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 with
known as wet Stigma 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 F1 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 F1 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 F2 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 σe2+ gσr2
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
pureline present in the original because most of the purelines included in
population. The pureline selection it will be inferior to the best pureline.
brings about the greatest improvement
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 than pureline variety.
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 be expensive yield trials are not necessary.
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. F2 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 F2 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 F4. This procedure is followed till F6 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 F6 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 F5 these single plants are
studied in progeny rows and best progenies are selected and harvested. In F6 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 the epiphytotics etc., may be used to assist
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 develop It takes much longer for the development
a new variety and to release it for and release of a variety. The bulk
cultivation. 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 from
plant selections have to be made and the breeder during the period of bulking.
pedigree records have to be maintained.
8. The segregating generations are space -
planted to permit individual plant
selection.
9. The size of population is usually smaller
than that in the case of bulk method.
The bulk populations are generally
planted at commercial planting rates.
Large populations are grown. This and
natural selection are expected to increase
the chances of the recovery of
transgressive segregants.
 PBG 301 Fundamentals of Plant Breeding 2+1
Theory notes after mid-semester
18. Backcross breeding – genetic basis –– procedures for transferring dominant and
recessive genes, back cross breeding – merits – demerits

Introduction
Crossing between a hybrid with one of its parents is known as Back cross. The hybrid
and its progenies in the subsequent generations are repeatedly back crossed to one of their
parent. As a result the genotype of back cross progeny becomes increasingly similar to that
parent to whom the back crosses are made. At the end of 6-8 back crosses, the progeny would
be almost identical with the parent involved in back crossing.

Objective

1. To improve one or two specific defects of a high yielding variety.

2. The characters lacking in this variety are transferred to it from a donor parent without
changing the genotype of this variety except for the genes being transformed.
Requirement of back cross breeding
1. Suitable recurrent or recipient parent must be available which lacks in one or two
characteristics. In back cross breeding, the parent to which one or few genes from the
donor parent are transferred is known as recurrent parent.
2. A suitable donor parent must be available, the character must be highly intense form.
The donor parent or non-recurrent parent - from which the character is transferred to
recurrent parent.
3. Character to be transferred must have high heritability controlled by one or two genes.
4. 6-7 back crosses are required for full recovery of recurrent parent.

Applications of back cross breeding

1. Inter varietal transfer of simply inherited traits

Characters governed by one or two genes like disease resistance are successful.
2. Inter varietal transfer of quantitative characters
Transfer of highly heritable quantitative characters like earliness, plant height, seed size
and seed shape.
3. Inter specific transfer of simply inherited characters
Disease resistance is transferred from related species to cultivated species.
Eg. Resistance to black arm disease in cotton from wild tetraploid species to G.hirsutum
Stem rust resistance from Triticum Timopheevi and Aegilops speltoides to T.aestivum.
4. Transfer of cytoplasm
Transfer of male sterility from wild species to cultivated species. The variety or species
from which the cytoplasm is to be transferred is used as the female parent. The parent
to which the cytoplasm is to be transferred is used as the male parent (recurrent parent)
 in the original cross and back cross. After 6-8 back crosses, the progeny would have the
nuclear genotype of the recurrent parent and the cytoplasm from the donor parent.
E.g. Sesamum malabariucum x S.indicum
Female parent Recurrent parent.
5. Transgressive segregation
F1 is back crossed to one or two times to the recurrent parent leaving much
heterozygosity for transgressive segregation to appear. In the second modification two
or more recurrent parent may be used in the back cross progeny to accumulate genes
from them into the back cross. Progeny of the new variety is not exactly like any one of
the recurrent parent.
6. Production of isogenic lines
Isogenic lines are identical in their genotype except for one gene
7. Germplasm conversion
When valuable germplasm cannot be utilized in breeding programmes and may be used
as recurrent parent in separate back cross programme. Popularly known as conversion
programme and these lines are called as converted lines. Eg.production of
photoinsensitive sorghum from photosensitive germplasm.

Genetic consequences of backcross

1. Reduction in heterozygosity
Leads to an increase in homozygosity and frequency of homozygotes
2. Increased similarity with the recurrent parent

3. Selection for the gene under transfer

4. Opportunity for breaking undesirable linkage
5.Selection for the recurrent parent type
Procedure for backcross method
Selection of parents
Recurrent parent: Most popular variety of the area - high yielding, desirable quality and
high adaptability

Non-recurrent parent or donor parent: Selected for the character that is to be improved in
the recurrent parent

The plan of backcross method would depend upon whether the gene being transferred is
recessive or dominant. The plan for transfer of dominant gene is simpler than that of recessive
gene.

Back cross method - Transfer of dominant gene

E.g. High yielding and widely adapted wheat variety A is susceptible to stem rust another
variety B is resistance to stem rust. Stem rust resistance is dominant to susceptibility.

1st year -Hybridization: Variety A is crossed to variety B. Generally variety A should be used
as female parent. This would help in identification of selfed plants.

2nd year- F1 Generation: F1 plants are back crossed to variety A. Since all the F 1 are
heterozygous for rust resistance, selection for rust resistance is not necessary.

3rd year- BC1 generation: Half of the plants in BC1 generation are resistant and the remaining
half would be susceptible to stem rust. Rust resistant plants are selected and back crossed
to variety A.
 4th year 7th year - BC2 to BC5 generation: Segregation would occur for rust resistance. Rust
resistant plants are selected and back crossed to variety A.
8th year- BC6 Generation: BC6 plants will have 99 percent genes from variety A Rust resistant
plants are selected and selfed, their seeds are harvested separately
9th year- BC6F2 Generation: Individual plants progeny from the selfed seeds of the selected
plants are grown. Rust resistant plant similar to the plant type of variety A are selected and
they are selfed. Seeds are harvested separately.
10th year- BC6F3 Generation: Individual plants progeny are grown. Progenies homozygous
for rust resistant and similar to plant type of variety A are harvested in bulk. Several similar
progenies are usually mixed to constitute the new variety.
11th year- Yield trials: Replicated yield trial along with the variety A as check.
12th year : Variety may be directly released for cultivation.
Back cross method -Transfer of a recessive gene

When Rust resistant is recessive, all the back crosses cannot be made one after the
other. For every two subsequent back crosses F2 generation must be grown to identify
the rust resistant plants. The F1 and the backcross progenies are not inoculated with
rust because they would susceptible to rust. Hence, only the F2 populations are tested
for rust resistance.
 1. Hybridization: The recurrent parent is crossed with the rust resistant donor parent;
the recurrent parent is generally used as female parent.
2. F1 Generation: F1 plants are back crossed to the recurrent parent; No rust resistant test
3. BC1 Generation: Since rust resistance is recessive, all the plants are rust susceptible. No
test for rust resistance and all the plants are self pollinated.
4. BC1 F2 Generation: Rust resistant test; rust resistant plants are selected and back
crossed to recurrent parent
5. BC2 Generation: No test for rust resistance and the plants are backcrossed to recurrent
parent.
6. BC3 Generation: No test for rust resistance and all the plants are self pollinated.
7. BC3 F2 Generation: Rust resistant test; Rust resistant plants are selected and back
crossed to recurrent parent.
8. BC4 Generation: There is no rust resistance test. Plants are back crossed to recurrent
parent.

9. BC5 Generation: There is no disease test. The plants are self pollinated to raise
F2. Selection is usually done for the plant type of variety A.
10. BC3 F2 Generation: Rust resistant test; resistant plants are self pollinated
11. BC3 F3 Generation: individual plant progenies are grown. Plants resistant to rust and
similar to variety A selected and composited
12. Yield trials are conducted along with variety A as check
13. Seed multiplication for distribution

Merits of Back cross breeding

1. The genotype of the new variety is nearly identical with that of recurrent parent except
for the genes transferred.
2. Useful for the transfer of disease resistance and incorporation of quality traits into a
variety
3. It is not necessary to test the variety developed in extensive yield trails because the
performance of the recurrent parent is already known.This may save upto 5 years time and
a considerable expense.
4. Back cross programme is not dependent upon environment. Therefore, off season
nurseries and green houses can be used to grow 2-3 generations each year. It would save
time.
5. Much smaller population is required compared to pedigree method.
6. Defects of a well adapted variety can be removed without affecting its performance
and adaptability.
7. This is the only method for the inter specific gene transfer and transfer of cytoplasm.
 8. It may be modified so that transgressive segregation may occur for quantitative
characters.

Demerits
1. New variety cannot be superior to recurrent parent except for the character
transferred
2. It involves lot of crossing work. 6-8 back cross is often difficult and time consuming.
3. Sometime undesirable gene linked with desirable also may be transferred.
4. By the time the back cross programme the recurrent parent may have been replaced
by other varieties superior in yield and other character.

Transfer of two or more characters to a single recurrent parent

1. Simultaneous transfer
❖ The genes for different characters may be transferred simultaneously in the same
backcross programme.
❖ The characters to be transferred are brought together in the hybrid by crossing each
of the donor parent to the recurrent parent (RP) and the hybrid thus produced
❖ A larger backcross population is needed
❖ Sometimes, the two genes under transfer may be linked. In such a case, the transfer
become very easy, and selection for only one gene may be necessary.
❖ Eg: favourable linkages are between the genes Lr 24 and Sr 24; Lr 19 and Sr 25; Lr
26 and Sr 31.

2. Stepwise transfer
❖ The RP is first improved for one character. The improved RP is then used as RP in a
backcross programme for the transfer of other character.
❖ If additional characters are to be transferred, they are transferred one time in a
stepwise fashion.
❖ This approach takes much longer time for the transfer of two or more characters.

3. Simultaneous but separate transfers

❖ Each character is transferred to the same RP in simultaneous but separate backcross
programme.
❖ The resulting improved varieties from the different programmes are then crossed
together.
❖ Homozygous lines for the characters being transferred are then selected from the
segregating generations using the pedigree method
❖ This approach appears to be the most suitable of the three methods.
Modifications of the Backcross Method
The backcross method may be modified in various ways to suit the needs of the breeder.
Following are the three common modifications of the backcross method.
1. Production of F2 and F3
The F2 and F3 generations are produced after the first and the third backcrosses. A rigid
selection for the character being transferred and for the characteristics of the recurrent
parent is done in the F2 and F3 generations. In the backcross progenies, selection need not be
done either for the character being transferred or for the characteristics of the recurrent
parent. The fourth, fifth and sixth backcross are made in succession. For the sixth backcross,
a relatively larger number of plants from the backcross progeny is used. This method may be
used for the transfer of both dominant and recessive genes. It is believed that an effective
selection in F2 and F3 generations is equivalent to one or two additional backcrosses.

2. Use of different recurrent parents

Often two or more good varieties with desirable quantitative characteristics used as RP in the
same backcross programme. Each variety is used as a recurrent parent for one or two
backcrosses. The objective of this approach is to combine in the new variety some good genes
from each of the recurrent parent (RP) with the genes from the non-recurrent parent.
Eg. Nobilisation of sugarcane
Noble canes (S. officinarum) x Indian canes (S. barberi): The resulting hybrids were
backcrossed to different varieties of noble canes to develop a large number of commercial
sugarcane varieties
Transfer of scab resistance in apples
Transfer of high vitamin C content from (Lycopersicon peruvianum) to (L. esculentum.

3. Backcross - Pedigree Method

In this method, the hybrid is backcrossed 1-2 times to the recurrent parent. Subsequently, the
backcross progenies and handles according to the pedigree method. This approach is useful
when one of the parents is superior to the other in several characteristics but the non
recurrent parent is not desirable agronomically. The superior parent is used as the recurrent
parent. The purpose of the one to two backcrosses is to make sure that the new variety would
get a majority of the superior genes from the recurrent parent. It also leaves enough
heterozygosity for transgressive segregants to appear. The varieties developed by this
method must be put to yield trials as those developed by the pedigree method. The same holds
true when two or more recurrent parents are used in the backcross programme. Eg. Wheat
programme in CIMMYT, Mexico

Application of the Backcross Method to Cross Pollinated Crops

The backcross method is equally applicable to cross-pollinated crops. The method is
essentially the same as in the case of self-pollinated crops. The only difference is that in cross
pollinated crops a large number of plants (100-300) from the recurrent parent must be used
in each backcross.
This is necessary so that the new variety has the same genetic constitution as the recurrent
parent. For example, wilt resistance was transferred to alfalfa variety California Common
from the variety Turkestan. Two hundred plants of California Common were used for each
backcross. The new variety Caliverde is exactly like California Common except for its wilt
resistance.

Comparison of back cross and pedigree method

Pedigree Back Cross

F1 and subsequent generation are allowed F1 and subsequent generation are back
for self pollinated crossed to recurrent parent

The new variety developed by this New variety is identical to recurrent parent
method is different from the parents in except for the character
agronomic and other characteristics

New variety has to be tested extensively Extensive testing is not required

before it is released

Aims at improving yielding ability and It aims at improving specific defects of a

other character well adopted variety

It is useful in improving both qualitative Useful for the transfer of both quantitative
and quantitative characters and qualitative characters provided they
have high heritability

Not suitable inter specific gene transfer Useful for the gene transfer from related
species

Hybridization is limited to F1 Hybridization is required for every back

cross

The F2 and the subsequent generations The backcross generation are small and
are much larger than those in the backcross usually consist of 20-100 plants in each
method generation

Breeding procedure same for dominant and The procedures for the transfer of dominant
recessive gene and recessive genes are different.
19. Multilines- types- procedure- merits and demerits

Generally, pureline varieties are highly adapted to limited area, but poorly adapted to
wider regions. Purelines have only one or few major genes for disease resistance, which offer
resistance to only some races of pathogen. New races are continuously produced which
overcome the resistance present in pureline varieties. Eg. Kalyansona wheat (T.
aestivum) originally resistant to leaf rust (brown rust), later on susceptible to new races.

To overcome these limitations, particularly, break down of resistance to disease, it was

suggested to develop multiline varieties. Multiline varieties are mixtures of several pure
lines of similar height, flowering time, maturity time, seed color and various other agronomic
characteristics but having different genes for disease resistance. At the same time, the
purelines constituting multiline variety must be compatible, i.e. they do not reduce the
yielding ability of each other when grown in mixture.

Multiline concept was first suggested in Oats by Jensen (1952). Later on Multiline was
applied in wheat by Norman Borlaug (1959).

Development of Multiline Variety

Norman Borlaug suggested that several purelines with different resistant genes
should be developed through back cross programme using one recurrent parent.
Kalyan Sona Wheat variety: It is the suitable example to explain the concept. This
variety was originally resistant to brown rust. Later on became susceptible to new races of
pathogen. Several pure lines with different resistance genes are produced through backcross
breeding using one recipient or recurrent parent. The donor parents are the one with
different genes for the disease resistance, every donor parent is used in separate back cross
program. Because of this each line receives different gene for disease resistance according to
the type of pathogen. Five to ten of such lines with different alleles for disease resistance are
mixed to develop multiline variety. The lines to be mixed are determined by the races of the
pathogen relevant to the area considered. If a line or lines become susceptible, they would be
replaced by resistant lines. New lines would be developed when new sources of resistance
become available. The breeder should keep several resistant lines in store for future use in
the replacement of susceptible lines of multiline varieties.
MLKS11 multiline variety developed by mixing 8 closely related lines
KML7404 - 9 closely related lines
Production steps for Multiline Variety
• Selection of recurrent parent
• Selection of donor parent
• Transfer of resistance
• Mixing of purelines

Merits of Multiline Variety

 1. All the lines are almost identical to the recurrent parent in agronomic characters,
quality etc., Therefore the disadvantages of pureline mixtures are not present in
multiline varieties.
2. At the time of disease outbreak, only one or few lines of the mixture get attacked,
others remain resistant. So the loss to the farmer is comparatively less.
3. The susceptible line would constitute only a small proportion of plants in the field.
Consequently the disease would spread more slowly than when the entire population
was susceptible. This would reduce the damage to the susceptible line also.
4. Multiline varieties are more adaptive to environmental changes than individual pure
line.
Disadvantages of Multiline Variety
1. Races of pathogen change as time goes on, so farmer has to change seeds every few
years which contains seeds of lines resistant to new pathogen races.
2. No improvement in yield or other characters
3. Production and maintenance of multiline is time taking job.
4. All the lines constituting multiline variety may get attacked by the new race of
pathogen.
5. Seed certification poses difficulties.

Population improvement approach in self-pollinated crops

Self-fertilization of F1 hybrids leads to a very rapid increase in homozygosity. After
several generations of self-pollination, about 94 per cent of the genes would become
homozygous. Even in F2, half of the genes are in homozygous state. As a consequence,
selection in such a segregating population only picks out the genes combinations present
in the population primarily as a result of recombination in F 2. This reduces the chance of
recombination between linked, especially tightly linked genes and of recovery of rare
transgressive segregants.
There is no opportunity for changing the genotype of the plant produced by
recombination in F1, F2 and to some extent, in F3.Thus the two obvious limitations of
breeding methods based on self-pollination of the hybrid (e.g., pedigree and bulk
methods) are: first, the recombination is limited to two or, at the best, three generations,
and second, there is no possibility for further changing the genotype of the segregants.
A population breeding approach has been suggested to overcome these problems.
In population breeding, outstanding F2 plants are mated among themselves in pairs or in
some other fashion. The intermating of selected F 2 plants restores heterozygosity in the
progeny, which provides for a greater opportunity for recombination. This also brings
together the desirable genes from different F 2 plants and would help in the accumulation
of favourable genes in the intermated population. Thus the chances of the recovery of
transgressive segregants would increase considerably. This process may be repeated one
or more times.
 This idea of population approach was first suggested by Palmer in 1953. The
population approach is akin to recurrent selection commonly used in cross-pollinated
crops.
The chief limitation of recurrent selection in self-pollinated crops is the difficulty in
making the large number of required crosses by hand (emasculation and pollination). This
difficulty may be overcome by using genetic or cytoplasmic male sterility. Thus the use of
male sterility effectively ensures intermating among the plants in the population and
eliminates the needs hand emasculation and pollination.
In 1970, Jensen proposed a comprehensive breeding scheme which provides for
the three basic functions of a versatile breeding programme.
Firstly, it allows the development of F2, F3 etc. (selfing series) at every stage of the
breeding programme, which permits the isolation of purelines for use as commercial
varieties. Secondly, it requires intermating among the selected plants/ lines in each stage;
the progenies from these intermatings form the basis for the next stage of the selfing
series in the breeding programme.
Thus the breeding programme progresses in two different directions:
(1) Vertically, through the selfing series leading to the isolation of commercial
varieties, and
(2) horizontally, through intermating among the selected plant / lines; this generates
the recurrent selection series.
Thirdly, new germplasm may be introduced at any stage of the programme by
intermating it with some of the selected plants of that stage. This permits the retention
and or the creation of large amounts of variability for effective selection through several
cycles, and the introduction of new genes in the breeding material, if so desired.
This breeding scheme is known as Diallel Selective Mating Scheme (DSM). This
scheme has not been widely used primarily due to the difficulties in making the large
number of crosses required in this scheme. Jensen has suggested the use of male sterility
to overcome this difficulty in the same way as in the recurrent selection scheme.
DSM is much more complicated than the simple pedigree method which still is the
favourite breeding method for self pollinated crops.

Merits of population Approach

1. The population approach provides for greater opportunities for recombination. This is
made possible by restoring heterozygosity through intermating of selected plants.
2. This approach helps in the accumulation of desirable genes in the population. This is
also brought about by the intermating of selected plants from segregating generation.
Demerits of Population Approach
1. The success of this approach depends upon the identification of desirable plants in
F2 and the subsequent segregating generations. This is very difficult for complex
characters like yield which show low heritability. This may be avoided to some extent by
using later generation (F3 or F4) progenies; replicated yield data may also be used.
2. Another draw back of this approach is the intermating of selected plants. This may
become a major limitation in some crops because crossing in many self-pollinated species
is difficult and time consuming.
3. The time taken to develop a new variety through population approach would be
always greater than that by the pedigree method.
4. There is no convincing evidence for the benefits from the population approach. It
has been argued that increased recombination may be detrimental, as it would break the
desirable linkage.
20. Genetic structure of a population in cross pollinated crop –
Hardy Weinberg law – gene frequencies in random mating population – principles in
population improvement.

