Experiment: Handing of segregation populations: Pedigree

method
Aim: To know about how to handle segregating population of pedigree method

Mass selection and pure line selection cannot be applied to segregating population E.g. F2, F3
etc. because in these methods selection made from the existing variability present in the
population. However, when there is limited genetic variability in a species than hybridization
acts as an important tool to create additional genetic variability for further improvement of the
species. The methods generally used for handling of segregation generation may be grouped
into following three categories.
a) Pedigree Method
b) Bulk Method and
c) Back Cross Method
The objectives of all these methods are to
1. Develop pureline
2. Develop new varieties.
3. Develop inbred line
4. Improve specific character of a well-adapted variety for which it is deficient.
Pedigree method
In pedigree method, individual plants are selected from F2 and the subsequent
generation and their progenies are tested. During the entire operation, a record of the entire
parent’s offspring relationship is kept, is known as pedigree record. The selection of individual
plant is continued till the progenies show no segregation. At this stage, selection is done among
the progenies, because there would be no genetic variation within progenies.
Pedigree record
The pedigree may be defined as a description of the ancestors of an individual and it
generally goes back to some distant ancestors. Thus, it describes the parents grandparents, great
grandparents so on of an individual.
Maintenance of pedigree record
Pedigree record may be kept in several ways, but it should be simple and accurate.
Generally, each cross is given a number. The first two digits of this number refer to the year in
which the cross was made, and the remaining digits denote the serial number of the cross in
that year. For example, the number 7911, denotes the cross number 11 of the year 79. In the
segregating generation one of the two systems of designation may be followed.
i) System I
In this system, the individual plant progenies in each generation are assigned row
number, corresponding to their location in the plot. In addition each progeny in F4 and the
subsequent generation is assigned the row number of the progeny in the previous generation
from which it was derived.
Table 9.1: Description of maintenance of pedigree record by System I
Generation Number Description
th
F3 7911-7 Progeny in the 7 row in the F3 plot.
Progeny in the 4th row in the F4 plot, selected from the progeny in
F4 7911-7-4
the 7th row of the F3 plot.
Progeny in the 14th row in the F5 plot selected from the progeny in
F5 7911-4-14
the 4th row of the F4 plot.
Progeny in the 3rd row in the F6 plot selected from the progeny in
F6 7911-14-3
the 14th row of the F5 plot.

ii) System-II
In this system, in each generation the selected plants are assigned serial numbers within
individual progenies. Each progeny or selected plant bear the serial number of all the plants in
the previous generation, related to it by direct descent. Thus, the plants selected in F2 are given
serial numbers of their parents (F2 plants). The plants selected from a progeny in F3 are given
the number of that progeny and in each generation the selected plant also given a serial number.
Table 9.2: Description of maintenance of pedigree record by System II
Generation Number Description
F3 7911-7 Progeny obtained from plant number 7 selected in F2
Progeny from plant No.4 selected from F3 progeny , derived
F4 7911-7-4
from the plant No.7 selected in F2
Progeny from plant No.2 selected from the F4 progeny derived
F5 7911-7-4-2 from plant no.4 , selected from the F3 progeny, obtained from the
plant No.7 selected in F2.
Progeny from plant No 8, selected from the F5 progeny, derived
F6 7911-7-4-2-8 from the plant N0.2 selected from the F4 progeny of the plant
No.4 selected from F3 progeny of the Plant No.7 selected in F2.
Procedure for pedigree method
1. Hybridization
Selection of parents to be used in a cross is the most important step in breeding program
based on hybridization. The selected parents are crossed to produce a simple or a complex
cross. For convenience we shall refer to the seed obtained from the width simple/complex
crosses as F1 seed.
2. F1 Generation
F1 seeds are space planted so that each F1 plant produces the maximum number of F2
seed ordinarily, 15-30 F1 plants should produce enough seeds for a good F2 population size.
3. F2 Generation
In F2, 2000-10000 plants are space-planted to facilitate selection. About 100-500 plants
are selected & their seeds are harvested separately. Usually the F2 population size should be
10-100 times the member of F2 plants that would be selected in the number may vary from
100-500 depending on the facilities available and objectives of the breeding program. When
the objective of breeding is to improve yield, a relatively large no of F 2 plants would be
selected. Selection in F2 is based on characteristics that are simply inherited, e.g. plant height,
head type, seed colour, disease resistance, presence of awns etc. Selection of vigour is generally
useless because vigour may be due to heterozygosis or environment or G×E interaction.
4. F3 Generation
Medical plant progenies having 30 or more plants each are space-planted. Medicinal
plants with desirable characteristics are selected, particularly from superior progenies may also
be selected. Diseased and lodging susceptible progenies are eliminated. The no of plants
selected in F3 should preferably be less than that at the F3 progenies. If the no of superior
progenies is small, the whole cross may be rejected.
5. F4 Generation
Individual plant progenies are space planted and desirable plants are selected mainly
from superior progenies. The no of plants selected in F4 is generally much smaller than that in
F3 progenies with defect and undesirable characteristics are rejected. If two or more progenies
coming from the same F3 progeny are similar and comparable, only on selection of desirable
plants from superior progenies.
6. F5 Generation
Individual plant progenies are generally planted according to the recommended
commercial seed rate. Often there are more rows grown for each progeny to facilitate
compression among many progenies would have in bulk. In progenies showing segregation,
individual plants may be selected. The no of progenies meant be reduced to a size manageable
in preliminary yield trials about 25-100 progenies.
7. F7 Generation
Individual plants are planted in multi row plots and evaluated visually; progenies are
harvested than bulk since they would have become almost homozygous. Progeny showing
outstanding, in such progenies individual plants may be selected, preliminary yields trait is
usually planted in F6 for those progenies, which are reasonably homozygous and have enough
seed, inferior progenies are eliminated based on yield from preliminary yield trial.
8. F8-F10 Generation
The superior lines are tested in replicated yield trails at several locations. These lines
are evaluated for yield diseases and lodging resistance, maturity time and quality etc. A line
that is superior to the test commercial variety, included in the trials as check. Yield & other
characteristics will be identified for release as a new variety.
9. F11 Generation
When a strain is likely to be released as a variety, the breeder usually multiplied its seed
during its last year in trail. The breeders have responsibility to supply the breeder seed for
production of foundation seed. Thus F11 to F12 the seed of the new variety will be multiplied
for distribution to the farmer.
 Fig.9.1: Breeding procedure of pedigree method
Application of Pedigree Method:
1) Selection of desirable plants from the segregating population in self- pollinated crops.
2) This method is commonly used to correct some specific weaknesses of an established
variety (Combination breeding).
3) It is also used in the selection of new superior recombinant type’s i.e. Transgressive
breeding.
4) This method is suitable for improving specific characteristics such as disease resistant,
plant height, maturity etc.
Experiment: Handing of segregation populations: Bulk method
Aim: To know about how to handle segregating population of bulk method

