Tissue Culture

by Abdul Hanan
 INTRODUCTION
 Tissue Culture
 Plant tissue culture is
collection of techniques
in which cells, tissues
or organs are used to
grown under sterile
condition on nutrient
medium of the known
composition.
Basics Steps In Tissue Culture:

Selection of Explant

Sterilization Of Explant

Culturing of the Explant (callus formation)

Regeneration & Multiplication

Rooting Hardening
Tissue culture Methods Used in
Sugarcane
 Micropropagation
 Rapid production of true to type plants
 Plants are identical
 Soma clonal variation
 Nuclear chromosomal rearrangements and losses
 Gene amplification and de-amplification
 Apparent point mutations
 Work Done In ABRI,AARI
 Media preparation
Sr.# Quantity(in one-liter dh2o)
1
Ingredients
1 MS media (Murashige and Skoog,1962) 4.43 g
2 Phytagel 1.75 g
3 Fe (Iron) 10.00 ml
4 D-sucrose 30.00 g
5 2,4-D 3g
#2 For Regeneration  
1    
Casein hydrolysate 480.00 mg
2
Kinetin 1mg
For Roots development  
#3
NAA 0.5-1mg
1
For rooting media Half MS media was used.
 Protocol for the sterilization of explants

Sr.# Chemicals Concentration (%)

0.1
1 Mercuric chloride
 

2 Sodium hypochlorite 10 to 15

3 Ethanol 70
Regeneration