Genetic structures of a population in cross pollinated crop

Cross pollinated crops are highly heterozygous due to the free inter mating among them
so these are random mating populations. Because each individual in a population has equal
opportunity of mating with any other individual. It is also known as Mendalian/panmictic
population. A Mendalian population may be thought of having a gene pool consisting of all
gametes produced by the population. So gene pool may be defined as the sum total of all
genes present in the population. A population consists of all such individuals that share the
same gene pool has an opportunity to inter mate with each other and contribute to the next
generation of the population.
Each generation of a Mendalian population may be considered to arise from a random
sample of gametes from the gene pool of previous generation. Hence, it is not easy to follow
the inheritance of a gene in a Mendalian population. It cannot be estimated by using the
techniques of classical genetics. So, to understand the genetic makeup of such population a
population genetics has been developed. Population genetics is the study of inheritance of
qualitative characters with frequency of genes and genotypes in natural populations. Hardy-
Weinberg law is the fundamental law of population genetics.

Hardy-Weinberg law
This law is independently developed by Hardy (1908) in England and Weinberg
(1909) in Germany. The law states that “the gene and genotype frequencies in a random
mating population remain constant from generation after generation if there is no selection,
mutation,migration or random drift”.
The frequencies of the three genotypes for a locus with two alleles A and a would be
P (AA), 2pq(Aa)and q2(aa)
2

Where, p= Frequency of ‘A’ allele in the population.

q= Frequency ‘a’ allele in the population
Such a population would be at equilibrium since the genotypic frequencies would be
stable, that is, would not change from one generation to the next. This equilibrium is known
as Hardy Weinberg equilibrium.
The sum of p+q is equal=1
p+q=1
p=1–q
q=1–p
Genotype frequencies occur in the population according to:
p2 + 2pq + q2 = 1

p2 - proportion of population that is homozygous for the first allele

(e.g., AA)
2pq- proportion of population that is heterozygous (e.g., Aa)
 q2 - proportion of population that is homozygous for the second
allele (e.g., aa)

Hardy - Weinberg assumes that:

Individuals never enter or leave a population
No mutations occur
The population is large
Individuals mate randomly
Selection does not occur

Factors affecting Hardy-Weinberg Equilibrium

1. Migration
Migration is the movement of individual into a population from a different
population. Migration may introduce new alleles into the population or may change the
frequency of existing allele. The amount of change in gene frequency ‘q’ will primarily
depend upon two factors.
a. Ratio of migrant individuals to those of the original population.
b. The Magnitude of difference between the values of q in the population and in
the migrants.
So, in plant breeding migration is by inter varietal crosses, poly crosses etc.,

2. Mutation
Mutation is a sudden heritable change in an organism and is generally due to a
structural change in a gene. It may produce a new allele not present in the population or may
change the frequency of existence allele.

3. Random drift or Genetic drift

It is a random change in gene frequency due to sampling error. Random drift is more
in small population than larger population. Ultimately the frequency of one of the allele
becomes zero and that of the other allele becomes one. The allele with the frequency one is
fixed in the population because there would be no change in the frequency. So, all genes
become homozygous. The genetic drift can be reduced by handling large population.

4. Inbreeding
• Mating between individuals sharing a common parent in their ancestry.
Inbreeding reduces the proportion of the heterozygosity and increase the frequency
of homozygosity and the rate of decrease in heterozygosity is equal to ½ N (N- Number
of plants in the population) per generation.
• In small population, even with strict random mating the frequency of homozygotes
increases while that of heterozygotes decreases due to inbreeding.
 6. Selection
Differential reproduction rates of various genotypes are known as selection. In
crop improvement, selection is important because it allows the selected genotypes to
reproduce, while undesirable genotypes are eliminated. Thus if in a random mating
population if we practice selection for the allele AA alone then its frequency in the
selected population will be one and the frequency of aa will be zero.
But in practice it is not possible to identify AA alone. So, we will not eliminate one allele
(aa) but instead the gene frequency will be changed. Thus selection in a random mating
population is highly effective in increasing or decreasing the frequency or alleles, but it is
unable to either fix or eliminate them.

SYSTEMS OF MATING

To change the genetic composition of a population, different systems of mating are available

1. Random mating
2. Genetic assortative mating
3. Genetic disassortative mating
4. Phenotypic assortative mating
5. Phenotypic disassortative mating

1. Random mating

Each female gamete is equally likely to unite with any male gamete. Here the rate of
reproduction of each individual is equal i.e. there is no selection. This random mating is
useful in plant breeding for the production and maintenance of synthetic and composite
varieties, production of polycross varieties.

2. Genetic assortative mating

It is the mating between individuals that are closely related by ancestry. It is other wise
known as inbreeding. It is useful for the development of inbreds. The genetic assortative
mating leads to
i) Increase in homozygosity ii) Fixation of characters and iii) Lethals will be eliminated

3. Genetic disassortative mating

It is mating between individuals that are not closely related by ancestry.
Eg. Intervarietal and interspecific crosses.

4. Phenotypic assortative mating

It is the mating between individuals, which are phenotypically more similar. This type
of mating leads to increase in homozygosity and division of population into two
extremes. i.e. there is highest and lowest phenotypes remain in the population and
there is no intermediate types. It is useful in isolation of extreme phenotype.
Eg. Recurrent selection
5. Phenotypic disassortative meting

❖ Mating between phenotypically dissimilar individuals belonging to the same

population.
❖ This system leads to maintenance of or increase in heterozygosity.
❖ It is useful in maintaining variability in small population as it reduces inbreeding.

Selection in cross pollinated crops

i) Change the gene and genotypic frequency

ii) Production of new genotypes due to changed gene frequencies.
iii) Cause a shift in the mean of population towards the direction of selection.
iv) Change in the variance of population to some extent.

Types of response to selection

1. Rapid gain followed by slow Progress

In some cases selection produces rapid gain for some generations. This is followed by a
period of slow gain. This type of response is seen in characters like plant height, days to
flowering. These characters will be governed by a few genes with major effect and several
genes with lesser effect. The major genes will give rapid gain and several genes having
lesser effect gives slow effect.

2. Slow progress for a long period

This is because, that these traits are governed by several genes, each having a small and
additive effect. So, progress under selection for such traits would be slow. Eg. Oil
content and protein content in maize.

3. Slow response for a shorter period only

Here the selection for some characters show slow gain for several generations and
afterwards there will be no response at all. This is due to many poly genes. Eg. Oil
content in maize

4. Lack of response to selection

This may be due to low heritability. Eg. Selection for yield maize

5. Rapid gain - plateau - Rapid gain

This is due to linked genes both positive and negative. eg. Selection for increased
abdominal bristle in Drosophila
21. Breeding methods of cross pollinated crops without involving artificial
hybridization: Mass selection in cross pollinated crops – modified mass selection –
Grid selection – progeny selection

Breeding Methods for Cross Pollinated Crops Populations of cross pollinated crops are
highly heterozygous. When inbreeding is practiced they show severe inbreeding depression.
So to avoid inbreeding depression and its undesirable effects, the breeding methods in the
crop is designed in such a way that there will be a minimum inbreeding. The breeding
methods commonly used in cross pollinated crops may be broadly grouped into two
categories.

A. Population improvement
1. Selection
a) Mass selection
b) Modified mass selection
i. Detasseling
ii. Panmixis
iii. Stratified or grid or unit selection
iv. Contiguous control
2. Progeny testing and selection
a) Half sib family selection
i) Ear to row
ii) Modified ear to row
b) Full sib family selection
c) Inbred or selfed family selection.
i) Sl self family selection
ii) S2 self family selection.
3. Recurrent selection
a) Simple recurrent selection
b) Reciprocal recurrent selection for GCA
c) Reciprocal recurrent selection SCA
d) Reciprocal recurrent selection
B. Hybrids, Synthetics and Composites

1. Selection
a. Mass selection
This is similar to the one, which is practiced, in self-pollinated crops. A number of plants
are selected based on their phenotype and open pollinated seed from them are bulked
together to raise the next generation. The selection cycle is repeated one or more times to
increase the frequency of favourable alleles.
Merits
i) Simple and less time consuming
ii) Highly effective for character that are easily heritable. Eg. plant height, duration.
 iii) It will have high adaptability because the base population is locally adapted one.
iv) Improved strain is similar to original population, hence extensive yield trials may
not required

Demerits
1. Selection is based on phenotype only which is influenced by environment
2. The selected plants are pollinated both by superior and inferior pollens present in
the population.
3. High intensity of selection may lead to reduction in population there by leading to
inbreeding.

b. Modified mass selection is proposed to overcome these defects and they are
a) Detasseling
This is practiced in maize. The inferior plants will be detasseled there by
inferior pollen from base population is eliminated.

b) Panmixis
From the selected plants pollen will be collected and mixed together. This will
be used to pollinate the selected plants. This ensures full control on pollen source.

c) Stratified or unit mass selection

Here the field from which plants are to be selected will be divided into smaller units
or plots having 40 to 50 plants / plot. From each plot, equal number of plants will
be selected. The seeds from selected plants will be harvested and bulked to raise
the next generation. By dividing the field into smaller plots, the environmental
variation is minimized. This method is followed to improve maize crop. It is also
known as Grid method of mass selection.
d) Contiguous control
In this method, checks are planted after every four plants. Yield of plant is
expressed as per cent of the yield of check plant

2) Progeny Testing and Selection

a) Half sib family selection

Half sibs are those, which have one parent in common. Here only superior progenies
are planted and allowed to open pollinate.
i. Ear to row method is the simplest form of progeny selection and is extensively used
in maize. This method was developed by Hopkins.
Procedure
a) A number of plants are selected on the basis of their phenotype.
They are allowed to open pollinate and seeds are harvested on single plant basis.
b) A single row of say 50 plants i.e. progeny row is raised from seeds harvested on
single plant basis. The progeny rows are evaluated for desirable characters and
superior progenies are identified.
c) Several phenotypically superior plants are selected from progeny rows.
Though this scheme in simple, there is no control over pollination of selected
plants. Inferior pollen may pollinate the plants in the progeny row.

ii. Modified ear to row method is followed to overcome these defects

 a) At the time of harvest of selected plants from base population on single plant basis,
part of the seed is reserved.
b) While raising progeny rows, after reserving part of the seeds, the rest are sown in
smaller progeny rows.
c) Study the performance of progenies in rows and identify the best ones.
d) After identifying the best progenies, the reserve seeds of the best progenies may be
raised in progeny rows.
e) The progenies will be allowed for open pollination and best ones are selected.

b) Full sib family selection: Full sibs are those which are produced by mating between
selected plants in pairs.
Here the progenies will have a common ancestry.
The crossed progenies are tested. A x B , B x A

c) Inbred or selfed family selection

Families produced by selfing
i. S1 family selection
Families produced by one generation of selfing. These are used for evaluation and
superior families are intermated (Simple recurrent selection).
ii. S2 family selection
Families obtained by two generations of selfing and superior families are intermated.

Merits of progeny testing and selection

1. Selection based on progeny test and not on phenotype of individual plants.
2. Inbreeding can be avoided if care is taken in raising a larger population for selection.
3. Selection scheme is simple.
Demerits
1. No control over pollen source. Selection is based only on maternal parent only.
2. Compared to mass selection, the cycle requires 2-3 years which is time consuming.

Applications
1. To maintain purity of the varieties
2. To develop new improved open pollinated variety (OPV)
22. Breeding methods of cross pollinated crops involving artificial hybridization:
Recurrent selection principles – types – merits and demerits

Recurrent selection: This is one of the breeding methods followed for the improvement of
cross pollinated crop.
The initial idea about recurrent selection was first suggested by Hayes and Garber in
1919 and independently by East and Jones in 1920. The Procedure of recurrent selection was
described by Jenkins in 1940. The term recurrent selection was coined by Hull in 1945.
Recurrent selection means “Selection generation after generation with interbreeding of
selected plants to get genetic recombination”
Here single plants are selected based on their phenotype or by progeny testing. The
selected single plants are selfed. In the next generation they are intermated (cross in all
possible combinations) to produce population for next cycle of selection.
The recurrent selection schemes are modified forms of progeny selection
programmes. The main difference between progeny selection and recurrent selection
i) The manner in which progenies are obtained for evaluation.
ii) Instead of open pollination, making all possible inter crosses among the selected
lines.

The recurrent selection schemes are of 4 different types.

1. Simple recurrent selection

In this method a number of desirable plants are selected and self pollinated. Separate
progeny rows are grown from the selected plants in next generation. The progenies are
intercrossed in all possible combination by hand. Equal amount of seed from each cross is
mixed to raise next generation. This completes original selection cycle. From this, several
desirable plants are selected and self pollinated. Progeny rows are grown and inter crosses
made. Equal amount of seeds are composited to raise next generation.
 i) Recurrent selection is effective in increasing the frequency of desirable genes
in the population
ii) Most suited for characters having high heritability
iii) Inbreeding is kept at minimum.

2) Recurrent selection for general combining ability (RSGCA)

General combining ability is the average performance of a strain in a series of cross
combinations. The GCA is estimated from the performance of F 1S from the crosses.
This method was suggested by Jenkins (1935). In this method, the progenies selected
for progeny testing are obtained by crossing the selected plants to a tester parent with
broad genetic base. A tester parent is a common parent mated to a number of lines. Such a
set of crosses is used to estimate the combining ability of the selected lines. A tester with
broad genetic base means an open pollinated variety, a synthetic variety or segregating
generation of a multiple cross.
 Recurrent selection for GCA can be used for two basically different purposes.
1. It may be used to improve the yielding ability and the agronomic characteristics of a
population. In this case the end product will be a synthetic variety.
2. It may be used to concentrate genes for superior GCA. Here the end product will be
superior inbreds. Such inbreds can be developed after a few cycles of RSGCA.

3) Recurrent Selection for Specific Combining Ability

This method was proposed by Hull (1945). This is similar to RSGCA except, that in the
case of tester. Here the tester will be an INBRED instead of open pollinated variety. The other
operations are similar to RSGCA. The objective of RSSCA is to isolate from population such
lines that will combine well with an inbred. These lines are expected to give best hybrids in
heterosis breeding

4. Reciprocal recurrent selection

It was proposed by Comstock, Robinson and Harvey (1949). The objective is to
improve two different populations in their ability to combine well with each other. In this
method, selection is made for both GCA and SCA. Basically two populations A and B are used.
Each serves as a tester for the other.
Original selection cycle
First year
1. Several plants selected in population A and B.
2. Selected plants are self pollinated
3. Selected plant from A is test crossed with plants in B and vice versa.
4. Harvest crossed plant on single plant basis each.
Second year
1. Separate yield trials conducted from test cross progenies of A and B
2. Superior progenies identified
Third year
1. Selfed seed from plants producing superior test cross progenies planted.
2. All possible inter crosses made
3. Seeds harvested and composited

First recurrent selection cycle

The same procedure is repeated as original selection cycle

Use of RRS
1. Two populations are developed by this method
2. They may be intermated to produce a superior population with broad genetic base.
This is similar to a varietal cross but in this case the populations have been subjected
to selection for combining ability (GCA and SCA)
3. Inbreds may be developed from populations A and B. These inbreds may be crossed
to produce a single cross or double cross hybrids.
Merits of recurrent selection
1) Efficient breeding method for increasing the frequency of superior genes in a
population for economic characters
2) This method also helps in maintaining high genetic variability in a population due to
repeated intermating of heterozygous plant
3) Selection is made on the basis of test cross performance and only selected plants are
allowed for intermating
Demerits of recurrent selection
1. This method is not used directly for the development of new varieties. The new
varieties are developed using the end product in hybridization.
2. This method involves more selection and crossing
23. Heterosis breeding – theories - genetic basis – hybrid vigour – estimation of
heterosis – inbreeding depression.

HETEROSIS
It is defined as the superiority of F1 hybrid over both the parents in terms of yield and
some other characters.
The term heterosis was first used by Shull (Father of Heterosis) in 1914.

HISTORY
Koelreuter (1673) : First artificial tobacco hybrid
Darwin (1876) : Hybrids from unrelated plants were highly vigorous
Beal (1877 & 1882) : yield advantage of 40 % in intervarietal hybrids of maize over the
parents.
Maize- most extensively studied crop for heterosis and development of hybrid maize
based on the work of East and Shull.

Genetic basis of heterosis

There are two main theories of heterosis and inbreeding depression
1. Dominance hypothesis
It was first proposed by Davenport in 1908. It was later expanded by Bruce, Keeble and
Pellow. According to this hypothesis “the dominant allele has favourable effect, while the
recessive allele has unfavourable effect at each locus”. In heterozygous state, the
deleterious effect of recessive alleles is masked by their dominant alleles. Inbreeding
depression is produced by the harmful effects of recessive alleles, which become
homozygous due to inbreeding.

Objections
a) Failure of isolation of inbreds as vigorous as hybrids
According to dominance hypothesis, it is possible to isolate inbreds with all the dominant
genes E.g. AA. This inbred should be as vigorous as that of hybrid. However in practice
such inbreds were not isolated.
b) Symmetrical distribution in F 2
In F2, dominant and recessive characters segregate in 3:1. According to dominance
hypothesis, quantitative characters should not show symmetrical distribution. Dominant
and recessive phenotypes segregate in the proportion of (3/4+1/4)n, n-number
of genes segregating. However, F2 nearly always show symmetrical distribution.

Explanation for the two objections

In 1917, Jones suggested that since quantitative characters are governed by many genes,
they are likely to show linkage. In such a case, inbreds containing all dominant genes
cannot be isolated. So also the symmetrical distribution in F 2 is due to linkage. This
explanation is often known as Dominance of Linked Genes Hypothesis.
2. Over dominance hypothesis
This hypothesis was independently proposed by East and Shull in 1908. It is also known
as single gene heterosis or super dominance theory. Heterozygotes at some of the loci are
superior to both the homozygotes. Thus heterozygote Aa would be superior to AA and aa.
In 1936, East proposed that at each locus there are several alleles at a1, a2, a3 & a4 etc,
with different functions Heterozygotes with more divergent alleles would be more
heterotic E.g. a1a4 superior to a1a2 or a2 a4.
Evidences for over dominance
i. In maize the maturity genes in heterozygous conditions are superior i.e. Ma ma
ii. The heterozygote Mama is more vigorous than MaMa or mama
iii. The human beings sickle cell anemia is caused by ss which is lethal. But heterozygote
with Ss have advantage of resistance against malaria compared to SS individuals

Comparison of dominance and over dominance hypothesis

Feature Hypothesis of heterosis

Dominance Over dominance

Similarities

Inbreeding leads to Reduced vigour and fertility Reduced vigour and fertility

Out crossing leads to Heterosis Heterosis

Degree Genetic diversity in parents Genetic diversity in parents

of heterosis increases with

Differences

Inbreeding depression is the Homozygotes for Homozygosity itself

result of deleterious recessive alleles

Heterosis is the result of Masking of the harmful Heterozygosity itself

effects of recessive alleles by
their dominant alleles

The phenotype of Comparable to that of the Superior to both the

heterozygote is dominant alleles homozygotes

Inbreds as vigorous as the F1 Can be isolated Cannot be isolated

hybrids

Molecular basis of heterosis

1. Intermediate amount of single gene product
 In certain cases the heterozygote produces an intermediate amount of a gene
product, which may lead to increased vigour and growth rate.
This is seen in case of bread mould - Neurospora crassa
Homozygote Pab+Pab+ - Produces more P-amino benzoic acid
Homozygote Pab Pab - Produces Less P-amino benzoic acid
Heterozygote Pab+ Pab - Intermediate amount of P amino benzoic acid which leads
to faster growth of the fungus.
2. Separate gene products
AA - produce protein
aa - Produce protein which is slightly different
Aa - will have proteins encoded by both the alleles
This may have many advantages by having more adaptation
Human beings
SS Resistant to sickle cell anemia
ss - Susceptible
Ss - Resistant to Sickle cell anemia + malaria, because the defective haemoglobin
produced by s allele not suitable for multiplication of malarial pathogen.
3. Combined gene product
The hybrid may produce an enzyme molecule which is different compared to enzymes
produced by homozygotes. Such heterozygote enzymes are termed as Hybrid Substance
which may be the reason for hybrid vigour
4. Effect in two different tissues
Both homozygotes produce high levels of an enzyme in two different tissues and
heterozygotes produce intermediate level in both tissues.
E.g. Maize Adh gene for enzyme alcohol dehydrogenase
AdhF - scutellar tissue of seeds; AdhS - pollen grains
Heterozygotes - AdhF AdhS - produce intermediate level in scutellar tissue and
pollen.