Bulk population method

Bulk population method of breeding in self –pollinated crop is also known as mass
method or population method of breeding. It was first used by Nilsson Ehle in 1908. It refers
to a species is grown in bulk plot (from F1 to F5) with or without selection, a part of the bulk
seed is used to grow the next generation and individual plant selection is practised in F6 or later
generation. In this method duration of bulking may vary from 6-7 to 30 generation.
Procedure of Bulk Population Method:
1) Hybridization:
Parents are selected according to the objective of the breeding programmer and crossed.
2) F1 Generation
The F1 generation ( 10 to 25 F1) is space planted and harvested in bulk.
3) F2-F6 – Generation
F2 to F6 generations are planted at commercial seed rate and spacing. These generations
are harvested in bulk. During these generations the population size should be as possible,
preferably 30 to 50 thousand plants should be grown in each generation.
4) F7 Generation
About 30 to 50 thousand plants are space planted and out of this only 1000 to 5000
plants with superior phenotypes are selected and their seeds harvested separately. Selection is
made on the basis of phenotypes of plants, grain characteristics etc.
5) F8 Generation
Individual plant progenies are grown in single or multi row plots. Most of the progenies
would be homozygous and are harvested in bulk. Weak and inferior progenies are rejected and
only 100- 300 individual plant progenies with desirable characters are selected.
6) F9 Generation
Preliminary yield trial is conducted along with standard variety as check. The
evaluation of progeny is done for important desirable characteristics. Quality test may be
conducted to reject the undesirable progenies.
7) F10- F12 Generation
Replicated yield trails are conducted at several locations using standard commercial
varieties as check. The lines are evaluated for important agronomic characteristics. If lines are
superior to the standard check than it is released as new variety.
8) F13 Generation
Seed multiplication of the newly released variety has done for distribution to the
farmers.
Application of Bulk Population Method
 This method is suitable and most convenient for handling the segregating generation of
cereals, smaller millet, grain legume and oilseeds. This may be used for three different
purposes.
1) Isolation of homozygous lines.
2) Waiting for the opportunity of selection.
3) Opportunity for natural selection to change the composition of the population.

