Principle and Application of Plant Tissue Culture

PRINCIPLE AND APPLICATION OF
PLANT TISSUE CULTURE
Mr. Diptendu Sarkar,

Lecturer,
Department Of Biotechnology
Acharya’s Bangalore B School
Lingerdianahalli,
Off Magdi Rd, Bangalore-91
INTRODUCTION
“Plant tissue culture” is a practice used to propagate

plants under sterile conditions, often to produce clones of a
plant. Different techniques in plant tissue culture may offer
certain advantages over traditional methods of propagation,
including:
• The production of exact copies of plants that produce
particularly good flowers, fruits, or have other desirable traits.
• To quickly produce mature plants.
• The production of multiples of plants in the absence of
seeds or necessary pollinators to produce seeds.
• The regeneration of whole plants from plant cells that have
been genetically modified.
• The production of plants in sterile containers that allows

them to be moved with greatly reduced chances of
transmitting diseases, pests, and pathogens.
• The production of plants from seeds that otherwise have

very low chances of germinating and growing, i.e.: orchids
and nepenthes.
• To clean particular plant of viral and other infections and to

quickly multiply these plants as 'cleaned stock' for
horticulture and agriculture.
NUTRIENT MEDIA
Nutritional composition varies according to the cells, tissue,

organ and protoplast and also with respect to particular plant
species.
 Basically media consists of:
Inorganic nutrients: e.g. N, Ca, Co, K , P, Zn, Fe, Cl etc.

Carbon and energy sources: e.g. Sucrose and Glucose
Phytohormones: e.g. Auxins, Cytokines, Gibberellins,
Ethylene
Organic supplements
CULTURE TYPES
Embryo Seed Meristem

culture culture culture
Cell Protoplast
PLANT TISSUE
culture CULTURE culture
Organ
Callus culture
Bud
culture
culture
1. CALLUS CULTURE
Explants can give rise to an unorganized growing and
dividing mass of cell known as callus.
During callus formation there is dedifferentiation both in

morphology and metabolism.
Callus culture is often performed in the dark as light can

encourage dedifferentiation of the callus.
Application of callus culture

Callus culture can be used to initiate cell suspension,
which is used in a plant transformation studies.
Used for the manipulation of the Auxins to cytokines

ratio.
2. EMBRYO CULTURE
Both mature and immature embryo can be used as
explants. Two types of embryo culture:
• Mature embryo culture
• Immature embryo culture/Embryo rescue
Applications of embryo culture

1. Prevention of embryo abortion in wide crosses:-
Hybrid plants raised by embryo culture have desirable
gene such as earliness, disease and pest resistance
such as
a) Lycopersicon esculentum × L.peruvianum
b) Solanum tuberosum × S.etuberosum
Embryo rescue technique has been successfully used for
raising hybrid embryos between Actidinia deliciosa ×
A.eriantha.
2. Production of haploids:- Kasha and Kao (1970) have
developed a technique to produce barley monoploids.
3. Overcoming seed Dormancy
4. Shortening of breeding cycle:-Flowering in some
plants can be brought early through embryo culture. E.g.
Rosa
5. Embryos are excellent materials for in vitro clonal
propagation. Mostly used for Conifers and members of
Gramineae family.
6. Germination of seeds of obligatory parasites without the
host is impossible in vivo but is achievable with embryo
culture.
7. Embryo culture has been practiced as a method in

horticultural crops including peach nectarine and plum.
“Goldcrest peach” and “Mayfire” have resulted from
embryo culture and are now commercially grown.
8. Macapuno coconuts are priced for their soft endosperm

which has been developed by embryo culture.
GOLDCREST PEACH
3. NUCELLUS CULTURE
 Rangaswamy (1958) studied for the first time

nucellar embryony in vitro in Citrus microcarpa.
 Pollination and Fertilization are generally

considered as essential prerequisite or induction of
nucellar ebmryogenesis.
Application of nucellus culture
1. Pseudo bulbils produced by nucellus culture of citrus

represent tissue banks capable of initiating clones of
adventive embryos.
2. Plants derived from nucellar embryos are free from
most viruses.
3. Many of the desirable plant vigor and fruiting

characteristics associated with juvenility are restored in
trees established from nucellar seedling.
4. ENDOSPERM CULTURE
 Attempt to grow endosperm tissue in culture began in

the 1930s.
 Both immature and mature endosperm of various
angiosperm taxa has been successfully cultured.
Application of endosperm culture
 Investigators have provided that triploid plant can be

raised through endosperm culture.
 Endosperm can be used as a nurse tissue for raising

hybrid embryos.
5. Application of CELL SUSPENSION CULTURE
 Embryogenic cell suspension offers the possibility for

