Molecular breeding is of very recent origin.

It developed in the 1980s with the evolution of DNA marker

technologies.
Molecular breeding is defined as:

a branch of plant breeding which utilizes molecular

genetic tools and approaches for genetic improvement of
crop plants.
genetic improvement of crop plants for various economic
traits using molecular marker and transformation
technologies is referred to as molecular plant breeding.
Points related to molecular plant breeding:
i. Molecular Plant Breeding (MPB) utilizes latest genetic
technologies to develop better varieties of crop plants. Two
molecular technologies, viz. molecular marker technology and
transformation technology are used in molecular plant
breeding.
ii. Molecular breeding is applicable to both plant breeding and
animal breeding. In other words MB can be used for genetic
improvement of both plants and animals.
iii. MPB is more precise, rapid and cost effective in
comparison to conventional phenotypic selection. MPB
reduces the time significantly required for the development of
new cultivars. It takes 4-5 years for development of new
cultivars against 10-12 years of conventional phenotypic
selection method.
iv. It offers possibility of achieving those goals which cannot
be achieved by conventional methods.
v. It permits gene transfer between unrelated organisms
through transformation technology or genetic engineering.
vi. It helps in identification of new genes of economic
importance.
vii. Now-a-days, molecular breeding is a vital part of plant
breeding programmes.
viii. MPB sometimes contributes novel traits of agronomic
value in crop plants.
In agriculture or crop improvement, molecular
breeding has two branches:
(i) Molecular marker technology
(ii) (ii) Transformation technology.
Q. 4. What are advantages of molecular markers?
Ans. A molecular marker is a unique sequence of nucleotide
located close to the gene of interest. In other words, molecular
markers are special segments of DNA which flag the presence
of genes that control particular trait. They are located very
close to the gene of interest and are easily identifiable.
The important advantages of molecular markers in
plant breeding are presented below:
(i) Molecular markers allow selection of desirable trait at any
stage of plant growth. There is no need to wait till maturity of
the plants.
(ii) They are not affected by environmental conditions. In
other words molecular markers are stable.
(iii) They eliminate the need for phenotypic scoring.
(iv) They provide uniform method for scoring.
(v) A very small sample of plant, leaf or grain is required for
scoring of molecular markers.
(vi) Much less numbers of breeding generations are required
for evaluation of molecular markers.
(vii) They provide information about the percentage of
genome contributed by each parent.
Q. 5. What are applications of molecular markers?
Ans. There are several useful applications of molecular
markers.
Some important applications of molecular markers
are listed below:
(i) Molecular markers help in better understanding of
quantitative trait loci (QTL).
(ii) They are highly reproducible and thus help in accurate
selection of desired genotypes.
(iii) They are useful in gene pyramiding i.e. in incorporation of
multiple target genes into a single cultivar.
(iv) They are useful in identification of cultivars and parents
of hybrid.
Q. 6. What is molecular marker technology?
Ans. A technology that utilizes molecular markers for genetic
improvement of crop plants is referred to as molecular marker
technology. It is also known as DNA marker technology.
Important points related to this technology are
briefly presented below:
i. It does not involve genetic transformation and hence does
not require biosafety and bioethics measures.
ii. It utilizes various DNA markers such as restriction
fragment length polymorphisms (RFLP), amplified fragment
length polymorphisms (AFLP), random amplified
polymorphic DNA (RAPD), simple sequence repeats (SSR),
single nucleotide polymorphisms (SNP), expressed sequence
tags (EST), sequence tagged site (STS), etc.
iii. It makes use of two major technologies, viz. marker
assisted selection (MAS) and DNA fingerprinting.
iv. Molecular marker technology helps in early detection of
cytoplasmic male sterility. By this technology, the cytoplasm
type of unknown plants can be identified.
v. It is highly accurate and rapid method of crop
improvement. Marker assisted selection (MAS) is a potential
tool of crop improvement which should be deployed along
with conventional breeding methods.
vi. It is effective technology in achieving new goals which
cannot be easily achieved by traditional breeding such as
pyramiding disease resistance genes with indistinguishable
phenotypes.
vii. The DNA fingerprinting is useful in identification of
cultivars, pure lines and parents of a hybrid.
viii. Molecular marker technology (MMT) is useful in mapping
desirable quantitative trait loci (QTL).
Q. 7. What is transformation technology?
Ans. The genetically engineered genes are known as
transgenes. The technology that utilizes transgenes in
developing new crop cultivars is referred to as transformation
technology. It is also known as gene technology or genetic
engineering.