Fig.10.1: Breeding procedure of bulk population method

Experiment: Handing of segregation populations: Backcross
method
Aim: To know about how to handle segregating population of backcross method

Backcross method
The plan of back cross method depend upon whether the gene being transferred is
recessive or dominant. The plan for transfer of a dominant gene is quite simple than for
recessive gene.
Requirements of a backcross programme
1. Existence of a good recurrent parent variety which requires improvement is some
qualitatively inherited character or a quantitative character with high heritability.
2. A suitable donor parent must be available possessing the character or characters to be
transferred in a highly intense form.
3. High expressivity of the character under transfer through several back crosses in the
genetic back ground of the recurrent parent.
4. The character to be transferred must have high heritability preferably determined by
one or few genes.
5. Simple testing technique for detecting the presence of the character under transfer.
6. Recoveries of the recurrent genotype in a reasonable number of back cross generations.
I. Transfer of Dominant Gene
Let us suppose that a high yielding and widely adopted variety ‘A’ is susceptible to
stem rust (rr) and another variety ‘B’ is poor yielding but resistant to stem rust (RR) i.e.
dominant to susceptibility. In this back cross programme rust resistance trait is transfer from
donor parent (B) into a recurrent parent (A).
1) Hybridization
Variety ‘A’ is crossed with variety ‘B’ in which variety ‘A’ is used as female parent
which is recurrent and variety ‘B’ is used as donor parent.
2) F1 Generation
During the second year F1 plants are backcrossed to variety ‘A’ since all the F1 plants
will be heterozygous for rust resistance. Selection for rust resistance is not necessary.
3) First Back Cross Generation
In the third year half of the plant would be resistant and remaining half would be
susceptible to stem rust, rust resistant plants are selected and backcross to variety ‘A’.
4) BC2 –BC6 Generation
In each backcross generation, segregation would occur for rust resistance. Rust resistant
plant are selected and backcrossed to the variety ‘A’ selection for plant type of variety ‘A’ may
be practiced particularly in BC2 and BC3 generation.

5) BC6 Generation
 On an average the plant will have 98490 genes from variety A rust resistant plants are
selected and selfed, their seeds are harvested separately.
6) BC6 F2 Generation
Individual plant progenies are grown from the selected plants. Rust resistance once
plant, which are similar to variety ‘A’ are selected and selected plants are harvested separately.
7) BC5 F3 Generation
Individual plant progenies are grown homozygous progenies resistant to rust and
similar to plant type of variety ‘A’ harvested in bulk. Several similar progenies are mixed to
constitute the new variety.
8) Yield Test
The new variety is tested in replicated yield trials along with the variety ‘A’ as a check.
Plant type dates of flowering date of maturity, quality, etc are critically evaluated. The new
variety would be identical to variety ‘A’ in performance. Therefore detail yield test are not
required, and the variety may be directly released for cultivation.

Fig. 11.1: Different steps involved in the back cross method when a dominant trait is
transferred from variety B to an otherwise improved variety A
II. Transfer of Recessive Gene:
When rust resistant is due to a recessive gene, all the backcross cannot make one after
other. After the first backcross and after every two backcrosses F2 must be grown to identity
the rust resistant plants. The F1 and the back cross progenies are not inoculated with rust
because they would be susceptible to rust. Only F2 is tested for rust resistant.
1) Hybridization:
The recurrent parent is crossed with rust resistant donor parent. The recurrent parent is
generally used as female. i.e. ( rr X RR).
2) F1 Generation:
F1 plants are backcrossed to the recurrent parent.
3) BC1 Generation:
If rust resistance is recessive all the plant will be rust susceptible. Therefore, there is no
test for rust resistance. All the plants are self- pollinated.
4) BC1 (F2) Generation:
Rust resistance plants are selected and backcrossed with recurrent parent. i.e variety
‘A’. Selection is made for the plant type and other characteristics of the variety ‘A’.
5) BC2 Generation:
No rust resistance test, plants are selected, which is identical to the recurrent parent (
A) and backcrossed with the recurrent parent.
6) BC3 Generation:
No disease resistance test. The plants are self – pollinated to raise F2. selection is made
for the plant type identical to variety ‘A’.
7) BC3 F2 Generation:
Plants are inoculated with stem rust. Rust resistant plant, similar to ‘A’ are selected and
backcrossed to variety ‘A’.
8) BC4 Generation:
No rust resistance test plants are backcrossed to variety ‘A’.
9) BC5 Generation:
No rust resistance test plants are self-pollinated to raise F2 generation.
10) BC5 (F2) Generation:
Plants are subjected to rust epidemic, resistance plant for rust and having similar
characteristic of variety. ‘A’ is selected and self-seed are harvested separately.
11) BC5 (F3):
Individual plant progenies are grown and subjected to rust epiphytotic selection is done
for rust resistance and for characteristics of variety ‘A’ seeds from several similar rust resistant
homozygous progenies are mixed to constitute new variety.
12) Yield Test:
Same as in case of dominant gene transfer.
Fig. 11.2: A generalised scheme for the transfer a recessive gene for disease resistant
(here rust resistance) through the backcross method in a self-pollinated crop.