1. Presented by
Surya Pratap Singh
M . Sc , Life Sciences
Central University of Gujarat

2. INTRODUCTION
 The aseptic culture of plant protoplasts, cells, tissues or
organs under conditions which lead to cell multiplication or
regeneration of organs or whole plants.

3. HISTORY OF PLANT TISSUE CULTURE
 The idea of the totipotency of plant cell was given by
Haberlandt in 1902.
 White first time established successful root culture of
tomato in 1937.
 Vanoverbeek used coconut milk for growth and
development of young Datura embryos in 1941.
 Skoog and Miller demonstrated the role of auxin and
cytokinin on root and shoot formation in tobacco – tissue in
1957.
 Murashige and Skoog introduced the medium for tobacco
culture in 1962.
 Guha and Maheshwari obtained for the first time haploid
Plants through anther culture of Datura.

4. TYPES OF PLANT TISSUE CULTURES
Plant tissue culture
Embryo culture Seed culture Meristem culture
Protoplast cultureCell culture
Callus culture
Bud culture
Organ culture

5. Callus – A tissue that develops in response to injury caused
by physical or chemical means, most cells of which are
differentiated although may be and are often highly
unorganized within the tissue. In nature, this wound tissue
produces a protective layer of cells to cover an injury, example.
Differentiated Cells - products of cellular maturation, i.e.
cell types with particular function, e.g. xylem tracheary
elements; large cells that are highly vacuolated with
relatively little cytoplasm
Undifferentiated Cells - meristematic; progenitors of
differentiated somatic cells, e.g. small, isodiametric, small
vacuoles.

6. Tobacco Callus

7. CELL SUSPENSION CULTURE
 Tissues and cells cultured in a agitated liquid medium
produce a suspension of single cells and cells clumps of few
to may cell, these are called suspension cultures.

8. 2nd Passage1st PassageFriable Callus

9. PROTOPLAST CULTURES
 Isolated protoplasts have been described as "naked" cells because the
cell wall has been removed by either a mechanical or an enzymatic
process.
 Protoplasts can be induced to reform a cell and divide if placed in a
suitable nutrient medium than form callus.

11. Isolation of Protoplast
(Separation of protoplasts from plant tissue)
1. Mechanical Method 2. Enzymatic Method

12. MECHANICAL METHOD
Plant Tissue
Collection of protoplasm
Cells Plasmolysis
Microscope Observation of cells
Cutting cell wall with knife
Release of protoplasm

13. ENZYMATIC METHOD
Leaf sterlization, removal of
epidermis
Plasmolysed
cells
Plasmolysed
cells
Pectinase +cellulase Pectinase
Protoplasm released
Release of
isolated cells
cellulase
Protoplasm
released
Isolated
Protoplasm

14. MERISTEM CULTURE
 Cultivation of auxiliary or apical shoot
meristems, particularly of shoot apical meristem ,is known
as meristem culture.

15.  Application
 Production of virus free germplasm.
 Mass production of desirable genotypes.

16. ANTHER CULTURE/POLLEN CULTURE
 Anther culture is a technique by which the developing
anthers from unopened flower bud are cultured on a nutrient
medium where the microspores within the cultured anther
develop into callus tissue or embryoids that give rise to
haploid plant.
Application
 Production of haploid plant
 Production of useful gametoclonal varition.

17. Figure Show the Anther culture and haploid plant

18. SOMATIC EMBRRYOGENESIS
 In somatic embryogenesis the embryos regenerate from
somatic cells, tissue or organs either de nove directly from
the tissues, which is the opposite of zygotic or sexual
embryogenesis.

19. TWO TYPES OF SOMATIC EMBRYOGENESIS
 Direct embryogenesis
 The embryos initiate directly from explants in the
absence of callus formation.
 Indirect embryogenesis
 Callus from explants takes place from which
embryos are developed.

20. APPLICATION
 Production of artificial seeds.
 As source materials for embryogenic protoplast.
 For genetic transformation.
 Production of primary metabolites specific to seeds
such as lipids in oil seeds.

21. OVARY/OVULE CULTURE
 Ovary or ovule culture involves development of haploid from
unfertilized cells of embryosac present in ovary.

22. Application
 Production of haploid plants.
 Achievement of in vitro fertilization.