Plant tissue culture is a collection of techniques used to grow plant cells, tissues, or organs in sterile conditions. It relies on the fact that many plant cells can regenerate a whole plant. There are several types of tissue culture including callus culture, organ culture, and suspension culture. Plant tissue culture has many applications including micropropagation, production of pharmaceuticals, and genetic engineering of plants. It is a valuable tool for producing disease-free plants and increasing crop yields.

1. Regeneration Of Plants & Pharmaceutical
Applications Of Plant Tissue Culture
Subject :- Pharmaceutical Biotechnology
DEPARTMENT OF PHARMACEUTICAL SCIENCES
Dr. HARISINGH GOUR VISHWAVIDYALAYA
Sagar ( M.P)- 470003, India
(A Central University )
SUBMITTED BY :-
Suruchi Dahiya
( M Pharm Sem I )
Roll No;- Y22254028
SUBMITTED TO :-
Prof. UMESH K PATIL
(Professor, DOPS)
DR. UDITAAGARWAL
(Guest Faculty, DOPS)

2. CONTENTS
•Introduction
•Regeneration Of Plants
•Types of regeneration in plant
•Plant tissue culture
•Types of tissue culture
•Application of tissue culture
•Advantages & Disadvantages of tissue culture
•References

3. Introduction
▪ Regeneration:- Regeneration is the natural process of replacing or restoring
damaged or missing cells, tissues, organs, and even entire body parts to full
function in plants and animals.
▪ The process of growing an entire plant from a single cell or group of cells.
▪ Regeneration is possible because plant cells can be made totipotent using
hormones.
▪ Differentiated tissue: stems, leaves, roots, etc.
▪ Undifferentiated (embryonic) cells are totipotent: can become a whole new plant
by differentiating into a whole new plant.

4. THE PLANTS CAN BE REGENERATED BY :-
• ORGANOGENESIS :- It refers to the formation of organs from the cultured
explants. The shoot buds or monopolar structures are formed by manipulating the
ratio of cytokinin's to auxins in the cultures.
• SOMATIC EMBRYOGENESIS :- In Somatic embryogenesis, the totipotent
cells may undergo embryogenic pathway to form somatic embryos which are
grown to regenerate into complete plants.

5. REGENERATION OF PLANTLETS
1. Preparation of Suitable Nutrient Medium: Suitable nutrient medium as per
objective of culture is prepared and transferred into suitable containers.
2. Selection of Explants: Section of explants such as shoot tip should be done.
3. Sterilization of Explants: Surface sterilization of the explants by disinfectants
and then washing the explants with sterile distilled water is essential.
4. Inoculation: Inoculation (transfer) of the explants into the suitable nutrient
medium (which is sterilized by filter-sterilized to avoid microbial contamination) in
culture vessels under sterile conditions is done.

6. 5. Incubation: Growing the culture in the growth chamber or plant tissue culture
room, having the appropriate physical condition (i.e., artificial light; 16 hours of
photoperiod), temperature (-26°C) and relative humidity (50-60%) is required.
6. Regeneration: Regeneration of plants from cultured plant tissues is carried out.
7. Hardening: Hardening is gradual exposure of plantlets to an environmental
conditions.
8. Plantlet Transfer :After hardening plantlets transferred to the green house or
field conditions following acclimatization (hardening) of regenerated plants

8. Plant tissue culture
• Plant tissue culture is a collection of techniques used to maintain or grow plant
cells, tissues or organs under sterile conditions on a nutrient culture medium of
known composition.
• It is widely used to produce clones of a plant in a method known as
micropropagation.
• Plant tissue culture relies on the fact that many plant cells have the ability to
regenerate a whole plant (Cellular totipotency)
• single cells, plant cells without cell walls (protoplasts), pieces of leaves, stems or
roots can often be used to generate a new plant on culture media given the
required nutrients and plant hormones.
• Preparation of plant tissue for tissue culture is performed under aseptic conditions
under HEPA filtered air provided by a laminar flow cabinet.

9. • It has proved beneficial for the production of disease-free plants and increase plant
yield in developing countries
• It only requires a sterile workplace, greenhouse, trained manpower, and a nursery.
Following are the main categories of Cultures:-
Primary Culture:- These model the natural function of the Tissue and are generally
mortal. They consist of natural Tissues excised from the living organisms by biopsy.
Culture of Established Cell Lines:- These are derived from tumor biopsies, or
from the primary cells that had undergone mutation and continued to replicate.

10. Types of Tissue Culture
• Some types of Tissue Culture techniques:-
Seed Culture:- In seed Culture, explants are obtained from an in- vitro derived
plant and hence are introduced into a laboratory where they proliferate. To prevent
the plants from Tissue damage it should be sterilized.
Embryo Culture:- Embryo Culture involves the in-vitro development of an
embryo. For this process, an embryo is isolated from and living organism, both a
mature and an immature embryo can be used. Mature embryos can be obtained from
ripe seeds whereas immature embryos are obtained from the seeds that failed to
germinate. The ovule, seed, or fruit has already been sterilized, hence there is no
need to sterilize them.

