MULTILINE BREEDING

Generally, pureline varieties are highly adapted to a limited area, but are generally
poorly adapted to wider regions. Further, their performance is not stable from year to year
because of changes in weather and other environmental factors. Purelines often have only one
or a few major genes for disease resistance, such as, rust resistance, which make them
resistant to some races of the pathogen. New races are continuously produced in many
pathogens which may overcome the resistance present in the pureline varieties. For example,
Kalyan Sona wheat (T. aestivum) originally resistant to brown rust (leaf rust), soon became
susceptible to the new races of the pathogen. To overcome these limitations, particularly the
breakdown of resistance to diseases, it was suggested to develop multiline varieties. Multiline
varieties are mixtures of several purelines of similar height, flowering and maturity dates,
seed colour and agronomic characteristics, but having different genes for disease resistance.
The purelines constituting a multiline variety must be compatible, i.e., they should not reduce
the yielding ability of each other when grown in mixture. The idea of multiline varieties was
put forward by Jensen in 1952 for use in cereals. In 1954, Borlaug suggested that several
purelines with different resistance genes should be developed through back- cross
programmes using one recurrent parent. This is done by trans- ferring disease resistance
genes from several donor parents carrying different resistance genes to a single recurrent
parent. Each donor parent is used in a separate backcross programme so that each line has a
different resistance gene or genes. Five to ten of these lines may be mixed to produce a
multiline variety. Which lines are to be mixed would be determined by the races of the
pathogen prevalent in the area. 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.

`DISADVANTAGES

(1) the utility of multilines is limited to high-risk regions for disease outbreaks;

(2) usually there is no genetic improvement for yield or agronomic traits;

(3) substantial labor is required to produce and maintain the component lines;

(4) release of an improved recurrent variety is delayed until the components are produced.

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 To quantify the performance of multilines, Jensen and Federer [34] applied the
concepts and computation of combining ability to competitive ability in wheat. In this
application, general combining ability (GCA) refers to the average performance of a line in
combinations, and specific combining ability (SCA) refers to the deviations in the expected
average performance of combinations. Jensen [32] outlined four different examples for
forming a multiline:

(1) using a single backcross to generate lines for use in the multiline;

(2) crossing unrelated lines;

(3) crossing to different selected recurrent parents; and

(4) making double crosses where each single cross has a common parent.

Marshall and Brown [35] used statistical models to determine the effect of intra-
populational genetic diversity on the stability of performance of mixtures as estimated by
their variance in yield across environments. Their models suggested that, in the absence of
intergenotypic interactions, the yield of a multiline will vary less than the least variant
component when the component lines perform differently in different environments.
Conversely, when there are intergenotypic interactions, the stability of a mixture will be more
stable than the best line only when each component responds differently to different
environments. They also predicted that, when mixtures are compared to their pure line
components, it is expected that improved stability is more easily attained than improved yield
because improvements in yield require net positive intergenotypic interactions whereas
stability does not.

In addition to stability, the use of multilines has the potential to improve resistance to
disease. Borlaug and Gibler [36] developed wheat lines for multiline cultivars at CIMMYT
using ‘donor parents’ selected from the International Wheat Rust Nursery and backcrossed to
recurrent parents. A number of studies have examined possible mechanisms for the observed
enhancements in disease resistance within multilines, and generally agree that the reduced
inoculum load results from both a lower frequency of initial infection when a spore lands on a
resistant component of the multiline and a lower rate of increase in inoculum. A review of
multilines for disease control was published by Mundt [37].

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 Given the changing climate and the need for greater protection of natural ecosystems
and sustainable agricultural practices, multilines and multiblends will likely play an important
role in the future agricultural production systems. Further research is warranted on durability
of resistance in multilines, experimental design, and design of mixtures.

MAJOR FEATURES OF PURE-LINE SELECTION:

Pure line selection is practised in heterogeneous populations such as introduced

materials,land races and mass selected varieties of self-pollinated species to isolate superior
genotypes.

The main features of pure-lines are briefly presented below:

1. Homogeneous:

All the genotypes of a pure-line are homogeneous i.e. genetically identical and
phenotypically similar. Thus pure-lines are homozygous and homogeneous.