Estimation of Heterosis

1. Average heterosis
 It is the heterosis where F1 is superior to mid parent value. In otherwords superior to
average of two parents.

F1 - MP
---------- x 100
MP
Where F1 = Mean of hybrid
MP = Mid parental value. (P1 + P2) where P1 = Parent 1; P2 = Parent 2

This type of heterosis is of no use in agriculture since the superiority is below the
better parent value
2. Heterobeltiosis
It is the superiority of F1 over the better parent.
F1 - BP
--------- x 100
BP
Where BP = Mean of Better Parent.
3. Economic heterosis (Standard heterosis)
Superiority of F1 compared to the high yielding commercial variety in a particular
crop.
F1 - CV
--------- x 100
CV
Where CV = Mean of Commercial Variety.
4. Negative heterosis
Performance of F1 inferior to better parent / mid parent value. - e.g. Duration.

Manifestation of heterosis
1. Increased yield: Heterosis is generally expressed as an increase in the yield of
hybrids. The yield may be measured in terms of grain, fruit, seed, leaf, tubers or the
whole plant.
2. Increased Reproductive Ability: More number of flowers/fruits/seeds. The hybrids
are generally more vigorous, i.e., healthier and faster growing and larger in size than
their parents.
3. Better Quality: In many cases, hybrids show improved quality. For example, many
hybrids in onion show better keeping quality, but not yield than open-pollinated
varieties.
 4. Earlier Flowering and Maturity: In many cases hybrids are earlier in flowering and
maturity than the parents. But earliness is highly desirable in many situations
particularly in vegetables.
5. Greater resistance to disease and pest: Some hybrids are known to exhibit a
greater resistance to insect or diseases than their parents.
6. Greater adoptability: Hybrids are generally more adapted to environmental
changes than inbreds.
7. Faster growth rate: In some cases, hybrids show a faster growth rate than their
parents. But the total plant size of the hybrids may be comparable to that of parents.
In such cases, a faster growth rate is not associated with a larger size.

Heterosis and Hybrid Vigour

Hybrid vigour is used as synonym of heterosis. In other words, hybrid vigour is the
manifested effect of heterosis. Generally hybrid vigour describes only superiority of
hybrids over the parents. The term hybrid vigour is used to distinguish the F 1 superiority
from negative heterosis.
Luxuriance
It is the increased vigour and size of interspecific hybrids. Difference lies in the
reproductive ability of the hybrid. Heterosis is associated with increasing fertility.
Luxuriance is expressed by interspecific hybrids which are generally sterile or poorly
fertile.

Inbreeding

It is mating between individuals related by descent or having common ancestry. The highest
degree of inbreeding is obtained by selfing.

Inbreeding depression

It is defined as reduction or loss in vigour and fertility as a result of inbreeding. Cross

pollinated species and asexually reproducing species are highly heterozygous. These species
show severe reduction in vigour and fertility when subjected to selfing or inbreeding .

History of inbreeding
Inbreeding depression has been recognized by man for a long time. Knowing the
consequences of inbreeding, many societies have prohibited marriages between closely
 related individuals. Darwin in 1876 published a book “cross and self fertilization in
vegetable kingdom” in which he concluded that progenies obtained from self fertilization
were weaker in maize. Detailed and precise information on maize inbreeding was
published by East in 1908 and Shull in 1909.

Effects of inbreeding
1. Appearance of lethal and sub lethal.
2. Reduction in vigour: Appearance of dwarf plants.
3. Reduction in reproductive ability - Less seed set, sterility.
4. Segregation of population in distinct lines.
5. Increase in homozygosity.
6. Reduction in yield.

Degrees of inbreeding depression

Various plant species exhibit different degrees of inbreeding depression. The

depression may be from very high to nil. Based on degree of depression, the plant species
can be grouped into 4 broad categories.
1. High inbreeding depression: E.g. Lucerne, Carrot. Inbreeding leads to severe
depression and exhibit lethal effects. After 3 or 4 generations of selfing it is hard to
maintain lines. These lines show very less yield i.e. <25% of open pollinated varieties
(OPV).

2. Moderate inbreeding depression: E.g. Maize, Jowar, Bajra. Though lethal effects
are there, lines can be separated and maintained. Lines can be maintained by self
pollination. Yield of inbred lines is 50% of OPVs. P
roduction and maintenance of inbred lines are relatively easier.
3. Low inbreeding depression: E.g. Cucurbits, Sunflower. Only a small degree
of inbreeding depression is observed. Small proportion of plants show lethal effects
Yield reduction is small or Nil, hence yield as that of OPVs.
4.No inbreeding depression: The self-pollinated crops do not
show inbreeding depression.
24.Heterosis breeding – procedure – development of inbreds- evaluation of inbred
lines – top cross method and single cross method- prediction of double cross
performance- hybrids – single cross-double cross- three way cross hybrids.
achievements – merits and demerits

Heterosis breeding

In production of hybrids, inbreds are preferred rather than open pollinated varieties (OPVs).

Advantages in using inbreds

❖ Inbreds can be maintained without a change in the genotype. Whereas OPV cannot be
maintained pure.
❖ The hybrids derived from inbreds will be uniform, whereas it may not be in case of
OPV.
❖ The inbreds are homogenous and their performance can be predicted where as OPV
are heterogeneous and their prediction in performance cannot be made

Operation in production of hybrids

1. Development of inbreds
2. Evaluation of inbreds
3. Production of hybrid seed

1. Development of inbreds

i. Development of inbreds through selfing

Inbred lines are developed from a genetically variable population through continued
inbreeding, selfing. Pedigree method is commonly followed for the development of
inbreds.
First year: A number of plants with desirable characters are selected from the source
population and self pollinated.
Second year: About 30-40 plants space planted from the selfed seed of each of the
selected plants. Best plants selected from the progeny rows are self pollinated.
Third to sixth year: Procedure of second year is repeated.
Seventh year: At this stage, individual plant progenies would be homozygous. The
inbred lines may be maintained by sib pollination.

ii. Development of inbreds from haploid plants

In maize, haploid plants occur at frequency of 10-3. Chromosomes doubling of haploids
to produce diploid progeny serve as instant inbreds.
2. Evaluation of inbreds: It involves four different steps

i. Phenotypic evaluation

Inbreds are evaluated for their phenotypic performance which is most suited for
simple inherited traits. The inbreds with poor performance are rejected.

ii. Top cross test

Top cross test provides a reliable estimate of Genral Combining Ability (GCA). The
selected inbreds are crossed to a tester parent with wide genetic base i.e. open
pollinated variety. The hybrid progenies are evaluated in replicated progeny rows and
based on results better inbreds are identified. About 50 % of the inbred lines are
eliminated.
iii. Single cross evaluation
The inbreds remaining after top cross teat are crossed in diallel manner to produce all
possible single crosses. The performance of single crosses is evaluated in replicated
trial. Outstanding hybrids tested over years in different locations, are released for
cultivation.
 iv. Prediction of double cross performance
The predicted performance of any double cross is the average performance of the four
non parental single crosses.
Inbreds: A, B, C, D.
Six possible single crosses = A x B, A x C, A x D, B x C, B x D, C x D.
From these 3 double crosses are produced
(A x B) x (C x D)
(A x C) x (B x D)
(A x D) x (B x C)
The performance of these double cross can be predicted from performance of the
four single crosses not involved in producing hybrid.
(A x B) x (C x D) = (A x B) x (C x D) = (A x C) + (A x D) +(B x C) +(B x D)/ 4

3. Production of hybrid seed

Hybrids are the first generation (F 1) progenies from crossing between two purelines,
inbreds, open pollinated varieties, clones or other populations that are genetically
dissimilar. Hybrids were first commercially exploited in maize.

Requirements for hybrid seed production

1. Easy emasculation

2. Effective pollen dispersal

Methods of hybrid seed production

I. Hand emasculation and dusting - Cotton, Tomato,Chillies, Bhendi

2. Use of male sterility
a) Cytoplasmic male sterility - ornamentals
b) Genic male sterility - Redgram, Castor.
c) Cytoplasmic - genic male sterility - Jowar, Bajra, Rice
3. Use of self Incompatibility
By planning cross compatible lines, hybrids are produced.
E.g. Brassicas
TYPES OF HYBRIDS

1. Single cross hybrids

Crossing two inbreeds or pure lines - (A x B)
2. Three way cross hybrid
A cross between a single cross hybrid and an inbred – (A x B) x C
3. Double cross hybrid
Cross between two Fl s. (A x B) x (C x D)
4. Top Cross hybrid
Cross between inbred and an open pollinated variety
5. Double Top Cross hybrid
Cross between Double Cross hybrid crossed with open pollinated variety

Success of hybrids depends

a) Easy hand emasculation
b) Abundant seed set to compensate cost of hand emasculation
c) Stable male sterile lines
d) Effective restorers
e) Effective pollen dispersal

MERITS OF HYBRIDS

▪ Hybrid varieties exploit both GCA and SCA, hence high yield
▪ More uniform compared to OPVs, synthetic or composite varieties
▪ Produced both in cross and self pollinated crops
▪ Hybrid varieties are maintained in the form of their parental inbreds, which ensures
that genetic constitution does not change
DEMERITS

▪ Farmers have to use new hybrid seed every year

▪ Hybrid seed production requires technical skill
▪ Exploiting full potential of hybrid requires adequate irrigation, fertilizer, weed
control etc.,
▪ In many cross pollinated species, the requirement of isolation are rigid and difficult
to fulfill except on large farms
Achievements
Maize
Ganga 1, Deccan, Ranjit- DC hybrids (1961)
Quality protein hybrids (opaque 2 )- Histarch Hybrid Makka, HM 4, Vivek QPM 9
Pearl millet
HB 1 –First hybrid developed during 1965.
PHB 10, PHB 11- resistant to downy mildew
Sorghum
CSH 1 - First hybrid in 1965
Latest : CSH 10, CSH 11
Red gram
ICPH 8 –First GMS based hybrid
GTH 1- First CGMS based hybrid; Popular hybrid ICPH 2671 (Pushkal)
25. Synthetics and composites - steps in development of synthetics and composites –
achievements – merits and demerits

Synthetic variety
A synthetic variety is produced by crossing in all combinations a number of
inbreds (4-6) that combine well with each other. The inbreds are tested for their
General Combining Ability (GCA). Once synthesised, a synthetic is maintained by open
pollination. The lines that make up a synthetic may be usually inbred line but open
pollinated variety (OPV), or other population tested for general combining ability are also
be used.
Synthetic variety was first reported by Hayes and Garber (1922) in maize. Synthetic
varieties are common in grasses, clover, maize and sugar beets.

Steps involved in the development of synthetics

1. Evaluation of lines for GCA
2. Production of synthetic variety
3. Multiplication of synthetic variety
1. Evaluation of lines for GCA
GCA of parental lines is usually estimated by top cross or poly cross test. Polycross refers
to the progeny of a line is produced by pollinating with random sample of pollen from
number of selected lines. The lines are evaluated for GCA because the synthetic varieties
that potion of heterosis which is produced by GCA.
2. Production of synthetic variety
Method I
Equal amounts of seeds from parental lines (Syno) is mixed and planted in isolation. Open
pollination is allowed to produce crosses in all possible combinations. The seeds are
harvested in bulk. The population raised from this seeds is Syn1 generation.
Method II
All possible crosses among the selected lines are made in isolation. Equal amount of seed
from all the crosses is composited to produce Syn1 generation.
3. Multiplication of synthetic variety
Once synthesised, the inbreds are generally multiplied in isolation for one or more
generation before its distribution. The open pollinated progeny from Syn1 generation is
termed as Syn2. The synthetic variety is maintained by open pollination.
Merits of Synthetic variety
1. Less costly compared to hybrids.
2. Farmer can reuse the seeds of synthetic variety for more seasons which is not
possible in hybrids.
3. Because of wider genetic base the synthetics are more stable over years and
environments.
4. Seed production is more skilled operation in hybrids where as it is not so in
synthetics.
5. Synthetics are good reservoirs of genetic variability
6. Means of exploiting heterosis in species where pollination control is difficult

Demerits
1. Performance is little bit lower compared to hybrids because synthetics exploit only
GCA while hybrids exploit both GCA and SCA.
2. The performance may not be good when lines having low GCA are used.
3. It is not possible in self pollinated crops
Composites

It is produced by mixing seeds of phenotypically outstanding lines and encouraging

open pollination to produce crosses in all possible combinations among mixed lines. The
lines used to produce a composite are not usually tested for combining ability. So the yield
of composite varieties cannot be predicted easily. Like synthetics, composites are
commercial varieties and are maintained by open pollination.
Composites were released in maize - Amber, Jawahar, Kissan.
Opaque 2 composites in maize: Shakti, Rattan, Protina
Composite in Cumbu : WCC 75

Comparison of Synthetic and composite varieties

Synthetic Composite

The parental lines are tested for their GCA The parental lines are not tested for their
GCA.

No of parental lines are limited to 4 - 6 No such limit.

inbreds

Synthetic varieties produced with inbreds It is not possible.

can be reconstituted

Yield performance can be predicted Cannot be predicted.

26. Genetic characters of asexual reproduction – clonal selection – hybridization and
clonal selection – merits and demerits – achievements

Asexual reproduction preserves the genotype of the individual indefinitely. Any genotype
is preserved and maintained.

Characteristics of asexual crops

• Majority of these are perennials like sugarcane and fruit trees. The annual crops are
mostly tuber crops e.g. potato and sweet potato.
• These are highly heterozygous indicating the evolutionary advantage of
heterozygote in these crops which leads to high degree of inbreeding depression on
selfing.
• Majority are polyploidy eg.sugarcane,potato,sweet potato
• Many crops are interspecific hybrids eg.banana, sugarcane
• These are highly cross pollinated.
• Most of these crops show reduced flowering and certain varieties do not even flower
at all. The seed set through normal sexual reproduction is usually low.
• Consists of large no of clones which is a progeny derived from a single plant through
asexual reproduction.
• Each variety of these crops is a clone.

CLONES
Clone is a group of plants produced from single plant through asexual reproduction.
Hence, asexually propagated crops are also known as clonal crops.

Characteristics of a clone

1. All the individuals of single clone are identical in genotype

2. Lack of genetic variation: Phenotypic variation is due to environment

3. .Immortality: can be maintained indefinitely.

4. Highly heterozygous and severe inbreeding depression

The genetical analysis is difficult due difficulty to induce flowering, high sterility and
perennial life cycle.
Origin of genetic variation
1. Mutation
It is known as somatic mutation or bud mutation. It occurs in low frequency (10-5
to 10-7). Dominant bud mutations are more frequent than recessive mutations. Bud
selection involves selection of mutant buds to establish new desirable clones
2. Mechanical mixture
3. Sexual reproduction
It leads to segregation and recombination. Old clone tend to become variable in
annuals and biennials
Clonal degeneration
The loss in vigour and productivity of clone with time is known as clonal degeneration.
Degeneration may result from
1. Mutation
2. Viral diseases
3. Bacterial diseases
Clonal selection
It is the procedure of selection of superior clones from the population of mixture of clones.
Procedure of clonal selection
First year
Few to several hundred superior plants are selected from the mixed variable population.
Second year
• Clones from the selected plants grown separately.
• inferior clones are rejected
Third year
• Preliminary yield trial along with standard check
• Selection for quality, disease resistance etc.,
• Few superior clones are selected.
Fourth to sixth year
• Multi location trial along with standard check
• Test for yielding ability.
• Quality and Resistance are evaluated.
• Superior clone identified for release.
Seventh year

• The best clone is released as a new variety

• Seed multiplication for distribution
Procedure for Hybridization and selection
Year 1 : Hybridization between clone A × clone B

Year 2 : Grow 10,000 seedling obtained through sexual reproduction and select 1000 -most
vigorous and propagate

Year 3 – Grow 1000 clonal rows and select 100 superior clones and propagate

Year 4 – PYT along with checks and select few superior clones

Year 5 to 7 – Grow selected clones in replicated field trials at different locations; Superior
clones identified for release

Year 8 –Outstanding clone released as variety

Merits
1. It is the only method of selection applicable to clonal crops; Avoids inbreeding depression
and preserves gene combination.
2. Useful in maintaining the purity of clones
3. Varieties evolved retain all the characters of parental clones
4. It can be combined with hybridisation to generate variability.
5. Varieties highly uniform as purelines
Demerits
1. It utilises natural variability and cannot create variability.
2. Sexual reproduction is required to create variability through hybridisation.
3. Varieties developed by clonal selection are highly to new disease.

Achievements
1. Clonal selection
Kufri red potato from Darjeeling round
Kufri safed potato from Phulwa
Bud selection
Bombay green banana from Dwarf Cavendish
Pidi Monthan from Monthan
High gate from Gross Michael
2. Hybridization
Potato- Kufri Alankar, Kufri Ashoka, Kufri Kundan etc.,
Kufri Jyoti – Blight resistant
Kufri Sheetman – Frost resistant
Mango - Mallika, PKM 1 & PKM 2
Sugarcane – CoS 109, CoS 510, Co 541
3. Inter specific Hybridization
Sugarcane – S. spontaneum / S. officinarum – Co 1148, Co 1158, CoC 86052
Potato- Kufri Kuber – (Solanum curtilobum / S. tuberosum ) S. andigena
27. Polyploidy breeding – classification – induction of polyploidy - achievements –
limitations.
Introduction
The change in number of chromosomes is an important source of genetic variation that
resulted in the evolution of a number of crop species. Each chromosome exists as a member
of the pair and the number of such pairs of chromosome is specific to a particular species. Eg.
Human beings have 23 pairs, maize has 10 pairs and Arabidopsis has only 5 pairs of
chromosomes. A basic set of chromosomes constitute a genome containing one number of
each pair of chromosomes.
Individuals having chromosome number more than two sets referred as polyploids. The
individual having two sets of chromosomes is called as diploid while individual with only
single set is called monoploid. The somatic chromosome number of an individual is always
designated as 2n and the chromosome number of the gametes as n whereas x is basic
chromosome number.
In true diploids e.g. maize x=n=10: 2n=20. But in Polyploids, the monoploid and haploid
chromosome number is different. For instance in common bread wheat which is polyploid,
the somatic chromosome number is 42 and haploid number 21 and the monoploid basic
number is 7 Thus wheat x=7, n=21, 2n=42 or n=3x and 2n=6x=42. The haploids of diploids
are called monohaploids, and the haploids of tetraploids are called as dihaploids and those
of higher polyploids are known as poly haploids. The doubling of chromosome number of
haploid results into a plant called doubled haploid.
Individuals carrying chromosome number other than the diploid (2x) number are known
as heteroploids, and the situation is known as heteroploidy. Thus numerical changes in
chromosomes (heteroploidy) can be mainly of two types.
1. Euploidy
2. Aneuploidy
Aneuploidy

Variation in chromosome number which do not involve whole set of chromosomes, but
only a part of a set is referred as aneuploidy. This condition can be expressed either as an
addition of one or more entire chromosome or as a loss of such chromosomes as compared
to the somatic chromosome number of that species. Therefore, the aneuploid is an
organism or a cell having one or few chromosomes more, or less than the normal
somatic number (2n) of the individual. It seems that the aneuploid changes in
chromosome number do not involve the whole genome. They relate only one or few
chromosomes of the genome.