11. • Callus Culture:- A callus can be defined as an unorganized, dividing mass of
cells. A callus is the explants are Cultured in a proper medium good. The growth
of callus is followed by organ differentiation. This Culture is grown on a gel-like
medium composed of agar and specific nutrients which are required for the growth
of the cells.
• Organ Culture:- In organ Culture, any organ of the plant such as a shoot, the leaf
can be used as an explant. Many methods can be used for the organ Culture such
as the plasma clot method, raft method, the grid method, and Agar gel method.
This method can be used to preserve the structure and functions of an organism.

12. • Protoplast Culture:- It can be defined as a cell without a cell wall. The
hanging-drop method or micro-Culture chambers can be used to Culture a
protoplast. A number of phases can be observed in protoplast Culture,
development of cell walls, cell division, regeneration of a whole plant.
• Suspension Culture:- suspension Culture can be defined as a form of Culture in
which single cells or small aggregates of cells multiply while suspended in an
agitated liquid medium. It can also be called cell Culture or cell suspension
Culture.

13. • Meristem Culture:- meristems have the main function of the production of new
cells and the synthesis of protoplasm. Shoot meristem consists of a group of
certain actively dividing cells that are being protected by the developing leaves.
Steps of Tissue Culture :-
• Following are the steps of Tissue Culture:-
Initiation Phase:-
• This is a stage when the Tissue is initiated into the Culture. To prevent the process
from any contamination the Tissue of interest is obtained, introduced, and
sterilized.

14. Multiplication Phase:-
• In the multiplication stage, The sterilized ex-plant is introduced into the medium
which consists of growth regulators and appropriate nutrients, they are responsible
for the multiplication of cells. Hence this undifferentiated mass of cells is known
as a callus.
Root Formation:-
• This is the stage when the root starts forming. To initiate the formation of root
plant growth hormones are added. Consequently, complete plantlets are obtained.

15. Shoot Formation:-
• For the formation of the shoot, plant growth hormones are added and growth is
observed for a week.
Acclimatization:-
• When the plant starts to develop, the plant is transferred to a greenhouse for it to
develop under controlled environmental conditions. Thereafter it is finally
transferred to the nurseries for its growth under natural environmental conditions.

16. • HORMONES USED IN PLANT TISSUE CULTURE :-
1. Auxins
2. Cytokinin's
3. Gibberellins
4. Abscisic Acid
5. Polyamines

17. • Application of plant tissue culture
• Plant tissue culture technology has been used in almost all the field of biosciences.
Its applications include
• Production of phytopharmaceuticals and secondary metabolites.
a) Biotransformation (Biochemical Conversion)
b) Plant cell immobilization)
c) Genetic transformation (Transgenic plant)
d) Elicitors
• Micropropagation (Clonal Propagation)
• Synthetic seed Protoplast culture and somatic hybridization

18. Extracts from black carrot tissue culture as potent anticancer agents.
Enhanced production of tropane alkaloids in transgenic Scopolia parviflora hairy
root cultures over-expressing putrescine N-methyl transferase (PMT) and
hyoscyamine-6B-hydroxylase (H6H).
Taxus globosa S. cell lines: initiation, selection and characterization in terms of
growth, and of baccatin III and paclitaxel production.
Production of camptothecin in cultures of Chonemorpha grandiflora.
Regeneration, in vitro glycoalkaloids production and evaluation of bioactivity of
callus methanolic extract of Solanum tuberosum L.
The influence of medium composition on alkaloid biosynthesis by Penicillium
citrinum.

19. A Differential production of tropane alkaloids in hairy roots and in vitro cultured
two accessions of Atropa belladonna L. under nitrate treatments.
Increased vincristine production from Agrobacterium tumefaciens C58 induced
shooty teratomas of Catharanthus roseus G. Don.
Enhancement of taxane production in hairy root culture of Taxus x media var.
Hicksii
An endophytic taxol-producing fungus from Taxus media, Cladosporium
cladosporioides MD2.
Optimized nutrient medium for galanthamine production in Leucojum aestivum L.
in vitro shoot system.
Salinity stress enhances production of solasodine in Solanum nigrum L.

20. REFERENCES
• D.Dumet, A.Adeyemi, O.B.Ojuederie (2008). Yam invitro genebanking. Genebank
manual. http://www.iita.org/genebank/manual
• D.Dumet, A.Adeyemi, O.B.Ojuederie (2008). Cassava in vitro processing and the
genebanking. Genebank manual. http://www.iita.org/genebank/manual
• HORT689/AGRO689 Biotechniques in Plant Breeding
• H.S Chawla .2002 Introduction to Plant Biotechnology 2nd edition. Oxford & IBH
Publishing C./ Pvt. Ltd New Delhi India
• Murashige T. and Skoog F. (1962). A revised medium for rapid growth and
bioassay with Tobacco tissue culture. Physiologia plantarum 15: 473-497.