2. Non-Heritable Variation:

The variation within a pure-line is entirely due to environmental factors. Thus the variation
isnon-heritable in the pure-lines.

3. Highly Uniform:

A variety developed by pure-line selection is highly uniform in quality due to absence

ofgenetic variation

TYPES OF MULTILINE:

1. Mixtures of isolines.

2. Mixture of closely related lines.

3. Mixture of unrelated or distinctly different genotypes.

THESE ARE BRIEFLY DISCUSSED BELOW:

PROCEDURE

(1) Selection of Recurrent Parent: The recurrent parent should be a high yielding popular
variety. The recurrent parent should be the best cultiva region.

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(2) Selection of Donor Parents: Parents with resistance to various races of a disease should
be chosen as donor parents. The resistance should be thoroughly examined under artificial
epiphytotic conditions before use of the donor parents in the crossing programmes. The donor
parents should be adapted varieties as far as possible. Because in un-adapted parents disease
resistance is sometimes linked with several un-desirable characters and transfer of resistant
genes from such parents to the recurrent parent becomes difficult task. Several donor parents
are selected to incorporate different resistant genes against various races.

(3) Transfer of Resistance: The resistant genes are transferred from donor parents to the
recurrent parent through a series of several separate backcross programmes. Generally 4-5
backcrosses are sufficient to retain the genotype of recurrent parent with added resistance in
the backcross derivatives. The backcross derivatives are evaluated for disease resistance
during backcrossing and also at the end of backcrossing. The desirable lines from each
backcross are mixed to form an isoline.

(4) Mixing of Isolines: The various isolines developed by various backcrosses are mixed
together to constitute a multiline cultivar. Generally 6-10 isolines are mixed to constitute a
multiline cultivar.

MERITS OF MULTILINE VARIETIES

1. All the lines are almost identical to the recurrent parent in agronomic characteristics,
quality etc. Therefore, the disadvantages of the pureline mixtures (Chapter 11) are not present
in the multiline varieties.

2. Only one or a few lines of the mixture would become susceptible of the pathogen in any
one season. Therefore, the loss to the cultivator would be relatively low.

3. The susceptible line would constitute only a small proportion of the plants in the field.

Therefore, only a small proportion of the plants would be infected by the pathogen.
Consequently, the disease would spread more slowly than when the entire population was
susceptible. This would reduce the damage to the susceptible line as well

DEMERITS OF MULTILINE VARIETIES

1. The farmer has to change the seed of multiline varieties every few years depending upon
the change in the races of the pathogen.

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2. There is a possibility that a new race may attack all the lines of a multiline variety.

ACHIEVEMENTS

Multiline variety appears to be a useful approach to control diseases like rusts where
new races are continuously produced. In India, three multiline varieties have been released in
wheat (T. aesti- vum). Kalyan Sona, one of the most popular varieties in the late sixties, was
used as the recurrent parent to produce these varieties. Variety 'KSML 3' consists of 8 lines
having rust resistance genes from Robin, Ghanate, K1, Rend, Gabato, Blue Bird, Tobari etc.
Multiline 'MIKS 11' is also a mixture of 8 lines; the resistance is derived from E 6254, E
6056, E 5868, Frecor, HS 19, E 4894 etc. The third variety, KML 7405 has 9 lines deriving
rust resistance from different sources.

POPULATION APPROACH TO BREEDING OF SELF-POLLINATED CROPS

Self-fertilization of F_{2} hybrids leads to a very rapid increase in homozygosity.

After only 4 generations of self-pollination, about 94 per cent of the genes would become
homozygous. Even in F, half of the genes are in homozygous state. Thus self- fertilization
quickly separates the progeny from a hybrid into a large nuraber of purelines. As a
consequence, selection in such a segre- gating population only picks out the gene
combinations present in the population primarily as a result of recombination in F_{2} This
reduces the chances of recombination between linked, especially tighty linked, genes and of
recovery of rare transgressive segregants. There is no opportunity for changing the genotype
of the plant produced by recombination in F_{L} F_{2} and, to some extent, in F_{s} 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 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 over-come these problems.

problems . In population breeding, outstanding F_{9} plants are mated among themselves in
pairs or in some other fashion. The intermating of selected F_{9} plants restores
heterozygosity in the pro-geny, which provides for 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 inter-mated population. Thus the chances of the
recovery of transgressive segregants would increase considerably. This process may be

5
repeated one or more time. This procedure is similar to recurrent selection in cross-pollinated
crops (Chapter 18). A variation of this approach would be to intermate Fs or later generation
progenies. This would allow a more effective selection of desirable progenies than in the case
of F. where individual plants have to be selected. As noted previously, selection in F. based
on individual plants is of little value, particularly for char acters like yield.

