..PLANTTISSUECULTURE.

INTRODUCTION TO PLANT TISSUE CULTURE

Plant tissue
Culture

Plant cell and tissue culture is a technique that involves growing plant
cells, tissues, or organs in an artificial, controlled environment. This
method allows the cultivation of plant material under sterile
conditions on a nutrient culture medium of known composition. It is
based on the concept of totipotency, which means that each plant cell
has the potential to regenerate into a whole plant, given the right
conditions. Plant tissue culture includes a wide range of techniques
like micropropagation (cloning of plants), somatic embryogenesis
(development of embryos from somatic cells), callus culture
(culturing of undifferentiated cells), and suspension cultures.

The process begins with the selection of plant material, known as the
explant, which can be any part of the plant, such as leaves, stems, or
roots. The explants are then sterilized to prevent contamination and
placed in a nutrient-rich culture medium. This medium typicallyY
contains essential nutrients like sugars, vitamins, minerals, and
growth regulators like auxins and cytokinins. Depending on the type
of tissue culture technique, the explants can develop into callus
(undifferentiated mass of cells) or directly regenerate into shoots and
roots.

Plant tissue cullure is widely used for plant breeding, conservation of

rare and endangered species, and the mass propagation of genetically
identical plants, known as clones. It is also a key tool in genetic
engineering, allowing the manipulation and insertion of specific genes
into plant cells before regenerating them into transgenic plants.
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TYPES OF CULTURE-
1. Callus Culture

Callus culture is one of the most basic forms of plant tissue

culture, where an undifferentiated mass of cells, called a callus, is
produced. The callus forms when plant tissues, such as leaf or
stem explants, are placed on a nutrient medium with appropriate
growth regulators like auxins and cytokinins. Callus cultures are
often used for genetic manipulation, studying cell differentiation,
and producing secondary metabolites.

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2. Suspension Culture
In suspension culture, cells or small clusters of cells are grown in a
liquid medium that is constantly agitated to prevent the cells from
settling. The constant agitation promotes aeration and nutrient
uptake. Suspension cultures are ideal for mass production of
secondary metabolites and are often used in industrial
applications, including pharmaceuticals and cosmetics.
PLANT SUSPENSION CULTURE

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 PLANT TISsUE CULTURE .

3. Protoplast Culture
Protoplast culture involves the isolation of plant cells devoid of
their cell walls, known as protoplasts. These are generated by
enzymatic digestion of the cell walls and are useful for somatic
hybridization, where two different species can be fused at the
protoplast level to produce hybrids. Protoplasts can regenerate cell
walls and eventually grow into whole plants under the right
conditions.
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4. Embryo Culture
Embryo culture involves the isolation and growth of plant embryos
in vitro. It is particularly useful for rescuing hybrid embryos that
would otherwise abort or fail to develop in nature. This technique is
employed in plant breeding programs to develop hybrids and
overcome breeding barriers between species.

5. Organ Culture
Organ culture is a method where an entire organ, such as a leaf,
shoot tip, or root, is cultured in vitro. Depending on the organ
cultured, it can regenerate shoots or roots. Shoot tip cultures are
commonly used for micropropagation and producing virus-free
plants. Root cultures can be employed to study root development
or to produce valuable root-derived compounds.
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APPLICATIon OF PLANT TISSUE CULTURE

Plant cell and tissue culture techniques have revolutionized the fields of
agriculture, horticulture, and biotechnology. They provide a controlled and
sterile environment where plant cels, tissues, and organs can be grown,
manipulated, and studied outside their natural environment. The wide range of
applications includes plant propagation, genetic manipulation, production of
bioactive compounds, and disease resistance breeding, among others.

Below are some major applications of plant cell and tissue culture:

1. MICRO-PROPAGATION -

Micro-propagation, also known as in vitro propagation, is

one of the most significant and widely applied techniques in
plant biotechnology. It involves the rapid multiplication of
plants using tissue culture techniques to produce a large
number of identical plants (clones) from a single parent
plant. This method has become indispensable in the
production of horticultural crops, ornamental plants,
forestry, and the conservation of endangered species.

SHOOT ESTABLISHMENT SHOOT MULTIPLICATION

Explant
ROOT ESTABLISHMENT

ACLIMATIZATION In-vitro

MARKETING Ex-vitro

MICROPROPAGATiON TECHNIQUE
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2. Production of Secondary Metabolites

Plants produce a wide range of secondary metabolites, which are

organic compounds not directly involved in the growth,
development, or reproduction of the plant. These compounds,
which include alkaloids, terpenoids, flavonoids, and phenolics, play
essential roles in plant defense, signaling, and stress responses.
Many of these secondary metabolites have significant economic
value as pharmaceuticals, flavors, fragrances, dyes, and pesticides.

Pigments Carotenoids, Anthocyanins,

etc.
Alkaloids Morphine, Codeine, etc.
Terpeenoides Monoterpenes, Diterpenes etc.
Essential oils L e m o n g r a s s oil. ete.
Toxins Abrin. Ricina
Lectins Concanavalin A

Drugs Vinblastin, curcumin. etc.

Polymeric Rubber. guums, cellulose
substances

3. Somatic hybrid and cybrid

Somatic hybrids
Created by fusing protoplasts (isolated plant cells without cell
walls) from two different species. Somatic hybrids can be
symmetric, asymmetric, or cytoplasmic. First inter genetic somatic
hybrids between Solanum tuberosome and Lycopersicon
esculetum and hybrids are known as pomatoes or topatoes.

