PLANT TISSUE CULTURE

Plant tissue culture is the growth of plant cell or

tissue in vitro under suitable conditions and in
solid, liquid or semi solid media to produce a
whole new plant. The process of plant tissue
culturing requires nutrient media, sterile conditions
to work and optimum environmental conditions.
The tissue culturing involves the preparation of
suitable media (having all organic and inorganic
nutrients, growth hormones, and vitamins), Figure 1 PLANT TISSUE CULTURE (Plant Tissue Culture, 2020)

sterilization of explant and other equipments and

incubators. (Sharma et al., 2015)

IMPORTANCE OF PLANT TISSUE CULTURE ON INDUSTRIAL LEVEL

In plant biotechnology, plant tissue culture is an essential component. It provides the opportunity
of producing large number of plant clones on large scale. Moreover, a novel plant or cell that is
genetically produced can also be multiplied into hundreds and thousands of cells, under aseptic
conditions. On industrial scale, the plant tissue culture can be used to produce the disease free
plants that are resistant towards bacterial or fungal pathogens, rapid and fast propagation of those
plants that are difficult to produce otherwise, to produce somatic hybrids by undergoing somatic
hybridization. Moreover, the commercial plants can be improved genetically, for the breeding
programs androgenic haploid plants can be produced; (Sharma et al., 2015) secondary
metabolites can be obtained. A very significant use of plant tissue culture is to save the
endangered, rare or threatened plant species by micropropagating the plants. (Hussain et al.,
2012)

Other than these benefits, plant tissue cultures can be useful in crop improvement by introducing
soma clonal or gametoclonal variations. The useful variants having superior genotypes are
isolated and propagated further. This is a rapid process which can also be used to produce virus
free or resistant plants, like Meristem tip culture of bananas was used to produce plants that were

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resistant against brome mosaic virus (BMV) and banana bunchy top virus (BBTV). (Hussain et
al., 2012)

HISTORY OF PLANT TISSUE CULTURE

History of plant tissue culture starts in 1838, when for the first time Schleiden and Schwann gave
the cell theory and declared cell as a functional and structural unit of life, they also studied the
totipotency of cells. Then in 1902, Gottlieb Haberlandt gave the concept of in-vitro culture of
plant cells. He cultured monocots; it was an unsuccessful attempt because the cells die after a
month but it gave a concept of totipotency and invitro culturing. In 1904, Hannig established
embryo culture of some Crucifers. Then in 1922, Robbins established invitro root tip culture.
(Sharma et al., 2015). In 1939, callus cultures were established by Gautheret, White and
Nobecourt. Different media were also developed with time to give plant cells and tissues
appropriate environment and nutrients to grow. In 1962, Murashige and Skoog developed the
MS media, in which the concentration of salt was high. (Hussain et al., 2012)

After that in 1964, Guha and Maheshwari two Indian scientist, used pollen grains to produce
haploid plants. After that in 1966, Steward uses the concept of totipotency by regenerating the
carrot plants. (Hussain et al., 2012).

In 1981, a very significant addition was done in tissue culture by Larkin, as he introduced the
term ‘soma clonal variations’ to culturing. Then in 1996, agrolistic method of plant
transformation was developed by Hansen. (Sharma et al., 2015).

TYPES OF PLANT TISSUE CULTURE

To perform culturing of plants, different explants can be used and based on the explant used;
there are different types of plant tissue culture. Some of them are callus culture, organ culture,
embryo culture, anther and pollen culture, protoplast culture and seed culture. (Sharma et al.,
2015).

Callus Culture:

Callus culture is the type of culture in which undifferentiated and unorganized cells are obtained
from the plant and are grown in nutrient medium under aseptic conditions to produce a whole

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new plant. (Sharma et al., 2015). In Biotechnology,
callus cultures are widely used cultures as their
proliferation rate is fast. As callus is unorganized mass
of cells, so by manipulating the ratio of auxin and
cytokinin hormones, the shoots and roots of the plant
can be induced and as a result due to organogenesis a
whole new plant regenerate from callus.
Figure 2 Callus Culture of Nicotiana tabacum
(Callus Culture of Nicotiana Tabacum, 2021)
Categories of callus culture

Callus culture is divided into two categories; 1) Compact callus and 2) Friable Callus. Compact
callus is the one in which the cells are densely aggregated together whereas on the other hand,
Friable callus is the one in which the cells are loosely attached and there is no compact
aggregation between the cells. Friable callus is soft and can break into cells easily. Friable callus
can be a good inoculum of suspension culture.