CONCEPT OF A MULTILINE

The concept of a multiline variety was proposed by Jensen [33] and defined as a
combination of pure lines chosen from a breeding program for uniformity of appearance,
especially for height and maturity, but also for other characteristics important for a desirable
agronomic type. The purpose is to combine different genotypes that have desirable attributes
but do not reduce the phenotypic uniformity. Performance data on the components are
necessary so that only compatible lines are blended. The individual component lines are
maintained separately so that the original blend can be recreated by mixing the seed stocks in
the correct proportions, and the breeder has the option of adding or removing individual lines
over time. Theoretically, a multiline variety could have a longer life because of enhanced
yield stability, broader adaptation, and resistance to diseases. The component lines could
have resistance to different races of the pathogen, thus avoiding a potentially devasting
disease outbreak that could occur if they were released individually.

In summary, the advantages of multiline cultivars include

(1) they provide a method to quickly develop a well-buffered, disease-resistant cultivar that
can employ several resistance genes;

(2) the useful life of a disease resistance gene is extended while a conventional breeding
program is ongoing;

(3) reduced losses due to disease should stabilize the cultivars deployed;

(4) an individual breeding program can distribute cultivars over a wide area without risk of
homogenizing the pathogen population.

Disadvantages

(1) the utility of multilines is limited to high-risk regions for disease outbreaks;

(2) usually there is no genetic improvement for yield or agronomic traits;

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(3) substantial labor is required to produce and maintain the component lines;

(4) release of an improved recurrent variety is delayed until the components are produced.

To quantify the performance of multilines, Jensen and Federer [34] applied the
concepts and computation of combining ability to competitive ability in wheat. In this
application, general combining ability (GCA) refers to the average performance of a line in
combinations, and specific combining ability (SCA) refers to the deviations in the expected
average performance of combinations. Jensen [32] outlined four different examples for
forming a multiline:

(1) using a single backcross to generate lines for use in the multiline;

(2) crossing unrelated lines;

(3) crossing to different selected recurrent parents; and

(4) making double crosses where each single cross has a common parent.

Marshall and Brown [35] used statistical models to determine the effect of intra-
populational genetic diversity on the stability of performance of mixtures as estimated by
their variance in yield across environments. Their models suggested that, in the absence of
intergenotypic interactions, the yield of a multiline will vary less than the least variant
component when the component lines perform differently in different environments.
Conversely, when there are intergenotypic interactions, the stability of a mixture will be more
stable than the best line only when each component responds differently to different
environments. They also predicted that, when mixtures are compared to their pure line
components, it is expected that improved stability is more easily attained than improved yield
because improvements in yield require net positive intergenotypic interactions whereas
stability does not.

In addition to stability, the use of multilines has the potential to improve resistance to
disease. Borlaug and Gibler [36] developed wheat lines for multiline cultivars at CIMMYT
using ‘donor parents’ selected from the International Wheat Rust Nursery and backcrossed to
recurrent parents. A number of studies have examined possible mechanisms for the observed
enhancements in disease resistance within multilines, and generally agree that the reduced
inoculum load results from both a lower frequency of initial infection when a spore lands on a

7
resistant component of the multiline and a lower rate of increase in inoculum. A review of
multilines for disease control was published by Mundt [37].

Given the changing climate and the need for greater protection of natural ecosystems
and sustainable agricultural practices, multilines and multiblends will likely play an important
role in the future agricultural production systems. Further research is warranted on durability
of resistance in multilines, experimental design, and design of mixtures.

MUTATION BREEDING

Physical Mutagens

Physical mutagens include various types of radiation, viz X-rays, gamma rays, alpha
particles, beta particles, fast and thermal (slow) neutrons and ultra violet rays. A brief
description of these mutagens is presented below:

Commonly used physical mutagens (radiations), their properties and mode of action.