Types of Aneuploids

Types Genomic constitution

Monosomic 2n – 1
Double monosomic 2n –1-1 Hypoploidy
Nullisomic 2n –2
Trisomic 2n +1
Double trisomic 2n +1+1 Hyperploidy
Tetrasomic 2n +2
Pentasomic 2n +3

Origin of aneuploidy
1. Non-disjunction: Generally during gametogenesis the homologous chromosomes of
each pair separate out (disjunction) and are equally distributed in the daughter cells.
The failure of separation of homologous chromosome is called non-disjunction. It
occurs during gametogensis. Here, one type contains 22 chromosomes, while other
will be 24.
2. Triploid plants : Irregularities of chromosome distribution during meiosis leading to
the production of different types of aneuploids in the progenies.
3. Asynaptic and desynaptic plants : In these plants, all chromosomes are present as
univalents at metaphase I of meiosis. In the progenies a large number of aneuploids
occur.
4. Tetrasomic plants: They produce n+1 gametes. When they crossed with normal
plants, they produce high frequency of trisomic plants.
Applications of aneuploids

1) Aneuploids have been used to determine the phenotypic effects of loss or gain of
different chromosomes.
2) They are used to produce chromosome substitution lines (replacing chromosome
of one variety with chromosome of another variety of same species). Such lines
provide information on the effect of different chromosomes of a variety in the same
genetic background.
3) They are also used to produce alien addition (addition of chromosome from related
species) and alien substitution (chromosome(s) of one species replaced by the
chromosome(s) of another species).
4) Monosomics are also used in transferring chromosomes with desirable genes from
one species to another.
5) Aneuploid analysis permits the location of a gene as well as of a linkage group on to
a specific chromosome. Monosomics and nullisomics are used for this purpose.
 6) Studies on nullisomic and tetrasomic combinations made it possible to establish
homoeology among the chromosomes of A, B and D genomes of wheat.
7) Aneuploids are also useful in identifying the chromosomes involved in
translocations (tertiary trisomics).
8) Aneuploids are also useful in the preparation of molecular maps.

Limitations
Production, identification and maintenance of aneuploids require elaborate cytogenetic
analysis, which is difficult, time consuming and requires more skill.

Euploids
❖ Euploid derived from Greek words; Eu = true ; ploidy = unit. The term euploidy
designates a change in chromosome number which involves entire set of
chromosomes (x).
❖ Euploids have one or more number of complete chromosome sets (genomes), which
may be identical with or distinct from each other.
❖ The somatic chromosome number of a euploid individual is exact multiple of basic
chromosome number of that species. Euploidy includes monoploids, diploids and
polyploids.

Origin and production of polyploids

1. Spontaneous
i. Chromosome doubling occurs in somatic tissues occasionally and unreduced
gametes are produced in low frequencies.
ii. Non – disjunction in meiosis leading to the production of unreduced gametes.
iii. Endomitosis i.e. chromosome doubling without cell division results into a high
frequency of polyploid cells/plants.

2. Production of adventitious buds

Decapitation in some plants leads to callus development at the cut end of the stem. Such a
callus has polyploidy cells and some shoot buds regenerated from the callus may be
polyploids Eg: Solanaceae

3. Physical agents
Heat, cold treatment, x-ray, gamma ray irradiation will produce polyploids. Tetraploids
branches produced in Datura by cold treatment. Heat treatment is successfully used in barley
and wheat.
4. Regeneration in vitro
Plants of various ploidy have been regenerated from callus cultures of Nicotiana, rice,
Datura.
5. Colchicine treatment
It is the most effective and widely used treatment for chromosome doubling. It is used
for both monocot and dicot plants. Colchicine is an alkaloid obtained from the seeds of
Colchicum autumnale. This chemical arrests the formation of a spindle and thus prevents
the migration of the daughter chromosomes to opposite poles which leads to a polyploid cell
and ultimately the plant. It is generally applied to the meristematic cells at the concentration
ranging from 0.05-0.1 percent using cotton pads or by dipping the tissues in the colchicines
solution for 2-10 hours. Treatment of presoaked seeds with 0.2 percent colchicines solution
for 2-8 hours has also been reported to be very effective in several crops.

Types of polyploidy
1. Autopolyploidy
2. Alloppolyploidy

Autopolyploidy
Autopolyploidy evolve through multiplication of one basic set of chromosomes or genome
derived from a single species
Eg. Auto triploids - 3x (Banana)=33
3x Water melon =33
Auto tetraploids - 4x (Potato) = 48 ; Grapes
Auto pentaploids - 5x
Auto hexaploids - 6x (Sweet potato)=90
Morphological features of autopoplyploids
1. They have larger cell size than diploids.
2. Leaves are fleshy, thicker, larger, deeper green in colour.
3. Flowers, pollen, seeds are larger than diploids
4. They show reduced fertility
5. Polyploids are slower in growth rate and late in flowering.
6. Higher water content
7. Low dry matter content
Applications of autopolyploids
 1. Triploids (3x=33)
a. Seedless Watermelon
Produced by crossing tetraploid (4x) as female and diploid as male (2x). The reciprocal
cross is not successful. Pollination is essential for good fruit setting in triploid. Diploid &
triploid lines are planted in 1:5 to effect pollination.

b. Triploid sugar beet

Triploids have larger roots and more sugar per unit area than diploids and tetraploids. 3x
is the optimum level of ploidy in sugar beets (3x=33). Seed production is done by planting
4x and 2x plants. It is largely cultivated in Europe.

c. Triploid banana (3x=33)

The domestic triploid banana plant (Musa acuminata), cannot undergo meiosis or
reproduce sexually. Triploids are maintained through vegetative propagation.
2. Tetraploids
Grapes : Developed in California which has large fruits and fewer seed than diploids.
Rye : Grown in Sweden and Germany which have large seeds and high protein content
Alfalfa : Better in yield and recovery after grazing
Berseem : Pusa Giant variety is cultivated for forage purpose.
Ornamental : Tetraploids are successful, because of increased flower size and longer
flowering period eg.snapdragon.

Limitations
1. Autopolyploids are successful in species with lower chromosome number.
2. Cross pollinated species are more responsive than self pollinating species.
3. Larger size of autopolyploids is generally accompanied with a higher water content. This
is not a desirable character in cabbage and turnip.
4. In crops grown for seed exhibit high amount of sterility though seed size is increased.
5. New autopolyploids cannot be used directly as crops because of some undesirable traits
e.g. poor strength of stem in grapes, irregular fruit size in water melon.
6. Effects of autopolyploidy cannot be predicted.

Alloployploids
 It is originated by combining complete chromosome sets from two or more
species. Individual having more than two genomes which are dissimilar.
Based on the origin, classified into
1. Natural allopolyploids
2. Artificial allopolyploids
Ex: Wheat, Oats, Cotton, Tobacco, Sugarcane, Mustard etc

Induction of allopolyploids
Hybridization between distinct species followed by chromosomal doubling under natural
conditions as well as through artificial means is the basic approach for the synthesis of
allopolyploids. Chromosomal doubling might have occurred in somatic tissues due to
irregular mitotic division or due to irregular meiosis leading to unreduced gametes.

Role of polyploids in evolution

Allopolyploids are more successful than autopolyploids. It has contributed to a great
extent in the evolution of crop plants. Many of the present day crop species are allopolyploids.
Natural allopolyploids has played important role in the evolution of our major field crops such
as wheat, oats, finger millet, groundnut, cotton, tobacco, Brassica species and sugarcane.
Raphanobrassica was developed artificially but unfortunately it possessed radish like
leaves and cabbage like roots. Triticale, a man made cereal is as an artificially synthesized
allopolyploid.
It is estimated that one third of angiosperms are polyploids and majority of them are
allopolyploids. It is possible to trace back the evolutionary history of allopolyploid crop
species and diploid progenitors. The identification of diploid parental species is primarily
based on pairing between the chromosome of diploid and the allotetraploid species.
Evolution is a slow process; but due to allopolyploids new species originate very quickly.
Polyploids have wider adaptation to different environmental condition.

1. Evolution of wheat
2. Evolution of cotton

3. Evolution of Brassica species

Application of allopolyploids in crop improvement

1. Utilisation as bridging species
Nicotiana tabacum x N. sylvestris
2n = 48 2n = 24
F1(2n=36)
(Chromosome doubling)
N. digluta (2n=72) (resistant to TMV)
Synthetic allohexaploid

N. digluta serves bridging species for transfer of tobacco mosaic virus from N. Sylvestris to
Nicotiana tabacum.
2. Creation of new crop species
Triticale-Man made cereal

Raphanobrassica

3. Widening the genetic base of existing allopolyploid

The genetic base of B.napus is narrow. To widen the genetic base it has been crossed with
B.compestris and B.oleraceae.

Limitations
1. Effects cannot be predicted. Eg. Raphanobrassica
2. Newly synthesized polyploids have defects like low fertility, undesirable features, etc.
3. The production of useful synthetic allopolyploids requires more time and albour.
4. In synthesized allopolyploids only a small proportion of them are useful.
28. Wide hybridization-importance-barriers and techniques for overcoming barriers-
utilization- Pre-breeding.

DISTANT HYBRIDIZATION
When crosses are made between two different species of same genus or between two
different genera, they are generally termed as distant hybridization (or) Wide
hybridization. It may be divided into
1. Interspecific Hybridization: Crosses made between different species of same genus
2. Intergeneric Hybridization: Crosses made between species of different genera
History of distant hybridization
i. First distant hybrid between Carnation and Sweetwilliam by Thomas Fairchild
(1717)
ii. Intergeneric hybrid- Raphanobrassica (Karpechenko, 1928) obtained from a cross
between radish (Raphanus sativus) and cabbage (Brassica oleraceae)
iii. Triticale - Intergeneric hybrid- cross between wheat and rye (Rimpau, 1890).
iv. Another example is Saccharum nobilisation involving three species
v. Most of the interspceific hybrids were no agriculture value. But many interspecific
hybrids, particularly in case of ornamentals, served as commercial varieties.

Interspecific hybridization
Crossing between two different species of the same genus is known as Interspecific
hybridization. Used when the desirable character is not found within the species of a crop.
It is the effective method of transferring desirable genes into cultivated plants from their
related cultivated species or wild species. It is more successful in vegetatively propagated
species like sugarcane and potato.
Interspecific hybridization gives three types of crosses
(a) Fully fertile ; (b) Partially fertile and (c) Fully sterile

(a) Fully fertile crosses

Interspecific crosses are fully fertile between those species that have complete
chromosomal homology. Chromosomes in such hybrids have normal pairing at meiosis and
as a result the F1 plants are fully fertile. Fully fertile interspceific crosses reported in cotton,
wheat, oats and soybean.
Cotton
G. hirsutum(2n=52) × G. barbadense(2n=52)
G.arboreum(2n=26) × G. herbaceum(2n=26)
Wheat
Triticum aestivum(2n=42) × Triticum compactum (2n=42)
Oats
Avena sativa(2n=42) × Avena byzantiana(2n=42)
Soybean
Glycine max(2n=40) × Glycine soja(2n=40)

(b) Partially Fertile Crosses

Partially fertile are those species which differ in chromosome number but have
some chromosomes in common. In such situation, the F1 plants are partially fertile and
partially sterile. It’s reported in wheat, cotton and tobacco.
Wheat
• Triticum aestivum(2n=42) × Triticum durum(2n=28)
Cotton
• Gossypium hirsutum(2n=52) × Gossypium thurberi(2n=26)
Tobacco
Nicotiana digluta(2n=72) × Nicotiana tabacum(2n=48)

(c) Fully sterile crosses

Fully sterile are those species which do not have chromosomal homology. In such
species chromosome number may or may not be similar. The lack of chromosomal homology
does not permit pairing between the chromosomes of two species during meiosis. The F1
plants are fully self sterile. Such hybrids can be made self fertile by doubling of
chromosomes through colchicine treatment. It’s reported in tobacco, wheat, cotton,
Brassica, Vigna.
Tobacco (Clausen and Goodspeed, 1928)
Nicotiana sylvestris (2n=24) × N. tomentosa (2n=24)
Cotton (Harland,1940)
Gossypium arboreum (2n=26) × G. thurberi(2n=26)
Brassica (Morinaga,1943; Olsson,1960)
B.nigra(2n=16) × B.oleracea(2n=18)
Vigna (Singh and Singh, 1975)
Vigna radiata(2n=22) × Vigna mungo(2n=22)

Intergeneric hybridization
It is the crossing between two different genera of the same family. It is used when the
desirable genes are not found in different species of the same genus. This method is rarely
used in crop improvement programmes. It has been used in asexually propagated species.
The F1 hybrids between two genera have been always sterile. The F1s are made into fertile
through chromosome doubling.
Eg. Triticale-Man made cereal

Raphanobrassica

Barriers to production of distant hybrids

In some cases, distant hybridisation may be obtained without an appreciable difficulty.
E.g. G.hirsutum x G.barbadense crosses. But in majority of cases, F1 hybrids may be
 obtained with variable degrees of difficulty and in many cases, hybrids may not be
obtained with currently available technique.
E.g. Corchorus olitorius x C.capsularis
The difficulties encountered in the production of interspecific hybrids may be grouped
into three broad cases viz.,

1. Failure of zygote formation (or) Hybrid sterility

2. Failure of zygote development (or) Hybrid inviability
3. Failure of F1 seedling (or) Hybrid break down
1. Failure of zygote formation
It may be due to failure of fertilization. Fertilization fails due to
a) Slow pollen tube growth E.g. Datura
b) Style may be longer than pollen tube - E.g. Maize and Tripsacum crosses
c) Pollen tubes of polyploid species may be thicker than diploid species and
growth of pollen tube will be slower.
d) The pollen tube may burst in the style of another species and thus fail to reach
the embryo sac e.g. Datura

2. Failure of Zygote development

In many cases, fertilization takes place and zygote is produced. But the development
of zygote is blocked at various stages. It may be due to
a) Lethal genes
Lethal genes cause the death of the zygote during early embryonic development. These
genes affect only the embryos and not the species carrying them. These lethal genes will
act only in interspecific crosses E.g. Aegilops umbellulata has 3 lethal genes. These will act
when crosses made between Ae.umbellulata and diploid wheat.

b) Genotypic disharmony between two parental genomes

The death of the embryos occurs due to genetic imbalance rather than action of specific
lethal genes. Genetic imbalance between two species may lead to death of the embryo. e.g.
Crosses in cotton involving Gossypium gossypioides as one of the parents. The
individuals will not survive owing to genetic disturbances
c) Chromosome elimination
Chromosomes are gradually eliminated from the zygote. This has been due to mitotic
irregularities. The resulting F1s are not true interspecific hybrids since they do not have
 the true parental genomes in full. e.g. crosses between T. aestivum and H. bulbosum, and
between T. aestivum and Z. mays.
d) Incompatability Cytoplasm
Embryo development blocked by an incompatibility between cytoplasm of the
species used as female and genome of the species used as male. This results in hybrid
weakness and male sterility in hybrids.

e) Endosperm abortion
Poor endosperm development leads to endosperm abortion e.g. cross between Triticum
and Secale to produce Triticale – endosperm abortion at much later stage- small
frequency of viable seeds are produced. H. bulbosum × H.vulgare - endosperm abortion at
an early stage- no viable seeds produced

3. Failure of hybrid seedling development

Some distant hybrids die during seedling development or even after initiation of
flowering. Interspecific hybrids in Melilotus are deficient in chlorophyll and fail to
germinate. Such plants are grown to maturity by grafting them on to normal plants or
growing them in vitro. An interspecific and intergeneric hybrid of wheat shows chlorosis and
necrosis which is often lethal.

TECHNIQUES FOR PRODUCTION OF WIDE HYBRIDS

There are several techniques which may be employed to circumvent the problems of
hybridization barriers.
1. Choice of parents: Genetic differences exist among parents in a species for cross
compatibility. More compatible parents should be selected for use in wide crosses. In general,
the species with shorter style should be used as a female parent. Where this is not possible
or desirable, a part of the style of the other parent may be cut off to make it shorter, egg part
of the stigma of maize is cut-off' when it is crossed as female with Tripsacum.

2) Reciprocal crosses: It is better to attempt reciprocal crosses when distant crosses are
not successful. Because reciprocal crosses are successful in some cases. For example
interspecific cross between V. radiata and V. mungo is successful only when former is used
as female and later as male parents.
3) Manipulation of ploidy: When species with different ploidy level % are crossed,
hybridization between them is relatively more difficult than when species with the same
ploidy level are mated. In case of direct hybridization of two species at the same ploidy level,
successes are often accomplished without difficulty. Greater difficulty is encountered when
interspecific hybridization is attempted between two taxa with different ploidy levels. Four
basic methods have been employed to overcome strerility arising from ploidy differences.

a) Direct hybridization: Direct hybridization at different ploidy levels is the most common
method of hybridizing diploids with their polyploidy crop counterparts as adopted in
tobacco, potato, peanut, cotton, wheat and others A tetraploid x diploid species hybrids
resulted in a partially sterile triploid F 1's. Fertility is restored by colchicine treatment, thus
producing hexaploid offsprings. Pollination of 2x or 4x with F l pollen will sometime restore
full fertility. Backcrossing the hexaploid with either the crop or wild species is a logical
method to re-establish chromosomally stable progenies at the same ploidy level as the crop.
b) Doubling chromosome number of the species at lower ploidy level: The chromosome
number of the wild species or of the interspecific hybrid (F l) may be doubled to overcome
sterility of hybrid. In most of the species colchicines is used to raise the chromosome number
of the diploids.
c) Doubling chromosome number at higher ploidy level: Doubling the chromosomes
number of a polyploidy cultivated species before crossing with a diploid species avoids
colchicines treatment of Fl sterile hybrids. The interspecific hybrids thus produced can either
be selfed or backcrossed to the cultivated species.
d) Reducing the chromosome number of species at the higher ploidy level: The
chromosome number of the higher ploidy level species may be reduced by employing anther
culture. Haploid plant production by reduced parthenogesnesis affords a unique and
successful method of creating polyploids in the genus Solanum.
iv. Bridge crosses: Sometimes, two species, say 'A' and 'C' do not cross directly, a third
species say 'B' which can cross with both 'A' and 'C' is chosen as a bridge species. First 'B' is
crossed with 'C' and then the amphidiploid is crossed with ‘A'. Bridge crosses have been used
in tobacco and wheat. In tobacco, Nicotiana repanda can cross with N.sylvestris but not with
N. tabacum. But N. sylvestris can cross with N. tabacum. For transfer of genes from N. repanda,
N.sylveslris is used as a bridge species. It is first crossed with N. repanda and the resulting
amphidiploid is crossed with N. tabacum.
4. Use of pollen mixtures: Cross incompatibility results due to unfavourable interaction
between the protein of pistil and pollen which inhibits normal germination and growth of
pollen tube. This problem has been overcome in certain interspecific crosses by using the
mixture of pollen from compatible (self) and incompatible parents.

5. Use of growth regulators: Many interspecific crosses, hitherto not possible, were
successful due to the use of growth regulators, ego, IAA, 2, 4-D, NAA etc. N. tabacum does not
hybridize with N. repanda, but this cross is possible when IAA is applied to the pedicel of
flowers in a lanoline paste. Similarly, application of 2,4-D prior to pollination, followed by GA
treatment has permitted intergeneric crosses between Hordeum vulgare and species of Avena,
Phleum, Dactylis, Triticum, Lolium and Festuca.

6. Large number of crosses: The success of seed set is generally very low in wide crosses.
Hence, sufficiently large number of flowers may be pollinated to produce interspecific
hybrids.

