
F 6 or F7 generation.
3. In this method there are chances
for reduction in population size
due to pest, disease or poor
germination.
 4. Rapid generation advancement (RGA) can be made with the use of glass house or off
season nursery

d) MODIFIED BULK METHOD

Here selection can be practiced in F2 and F3 and subsequent generations. There will
not be any pedigree record but superior plants are selected bulked and carried forward. In F 4
superior plants are selected and harvested on single plant basis. In F 5 these single plants are
studied in progeny rows and best progenies are selected and harvested. In F 6 PYT can be
conducted to select best families. In subsequent generations regular trials can be conducted.
This modification of the bulk method provides an opportunity for the breeder to
exercise his skill and judgment in selection. Further there is no maintenance of pedigree
record which is another advantage.
e) MASS PEDIGREE METHOD
This was proposed by Harrington. It is a solution to one of the deficiencies in the
pedigree method of breeding. For e.g. if the population is to be subjected to disease
resistance screening like YMV and if there is no method to create artificial epiphytotic
conditions, it is wasteful to study the population in pedigree method. Instead we can carry
the population as a mass and test them when there is occurrence of the disease. When
conditions are favourable for the disease, we can terminate the bulking and resort to single
plant selection.
Comparison between bulk and pedigree methods
Pedigree method Bulk method
1. Individual plants are selected in F2 and F2 and the subsequent generations are
the subsequent generations and maintained as bulks.
individual plant progenies are grown.
2. Artificial selection, artificial disease Artificial selection, artificial disease
epidemics etc., are an integral part of epiphytotics etc., may be used to assist
the method. natural selection. In certain cases,
artificial selection may be essential
3. Natural selection does not play any role Natural selection determines the
in the method. composition of the populations at the
end of the bulking period.
4. Pedigree records have to be maintained No pedigree record is maintained.
which is often time consuming and
laborious
5. It generally takes 14-15 years to It takes much longer for the
develop a new variety and to release it development and release of a variety.
for cultivation. The bulk population has to be
maintained for more than 10 years for
natural selection to act.
6. Most widely used breeding method. Used only to a limited extent.
7. It demands close attention from the It is simple, convenient and inexpensive
breeder from F2 onwards as individual and does not require much attention
plant selections have to be made and from the breeder during the period of
pedigree records have to be maintained. bulking.
8. The segregating generations are space - The bulk populations are generally
planted to permit individual plant planted at commercial planting rates.
selection.
9. The size of population is usually Large populations are grown. This and
smaller than that in the case of bulk natural selection are expected to
method. increase the chances of the recovery of
transgressive segregants.