Application of Tissue

culture in
Plant Breeding
BY;

Zulfiqar
Nasir Hussain
(M.phil Students)

National University of Agricultural Sciences (NUAS),

Islamabad
What is Plant Tissue Culture?
 Planttissue culture is a form of
asexual propagation of plants
under laboratory conditions.

 Itis grown on a special culture

medium which supports its growth
and development.

 The medium can be either

semisolid, such as agar, or liquid,
such as purified water.
Microorganisms and Plant Tissue Culture
 In the practice of plant tissue culture,
microorganisms are called “contaminants”
because of their harmful effects on plant
growth in vitro.
 Six potential sources of contamination in the
plant tissue culture lab are:
 Air
 Water
 Growth Media
 People
 Equipment
 Plant Material
Why is Plant Tissue Culture Important?

 It has made significant

contributions in:
 The production of plant material

 Plant breeding

 Gene banks

 The production of chemical

compounds
What is Plant Breeding?

 Isa genetic improvement in a

heredity material of a plant in such
a way that newly formed plants
are better than existing one.
 A plant breeder may use tissue
culture to screen cells rather than
plants for advantageous
characters e.g. herbicide
resistance/tolerance
Applications to Plant Breeding

 Haploid Production
 Triploid Production
 In Vitro Pollination and Fertilization
 Somatic Hybridization &
Cybridisation
 Genetic Transformation
 Somaclonal and Gametoclonal
Variant Selection
Application to Horticulture and
Forestry

 Production of Disease-Free Plants

 Clonal Propagation

General Applications
 Secondary Metabolite Production
 Germplasm Conservation
Haploid Plants from Tissue Culture
 New Plant Varieties in a Shortened Time
Frame
 Production of haploid plants by anther
and microspore culture techniques
 Production of homozygous diploid lines
through chromosome doubling, thus
reducing the time required to produce
inbred line
 Uncovering mutations or recessive
phenotypes
Triploid Production by Plant Tissue Culture

 Triploid plants, with three sets of

chromosomes, are seed-sterile due to
disturbance in gamete formation
 Tissue culture to morphogenic
response and production of triploid
plantlets have done by endosperm
culture.
 Endosperm tissue often shows a high
degree of chromosomal variations and
polyploidy
 Triploids are usually seed sterile
 Seed-less ness is improve the quality of
fruits
In vitro pollination and Fertilization

 Male and female gametes are isolated

and introduced to each other under
conditions suitable for zygote formation
 In plant breeding embryo culture have
been valuable tools, especially for the
transfer of disease resistance genes
from wild relatives into crop plants
 It is used for recovery of hybrid plant
 To protect crops from weather, pests,
and diseases.
Somaclonal and Gametoclonal Variant
Selection

 Somaclonal variation and gemetoclonal

variation are the important source of
introducing genetic variation
 These variations include chromosome
aberrations, DNA amplification, and the
occurrence of transposable elements.
 Applicable to agriculture and industry
specially resistance to herbicide,
pathotoxin, salt or aluminium, useful in
the synthesis of secondary metabolites
on a commercial scale,
Somatic Hybridization and Cybridisation
through Tissue Culture

 Non-conventional genetic procedure

involving fusion between isolated
somatic protoplasts under in vitro
conditions and subsequent development
of their product (heterokaryon) to a
hybrid plant is known as somatic
hybridization
 Protoplast technology
 Cybridization.
 In cybridization, heterozygosity of extra
chromosomal material can be obtained,
which has direct application in plant
breeding.
Genetic Transformation

 Tissue culture is an essential part

of many genetic transformation
protocols like; Biolistics,
Agrobacterium tumefaciens,
Electroporation & Microinjection
 In plant breeding many different
explants can be used, depending
on the plant species and its
favored method of regeneration
Application to Horticulture and
Forestry

 Production of Disease-Free Plant

 Clonal
Propagation or Micro-
propagation
Production of Disease-Free Plant

 Meristem Culture
 Is a zone of cells with intense divisions;
about 0.1 mm in diameter, situated in
the top of buds, and extremities of roots
Meristem and usually involves the
removal of the meristem and
subsequent culture on a nutrient
medium
 Pioneered Morel (1960)
 Large numbers of disease-free plants
may be produced from meristematic
explants
 stem-disc dome (SD dome) culture
Clonal Propagation or Micro-propagation

 A method of asexual propagation used

by commercial growers to produce
clones of a particular plant in large
quantities
 Major Benefits:
 Rapid multiplication of superior clones
and maintenance of uniformity;
 Multiplication of disease free plants
and
 Multiplication of sexually derived
sterile hybrids
Stages in clonal propagation:
 Stage I involves establishment of
tissue in vitro.
 Stage II involves multiplication of
shoots (often media are not changed
between stage I and stage II).
 Stage III concerns root formation and
conditioning of propagates prior to
transfer to the green house; this stage
requires alteration of media for
promotion of root formation.
 Stage IV involves growth in pots
followed by field trials.
General Applications

Secondary Metabolite Production

 Produce a proliferation of shoots and

leafy material, and harvesting the leafy
material and shoots while in a green,
actively-growing, non-senescent stage
 Particular interest is plant material that
produces alkaloids, especially material
from poppy (Papaver sp.)
Germplasm Conservation:
 Genetic resources of recalcitrant-
seed and vegetatively propagated
species as well as of endangered
species
 elite genotypes which are
multiplied on a large scale in
production laboratories
 cultures with special attributes,
e.g. Metabolite-producing cell lines
and genetically engineered
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