7. Protoplast fusion: Protoplast fusion may be employed to obtain the wide crosses when is
not possible through sexual fusion. However, this requires perfection and refinement for
adoption in practical plant breeding.

8. Embryo culture: This technique is being used widely to obtain viable interspecific or
intergeneric hybrids. This is used when hybrid zygote is unable to develop. This has been
successfully used in Triticum, Hordeum, Nicotiana, Gossypium and Cucurbita.

9. Grafting: Grafting of interspecific hybrid on to the cultivated species helps in making the
cross successful. Grafting has helped in survival of interspecific hybrid in sugarbeetand
Trifolium and induced flowering in interspecific Glycine hybrids.

Role of wild species in crop improvement

Many of our important crop species are allopolyploids. These crops evolved through
distant hybridization. E.g. Groundnut, Ragi, Sugar cane, Cotton, Wheat, Brassicas, Tobacco,
Potato. Many of the wild species are having resistance against both biotic and abiotic stresses
which are successfully utilized for crop improvement.

Application of wide hybridization in crop improvement

1. Alien - Addition lines
An alien addition line carries one chromosome pair from a different species in addition to
normal diploid chromosome complement of the parent species. Alien addition lines have
been produced in wheat, oats, tobacco and several other species. This is generally used to
transfer disease resistance from wild species. e.g., transfer of mosaic resistance from
Nicotiana glutinosa to Nicotiana tabacum.

2. Alien - Substitution lines

An alien-substitution line has chromosome pair from a different species in the place of one
chromosome pair of the recipient species. Alien substitution lines have been developed in
wheat, cotton, tobacco. In tobacco, mosaic resistance gene N was transferred from
N.glutinosa to N.tabacum. The resistant N. tabacum line had 23 pairs of N. tabacum
chromosomes and one pair (chromosome H) of N. glutinosa chromosomes.

3. Transfer of small chromosome segments

Transfer of small chromosome segments carrying specific desirable genes have been widely
used in crop improvement programmes. E.g. Transfer of black arm resistance from
G.barbadense to G.hirsutum.

4. Quality Improvement
Genes for increased protein content have been transferred to rice, soybean, oats and rye.
Increased soluble solids (from green fruited species) and carotenoid content (the B gene
from L. hirsutum) in tomato; improved leaf quality in tobacco (from Nicotiana debneyi) and
oil quality in oil palm etc.

5. Mode of reproduction
In some cases, incompatability alleles from wild species can be transferred to
cultivated species for hybrid seed production.
Eg. Self-incompatibility alleles from B. campesiris have been transferred to the self-
compatible B. napus for the production of hybrid seed.
Genes for apomixis have been transferred to maize from Tripsacum and sugarbeet from
wild Beta sp.

6. Yield
 Wild relatives of many crop species are excellent sources of the much needed 'yield genes'.
Increased yield through introgression of yield gene from a related wild species into
cultivated species
E.g. Avena sativa was crossed to A. sterilis. The resulting F1 was backcrossed four times to A.
sativa. In BC4 and the subsequent selfed generations, selection for high yield resulted in the
isolation of purelines, which gave 25-30% more yield than the recurrent parent.

7. Wider adaptation
Wild species have served as useful sources of genes for earliness and wider adaptation. Cold
tolerance has been transferred from wild relatives to wheat, onion, potato, tomato etc.,
Earliness has been transferred to cultivated species of Brassica and soybean from their
wild relatives. Wild species of wheat and peas have contributed genes for drought and heat
tolerance. Other similar gene transfers are, salt tolerance to tomato, tolerance to
calcareous soils in grape, and lack of photosensitivity of Pennisetum.

8. Transfer of cytoplasm
It is desirable to transfer cytoplasm of one species into another species. This is
achieved through repeated backcrossing of the F 1 hybrid with the species to which the
cytoplasm is to be transferred. The most common example of the transfer of cytoplasm is
provided by the production of male sterile lines.CMS lines of N. tabacum (tobacco) are
produced by transferring the cytoplasm from N.debneyi.

9. Development of new crop species

E.g. Raphano brassica, Triticale

Limitations of Distant hybridization

1. Incompatible crosses
2. Fl sterility
3. Problems in creating new species
4. Lack of homoeology between chromosomes of the parental species
5. Undesirable linkages
6. Problems in the transfer of recessive oligogenes and quantitative traits
7. Lack of flowering in Fl
8. Problems in using improved varieties in distant hybridization
9. Dormancy

Achievements
 • Upland cotton — MCU2, MCU5, Khandwal, Khandwa2 etc are derivatives of
interspecific hybridization.
• Hybrid between Pearl millet x Napier grass- Hybrid Napier which is very popular for
its high fodder yield and fodder quality e.g. Jaywant and Yashwant.
• Interspecific hybrids in cotton- Varalaxmi, Savitri, DCH.32, DH 7, DH 9 etc.
Parbhani Kranti variety of bhendi.

Pre-breeding

Pre-breeding is the step before practical breeding. Land races not used directly in
breeding programme. Hence, land races are crossed with modern varieties to develop
lines with improved agronomic features. These derived lines are used in breeding and this
phase is prebreeding. Its aim is “To introduce new desirable traits/genes into an adapted
genetic background. This will broaden the genetic base in a breeding material”
It is vital link between conservation of PGR in gene bank and utilisation of these resources
in agriculture and horticulture.
29. Mutation breeding: mutation – types – mutagens – breeding procedure –
achievements – limitations

Mutation breeding
Introduction
Mutation Sudden heritable change in a characteristic of an organism due to change in a gene, in
chromosome that involves several genes or in a plasma gene. The term was coined by Hugo de
Vries in 1900 from the Latin word “mutare” - ‘to change’. Mutagenic action of X rays by
Muller (1927) in Drosophila and Stadler in barley and maize

Mutation is a sudden heritable change in a characteristic of an organism. It is the results in gene

changes in chromosome or in plasma gene.
Gene or point mutations.
Mutation produced by changes in the base sequence of genes as a result of transition,
transversion, deletion, duplication or inversion etc.
Chromosomal mutation
Mutation produced by changes in chromosome structures and chromosomal numbers.
Cytoplasm mutation is occurs in cytoplasmic genes.
History
Term mutation was first introduced by Hugo de varies in the year 1900. The discovery of the
mutagenic effects of X-rays on the fruit fly (Drosophila) by H. Muller in the 1920. And gamma
rays and x-rays by L. J. Stadler, later Muller got Nobel Prize In 1946.
• Spontaneous mutation: Mutations occur in natural population at a 10 6 rate. The frequency
of spontaneous mutation is one in 10 lakhs.
• Induced mutation: Mutation may be artificially induced by a treatment with certain physical
or chemical agents. The chemicals used for producing them are termed as mutagens. The
utilization of induced mutation for crop improvement is known as mutation breeding.
Induced mutations are used more often in a supplementary role as a source of new alleles.
However, it is still important in breeding vegetatively propagated species, including field
crops, ornamentals, fruit and forest species. It is especially useful in ornamental plant
breeding
Characteristics of mutation
1. Mutations are generally recessive, but dominant mutations do occur.
2. Mutations are generally harmful to the organism, 0.1 percent are beneficial.
3. Mutations are random that they may occur in any gene.
4. Mutation are recurrent, the same mutation may occur again and again.
5. Induced mutation commonly shows pleotropic, due to linked genes.
Types of Mutations
Transition
Mutation by transition entails the conversion of one purine base to another purine (or a pyrimidine
to another pyrimidine) When a purine is replaced by purine. E.g. Adenine is replaced by guainine
or pyrimidine by pyrimidine. E.g. Thymine is replaced by cytosine.
Transversion
A transversion involves the substitution of a purine by a pyrimidine and vice versa.
• Pyrimidine is replaced by purines. A to Z, A-C, G-C G-T Transition, Trans version are
relatively also deleterious, but base substitution may generate such codon that codes for any
amino acid (sense codons) whenever the nonsense codon is produced it acts on terminator
of the poly peptides changes.
Base addition/ deletion
A variety of alkylating agents (E.g., sulfur and nitrogen mustards) can act on the DNA molecule,
reacting mainly with guanine (G) to alkyl ate and remove it from the DNA chain.
• The missing spot may be occupied by any one of the four bases to create mutations, usually
by transition.
• Acridine is also known to express its mutagenic effect through the addition of deletion of
bases.
• An insertion of one or more bases in a DNA molecule is called based addition.
Frame shift mutation
• Insertion-deletion mutations may cause significant changes in the amino acid composition
of a protein and hence its function. E.g. GAG-CCG-CAA-CTT-C (corresponding to Glu-
Pro-Glu-Leu) may be altered by a deletion of G that shifts the reading frame to the right by
one nucleotide to produce AGC-CGC-AAC-TTC (corresponding to Ser-Arg-Asi-Phe).
Mutagens: Physical mutagens
Mutagens: Agents which induce mutation are known as mutagens.
There are two types of mutagen
• Physical Mutagens
• Chemical Mutagens

Mutagen Characteristics
 X- rays Electromagnetic radiation; penetrate tissues from a few millimeters
to many centimeters first discovered by Roentgen 1895 wavelength
varies from 10-11 to 10-7. Highly penetrating they break
chromosomes and produce all types of mutation in the nucleotides
like addition, deletion, inversion, transition and transversion.

Gamma rays Electromagnetic radiation produced by radioisotopes and nuclear

reactors; very penetrating into tissues; sources are Co60 and Ce137
largely used in crop plants

Neutrons A variety exists (fast, slow, thermal); produced in nuclear reactors;

uncharged particles; penetrate tissues to many centimeters; source
is U235

Beta Produced in particle accelerators or from radioisotopes; are

particles electrons; ionize; shallowly penetrating; sources include P32 and
C14

Alpha Derived from radioisotopes; a helium nucleus capable of heavy

particles ionization; very shallow penetrating

Protons Produced in nuclear reactors and accelerators; derived from

hydrogen nucleus penetrate tissues up to several centimeters

Non ionizing Non-ionizing radiation produced from mercury vapor lamps or

radiations tubes. They penetrate one or two cell layers eg. Pollen.
Chemical Mutagen
The material is soaked in a solution of the mutagen to induce mutations.
• Chemical mutagens are generally carcinogenic and must be used with great caution.
• One of the most effective chemical mutagenic groups is the group of alkylating agents (these
react with the DNA by alkylating the phosphate groups as well as the purines and
pyrimidines).
• Another group is the base analogues (they are closely related to the DNA bases and can be
wrongly incorporated during replication); examples are 5-bromo uracil and maleic
hydrazine. (Chemical mutagens commonly used are ethyl methane sulfonate (EMS) and
diethylsulfonate (DES).
• The half-life (time taken for degradation of the initial amount of alkylating agent) for EMS
in water is about 3 hours at 20°C but only 10 hours at 37°C.
• Consequently, chemical mutagens are best freshly prepared for each occasion.

Mutagen group Examples

 Base analogues 5-bromouracil, 5-bromodeoxyuridine

Related compounds Maleic hydrazide, 8-ethoxy caffeine

Antibiotics Actinomycin D, Mitomycin C, Streptonigrin

Alkylating agents Sulfur mustards Ethyl–2-chloroethyl sulfide

Nitrogen mustards 2-chloroethyl-dimethyl amine
Epoxides Ethylene oxide
Ethyleneimines Ethyleneimine
Sulfonates, etc. Ethyl methane sulfonate (EMS),
Diethylsulfonate (DES)

Diazoalanes Diazomethane
Nitroso compounds N-ethyl-N-nitroso urea
Azide Sodium azide
Hydroxylamine Hydroxylamine
Nitrous acid Nitrous acid
Acridines Acridine orange

Situation of Mutation Breeding

1. When the desired variability is not found its cultivated varieties or in the germplasm
2. When a high yielding variety has a defect, the defect can be also being changed without
much alteration in the genetic background.
3. To break the undesireble linkage
4. In crops sexual reproduction is absent and creation of variability through recombination is
not possible
5. Generation cycle is very long. E.g: Plantation crops.
6. When attractive flowers and of foliage colour have to be developed in ornamental crops
Steps in mutation Breeding
• Objectives: Well defined clear cut objective
• Selection of Varieties: Best variety available in the crops
• Part of the plant : Seeds, pollen grains, vegetative Propagates
• Doses of the mutagen: Dose which produce maximum frequency of mutation
• LD-50- the dose at which 50% of the treated individuals are killed.
• Seeds are presoaked in water to initiate the mutagen activity and exposed followed
Chimera: A chimera is an individual with one genotype in some of its parts and another genotype
in the others
Procedure for oligogenic traits
• M1- Several hundred seeds are treated with mutagen and are space planted M1 plants are
selfed and harvested separately.
• M2-About 2000 progeny rows are grown. Oligo genic mutants are selected and the seed are
harvested separately and advanced to M3 , M3 are grown as individual plant progeny
• M3 -Progeny rows from individual selected plants are grown. Poor and inferior mutant rows
are eliminated if the mutant progenies are homogenous two or more M3 progeny may be
bulked. M3 are harvested in bulk and are advanced to preliminary yield trail in M4.
• M4 – Preliminary yield trial with suitable checks, promising mutants are selected for
replicated multi location trial.
• M5-M7 Replicated multi location trials are conducted. Outstanding variety may be retained
for hybridization.
Procedure for Poly genie traits
Mutagenesis does produce genetic variation in polygenic traits
1. M1 and M2 : M1 and M2 are grown, M2 vigouros fertile and normal looking plants that alone
exhibit a mutant phenotype are selected and their seeds are harvested separately to raise
individual plant progeny rows in M3.
2. M3 Progeny rows from individual selected plants are grown. Careful observation is made
on M3 rows for small deviation in phenotype from the parent variety. Few rows which are
homogenous and would be harvested in bulk. Selection is done in M3 rows showing
segregation.
3. M4: Bulked seed from homogenous M3 rows may be planted in preliminary yield trial with
suitable check. Superior progenies are selected for replicated multilocation yield trial
superior homogenous progenies are harvested in bulk for preliminary yield trial in M 4.
4. M5- M8: Preliminary yield trials and MLT are conducted depending upon the when
progenies become homogenous. Outstanding progeny may be released as new variety.
Mutation breeding Achievements
• Totally 1029 mutant varieties are released in India.
• Disease resistance: Verticilium wilt resistance in peppermint, victorial blight resistance in
barley, downy mildew resistance in pearl millet.
• Modification of plant structure: E.g., bush habit in drybean, dwarf mutants in wheat and
other cereals.
• Nutritional quality augmentation: E.g. Opaque and floury endosperm mutants in maize
 • Chemical composition alteration: E.g., low ericic acid mutants of rape seed
• Male sterility: for use in hybrid breeding in various crops
• Horticultural variants: development of various floral mutants
• Breeding of asexually propagated species: numerous species and traits
• Development of genetic stock: various lines for breeding and research
• Development of earliness: achieved in many species
Merits
It can be used in inducing male sterility. eg: Ethidium di bromide
• Cheap and rapid method as compared to other methods.
• More effective for oligogenic traits.

Somaclonal variation - utilization in crop improvement; In vitro selection techniques –– Use

of doubled haploids in crop improvement.
Tissue culture
It refers to growth of living plant tissues in a suitable culture medium.
• It is defined as a technique of growing plant tissues on synthetic medium under controlled
and aseptic conditions.
G. Haberlandth: German plant physiologist is considered to be the father of plant tissue culture,
who conceived the idea of Totipotancy in 1902.
Totipotancy: It is the ability of an individual cell or plant part to produce whole plant. The
concept of tissue culture depends on totipotency.
Explant: They are the organs excised from plants such as roots, hypocotyls, cotyledons.
Commonly used media in tissue culture
• Murashige and Skoog's media (1962) - MS media.
• White media (1963)
• Gamborg et.al (1968) -B5 media
• Chu media (1978) -N6
Plant tissue culture areas
• Micropropagation
• Somatic embryogenesis
• Somaclonal variation
• Anther culture
• Pollen culture
• Meristems culture
 • Embryo culture
• Protoplast culture
• Cryopreservation of germplasm
• Production of secondary metabolites from plant tissue cultures.
Methods which are extensively used in plant breeding
Micro-propagation
Involves the production of plant from very small plant parts through tissue culture techniques.
Advantages:
It is independent of seasonal constraints hence ensures year round rapid propagation.
• Micro propagated plants are usually true to type.
• Production of disease free plants.
• Micro-propagated field grown plants usually exhibit vigorous growth, better quality
and higher yields.
Problems encountered in micro-propagation.
• Microbial contamination.
• Browning of the culture medium.
• Callusing: Extensive callusing leads to variability among the regenerated plants.
• Vitrification: Abnormal looking plants. It is because of liquid medium, it can be over
come by conditioning.
• Vulnerability of micro-propagated plants to transplanting shock.
• Tissue culture induced variation.

Somaclonal variation

It is the variation among the tissues or plants derived from the In-vitro somatic cell culture
i.e. Callus and suspension culture.
Variations are of two types:
1. Gamatoclonal variation
2. Protoclonal variation
Factors effecting soma clonal variation
1. Degree of departure from organized growth.
2. Genetic constitution of the donor plant
3. Culture environment.
4. Tissue source.
Significance of somaclonal variation in crop improvement.
1. It is an important alternative for creation of variation in such crops, which are
extensively propagated by tissue culture.
2. This is help full in breaking linkages between certain undesirable genes.
3. New varieties are developed in tomato, sugarcane, celery, brassica and sorghum.
4. It has greater advantage for crop improvement in apomictic and vegetatively
propagated crops and also in cross-pollinated crops with narrow genetic base.
5. In India somaclonal variants of medicinal plant Citronella javaI has been
released as a commercial variety B-13, which gives higher yield and oil content.
6. Pusa Jai Kisan- Brassica juncea as somaclonal variant of Varuna variety.
• Protoplast culture or somatic hybridization
Protoplast is a nacked cell without cell wall surrounded by plasma membrane and potentially
of cell wall regeneration growth and division.
It includes following steps.
1. Isolation of protoplast.
2. Fusion of the protoplast of the desired species or varieties.
Selection of somatic hybrid cells.
1. Culture of the hybrid cells and regeneration of hybrid plants from them.
Applications:
1. Used to produce symmetric hybrids and asymmetric hybrids.
2. Hybrids can also be produced
3. Used for anther or pollen cultures
Pollen culture
Haploids plant can be obtained from pollen grains by planting an anther or isolated pollen grains
on a suitable culture medium, this is known as anther or pollen culture.
Applications of Pollen culture:
• It is useful in development of haploids.
• Haploids have been obtained by pollen culture in wheat, barley, rice. By doubling the
chromosome number of haploid we get homozygous diploids.
Ovule culture
Regeneration of whole plant from the ovule in the nutrient medium is called ovule culture/ovary
culture. Here we can use two types of ovules.
• 1. Fertilized
 • 2. Unfertilized
Application of tissue culture in crop improvement
• 1. Generation of variability.
2. Development of haploids.
3. Embryo rescue.
4. Somatic hybridization.
5. Selection for diseases resistance.
6. Selection for salinity and metal toxicity.
7. Selection for drought resistance.
8. Micro-propagation.
9. Preservation of germplasm.

Application of Haploids in Plant Breeding

In Vitro production of haploids can solve some problems in genetic studies since gene action is
readily manifested due to a single allelic gene present in chromosome of entire genome.

1. Releasing New Varieties through F1 Double –haploid System:

Haploid breeding technique usually involve only one cycle of meiotic recombination. However,
many agronomic traits are polygenically controlled. One cycle of recombination is usually
insufficient for the improvement of such quantitative traits since linkage between Polygenes will
not release all potential variations available in the cross. To overcome these disadvantages, the
Chinese developed a method combining anther culture with sexual hybridization among different
genotypes of anther derived plants. The anthers of the hybrid (F1) progeny are excellent breeding
material for raising pollen-derived homozygous plants (Double –haploids) in which complementary
parental characteristics are combined in one generation.