Genetic engineering is the process of removing, modifying or
adding genes to a strand of DNA for specific purposes. For
example rye grass produces a protein in its pollen that causes
an allergic reaction in humans. By switching off the gene
responsible for producing allergen, we can develop allergy free
rye grass.
Main points related to transformation technology are
briefly presented below:
i. It involves transgenes and hence biosafety and bioethical
measures are essential.
ii. It leads to development of transgenic cultivars which are
not permitted for organic farming. Moreover, development of
transgenic cultivars is not permitted under organic plant
breeding.
iii. Transformation technology is useful in developing crop
cultivars resistant to biotic (insects, diseases and parasitic
weeds) and abiotic stresses and also in improvement of
quality traits.
iv. It is an accurate and rapid method of developing superior
crop cultivars.
v. It utilizes mainly two methods of gene transfer, viz. biolistic
method and agrobacterium mediated method. These methods
are being applied to various field crops for developing
transgenic cultivars.
vi. This technology leads to over expression of novel genes
conferring novel traits. It leads to inhibit expression of
harmful genes.
vii. It is useful in evaluation of gene promoters and also of the
function of unknown genes.
Q. 8. Give a comparison of molecular marker
technology and transformation technology.
Ans. There are some similarities and some dissimilarities
between marker technology and transformation technology
which are presented in Table 24.1.
Q. 9. What are practical applications of molecular
marker technology?
Ans. Molecular marker technology has four principal
applications in crop improvement programmes, viz.:
(i) Molecular selection,
(ii) Molecular backcrossing,
(iii) Molecular mapping, and
(iv) Molecular genotyping.
These are briefly discussed below:
(i) Molecular Selection:
This method is used in the segregating populations (F2 or F3)
for selection of plants carrying the gene of Interest. It is used
to select those genes which are difficult to select on the basis
of phenotypes. It is very useful in selection of genes which are
highly influenced by environmental conditions. It allows easy
stacking of disease resistant genes. The selection can be
practised at an early stage of plant growth.
(ii) Molecular Backcrossing:
This is the most common application of molecular marker
technology in crop improvement. It requires less material and
less space for conducting DNA marker test. It allows recovery
of the highest portion of the recurrent parent, with desired
genes, in the shortest number of generations. It is possible to
identify between homozygous and heterozygous plants
through DNA markers, Thus it helps in rapid transfer of
desirable gene.
(iii) Molecular Mapping:
DNA marker technology plays important role in gene
mapping. It is useful in mapping of both qualitative and
quantitative traits. It identifies molecular markers linked to
chromosomal regions affecting the desirable trait and thus
helps in gene mapping. The gene mapping is possible even
during early stages of plant growth. Various breeding
populations, viz., recombinant inbred lines (RILs), near
isogenic lines (NILs), bulk segregant populations,
introgressed lines etc., are used for gene mapping,
(iv) Molecular Genotyping:
It refers to molecular characterization of breeding material. It
fulfills DNA fingerprinting needs. It is useful to identifying
differences between varieties and hybrids. It is used for
varietal verification, varietal purity and hybrid purity and thus
is useful for patent protection.
The gene technology or genetic engineering has four
principal applications in crop improvement which
are listed below:
i. Development of transgenic crop cultivars in different crop
plants resistant to biotic and abiotic stresses.
ii. Development of herbicide resistant cultivars in different
crop plants.
iii. Improvement in nutritional quality, keeping quality and
industrial quality in food, vegetable, fruits and other
commercial crops.
iv. Identification of new genes and gene systems. It enables
the introduction of novel traits in crop plants such as golden
rice and longer blooming flowers in flower plants such as
petunia.
Q. 10. What are practical achievements of molecular
plant breeding?
Ans. The molecular plant breeding has been used for genetic
improvement of various field crops (maize, barley, wheat, rice,
sorghum, soybean, chick pea, pea, mungbean), vegetable
crops, forage crops and fruit crops for various economic
characters.
The notable achievements and briefly presented
below:
i. Bacterial blight and blast resistant cultivars have been
developed in rice through marker assisted selection.
ii. In soybean, cyst nematode resistant cultivars have been
developed through MAS.
iii. In maize, quality protein maize (QPM) lines have been
developed through MAS.
iv. In rye, bread making quality has been significantly
improved by transferring glutelins gene of wheat through
genetic engineering.
v. In wheat and rice, yield has been increased by transferring a
gene from maize through gene technology.
vi. In cotton, bollworms and herbicide resistant cultivars have
been developed through transgenic technology by using Bt.
gene from soil bacterium. Several other examples can be cited
from vegetable and forage crops.