Type of Radiation Main properties

X – rays S.I., penetrating and non-particulate
Gamma rays S.I., very penetrating and Non-particulate
Alpha Particles D.I., particulate, less penetrating and positively
charged.
Beta Rays Particles S.I., particulate, more penetrating than alpha
particles and negatively charged.
Fast and Thermal D.I., particulate, neutral particles, highly penetrating.
Neutrons
6. Ultra Violet Rays Non-ionizing, low penetrating

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 X-rays

X-rays were first discovered by Roentgen in 1895. The wavelengths of X-rays vary
from 10-11 to 10-7. They are sparsely ionizing and highly penetrating. They are generated in
X-rays machines. X-rays can break chromosomes and produce all types of mutations in
nucleotides, viz. addition, deletion, inversion, transposition, transitions and transversions. X-
rays were first used by Muller in 1927 for induction of mutations in Drosophila. In plants,
Stadler in 1928 first used X-rays for induction of mutations in barley.

Gamma rays

Gamma rays have shorter wave length than X-rays and are more penetrating than
gamma rays. They are generated from radioactive decay of some elements like 14C, 60Co,
radium etc. Of these, cobalt 60 is commonly used for the production of Gamma
rays. Gamma rays cause chromosomal and gene mutations like X-rays.

CHEMICAL MUTAGENS

Many chemical compounds are known to increase the mutability of genes.

Auerbach and Robson’s experiment with male Drosophila melanogaster in 1947,

utilizing mustard gas and related substances such as nitrogen and Sulphur mustards, mustard
oil, and chloracetone, was the first to show that chemicals can cause mutations.
Chemical mutagens affect the chromosomal DNA in the following two ways:

1. Direct gene change

2. Copy error
Chemical mutagens can be classified into different categories, such as:

1. Base analogs
2. Intercalating agents
3. Metal ions
4. Alkylating agents
A. Base analogs
 These substances have structural characteristics with bases like purines and pyrimidines.
 5-Bromouracil and aminopurine are the two most prevalent base analogs that are
considered to be chemical mutagens.

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 Base analogs are integrated into the DNA structure during replication because of the
structural resemblances between these agents and DNA bases.
 Like adenine, aminopurine can pair up with either C or T to form a base pair (though base
pairing with C is rare).
 Some additional base analogs, including urethane triazine, caffeine (found in coffee, tea,
and soft beverages), phenol and carcinogens, acridines (proflavin, etc.), and others, are
similarly mutagenic.
B. Intercalating agents
 Intercalating molecules have a hydrophobic heterocyclic ring structure that resembles the
base pair ring structure.
 These agents embed themselves in the DNA helix, causing interference with transcription,
replication, and mutation, most frequently a frameshift mutation.
Some of the common intercalating agents are:

 Ethidium bromide
 Proflavine
 Acridine orange
 Actinomycin D
 Daunorubicin
C. Metal ion
 Reactive oxygen species (ROS) are produced by mineral ions, such as nickel, chromium,
cobalt, cadmium, arsenic, chromium, and iron, that lead to DNA hypermethylation. This
promotes DNA damage and obstructs the DNA repair process.
 For many organisms, certain inorganic chemicals like manganese chloride are mutagenic
because they bind calcium and interfere with the integrity of the chromosome structure.
D. Alkylating agents
These chemicals cause DNA damage by inducing alkyl groups.

Alkyl group introduction boosts ionization, leading to base-pairing mistakes that

eventually cause holes in the DNA strand and have a direct mutagenic effect on the DNA
molecule.

Some of the common alkylating agents are:

 Nitrous acid
 Ethylnitrosourea

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 Methylhydrazine
 Dacarbazine
 Formaldehyde
 Vinyl chloride
 Epoxides
 Dimethyl and diethyl sulphonate
 Methyl and ethyl methanesulphonate (MMS and EMS)
 Nitrosoguanadine (NG)
However, these substances can be eliminated from the DNA by the depurination process
during the DNA repair process.

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REFERENCE :

B.D. Singh ,Plant Breeding Principles And Methods, 12th edition, page no:264 -272

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