Double –haploids are also useful in studies related to inheritance of quantitative traits. Using double
–haploid technique new varieties have been developed in barley, Brassica, rice, maize , rye, potato,
pepper and asparagus.

2. Selection of Mutants Resistance to Diseases:

Mutants with resistance to disease is of prime importance in crop improvement. Haploids provide
a relatively easier system for the induction of mutations. Some examples of using anther culture
technique in mutant successfully are tobacco mutants resistant to black shank disease and wheat
lines resistant to scab. (Fusarium graminearum).

3. Developing Asexual Lines of Tree Perennial Species:

Chinese workers obtained pollen –derived rubber tree taller by sic meters which could then be
multiplied by asexual propagation to raise several clones. Another example of pollen –haploids in
plant improvement is popular.

4. Transfer of Desired Alien Gene:

Chromosomal instability in haploids makes them potential tools for introduction of alien
chromosomes on genes during wider crossing programmes. In rice , developing a resistance to blast
requires about 12 years by conventional breeding through back crossing. Through hybridization
and anther culture, this can be achieved in two years (Examples: cv . Zhonghua No.8 and 9 released
by the institute of crop Breeding and cultivation in china.

5. Establishment of Haploids and Diploid Cell Lines of Pollen Plant:

The anther culture technique was used to establish both haploid and diploid somatic cell lines of
pollen plants in wheat and maize. Similarly, a haploid tobacco line resistant to methionine
sulfoxomide was selected which turned out to be identical in phenotype and effect to the toxin
produced by the pathogen Pseudomonas tabaci.

Introduction to markers – morphological – biochemical- DNA markers – uses of marker

assisted selection - major genes – merits – demerits – achievements.
Introduction
The new tools of molecular biology have broadened the scope of gene manipulations at the level
of specific DNA segments to produce novel genomes with enhanced levels of resistance to biotic
and abiotic stresses, physiological efficiency, quality of product and yield potential.
• The current plant breeding programme are being complimented by biotechnology which
involves the utilization of biological agents such as micro organisms, cells of higher plants
and animals for the isolation of useful products.
• The molecular biology is applied in the plant breeding for the identification, location,
isolation, characterization, cloning and transfer of specific genes in crops plants.
 • Molecular marker systems:
1. Restriction fragment length polymorphism (RFLP)
2. Randomly amplified polymorphic DNAs (RAPDs)
3. Sequence tagged sites (STS)
4. DNA Amplification Fingerprinting (DAF)
5. Amplified fragment length polymorphism (AFLP)
Randomly amplified polymorphic DNAs (RAPDs)
This is a PCR (Polymerase chain reaction) based technique where a single short oligonucleotide
primer which binds to many different loci, is used to amplify random sequences from a complex
DNA template such as plant genome.
• For most plants the primers that are 9-10 nucleotide long are expected to generate 2-10
amplification products.
• The primers are generally of random sequence, biased to contain at least 50% GC content
and to lack internal repeats.
• The products are easily separated by standard electrophoretic technique and visualized by
UV illumination of ethidium bromide stained gels.
Construction of molecular maps
It involves 5 major steps:
1. Identification of divergent parents
2. Generation of a suitable mapping population
3. Identification of polymorphic probe-enzyme combination and informative primers
4. Analysis of marker segregation in the mapping population
5. Establishment of linkage.
Identification of divergent parents:
A survey of the available germplasm is made for existing morphological diversity.
• A preliminary survey can be carried out using a few DNA markers to identify divergent
parents among the available lines.
• It is advantageous to select parents which posses constricting traits of agronomic
importance, so that genes for those traits can also be simultaneously localized on map
Analysis of marker segregation in the mapping population
The individual plants of the mapping population are analyzed using the same primers which
detect polymorphism between parents.
• The plants are categorized as parent 1 type, parent 2 type or F1 depending on the size of the
polymorphic fragments.
 • The observe frequency of the plants based on the marker locus genotypes is compared with
the expected segregation ratio of 1:2:1 in case of F2 and 1:1 in case other populations. Based
on the chi-square analysis, the observed segregation pattern is tested for possible distortion.

Generation of a suitable mapping population

• Selected plants are crossed to obtained f1 hybrids from which four different mapping
population can be generated: F2 (by Selfing a single F1 plant ) , BC (by crossing F1 with
either of parents) ,Recombinant Inbredlines (by Selfing individual F2 plants for 7-8
generations), and doubled haploids (by invitro culture of anthers or pollens from the F1
followed by chromosomes doubling).50-100 F2 plants are required for construction of
linkage map.

Identification of polymorphic probe-enzyme combination and informative primers

• For RFLP analysis, several DNA probes should be tried in combination with set of
restriction enzymes to find the polymorphic probe-enzyme combinations for the
parents.
• One or two enzymes such as Eco RI and Hind III were sufficient to detect high
level of DNA polymorphism
Application Of DNA-markers In Plant Breeding:
Estimation of Genetic Diversity
• Characterization of Germplasm Resources
• Identification of crop varieties
• Marker aided selection
Estimation of Genetic Diversity
Divergent parents carrying different favorable genes are used in crossing programme for realizing
trsangressive segregants in self-pollinated crops and for creating heterotic hybrids in cross
pollinated crops.
1. DNA selected for diversity analysis should preferable belongs to non-overlapping expressed
part of the genome. The RFLP markers, if included in the analysis should be generated with
cDNA probes.
2. For genotyping large number of collections available in the gene bank, a PCR based marker
system will serve better by way of saving time and efforts.
Identification of crop varieties:
Unambiguous differentiation and identification of crop varieties is essential for their registration,
protection, utilization and quality control of seeds.
1. DNA fragment profiles obtained with different marker systems should provide fingerprints
highly characteristic of varieties.
2. Molecular markers systems have been extensively used for identification of crops cultivars,
somatic hybrids and cybrids. Genetic stocks such as somaclones and cytoplasmic male
sterile lines in different crops.
Marker aided selection:
Selection of a genotype carrying desirable gene or gene combination via linked marker is called
MAS.
1. Breeders practice MAS when an important trait, that is difficult to assess, is tightly linked
to another Mendelian trait which can easily scored.
2. MAS involve scoring for the presence or absence of a desired plant phenotype indirectly
based on DNA banding pattern of linked markers on gel or on autoradiogram depending on
the marker system.
3. The rationale is that the banding pattern revealing parental origin of the bands in segregants
at a given marker locus indicates presence or absence of a specific chromosomal segment
which carries the desired allele. This increases the screening efficiency in the breeding
programmes in a number of ways such as.
4. Gene introgression and elimination of linkage drag
5. Molecular markers can aid introgression and impart greater precision. With the use of
isozyme marker it has been possible to introgress nematode resistance in to the cultivated
tomato from one of its wild relatives.
6. It is often observed that the desirable genes such as those for disease resistance remain
linked with undesirable weedy characteristic of the alien species. During gene introgression
by back crossing, the linked undesirable genes are also get transferred to the recipient parent.
This has been referred as linkage.
Gene pyramiding:
Pyramiding of genes has been suggested as an effective way to provide durable form of disease
and insect resistance in crop plants. Because of difficulty involved and number of years it
requires, development of lines carrying genes for resistance against several races of a single
pathogen/insect or different biotic stresses has not been very successful.
 Recently successful pyramiding of four genes, Xa4, Xa5, Xa13 and Xa21 conferring
resistance to four different races of bacterial blight has been achieved in rice by using molecular
markers.
Development of heterotic hybrid:
Molecular markers based analysis can be utilized to identify specific genomic regions containing
QTLS for yield showing dominance or over-dominance gene action in f2 of a cross between two
inbred lines.
• This will enable analysis of an existing cross in terms of the number of QTLS involved and
the magnitude of there effect on the measured trait. Subsequently additional Inbredlines can
be tested against standard lines for the presence of additional dominant or over-dominant
QTLS. These could then be incorporated into the standard lines by marker based
introgression. In this way existing inbred can be improved first and then used to develop
hybrids for realizing higher heterosis than in the existing ones.
Limitations in the use of molecular markers
The cost and time required to undertake genetic analysis, besides lack of detectable
polymorphism in certain crops also restricts the use of such markers.
• High density maps have been developed using very wide crosses but the extent of
polymorphism is much less in routine breeding populations particularly in self pollinated
crops.
 32. Procedure for release of new varieties-stages in seed multiplication-steps in
nucleus and breeder seed production.

Varietal release
Before a variety is released and reaches to the farmer, the All India Co-ordinated Crop
Improvement Project identifies the variety for release in its workshop as per the established
norms for testing for its value for cultivation.
Presently, All India Co-ordinated Crop Improvement (AICCPs) Projects have been created for
almost for all the crops or groups of crops (Paroda, 1992).
AICCIPs follow a three tier system of multilocation evaluation spread over a minimum of
three years involving the following stages:
• First year - (IET, Initial evaluation trial)
• Second year — Advance varietal trial — I (AVT - 1)
• Third year — Advance varietal trial — II (AVT-II)
The initial evaluation trial includes the following entries:
The entries to be nominated must have undergone critical evaluation/screening in the station/
station trials conducted by the sponsoring breeder. Secondly, the entries to be Nominated must
have high degree of phenotypic uniformity and genotypic stability.
• The IET is conducted across the zones of all over the country along with check varieties. A
minimum of three check varieties comprising of the following is used. i. National check, ii.
Zonal check, iii. Location check, IV. Qualifying check
• All the trials are monitored by a team of scientists deputed by project coordinator to record
the observations on the uniformity of crop stand, disease and insect-pest, resistance, bird
damage etc.
• The main composition of the monitoring team is:
a) Project Director/Project Co-coordinator/PI/Zonal Co-ordinator Team leader
b) Breeder Member
c) Agronomist Member
d) Pathologist/Entomologist Member
e) Scientist from any other specified discipline Member
• The observations recorded according to guidelines on the data books for further supply to
the co-ordinators
• The data received at the co-coordinator’s cell is critically examined for the inclusion in the
Annual Report.
 • Outstanding performance for yield of a variety by a margin of 10% over the best
performing check is promoted to advance varietal trial.
Advance varietal trials (AVT-1)
Advance varietal trial is constituted by the entries promoted from IVT on the criteria specified
above.
• Limited number of entries in AVT-1 (not exceeding 16) is tested along with a minimum of
three checks comprising of national check, zonal check and local check.
• All these entries are evaluated in a randomized block design with 3-4 replications at the
different locations.
• The monitoring is done by the same team as given for IVT.
• Besides the agronomic and morphological observations, the additional data may be
generated by the co-operators on disease and insect-pest resistance, and quality.
• Again, if a variety gives significant superior performance by a margin of 10% over the, best
performing check in combination with other attributes is promoted to next stage
Advance varietal trials (AVT-II)
Same steps are followed as mentioned under AVT-1. However, the additional data to be
generated at this stage.
• Response to agronomic variables such date of sowing, population densities and weedicide
may be recorded.
• Data on diseases, insect-pests, quality parameters and abiotic stresses may also be recorded
and discussed during the workshop.
• If the variety gives outstanding performance over the check (by a margin of 10%) besides
having all the favourable attributes, then the proposal for identification of a given variety is
submitted by the concerned breeder on a variety identification proforma specified by the
ICAR
Variety Identification System
The proposal containing all available data for the variety is considered by the variety
identification committee constituted by the ICAR which meet during each AICCIP workshop.
• Recommendations are made for country-wide release or for a specific zone or states.
• Afterwards, the sponsoring agricultural university/research institute then submits the
proposal for its release and notification to central sub-committee with the support of the
Project.
 • Once the Central sub-committee accepts the proposal, the variety/hybrid is released for the
specific state or zone. The release proposal also ensures the availability of enough seed
stock.
State Varietal Identification System
The State Seed Sub-Committee (SSSCs) is constituted by Central Seed Committee and these
SSSCs have been given responsibility to set up a State, Seed Certification Agency (SSCA), a
State Seed Laboratory, and to appoint a seed analyst and seed inspectors.
• The SSCC is responsible for the release of a variety in its own state on the basis of data
generated by State Agriculture University.
• The concerned breeder along with agronomist, pathologist, entomologist and biochemist
generate sufficient data (usually more than three years).
• Secondly, sample variety must be evaluated in All India Co-ordinated crop improvement
projects trials.
• Thirdly, on-farm trial data for a year or two are collected by extension agencies of State
Department of Agriculture.
• After having all the above information, the State Agriculture University deliberates on the
release proposal of a variety in a series of meetings before recommending to the SSC.
• Once approved by the SSSC for release in a state, the variety is requested to be notified for
seed production purpose by the Central Sub-committee.
Notification of varieties
CLASSES OF SEED :
The four generally recognized classes of seeds are:
Breeder's seed, Foundation seed, Registered seed and Certified seed.
The Association of Official Seed Certifying Agencies (AOSCA) has defined these seed
classes as follows:
Breeder seed: The seed or vegetatively propagated material directly controlled by the originating
or the sponsoring breeder or institution which is the basic seed for recurring increase of foundation
seed.
Foundation seed: It is the progeny of breeder seed. The seed stock handled to maintain specific
identity and genetic purity, which may be designated or distributed and produced under careful
supervision of an agricultural experiment station. This seed is the source of all other certified seed
classes either directly or through registered seed.
Registered seed: The progeny of the foundation seed so handled as to maintain its genetic identity
and purity and approved and certified by a certifying agency. It should be of quality suitable to
produce certified seed.
Certified seed: It is the progeny of the foundation seed. Its production is so handled to maintain
genetical identity and physical purity according to standards specified for the crop being certified.
It should have the minimum genetical purity of 99%. Certified seed may be the progeny of certified
seed , provided this reproduction does not exceed two generations beyond foundation seed and
provided that if certification agency determines the genetic and physical purity, if not be
significantly altered. In case of highly self pollinated crops certification of one further generation
may be permitted.
Certified seed produced from certified seed shall be eligible for further seed increase under
certification, except in case of highly self-pollinated crops, where certification of one further
generation may be permitted. Certification tags issued once for certified seed not eligible for further
seed increase under certification.
For paddy and wheat , certified seed produced from certified seed is eligible for
certification by NSC up to two generations from foundation seed Foundation seed - Certified seed
(I) - Certified seed (II). For barley, garden pea ,ground nut, soyabean, certified seed produced
from certified seed is eligible for certification up to 3 generations from foundation seed
Foundation seed - Certified seed (I) - Certified seed (II) - Certified seed (III) Certification of
certified seed produced from certified seed is not permitted for crops other than those listed above.
Differences between certified seed and truthful labelled seed
Certified seed Truthful labelled seed
Certification is voluntary Truthful labelling is compulsory for notified
kind of varieties
Applicable to notified kinds only Applicable to both notified and released
varieties
It should satisfy both minimum field and seed Tested for physical purity and germination
standards
Seed certification officer seed inspectors can Seed inspectors alone can take samples for
take samples for inspection checking the seed quality.

GENERATION SYSTEM OF SEED MULTIPLICATION

 Generation system of seed multiplication Generation system of seed multiplication is
nothing but the production of a particular class of seed from specific class of seed up to certified
seed stage. The choice of a proper seed multiplication model is the key to further success of a seed
programme. This basically depends upon
• The rate of genetic deterioration
• Seed multiplication ratio and
• Total seed demand
Based on these factors different seed multiplication models may be derived for each crop and
the seed multiplication agency should decide how quickly the farmers can be supplied with the
seed of newly released varieties, after the nucleus seed stock has been handed over to the
concerned agency, so that it may replace the old varieties. In view of the basic factors, the chain
of seed multiplication models could be.,
a. Three - Generation model
Breeder seed - Foundation seed - Certified seed
b. Four - Generation model
Breeder seed - Foundation seed (I) - Foundation seed (II) -Certified seed
c. Five - Generation model

Breeder seed - Foundation seed (I) - Foundation seed (II) - Certified seed (I)
Certified seed (II)
For most of the often cross pollinated and cross pollinated crops 3 & 4 generation models
is usually suggested for seed multiplication.
Ex: Castor, Redgram, Jute, Greengram, Rape, Mustard, Sesame, Sunflower and most of the
vegetable crops.

GENERATION SYSTEM OF SEED MULTIPLICATION AND QUALITY

CONTROL (NOTIFIED VARIETIES AND HYBRIDS)
Nucleus and Breeders seed production
The initial handful of seeds obtained from selected individual plants of a particular variety,
for the purposes purifying and maintaining that variety by the originating plant breeder and its
further multiplication under his own supervision, or the supervision of a qualified plant breeder, to
provide Breeder’s Seed constitutes the basis for all further seed production. The varietal purity of
subsequently multiplied foundation, registered and certified seed largely depend upon the quality
of the nucleus/breeder’s seed. Unless the nucleus/ breeder’s seed is of highest purity and quality the
seed multiplied from it cannot be regarded as of satisfactory genetic purity. Unsatisfactory genetic
purity, especially in cross pollinated crops, could ultimately severely affect the performance of a
variety. It is therefore, of utmost importance that the nucleus/breeder’s seed is produced in such a
manner that satisfactory genetic purity, identity and the other good qualities of seed are maintained.
Methods of maintenance of nucleus and breeder’s seed in self fertilized crops.
Methods of maintaining nucleus seed/breeder’s can be conveniently divided into the
following two groups:
1. Maintenance of newly released varieties
 2. Maintenance of established varieties
Maintenance of Nucleus Seed of Pre-released or Newly Released Varieties
The procedure outlined by Harrington (1952) for the maintenance of nucleus seed of pre-released
or newly released varieties is described below:
a. Sampling of the variety to obtain nucleus seed. New numbers, lines or selection which are highly
promising, on the basis of performance in breeding nurseries and yield trials, should be sampled for
seed purification. These samples provide a beginning for purifying new varieties and for possible
increase and distribution to farmers. Not more than fifteen new varieties in any one crop at a station
should be sampled in one year.
b. Table examination of samples: The two hundred plants of each sample should be threshed
separately and the seed should be examined in piles on the table. Discard any pile appearing
obviously off type, diseased or otherwise unacceptable. The seeds of each two hundred plant
samples or less is now ready to be sown in a variety purification nursery called as nucleus.
c. Locating and seeding of nucleus: Each nucleus seed should be grown on clean fertile land at an
experiment station in the region or in area in which this new variety could be grown, in the event
of its release. The land must not have had a crop of the same kind in the previous year.
d. Inspection of nucleus two-row plots and removal of off types: Throughout the season of growth,
from the seedling stage until maturity, the nucleus plot should be examined critically. Differences
in the habit of early plant growth, leaf colour, rate of growth, time of heading, height head
characteristics and diseases reactions should be looked for. If a plot differs distinctly from the
average in the preheading stages of growth, it should be removed before heading.
e. Harvesting and threshing of nucleus; each remaining plot, of which there should be at least 180
out of the original 200. Should be harvested individually with a sickle and tied in a bundle. The
total bundles of each nucleus should be labelled and stored until the current years yield rests for
trials are obtained. The nucleus bundles of any new variety should be discarded, if it is found
unworthy of being continued.
Later the seed should be cleaned in a fanning mill or by hand methods, the grain from each
nucleus plot being placed in a pile on the seed table. The 180 or more piles of seed of one nucleus
must be examined for approximate uniformity of seed appearance, and any pile, which appears to
be off type discarded. All the remaining piles of the seed should be masked together in one lot. This
should treated with fungicide and insecticide, bagged, labelled and stored as "Breeder’s Stock Seed"
for use in the next year. Breeder’s stock seed is the original purified seed stock of a new variety in
the hands of the plant breeders.
Maintenance of Breeder’s Seed of Pre-released or Newly Released Varieties
 The following steps are involved in the maintenance of breeder’s seed.
a. Breeder’s stock seed from the nucleus should be sown on the clean, fertile land, which
did not grow a crop of the same kind in the previous year. The space required for the seeding the
breeder’s stock is about 1.2 ha in the case of wheat and as much as 3 ha in the case of transplanted
rice.
b. The field should properly isolated.
c. The best farm procedures should be used in the sowing, raising and harvesting of
breeder’s stock.
d. It should be produced at the experiment station in the area in which the new variety has
been bred.
e. The seeding should be done in such a way as to make the best use of the limited amount
of seed available and to facilitate roguing. The row spacing should be sufficient to permit
examination of plants in rows for possible mixture or off types.
f. Roguing: All plants not typical of the variety should be pulled and removed. There should
be very few plants to rogue out if the previous years nucleus breeder’s stock seed was well protected
from natural crossing and careful roguing was done and there were no impurities during cleaning
etc. The rouging should be done before flowering, as was done for the nucleus/breeder’s stock seed.
g. Harvesting the breeder’s stock: In the breeder’s stock is harvested and threshed, the
equipment used must be scrupulously clean and free from seeds of any other varieties. This
cleanliness should be extended to cards and bags as well as threshing machine it self. The seed
should now be about 99.9 per cent pure as to variety. These breeder’s seed is ready now for increase
of foundation seed. A portion of this breeder’s seed should be retained by the breeders to sown a
continuation breeders seed of the variety.

