1. Plant Tissue Culture
By Dr. S.D Randive-Aherkar

2.  What is tissue culture
 History of tissue culture
 Applications of tissue culture
 Ex-plant
 Infrastructure of tissue culture laboratory
 Types of media
 Composition of media
 Micro propagation - Process
 Applications of micro propagation

4.  Habarlandt (1902): Origin concept of tissue culture.
 Robbin (1922): Technique of root culture
 Robbins & Maneval (1923): Subculturing technique
 White (1939): Callus culture
 Skoog & Miller (1957): Shoot & root initiation
 Torrey & Shigomura (1957): Cell suspenssion culture
 Reinert (1959): Somatic embryogenesis
 Vasil & Hilderbrandt (1965): Totipotency
 Murashinge & Skoog (1962): MS media
 Guha & Maheshwaari (1966): Haploids
 Gamborg (1968):B5 medium
 Nitsch & Nitsch (1969): NN medium

5.  Rapid multiplication of plants
 To produce disease resistant plant
 To produce virus resistant plants
 To produce genetically modified plants
 To isolate photochemical
 To preserve RET plants
 To produce high yield
 To produce high vigour & vitality
 This process is applicable in any season.

6.  To produce haploids
 Applicable for plants which are sterile
 Anther culture
 Embryo culture
 Organ culture
 Shoot culture
 Somatic embryogenesis
 Somaclonal variations
 Germplasm conservation
 Phytoremedation

7.  Ability of the plant cell to divide and redivide to form a
new plant.

8. The ex-plant is any plant cell, tissue or organ which we
select for tissue culture.

10.  Sterilization room
 Glassware washing room
 Media preparation room
 Inoculation room
 Incubation room
 Growth room
 Green house

11.  Fumigation
 Surface sterilization
 Ex-plant sterilization
 Media sterilization
 Glassware sterilization

12.  MS media
 MS basal media
 MS ½ basal media
 Black media
 Whites media
 Nitsch Nitsch media
 B5 media
 N6 media
 LS media
 WPM media

13.  Macronutrients: Calcium, Magnesium, Potassium,
Nitrogen, Ammonium, Phosphorous, Sulphur
Role?
 Protein synthesis: N & S
 Nucleotide synthesis: P, N & S
 Cell wall synthesis: Ca
 Enzyme co-factors: Mg
 Membrane integrity: Mg

14.  Micronutrients: Boron, Cobalt, Iron, Manganese,
Molybdenum, Copper, Zinc, Iodine.
Role?
 Photosynthesis: Fe & Mn
 Cofactor of enzymes: Mo, Zn, Cu
 Lignin biosynthesis: B

15.  Growth regulators: Auxin, Cytokinene, Gibrellic acid,
ABA, Brassinostroides
 Vitamines: Thiamine- Carbohydrate metabolism
 Vit.c – Antioxidant
 Inositol – Membrane stabilization
 Other organic suppliments: Coconut milk, tomato juice,
polyamines, polyphenols

16.  Carbon source: Sucrose, fructose, maltose, glucose,
sorbitol, raffinose
 Activated charcoal
 Agar

17.  Propagation of plant by tissue culture under aseptic
conditions.
OR
 Rapid vegetative propagation of plants under in-vitro
condition.
 Selection of ex plant
 Multiplication
 Rooting & acclimatizing
 Hardening

18. In Direct Method of Propagation explants directly
developed in to meristomers without formation of callus.

19. In Indirect method of
propagation ex-plant first
develop in to callus & callus
after sub culturing gives rise
to new plantlet.

21.  Ex-plant
 Wash with D/W
 90% ethanol
 1% Hgcl2
 Tween 20/80
 Cut in to small pieces (1cm)
 Inoculate on ms media
 Callus formation
 Sub culturing
 Inoculation on same media
 Plantlet
 Hardening

22.  Production of millions of copies of plants.
 Large production in small space.
 Disease free plant
 High vigour & vitality
 To isolate photochemical
 Yield increases
 Carried out throughout the year