Q. 11. What are advantages of molecular marker
technology?
Ans. Some of the advantages of molecular marker
technology are presented below:
i. It helps in better understanding of quantitative trait loci and
as a result in more effective breeding,
ii. Rapid Introgression of simply inherited characters. The
number of backcrosses required can be reduced drastically
once the marker for the character to be introduced is
identified.
iii. Easy or early indirect character selection It is useful for
those genes that cannot be detected at an early stage of plant
growth. For example, lysine and tryptophan genes in maize.
iv. It is useful in achieving new goals that cannot be achieved
through conventional breeding such as pyramiding of disease
resistance genes which cannot be detected by phenotypes. In
other words, it offers possibility of achieving unattainable
goals.
v. It is accurate, rapid and more reliable method of crop
improvement.
vi. The DNA marker test can be conducted at any stage of
plant growth even for those traits which express at the
maturity.
vii. A very small amount of plant tissue i.e. leaf, stem, flower,
seed etc. is required to conduct DNA marker test.
viii. The DNA test is the only available practical technique for
ensuring the presence of multiple beneficial genes in a single
variety.
ix. DNA based fingerprinting allows the reliable tracking of
beneficial traits during varietal selection.
x. DNA based fingerprinting is the best currently available
technology to differentiate among varieties for patent
protection and plant variety protection Act Certification.
xi. DNA based marker systems are useful in selection of
disease or insect resistant material even in the absence of pest
incidence.
Q. 12. What are advantages of gene technology?
Ans. Some of the advantages of gene technology are
given below:
i. It involves transgenes and leads to development of
transgenic cultivars in different self and cross pollinated
species.
ii. It permits transfer of desirable gene from unrelated species
i.e. from micro-organisms to crop plants and even from
animals to plant.
iii. It enhances the expression of novel genes conferring novel
traits and switch off the expression of harmful traits such as
allergen in rye grass.
iv. It is precised and rapid method of crop improvement.
Moreover, it requires less space than conventional breeding
methods.
v. It is useful method of improving crop plants for resistance
to biotic and abiotic stresses and quality traits.
vi. It helps in evaluating the functionality of unknown genes.
Q. 13. What are disadvantages of molecular plant
breeding?
Ans. However, it has some disadvantages which are
briefly presented below:
i. The initial cost of both the branches of molecular plant
breeding i.e. molecular marker technology and gene
technology is very high. Both these technologies require costly
equipment, glass wares and chemicals.
ii. Molecular breeding required team of well-trained
manpower for handling of costly equipments, chemicals and
glassware.
iii. Gene technology involves transgene and thus requires
biosafety and bioethical measures,
iv. The scoring of DNA or molecular markers is a laborious
and time consuming task.
v. In gene technology, the frequency of desirable
transformants is very low which restricts the use of this
technology.
Q. 14. What are restriction enzymes?
Ans. A group of endonuclease enzymes, that cuts DNA only at
specific sites or base sequences.
Q. 15. What are endonuclease enzymes?
Ans. The enzymes which are able to make Internal cut in the
DNA molecule are called endonucleases. Most of the
endonucleases cut DNA molecule at random sites, but some
cut only at specific sites. Endonucleases are found. In micro-
organisms and are also referred to as biochemical scissors.
Endonucleases do not cut DNA of their own cell and protect it
by methylation process.
Q. 16. What is recognition site?
Ans. The specific base sequence or site recognized by
restriction enzyme in DNA molecule to make a cut or cleavage
Is called recognition site; also called recognition sequence.
Q. 17. What is DNA Finger Printing?
Ans. A method of genetic engineering in which a DNA probe
is hybridized with DNA of plants or animals to determine
similarities: and differences between two DNA molecules.
Q. 18. What are DNA probes?
Ans. Small segments of DNA with known base sequences,
origin and function are called DNA probes. DNA probes are
useful in marker aided selection.
Q. 19. What is DNA library?
Ans. A set of DNA probes is called DNA library.
Q. 20. Define gene amplification.
Ans. An increase in the copy number of a gene/DNA
sequence is called gene amplification.
Q. 21. What is restriction fragment?
Ans. A fragment of DNA created by cleavage at specific sites
by a restriction endonuclease is called restriction fragment.
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