Maintenance of breeder’s seed of established varieties

The breeder’s seed of established varieties could be maintained satisfactorily by any one of
the following methods
a) By raising the crop in isolation: The breeder’s seed of local varieties could be
maintained by growing them in isolated plots and by very rigorous roguing during various stages
of crop growth, where the various plant characters are observable. The method of handling the
breeder seed crop is the same as described earlier for breeder’s seed of newly released varieties.
b) By bulk selection: The genetic purity of established varieties could be satisfactorily
improved by bulk selection. In this method 2,000 to 2,500 plants typical of the variety are selected,
harvested ,and threshed separately. The seeds from each plant are examined and any pile which
shows any obvious off-types, or otherwise appears dissimilar, are discarded. The remaining piles
of seed are bulked to constitute the breeder’s seed. The other practices of handling remains the
same.
Carry-over Seed The breeder must carry-over at least enough seed to safeguard against,
the loss of variety if there is a complete failure during the foundation seed multiplication phase. In
addition, the breeder should further safeguard variety by arranging to have a portion of the seed
originally released stored under the ideal conditions.

Intellectual Property Rights

Intellectual property is a legal field that refers to creations of mind such as musical literacy,
artistic work, inventions symbols, names, images, and designs used in commerce,
including copyright, trademarks, patents & related rights. Under Intellectual Property Law the
holders of one of these abstract properties have certain exclusive rights to the creative works,
commercial symbols or inventions which is covered by it.

• Depending on the nature of innovation, the subject matter involved and the manner
in which it is used for gaining economic benefit, the intellectual property is divided
into two basic groups; the industrial property and copyrights.
• Copyright automatically assigns the exclusive right to the creative owner of a
literary, all performing art like dramatic, musical or artistic work, paintings,
photographs, sculptures advertisements, maps, technical drawings, cinematograph
film, photography, sound or video recording, broadcast/telecast, phonograms, a
computer programme, etc to reproduce the work, to issue copies to or perform or
communicate in public, make any translation or an adaptation of the work, offer on
sale or rental and to authorize others for doing so.
• The industrial property includes patents, trademarks, industrial designs, trade
secrets and geographical indications. Among these, patent assumes highest
importance in view of its impact on technological and economic aspects of peoples
and nations. The earliest industrial property act was enacted in Europe.
• This was followed by legislation of US Patent Act in 1793. In 1883, eleven
western countries joined together to establish the Paris Convention for the
Protection of Industrial Property, which was aimed to harmonize the IPR law and
practices across member countries. Currently 169 countries are its members. Pre-
Colonial India had an IPR regime that was practiced by the Britain.
 • India is a signatory to the TRIPS Agreement hence it modified its patents law in
conformity with TRIPS Agreement. We should also consider the use of
Information and Communication Technology (ICT) for an effective enforcement
of the Intellectual Property Rights in India. It is desirable that more initiatives on
the lines of ICT HELPDESK must be undertaken so that contemporary
International Standards can be adopted in India too.
The protective umbrella of TRIPS covers;
1. Copyright and Related Rights
2. Trademarks
3. Geographical Indications
4. Industrial Designs
5. Patents
6. Layout designs of Integrated Circuits and
7. Protection of Undisclosed Information.
• Intellectual property rights give creators exclusive rights to their creations, thereby
providing an incentive for the author or inventor to develop and share the
information rather than keep it secret. The legal protections granted by Intellectual
property laws are credited with significance contributions toward economic growth.
• Economists estimate that two third of the value of large businesses in the USA can
be traced to intangible assets, Likewise industries which rely on intellectual property
protections are estimated to produce 72% more value per added employee than non
intellectual property industries.
• Intellectual Property (IP) has emerged as a strong tool for building and sustaining
competitive advantage in the contemporary knowledge economy. Both domestic and
international competition is boosted by the scientific and technological progress in
a sustainable manner by utilizing the Intellectual Property Rights (IPRs).
• There is a direct rationale for providing a healthy and global system for protection
of IPR, due to their role in wealth creation. Moreover, the technology, investment
and trade, which are key drivers of economic growth, are imbedded in IPR
protection.
• The Indian Intellectual Property Office is dedicated to mobilize the use of
intellectual property for economic and social development by creating an IP
(Intellectual Property) culture and enhancing knowledge & competencies is
synchronizing with the global environment.
Orthodox or conventional IPRs
These are the conventional IPRs used by individuals, companies, association of persons etc.
• They are further segregated in different types like Copyright, Patent and Trademark.
Copyright
Copyright is an IPR deals with expression of idea not an idea itself. Copyright comes into
existence immediately after expression and requires no formal registration.
• Copyright is an expression and manner of expression.
• It protects original literary works, ie books, novels, lyrics, songs, computer programmes,
etc.
• Whenever these are created, they become the property of the creator.
Copy right law is used in creative and artistic works (For example: books, movies, music,
paintings, photographs and software) and gives the holder the exclusive right to control
reproduction or adaptation of such works for a certain period of time (historically, a period of
between 10-30 years depending upon jurisdiction, but more recently the life of the author, plus
several decades)

Patents
• Patent is a right exclusively used for ideas but it is of commercial use. Patents are
generally of three types viz. Novelty, non-obviousness and novel art.
• A patent is the right of an individual or of a company/ organization to gain profit
from a particular invention or unique manufacturing process.
• A patent is an intellectual property relating to scientific and technological
inventions.
• It is granted by the government of the country to the applicant and gives the inventor
the right for a limited period to prevent others from using that invention in any form
without permission.
• Like any other form of property, a patent can be transacted purchased, sold or even
mortgaged.
• Patent law protects inventions it gives the patent holder a right to prevent others
from participating and practicing the inventions without a license from the inventor
for a certain period of time.
• Patents can protect the functional features of a process, machine, manufactured item,
asexually reproduced plant or composition of matter.
 • The main Amendments to Indian Patent Act are; new chemical entity will not be
treated as patent, they must pass the test to complete patent and registration in any
member country is applicable to all members of WTO.
Trademark
• Association of traders name with the commodity traded gives rise to trade mark.
• A trademark is a distinctive sign, which is used to distinguish the products or
services of different businesses.
• Trade mark law protects words, phrases, logos, symbols, sound or even smell or
combinations used to distinguish one product from another, which gives a distinct
identity to a good.
• In circumstances where a competitor uses a protected trademark can go to court and
obtain an injunction to stop the use.
• Trade secret protection helps protect the secrets of a product or work.
Unorthodox or Innovative IPRs
• These are the new entrants in the field of IPRs.
• All due to globalization and whole world has become a single platform, so do these
gained importances.
• They include Industrial Design, Geographical Indications, Plant Varieties, Semi
conductor layout design (SCLD).
• Out of these four semi conductor layout design is more vulnerable to copying. SCLD
law was brought in reference to agreements of TRIPs.
• A design is the presentation of a product resulting from features of colour, size,
shape, texture or materials of a product or packaging.
• An industrial design right protects the form of appearance, style or design of an
industrial object for example: Spare parts, Furniture, textiles etc.
Cyber Law
• It is a term used to describe the legal issues related to use of communications
technology, particularly cyberspace i.e. the internet.
• It is an interaction of many legal fields, including intellectual property, privacy,
freedom of expression and jurisdiction.
• In essence cyber laws are designed for the physical world to human activity on the
internet
Geographical Indications of goods
 • In recent years, the Indian government has become aware of the importance of GI,
following the debacles related to Basmati and Turmeric GI is a notice of a definite
product have been produced in a particular place.
• The producer can use this sign only for products from the specified region.
A geographical indication (GI) is a sign used on products that have a
specific geographical origin and possess qualities or a reputation that are due to
that origin. ... Geographical indications are typically used for agricultural
products, foodstuffs, wine and spirit drinks, handicrafts, and industrial products.
A geographical indication (GI) is a name or sign used on certain products which
corresponds to a specific geographical location or origin (e.g. a town, region, or
country). ... Darjeeling tea became the first GI tagged product in India, in 2004-05,
since then by oct 2018, 321 had been added to the list.
• Unlike a trademark however the GI is not an individual property for use by the
geographical indication of that region which any producer may use.
Organizations involved in IPR in India
The Ministry of Commerce and Industry, Ministry of Human Resource development, Ministry of
Information and Broadcasting, CII – Confederation of Indian Industries, FICCI- Federation of
Indian Chambers of Commerce and Industries, ASSOCHAM- Associated Chambers of
Commerce and Industries.
• Department of Science and Technology (TIFAC-Technical Information Forecasting and
Assessment Council)
• World Intellectual Property Organization (WIPO) of the UN, based in Geneva is the apex
intergovernmental body dealing with IPRs.

WTO-a Major Shift in IPRs in Agriculture

As Indian IP act does not permit the patenting of plants and animals which are existing in nature,
Govt. of India has developed its own sui generis (a Latin phrase meaning ‘of their own kind’)
system to provide a frame work for Plant Variety Protection and Farmers Right (to comply with
WTO).
Plant Varieties Protection and Farmers Rights Act (2001)
The Plant Varieties Protection and Farmers’ Rights Act 2001 enacted by Government of India has
several unique features.
• The act ensures Plant Breeder’s Rights on New varieties of seeds.
 • The law also grants Farmer’s Rights.
• The act also include for setting up of a Plant Varieties Protection and Farmers’ Rights
Authority and its Regional Centres, National Community Gene Fund, Compulsory
Licensing and Protection of Public Interest Appellate Board.
Plant Varieties Protection and Farmers Rights Act (2001)(PPVFR)
Criteria for registration of varieties UPOV1991
• Registration of the variety will not be done in case the variety does not fulfill the criteria.
• Novelty: A variety should not have been commercially exploited more than one year.
• Distinctiveness: Distinguishable for characters.
• Uniformity: uniform in appearance in specified environment.
• Stability. Stable in appearance over successive generations in specified environment.
Rights of Breeder or his Successor
A certificate of registration for a variety shall confer an exclusive right on the breeder or his
successor, his agent or license to Produce, Sell, market, Distribute and Import or export the
variety
Farmers’ Right
• Farmer who has bred or developed a new variety is entitled for protection as a breeder of a
variety at free of cost.
• Farmer, who is engaged in conservation of genetic resources of land races and wild species
entitled for recognition and reward from the National Gene Fund.
• Farmers will be entitled to save, use, sow, re-sow, exchange, share, store and sell his farm
produced seed of a variety, protected under this act.
• However the farmer is not entitled to sell as branded seed of a protected variety.
• The pivotal importance of the farmer having the right to sell seed has to be seen in the
context of seed production in India, where the farming community is the largest seed
producer providing about 80 per cent of the country’s annual requirement.
VARIETAL RUNDOWN AND RENOVATION

1) Causes for varietal run down or Genetic deterioration in released varieties Normally the
farmers are advised to renew the cultivars once in three years.
The main reason is that a variety may undergo genetic deterioration by a number of ways.
They are : 1. Presence of crossable genera or species in the near by field or bunds
E.g. (1) In the rice field there may be other graminaceous grasses which can hybridise with
rice./ Presence of red rice in varieties is due to this.
(2) Presence of Johnson grass (Sorghum halepense) as weed in sorghum (S. bicolor) field
will lead to varietal contamination due to natural crossing.
2. Lack of isolation distance in the seed production plots Each crop variety requires proper
isolation distance for maintenance of varietal purity. For Eg. Sorghum : 400m Red gram :
200m Sunflower : 600m Lack of isolation distance lead to natural crossing and genetic
deterioration
3. Genetic drifft due to sampling error
The genetic equilibrium in a variety will be disturbed due to improper selection. This
is high in case of small populations. This can be prevented by adopting proper selection
procedure and following phenotypic disassortative mating.
4. Natural mutation : Though the frequency of natural mutation is very low, it is also one
of the causes of varietal rundown. Micro mutations which cannot be detected easily will
lead to genetic deterioration in crop plants.
5. Admixture due to Farm machinery : Improper cleaning of farm tools and machinery
like threshers will also lead to varietal admixtures, natural crossing and rundown.
6. Threshing floor admixtures : Threshing floor must be free from cracks and crevices so
that while threshing and drying there is no chance for left over seed in threshing floor.
Otherwise some seeds may be caught up in cracks and get admixed with other varieties.
7. Store room admixtures : The gunny bags and other container used for seed storage must
be properly cleaned; otherwise it will also lead to admixture.
8. Physiological stresses: Extreme drought conditions will prevent panicle exertion in full
e.g. sorghum. Growing rice in colder months may lead to physiological awning.
9. Not following proper crop rotation practices : The left over seeds may germinate and
contaminate the subsequent crop varieties.
Eg. groundnut after groundnut.
2) Steps to prevent genetic deterioration
1. Nucleus seed production and maintenance Cent per cent purity is to be maintained in
nucleus seed production plot. Different methods are advocated for different crops in
maintenance of nucleus seeds.
For eg. in cotton mass pedigree method is followed for maintenance of nucleus seed. In
this method 1000 to 2000 single plants are raised in replicated progeny row trial. Each and
every single plant is examined for pollen colour and petal colour to maintain genetic purity.
If off types are seen, then the whole line in all the replication will be rejected.
 Selfing is done to prevent contamination Harvest is done on single plant basis and
progenies are selected on single norm.
2. By providing proper isolation distance for seed multiplication plots For
eg. for sorghum nucleus seed production plot 800 metre isolation distance is maintained.
The single plants are raised and allowed for sibmating.
3. Removal of all grasses from field as well as bunds : This is to be followed especially
in case of rice.
4. By following proper crop rotation
5. By proper cleaning of farm equipments, tools, threshing floor, gunny bags and store
room
6. By following proper selection procedures in seed production plots For eg. in groundnut
seed production plot, the plot mean for yield will be worked out. Then SE and CD will be
worked out. The single plant yield which are around = 2 SE is to be selected for further
maintenance.
7. By following the proper varietal maintenance technique E.g. In sunflower, varietal
renovation technique as advocated by Pustovoit will have to be followed.
3) Varietal renovation in sunflower Russian scientist Pustovoit has given the method of
varietal renovation.
It is called as Pustovoit method of renovation. Sunflower varieties all called as population.
Due to heterozygous nature, the variety to be renovated is raised under isolation of 600m.
Rouging should be done. About 10,000-12,000 plants are selected based on head size, seed
size, seed yield and oil content. The mean and standard deviation is calculated for each
character. The average was taken. In all the characters value for an individual must exceed
the value of mean +2 SD. Then that individual is selected. Then the selected plants are
studied for disease resistance and progeny row testing. Progeny row testing is replicated
twice. In each time the plants are selected and the characters are recorded and Standard
Deviation (SD) and mean are worked those individuals whose character value exceeding
mean + 2 SD are selected. While using for progeny row testing only half of the seeds are
reserved. After selecting the plants the remnant, seeds of the selected plants are used for
raising super elite seeds at 600m isolation. Rouging should be done before and after
flowering. Super elite seeds are used for raising the elite seeds or Stage I foundation seed.
These seeds are used for raising Stage II foundation seed. These seeds are used for raising
certified seeds and then for commercial cultivation. This seed renovation method maintain
yield and oil content and also sometimes upgrade them.
BREEDING FOR BIOTIC AND ABIOTIC STRSSES
Stress: Constraining influence, force, pressure or adverse conditions for crop growth caused by
biological or environmental factors.
Biotic (living) : Adverse effects due to pests and diseases abiotic stresses
Abiotic (non living) : Adverse effects on host due to environmental factors eg: Drought, water
logging, heat, cold, salinity, alkalinity and air pollution etc.
Host : Plant effected by a disease or which can accommodate pathogen.
Pathogen : An organism that produces the disease
Disease : an abnormal conditions in the plant caused by an organism (pathogen)
Pathogenicity : The ability of a pathogen to infect a host strain
Virulence : Capacity of a pathogen to incite a disease
Avirulence : The inability of a pathogen to cause or incite a disease
Physiological race : Strains of a single pathogen species with identical or similar morphology but
differ in pathogenic capabilities.
Pathotype : Strains of a pathogen classified on the basis of their virulence to known resistance
genes present in the host.
Epidemic : Severe and sudden out break of disease beginning from a low level of infectio

MECHANISMS OF DISEASE RESISTANCE:

There are different ways of disease resistance viz., disease escape, disease endurance or
tolerance disease resistance and immunity
1. Disease escape : The ability of susceptible host plants to avoid attack of disease due to
environmental conditions factors, early varieties, charge in the date of plating, change in the site of
planting; balanced application of NPK etc.
Eg. Early varieties of groundnut and potato may escape ‘Tikka’ and ‘Late blight’ diseases
respectively since they mature before the disease epidemic occurs. Changing planting season in
sugarcane from June to October has successfully escaped leaf-rust.
Virus free seed potato is produced by sowing the crop in October in Jullundher and other
places instead of November, the normal planting time.
2. Disease endurance or tolerance : The ability of the plants to tolerate the invasion of the
pathogen without showing much damage. This endurance is brought about by the influence of
external characters. Generally, tolerance is difficult to measure since it is confounded with partial
resistance and disease escape. To estimate tolerance the loss in yield and some other trait of several
host varieties having the same amount of disease eg., leaf area covered by disease etc., is compared.
Eg. In Barley the variety Proctor shows 13% yield loss as compared to 20% loss in the varieties
Zephy and Sultan. Wheat varieties when fertilized with potash and phosphorus are more tolerant
to the rust and mildew infection.
· The Rice crop fertilized with silicate is resistant to blast infection in Japan.
3. Disease Resistance : The ability of plants to withstand, oppose or overcome the attack of
pathogens. Resistance is a relative term and it generally refers to any retardation in the development
of the attacking pathogen. In case of resistance, disease symptoms to develop and the rate of
reproduction is never zero i.e., r? o but it is sufficiently lower than 1 (the rate of reproduction on
the susceptible variety) to be useful. The inhibition of growth of the pathogen is believed to be
nutrional in nature and in some cases chemical growth inhibitors may be involved.
Resistance is largely controlled by inherited characters i) may be controlled by single dominant
gene in Ottawa 770 B, Newland flax variety, wheat all rusts NP 809
4. Immunity: When the host does not show the symptoms of disease it is known as immune
reaction. Immunity may result from prevention of the pathogen to reach the appropriate parts of the
host e.g. exclusion of spores of ovary infecting fungi by closed flowering habit of wheat and barley.
It is more generally produced by hypersensitive reaction of the host usually immediately after the
infection was occurred. In immune reaction the rate of reproduction in zero i.e. r = 0
5. Hypersensitivity: Immediately after the infection several host cells surrounding the point of
infection are so sensitive that they will die. This leads to the death of the pathogen because the rust
mycelium cannot grow through the dead cells. This super sensitivity (hypersensitivity) behaves as
a resistant response for all practical purposes.
Phytoalexins are specific polyphenolic or terpenoid chemicals and are produced by the host in
response to the infection by a pathogen. More than 30 different phytoalexins have been identified.
Phytoalexins are either fungicidal or fungistatic. Eg. Rust fungi and virus attack.
Factors for disease resistance (Causes of Disease resistance)
The disease resistance may be caused due to
1. Morphological, structural and functional characteristics which prevents the entrance of the pathogen
i.e. prevents the first stage of infection.
2. Biochemical or anatomic al properties of tissue which prevent the establishment of parasitic
relationship.
a. Morphological characters
Certain morphological features of the host may prevent infection.
Eg. Resistance to Jassid attack in cotton has been shown to be correlated with the hariness of varieties
: hairy type resists the attack more, than glabrous types.
Failure to germinate rust spores on the leaves of the barley due to waxy coating.
Young sugarbeet leaves practically immune to attack of the circos pora because the stomata size is very
small.
b. Physiological characters
Protoplasmic factors or chemical interactions :
By virtues of its chemical composition the protoplasm may exert an inhibitory influence on the
pathogen bringing about the desired resistance in the plant.
Eg. : Resistance of grape to powdery mildew is highly correlated with the acidity of cell sap.
Presence of toxic substance in the red pigment in the coloured onions. The outer scales resist
the smudge fungus attack when the scales are removed they become susceptible.
c. Anatomical: More secondary thickening of the cell walls of resistant potato varieties which
resists the mechanical puncture of the invading Pythium pathogen.
d. Nutritional factors : Reduction in growth and in spore production is generally supposed to be
due to unfavourable physiological conditions within the host. Most likely a resistant host does not
fulfill the nutritional requirements of the pathogen and thereby limits its growth and reproduction.
e. Environmental factors : In addition to the above the environmental factors have marked effect
on the pathogen attack. Temperature, moisture, humidity and soil PH and fertility status of the soil effects
the pathogen reaction greatly.
Genetic basis of disease resistance
The first study on genetics of disease resistance was done by Biffen in 1905. He reported the inheritance
of resistance to leaf rust of wheat variety Rivet in crosses with some susceptible varieties. In F 2 there
were 3 susceptible : 1 resistant plants indicating that resistance was controlled by a single recessive
gene. Most of the earlier studies were conducted without taking into consideration the physiological
specialization (pathotype differentiation) of the pathogen which can materially influence the
conclusions drawn. It is now recognized that disease resistance may be inherited in three different ways
:
Oligogenic
Polygenic and
Cytoplasmic inheritance
Oligogenic inheritance:
The disease resistance is governed by one or few major genes and resistance is generally dominant to
the susceptible reaction. The action of major resistance genes may be altered by modifying genes in
many cases. Eg. bunt resistance in Wheat. Oligogenes generally produce immune reaction. The chief
characteristic of the oligogenic disease resistance is pathotype specificity,
i.e. resistant gene is effective against some pathogens, while it is ineffective against the others. In most
cases, there are a number of major genes that determines resistance to a particular disease Eg. more than
20 different resistance genes are known for leaf rust of wheat, while those for stem rust resistance exceed
30. The genetics of oligoganic resistance has advanced by two events viz.,
1. Discovery of a resistance gene to the prevalent pathotype and
2. Evolution of a pathotype virulent to the new resistance gene.
Oligogenic resistance is synonymous to vertical resistance.
Gene for gene hypothesis:
The concept of gene for hypothesis was first developed by Flor in 1956 based on his studies of
host pathogen interaction in flax rust caused by Malampsora lini. The gene for gene hypothesis states
that for each gene controlling resistance in the host, there is a corresponding gene controlling
pathogenicity in the pathogen. The resistance of host is governed by dominant genes and virulence of
pathogen by recessive genes. The genotype of host and pathogen determine the disease reaction. When
genes in host and pathogen match for all the loci, then only the host will show susceptible reaction. If
some gene loci remain unmatched, the host will show resistant reaction. Now gene-for -gene
relationship has been reported in several other crops like potato, Sorghum, wheat etc.
The gene for gene hypothesis is known as “Flor Hypothesis”.

Vertifolia Effect : Vander plank introduced the term vertifolia effect and refers to epidemic
development in a variety carrying vertical resistance genes (oligogenes) leading to heavy economic
losses. Total failure of vertical resistance leading to a disease epidemic is known as
vertioalia effect. This failure occurs because of two reasons :
1. The level of horizontal resistance in varieties carrying oligogenes is usually low and
2. The pathogen is able to evolve new virulent pathotypes.
Polygenic inheritance
In this type the disease resistance is governed by many genes with small effects and a continuous
variation for disease reaction is produced. The genes show additive and non additive effects and the
environmental effect is also observed. The polygenic resistance does not show pathotype-specificity
as against the oligogenic resistance. It is almost same as horizontal resistance. In some cases the
polygenic inheritance may have a oligogenic component, the oligogenes acting in an additive
manner eg. bacterial blight resistance in cotton
Cytoplasmic inheritance :
Resistance in some cases is determined by cytoplasmic genes or plasma gene(s). Eg. The T-
male sterilizy cytoplasm (cms-T) in maize is extreamly susceptible to Helminthosporium
leafblight, while the non-T cytoblasms are resistant to this disease.

Vertical and Horizontal Resistance (Van der plank)

Vertical Resistance is generally d etermined by major genes and is characterized by
pathotype specificity. Clearly immune or susceptible response in the case of vertical resistance
depends on the presence of virulent pathotype. When virulent pathotype becomes frequent,
epidemics are common in the cases of vertical resistance. Thus an avirulent pathotype will produce
an immune response i.e. r=0 or close to 0 but the virulent pathotype will lead to susceptible reaction
i.e. r=1. It is also known as race specific, pathotype specific or simply specific resistance.
Horizontal Resistance
Race non-specific, pathotype -nonspecific and partial, general or field resistance.
Horizontal resistance is generally controlled by polygenes i.e. many genes with small effects and it
is pathotype nonspecific. In this case, the reproduction rate is not zero but it is less than one. Poly
genes, govern horizontal resistance.
Methods of Breeding for Disease Resistance
The methods of breeding for disease resistance are essentially same as those used for
other agronomic traits. They are :
1. Introduction
2. Selection
3. Hybridization
4. Budding & Grafting
5. Mutation Breeding
6. Biotechnological methods.
1. Introduction : Resistant varieties may be introduced for cultivation in a new area.
Eg. · Early varieties of groundnut introduced from USA have been resistant to leaf spot
(Tikka), · Kalyanasona and Sonalika wheat varieties originated from segregating material
introduced from CIMMYT, Mexico, were rust resistant. African bajra introductions have been used
in developing downy mildew resistant cms lines.
2. Selection : Selection of resistant plants from commercial varieties is easiest method.
Eg. · Kufri Red potato is selection from Darjeeling Red round
· Pusa Sawani behind (yellow mosaic) selection from a collection obtained from
Bihar, · MCU I was selection from CO4 for black arm resistance in cotton
3. Hybridization : Transfering disease resistance from one variety or species to the other.
a.Pedigree method is quite suitable for horizontal resistance. Artificial disease epiphytolics are
produced to help in selection for disease resistance.
Eg. In wheat Kalyana Sona, Sonalaka, Malvika 12, Malvika 37, Malavika 206, Malavika 234
Laxmi in Cotton (Gadag 1 x CO2) for leaf blight resistance
b. Backcross method is used to transfer resistance genes from an undersirable agronomic variety to
a susceptible, widely adoptable and is agronomically highly desirable variety.
If the resistant parent is a wholly unadapted variety, backcross method is a logical choice.
If resistant variety also possess some good qualities then chose pedigree method of handling
segregating material.
4. Budding & Grafting : The disease resistance in vegetatively propogated material is transferred
by adopting either by budding or grafting. By grafting or budding the resistant
material, the resistance can be transferred.
5. Mutation Breeding : When adequate resistance is not available in the germplasm ; Mutation
breeding is resorted to induce resistance. This is also us ed to break the linkages
between desirable resistant genes and other desirable genes.
Precautions
1. The donor parent must possess the required amount of resistance
2. It must be simply inherited without any linkage
3. The recovery in the recipient parent should be more
4. Proper condition for full expression of the resistant genes has to be provided
Advantages with breeding for disease resistance
1. Helps in reducing the losses caused by patogens
2. Reduces the high cost of disease control by chemical treatment
3. Helps to avoid the use of poisonous fungicides
4. Only method available to some specific diseases like viruses, wilt etc.
Limitations
1. Linkage of resistant genes with genes of inferior quality
2. Occurrence of physiological races of varying capacities
3. Self sterility in host plants
Utilization and achievements
1. Rice ADT 10 x CO4 (resistant to blast)
2. Potato Solanum tuberosum x Solanum demissum
(susceptible to late blight) (wild resistant to late blight)
F1 backcrossed with Sol. tuberosum
Resistant variety
Varieties resistant to different diseases
Rice : Blast Co25, Co26,
Wheat : all three rusts : NP 809
Yellow rust : NP 785, NM86
Black rust : NP 789
Brown rust : NP 783, NP 784
Sugarcane : Red rot Co 419, Co 421, Co 527
Cotton : Wilt Vijay, Kalyan, Suyog
Groundnut : Tikka, leafspot, Ah 45

INSECT RESISTANCE BREEDING

Biotypes : Strains of a species of an insect pest, differing in their ability to attack different
varieties of the same host species (syn: Physiological races)
Host Habitation :
1. Polyphagy 3. Seasonal Oligophagy
2. Oligophagy 4. Monophagy
1. Polyphagy : Insects feed on a vide range of hosts avoiding few plant species. Eg. Scales
& moths.
2. Oligophagy : Live on one taxonomic unit only. Eg. Hessianfly on wheat
3. Seasonal oligophagy : Insects may live on many species in one part of the year and on
few in another part of the year. Eg : Aphids.
4. Monophagy : Avoid all hosts except one particular species or variety Eg. Boll weevil on
cotton.
Mechanism of Insect Resistance :
Insect resistance is grouped into four categories :
1. Non preference 2. Antibiosis
3. Tolerance 4. Avoidance
1. Non preference : Host Varieties exhibiting this type of resistance are unattractive or unsuitable
for colonization, oviposition or both by an insect pest. This type of resistance in also termed as non-
acceptance and anti-xenosis. Non preference involves various morphological and biochemical
features of host plants such as – color, hairness, leaf angle, taste etc.
2. Antibiosis : Antibiosis refers to an adverse effect of feeding on a resistant host plant on
the development and/or reproduction of the insect pest. In severe cases, it may even lead to the
death of the insect pest. Antibiosis may involve morphological, physiological or biochemical
features of the host plant; some cases of insect resistance involve a combination of features. Eg.
Resistance to BPT is due to antibiosis & non preference
3. Tolerance : An insect tolerant variety is attacked by the insect pest to the same degree as
a susceptible variety. But at the same level of infestation, a tolerant variety produces a higher yield
than a susceptible variety. Ability of the host plant to withstand the insect population to a certain
extent which might have damaged a more susceptible host.
Tolerance is mainly a host character and it may be because of greater recovery from pest
damage. Eg. Rice varieties tolerant to stem borer/gall midge produce additional tillers to
compensate yield losses (as in stem borer in sorghum) or due to the ability of host to suffer less damage
by the pest eg. aphid tolerance in Sugarbeet & Brassica sps. And green bugs tolerance in cereals.
Inheritance of tolerance is complex in many cases and is supposed to be governed by polygenes.
4. Avoidance : Pest avoidance is the same as disease escape , and as such it is not a case of true resistance
Mostly insect avoidance result from the host plants being at a much less susceptible developmental
stage when the pest population is at its peak.
Eg. 1. Early maturing cotton varieties escape pinkboll worm infestation, which occurs late in the season.
Nature of Insect Resistance / Factors for insect -resistance
Insect resistance may involve :
1. Morphological
2. Physiological (or)
3. Biochemical features of the host plant
1. Morphological features : Morphological factors like, hairiness, colour, thickness and toughness of
tissues etc. are known to confer insect resistance.
a) Hairiness of leaves is associated with resistance to many insect pests leaf beetle in cereals, in cotton
to Jassids , in turnip to turnip aphid.
b) Colour of plant : Color may contribute to non preference in some cases.
For example : Red cabbage, Red leaved brussel’s sprouts are less favored than green varieties by
butterflies and certain Lepidoptera for oviposit ion. Boll worms prefer green cotton plants to red ones.
c) Thickness and Toughness of plant – Tissues prevent mechanical obstruction to feeding and
oviposition and thereby lead to non-preference as well as antibiosis.
Eg.
1. Thick leaf lamina in cotton contributes to Jassid resistance
2. Solid stem in wheat confers resistance to wheat stem sawfly
3. Thick and tough rind of cotton bolls makes it difficult for the boll worm
larve to bore holes and enter the bolls.
Other characters : also contribute to insect resistance.
Eg. 1. Gossypium arboretum varieties with narrow lobed and leathery leaves are more
resistant to Jassids than are those with broad lobed and succulent leaves.
2. Cotton varieties with longer pedicels are more resistant to boll worms.
2. Physiological Factors : Osmotic concentration of cell sap, various exudates etc; may be
associated with insect resistance.

Eg.
1) Leaf hairs of some solanum sps. secrete gummy exudates. Aphids and coloradobeetles get trapped in
these exudates.
2) Exudates from secondary trichomes of Medicago disciformis leaves have antibiotic effects on alfalfa
weevil.
3) Cotton- High osmotic concentration of cell sap is associated with Jassid resistance.
3. Biochemical Factors : Several biochemical factors are associated with insect resistance in many
crops. It is believed that biochemical factors are more important than morphological and physiological
factors in conferring non-preference and antibiosis.
Eg. 1) High concentrations of gossypol is associated with resistance in several insect pests
in cotton.
2) In rice – high silica content in shoots gives resistance to shoot borer
Genetics of Insect Resistance
Insect resistance is governed by -
1. Oligogenes 2. Polygenes 3. Cytoplasmic genes
1. Oligogenic Resistance : Insect resistance is governed by one or few major genes or ligogenes, each
gene having a large and identifiable individual effect on resistance.
Oligogenic resistance may be conditioned by the dominant or the recessive allele of the oncerned
gene. The differences between resistant and susceptible plants are generally large and clear-cut. In
several cases, resistance is governed by a single gene (monogenic resistance)
Eg. In wheat to green bugs In cotton to Jassids
In apple to woolly aphis In rice to plant & leaf hopper
2. Polygenic Resistance : It is governed by several genes, each gene producing a small and
usually cumulative effect. Such cases of resistance.
1) Involve more than one feature of the host plant
2) Are much more durable than the cases of oligogenic resistance.
3) Difference between resistance & susceptible plants are not clear cut
4) Transfer of resistance is much more difficult
Examples for polygenic resistance
1) In wheat to cereal leaf beetle
2) In alfalfa to spotted aphid
3) In rice to stem borer
4) In maize to ear worm and leaf aphid
Evolution of resistance breaking biotypes is almost rare.
3. Cytoplasmic Resistance : governed by plasmagenes
Eg. 1. Resistance to European corn borer in maize
2. Resistance to root aphid in lettuce
Sources of Insect Resistance
1. A cultivated variety 3. A related wild species
2. Germplasm collections. 4. An unrelated organisms
1. Cultivated variety : Resistance to many insect pests may be found among the cultivated
varieties of the concerned crop.
Varieties - SRT 1, Khand waz ; DNJ 286 and B 1007 of G. hirsuturn are good sources of
resistance to Jassids.
2. Germplasm collection :
Eg.
1) In apples for rosy apple aphid, green apple, apple maker and apple saw-fly.
2) In cotton, several strains resistant to Jassids.
3. Related wild species :
Eg.
1) Resistance to both the species of potato nematodes has been transferred fro
Solanum vernei to potato
2) Jassid resistances is known in wild relatives of cotton G. tomentosum;
G.anomalum and G.armourianum
4. An unrelated organism : It is done through recombinant DNA technology
a) T he ‘Cry’ gene of Bacillus thuringiensis is the most successful example.
Other genes of importance are the
b) Protease inhibitor encoding genes found in many plants eg. the cowpea pea, trypsin
inhibitor (cp TI) gene.
Breeding Methods for Insect Resistance
1. Introduction 2. Selection
3. Hybridization 4. Genetic Engineering
1. Introduction :
Eg. Phylloxera vertifoliae resistance grape root-stocks from U.S.A. into france.
2. Selection :
Eg.
1) Resistance to potato leaf hopper
2) Resistance to spotted alfalfa aphid
3. Hybridization : Pedigree oligogenic characters
Back cross Polygenic characters
4. Genetic Engineering : B.theningiensis (cry gene) resistance in maize, soybean, cotton etc.
Screening Techniques for determining resistance
The most crucial and, perhaps , the most difficult task in breeding for insect resistance is the
identification of insect resistant plant during segregation generations. There are two types of
screeings
1. Field Screening 2. Glass house screening
Field Screening :
The techniques designed to promote uniform infestation by an insect pest in the field are
1. Inter planting a row of known susceptible variety between two rows of testing material.
2. Screening in highly prone areas
3. in case Soil insect pests to be tested in sick plots only
4. Testing in a particular season when the infestation is very high.
Eg. Rice stem borer in off season.
5. Transferring manually equal number of eggs or larvae to each test plant.
Glass house screening
Result from glass house tests are much more reliable than those from field tests since
both the environment and the initial level of infestations are more or less uniform for all the
plants being tested.
Problems in Breeding for Insect Resistance :
1. Breeding for resistance to are insect pest may leads to the susceptibility to another pest.
Eg. Glabrous strains of cotton are resistant to bollworms but susceptible to Jassids.
2. Reduction is quality or make unfit for consumption.
3. Linkage between desirable & undesirable genes. Inter specific varieties are generally low
yielding and their produce is often of inferior quality.
4. Screening for resistance is the most critical and difficult step is a breeding programme it
necessitates a close co-ordination among scientists belonging to different disciplines.
5. It is a long term programme
Achievements
INDIA
1. India – cotton varieties – G 27, MCU 7, LRK 516 – resistant to boll worms.
2. Rice – variety Vijaya – resistant to leaf hopper
 Rice – TKM 6, Ratna – Stemborer

Transgenic crops
Transgenic plants are developed by transferring or modifying genes from another
organism by a diverse technique like physical, chemical and biological methods. Transgenic
plants are obtained by introducing a gene into its genome with the help of vectors in order to
develop a plant with new characteristics. This process of recombinant DNA technology is used
for developing genetically modified plants to embolden the variety against pests, diseases and
stress or to improve the quality of the product. These can also be achieved by conventional
breeding if the genes for the desired characters are
present in close or wild relatives else transgenesis is the only viable alternative. Apart
from this, genetic engineering can overcome the other limitations of conventional breeding such
as large space requirement, time consuming, and uncertainty of the result. It is not mandatory to
opt for transgenics if the problem can be addressed by conventional breeding.
Tissue culture plant seed certification
Government of India has established the “National Certification System for Tissue
Culture Raised Plants(NCS-TCP) ” authorizing Department of Biotechnology, Ministry of
Science & Technology as the Certification Agency vide the Gazette Notification dated 10th March
2006 under the “Seeds Act, 1966” for ensuring production and distribution of quality tissue culture
planting materials.
The purpose of NCS-TCP is to ensure production and distribution of quality tissue culture
planting materials. NCS-TCP is a unique quality management system, first of its kind in the world,
which ensures recognition of Tissue Culture Production Facility for the production of quality
planting material and certification of end products.
Requirements for Certification of Tissue Culture Raised Plants
• The mother plant tissue/stock culture must be tested for freedom from all known viruses from ATLs
(Accredited Test Labs) or any reputed Government institutions. (eg. NRCB,
• The respective batch (lot) of tissue culture plants should be derived from tested stock culture as
stated above.
• Recognized TCPFs should assign 4 digits batch number to the said batch of tissue culture plants.
This above number should be provided to ATLs while sending samples for certification.
• Tissue culture raised plants ready to dispatch to the farmers (ideally secondary hardened) will be
tested for all known viruses and true to type in order to certify under NCS-TCP