This document discusses various tissue culture techniques used in plant biotechnology, including organogenesis, embryogenesis, synthetic seed production, somaclonal variation, protoplast fusion, hairy root culture, micropropagation, and gene transfer. Organogenesis involves inducing organ development from cultured plant tissues. Embryogenesis refers to embryo formation, which can occur somatically from undifferentiated callus cells. Synthetic seeds are artificially encapsulated plant materials that can be used for propagation. Somaclonal variation results from phenotypic changes during long-term culture. Protoplast fusion combines genetic material from two distinct plant species. Hairy root culture uses transformed roots induced by Agrobacterium rhizogenes for metabolite production. Micropropagation is high

1. Different
Tissue Culture
Techniques
BY- DURGASHREE M D
MEDICINAL PLANT BIOTECHNOLOGY
DEPT OF PHARMACOGNOSY
KLE COLLEGE OF PHARMACY, BENGALURU
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2. Introduction
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14. Organogenesis
The process of initiation
and development of an
organ is called
organogenesis.
In-plant tissue culture,
inducing organogenesis is an
important way to regenerate
plants from the culture.
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18. Embryogenesis
The process of formation of an embryo is called
embryogenesis.
Embryogenesis starts from a single embryogenic cell,
which can be a zygote (the product of the fusion of an
egg and a sperm during fertilization),
Embryogenesis from an undifferentiated callus cell is
termed Somatic Embryogenesis.
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25. Synthetic Seed
Synthetic seeds are artificially
encapsulated plant propagation
material.
This material could be somatic
embryos, shoot buds, cell aggregates,
or any other tissue that we can use as
a seed for propagation.
Synthetic seed technology primarily
involves encapsulating somatic
embryos in a protective coating5.
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28. Advantages of Synthetic Seed
They can easily be stored for up to a year without loss of
viability.
They are easy to handle and useful as units of delivery.
They can be directly sown in the soil like a natural one.
Do no need hardening in the greenhouse.
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30. Monoclonal Variation
Generally called Somaclonal
Variation.
Plant regeneration from
callus, leaf explant, or plant
protoplast leads to the
generation of considerable
variation- Somaclonal variation.
This variation includes
Aneuploidy, sterile plants,
morphological variants, etc.
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31. Monoclonal Variation
This was ignored before, but in the last decade, it has
received increased attention, in view of potential
improvement programs.
This is due to phenotypic variability and even changes in
chromosomal number during long-time culture.
This is exhibited in callus and in plants generated from the
callus1.
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32. Reported Variation of MV
Species Characters modified
Allium sativum Bulb size, clove no, aerial bulbil
Oryza sativa Plant height, seed fertility, grain no, etc.
Lolium hybrids Leaf size, flower, survival, etc.
Lactuca sativa Leaf weight, length, width, color, etc.
Solanum tuberosum Tuber shape, maturity date, color, etc.
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34. Application of MV
Plants with
Dieasese resistant
Abiotic stress resistance
Salt tolerance
Herbicide resistance
Insect resistance, etc.
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35. Protoplast Fusion
Somatic fusion, also called protoplast
fusion, is a type of genetic
modification in plants by which two
distinct species of plants are fused
together to form a new hybrid plant
with the characteristics of both, a
somatic hybrid6.
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Protoplast fusion of Asteraceae family

41. Hairy Root Culture
Hairy roots are differentiated cultures of transformed roots
that are developed by the infection of wounded higher
plants with Agrobacterium rhizogenes and are an
exceptional source of advantageous metabolites with high
medicinal value for drug development7.
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42. Hairy Root Culture
The term was first coined by
Steward in 1900.
In 1930, Ricker first named the hairy
root-causing organism- Phytomonas
rhizogenes.
Later it is known as Agrobacterium
rhizogenes.
The ability of root induction lies in
the typical genome structure of
A.rhizogenes.
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44. Hairy Root Culture
Hairy roots can produce secret complex active glycoproteins from
a large spectrum of organisms.
They are also adequate to express plant natural biosynthesis
pathways required to produce specialized metabolites.
This adaptability has positioned hairy root platforms as major
biotechnological tools8.
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48. Multiple shoot culture
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 Induction of multiple-shoot culture (MSC) formation from hemp
(Cannabis sativa L., cv. USO-31) isolated apical meristems using
two different induction media: a IMB4-T-AS medium containing
BAP9THP and adenine hemisulphate; b IMB4-T-P medium
containing BAP9THP and the auxin inhibitor PEO-IAA (seventh
experiment). Apical dominance was suppressed under both
conditions

49. Micropropagation
Micropropagation is the rapid
vegetative propagation of plants
under in vitro conditions, of
high light intensity,
controlled temperature, and
a defined nutrient medium.
The technique has been applied to a
substantial number of commercial
vegetatively propagated plant
species9.
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50. Stages of Micropropagation
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53. Application in
Aromatic Plants
1. Black pepper
regeneration from
leaf and shoot tip.
2. Vanilla from root
meristem, axillary
bud, shoot tip, etc.
3. Strawberry from
apical meristem, tip,
leaf disc, etc.
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54. Sterilization
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60. Gene Transfer in Plants
Gene transfer pertains to the transfer of genes between organisms.
It may be a horizontal gene transfer or a vertical gene transfer.
The transfer of genes is horizontally when a segment of DNA is copied
and inserted from one site to another of the same or of a different
chromosome.
It is also referred to as transposition.
The genes, referred to as transposons or jumping genes, are
transferred from the donor organism to the recipient organism through
gene copying and insertion10.
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61. Gene Transfer in Plants
Plant transformation is a way to insert DNA from another
organism- normally another plant, into the genome of a
plant of interest.
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Agrobacterium-mediated gene transfer.

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67. References
1. Elements of Biotechnology- P K Guptha
2. Plant tissue culture- M K Razdan
3. Textbook of Medicinal Plant Biotechnology- Dr. Rageeb M D.
4. Plant Tissue Culture ppt by Sanjitha P
5. Application of synthetic seed for propagation.
(https://link.springer.com/chapter/10.1007/978-3-030-24631-0_13)
6. Somatic fusion.
(https://en.wikipedia.org/wiki/Somatic_fusion#:~:text=Somatic%20fusion%2C
%20also%20called%20protoplast,of%20both%2C%20a%20somatic%20hybrid.)
7. Hairy root culture- An Overview.
(https://www.sciencedirect.com/topics/immunology-and-microbiology/hairy-
root-culture)
8. Hairy root cultures.
(https://www.frontiersin.org/articles/10.3389/fpls.2020.00033/full)
9. Micropropagation- An Overview.
(https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/micropropagation)
10. Gene Transfer. (https://www.biologyonline.com/dictionary/gene-transfer)
11. Induction of multiple-shoot culture (MSC) formation from hemp.
(https://www.researchgate.net/figure/Induction-of-multiple-shoot-culture-
MSC-formation-from-hemp-Cannabis-sativa-L-cv_fig3_335752495)
12. Plant Biotechnology- Slater, Scott, Fowler.
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