large scale clonal propagation.
 Embryogenic suspension cultures present an excellent

tool for both theoretical studies and practical applications.
 Embryogenic cell suspension cultures and population of

somatic embryos are a source for the production of
important chemicals,cellular enzymes and many
secondary metabolites.
6. Application of PROTOPLAST CULTURE
 Production of novel intrespecific and intergeneric crosses

between plants. E.g. Pomato
 Protoplast of sexually sterile plants can be fused to

produce fertile diploids and polypoids.
 Hybridization becomes possible between plants that are

still in the juvenile phase.
 Used in the study of cytoplasmic genes and their

activities that can be applied in plant breeding
experiments.
 Somatic hybridization for gene transfer:-Disease

resistance, Abiotic stress resistance, Quality
characters,Cytoplasmic male sterility
Production of heterozygous lines within a single species
that normally could only be propagated by vegetative
means. e.g. Potato
In-vitro tissue culture potato explants

POMATO
OTHER APPLICATIONS
Micropropagation is widely used in forestry and in floriculture.
Micropropagation can also be used to conserve rare or

endangered plant species.
A plant breeder may use tissue culture to screen cells rather

than plants for advantageous characters, e.g. herbicide
resistance/tolerance.
Large-scale growth of plant cells in liquid culture inside

bioreactors as a source of secondary products, like
recombinant proteins used as biopharmaceuticals.
To cross-pollinate distantly related species and then tissue

culture the resulting embryo which would otherwise normally
die (Embryo Rescue).
For production of doubled monoploid (dihaploid) plants
from haploid cultures to achieve homozygous lines more
rapidly in breeding programmes, usually by treatment with
colchicines which causes doubling of the chromosome
number.
As a tissue for transformation, followed by either short-

term testing of genetic constructs or regeneration of
transgenic plants.
Certain techniques such as Meristem tip culture can be

used to produce clean plant material from virused stock,
such as potatoes and many species of soft fruit.
Micropropagation using Meristem and shoot culture to
produce large numbers of identical individuals.
Screening programmes of cells, rather than plants for

advantageous characters.
Crossing distantly related species by protoplast fusion and

regeneration of the novel hybrid.
Plant tissue culture
REFERENCES:-
Adrian Slater (2003) PLANT BIOTECHNOLOGY De
Mont Fort University.
H.S. Chawala, INTRODUCTION TO PLANT

BIOTECHNOLOGY ( II edition) New Delhi.
http://en.wikipedia.org/wiki/User:Plant tissue culture

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PRINCIPLE AND APPLICATION OF

PLANT TISSUE CULTURE

Mr. Diptendu Sarkar,

“Plant tissue culture” is a practice used to propagate

• The production of plants in sterile containers that allows

• The production of plants from seeds that otherwise have

• To clean particular plant of viral and other infections and to

Nutritional composition varies according to the cells, tissue,

 Basically media consists of:

Inorganic nutrients: e.g. N, Ca, Co, K , P, Zn, Fe, Cl etc.

Embryo Seed Meristem

During callus formation there is dedifferentiation both in

Callus culture is often performed in the dark as light can

Application of callus culture

Used for the manipulation of the Auxins to cytokines

Applications of embryo culture

7. Embryo culture has been practiced as a method in

8. Macapuno coconuts are priced for their soft endosperm

 Rangaswamy (1958) studied for the first time

 Pollination and Fertilization are generally

Application of nucellus culture

1. Pseudo bulbils produced by nucellus culture of citrus

3. Many of the desirable plant vigor and fruiting

 Attempt to grow endosperm tissue in culture began in

 Investigators have provided that triploid plant can be

 Endosperm can be used as a nurse tissue for raising

 Embryogenic cell suspension offers the possibility for

 Embryogenic suspension cultures present an excellent

 Embryogenic cell suspension cultures and population of

 Production of novel intrespecific and intergeneric crosses

 Protoplast of sexually sterile plants can be fused to

 Hybridization becomes possible between plants that are

 Used in the study of cytoplasmic genes and their

 Somatic hybridization for gene transfer:-Disease

In-vitro tissue culture potato explants

Micropropagation can also be used to conserve rare or

A plant breeder may use tissue culture to screen cells rather

Large-scale growth of plant cells in liquid culture inside

To cross-pollinate distantly related species and then tissue

As a tissue for transformation, followed by either short-

Certain techniques such as Meristem tip culture can be

Screening programmes of cells, rather than plants for

Crossing distantly related species by protoplast fusion and

H.S. Chawala, INTRODUCTION TO PLANT

http://en.wikipedia.org/wiki/User:Plant tissue culture

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