Anther and Pollen Culture

Anther and pollen culture involves the growth of a

haploid plant from pollen grains of the flower by going
many cell divisions and pathways. Anther culture is also
known as Androgenesis. In anther culture, unopened bud
is taken as whole where as in pollen culture the pollen
grains are extracted and cultured. (Sharma et al., 2015).
The plants that are produced as a result of pollen or
anther culture are Haploid, means they have a single set
of chromosomes. Such plants are of great importance to
Figure 3 Fig 3: Anther culture (Anther Culture,
geneticist and plant breeders. In 1964, Gahu and 2020)
Maheshwari produce a haploid plant from pollen grains.

Haploid plants are weaker, difficult to maintain and highly sterile as compared to normal diploid
plants. To identify or verify whether a plant is haploid or not a number of methods are used
including, biochemical studies, cytological studies or marker genes that are associated with
haploidy can be studied.

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Embryo Culture:

Embryo culture involves the growing of embryo regardless of shape, developmental stage or size
in invitro environment. As the embryo develops by the fusion of egg and sperm so in 1925
Laibach for the first time isolated an inter-specific embryo of Linum perenne x Linum
austriacum. He excised embryo from the seed of plant and
grow it on moist cotton that was soaked in 15% glucose
and as a result a whole new plant was produced. He
Figure 4 Embryo culture (Embryo Culture, 2017)
provides the base of embryo culture. So now the plants or
seeds that have threat to have embryo abortion are grown
using this technique.

Embryo culture is used to overcome seed dormancy, to test the viability of the seeds, to avoid
embryo abortion and to save the threatened or endangered plant species. Recently,
Khayagrandifoliola is propagated by taking the embryo out of the mature seed and culturing it.
(Hussain et al., 2012).

Types of Embryo Culture:

Basically, there are two types of embryo culture. 1) Mature Embryo Culture and 2) Immature
Embryo Culture. In mature embryo culture, the embryo is derived from the ripe seeds. This
type of embryo culture requires simple media. The culture is established due to certain reasons
like when embryo is unable to grow invitro or when it becomes dormant for longer period of
time. Some plant species like orchids or iris produce sterile seeds due to incomplete development
of embryo, so these embryos can be cultured and viable seeds can be generated. In immature
embryo culture, the culture is developed to rescue the wild crosses to produce viable seed by
avoiding embryo abortion. This type of culture needs complex media for the growth of embryo.
The media should have amino acids, hormones, mineral salts and coconut milk. (Hussain et al.,
2012).

Protoplast Culture

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Protoplasts are the naked plant cells without cell walls and are surrounded by plasma membrane.
The plasma membrane has the potential to regenerate
cell wall, cell growth and division and ultimately to
regenerate a whole new plant. (Sharma et al., 2015). The
protoplast culture is mainly used for the somatic
hybridization in which protoplasts of two cells are fused
together to generate a somatic hybrid. It involves the Figure 5 Protoplast culture (Protoplast Culture,
2020)
gene transfer from one specie to specie to have desire
traits in new one. (Hussain et al., 2012). Vegetatively propagated plants like potato, sweet potato,
banana and sugar cane, can be produced by using this technique.

Meristem culture

Meristem culture involves the culturing of

apical shoot meristem. In 1952,
meristem culture was used to generate virus-
free plants. Morel and Martin took 100m
long meristem of a plant that was infected
from virus and cultured it to generate
virus free plant. Since then this type of
culture is used to Figure 6 Schematic representation of production of hybrid
obtained plants that are
plant via protoplast fusion (Schematic Representation of
free from or resistant Production of Hybrid Plant via Protoplast Fusion, 2012)
against viruses,
bacteria, mycoplasma, viroids and fungi etc.
The method can also be used for clonal propagation of plants. (Sharma et al., 2015).

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