Cybrids-Also known as cytoplasmic hybrids, cybrids are created

by fusing the cytoplasm of one species with the nucleus of another
 . PLANT TISSUE CULTURE.

species. Cybrids have the genetic makeup of both parents in their

cytoplasm, but only the nucleus of one parent.
TRANSGENIC PLANTS

Transgenic plants, also known as genetically modified (GM) plants,

are developed using modern biotechnology to introduce new traits
such as pest resistance, herbicide tolerance, or enhanced
nutritional content.

Bt cotton is genetically modified to express a toxin from the

bacterium Bacillus thuringiensis (Bt). This toxin targets specific
insect pests, notably the bollworm, a major cotton pest. The
gene inserted into Bt cotton produces Cry proteins, which, when
ingested by pests, bind to their gut cells, causing paralysis and
death.

Golden Rice is a transgenic crop engineered to produce

beta-carotene, a precursor to vitamin A. Rice does not naturally
contain beta-carotene in its edible parts. The modification aims
to address vitamin A deficiency, which is a major health issue in
many developing countries.

Herbicide-resistant crops, such as Roundup Ready

soybeans, have been genetically modified to tolerate
specific herbicides, such as glyphosate. These crops allow
farmers to apply herbicides to control weeds without
harming the crop itself.
 .PLANT TISSUE CULTURE.

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BENEFITS
Stress tolerance plants encounter both biotic and abiotic
stresses and these stresses cause a clolossal loss of crop yield
and quality . Biotechnological strategies can be used to create
transgenic plants with increased resistance to disease and
pests.
Delayed fruit ripening Ripening can be slowed by reducing
ethylene production.This can be achived by introducing
ethylene forming gene in a way that will suppress its own
expression in the crop plant. Flavr-Savr is transgenic tomato
used in USA.
Male sterility is used to prevent unnecessary polination.
These are created by introducing a bacterial gene from
Bacillus amyloliquefaciens that encode enzyme Barnase.
Male fertility is restored by Brastar ,protein.
Edible vaccines Crop plants offer cost effective bioreactorto
express antigens which can be used as edible vaccines. Some
of antigenic determinants on surface of viruses and bacteria are
proteins made by pathogen. The expression of such antigenic
proteins in crops like banana and tomato are useful for
immunization of humans since these can be eaten raw.
Biodegradable plastic The biodegradable plastics, e.g
polyhydroxybutyrate ,is being commercially by fermentation-
with the bacterium Alcaligenes eutrophus. The large-scale
may be achieved by tree plants like populus.
 PLANT TISSUECULTURE.

ADVANTAGES OP PLANT TISSUE CULTURE

The plantlets are obtained in a very shorttime with a small

amount of plant tissue.

The new plants produced are disease-free.

The plants can be grown throughout the year, irrespective of

the season.

A large space is not required to grow plants by tissue culture

technique.
The production of new varieties in the market place speeds
up.

This technique is being used for the production of ornamental

plants such as dahlia, chrysanthemum, orchids, etc.

DISADVANTAGE8 OP PLANT TISSUE

CULTURE

Tissue culture can require more labor and cosst

more money in building the facility and equipping
the lab with all the instruments and chemicals.
There is a chance that the propagated plants will be
less resilient to diseases when grown in outside
conditions due to the type of environment they are
grown in.

Emergence of superpest, loss of biodiversity.

 .PLANT TISSUE CULTURE.

Contamination is the major issue in tissue culture

setting. Plants can get infected by bacteria, fungi,
and viruses
Tissue culture is an advanced technique and
require some advanced knowledge and practice for
anyone to get started in the field.
 .PLANT TISSUE CULTURE.

BIOSAFETY ISSUES

Biosafety in plant genetic engineering is a critical issue that

addresses the potential risks associated with the development,
use, and commercialization of genetically modified organisms
(GMOs), particularly transgenic plants. The growing adoption of
transgenic plants for agricultural purposes has sparked debates
over their safety for humans, the environment, and biodiversity.
Governments, international organizations, and researchers have
developed biosafety frameworks, guidelines, and risk assessments
to ensure that the benefits of transgenic plants are realized while
minimizing potential hazards.

O The antibiotic resistance genes used to produce transgenic

crops may horizontally transfer into microbes and thus
exacerbate problems of antibiotic resistance in human and
animal pathogens.
O The GM crops may lead to the change in the evolutionary
pattern.
The GM crops may changes the fundamental vegetable nature
of plants as the genes from animals are being introduced into
crop plants.
 ..PLANT TISSUE CULTURE .

CONCLUSION
The development of transgenic plants has
revolutionized agriculture by introducing
beneficial traits such as pest resistance, herbicide
tolerance, and enhanced nutritional content.
These crops have the potential to address global
food security challenges, improve environmental
sustainability, and boost farmers' economic
prosperity. However, the widespread adoption of
transgenic plants is accompanied by ethical,
ecological, and public concerns that need to be
carefully managed. Balancing the benefits of
genetic engineering with the need for safety,
transparency, and biodiversity conservation will
be crucial as the world continues to explore the
possibilities of biotechnology in agriculture.
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BIBLIOGRAPHY
o Food and Agriculture Organization (FAO). (2019). 1 tate d
Food Securiy and Nutrition in the World 2019. Retrieved from
FAO.org.
o World Health Organization (WHO). (2020). Genetically Modified
Foods: A Guide for Consumers. Retrieved from WHO.int.
National Academy of Sciences. (2016). Genetically
o
Engineered
Crops: Experiences and Prospects. Retrieved from nap.edu.
o McHughen, A., & Smyth, S. J. (2008). The Role of Biotechnology
in Global Food Security. Trends in Biotechnology, 26(7), 363-368.
DOI: 10.1016/j.tibtech.2008.03.006.
o
Zhang, Q, & Huang, J. (2015). Impacts of Transgenic Technology
on Agriculture in China. Transgenic Research, 24(6), 1039-1050.
DOI: 10.1007/s11248-015-9924-4.