Plant Tissue Culture By Rafeeq Begum

PLANT TISSUE CULTURE RAFEEQ BEGUM, Lecturer in BOTANY, Govt. Degree College for Women, KOLAR.

INTRODUCTION  Did you ever had a plant that was so unique or beautiful that you wished in plenty to enjoy or sell it.  Plant Tissue culture is thus a Biological Tool to protect, improve and safe guard the plants for us.  Plant Tissue Culture is defined as In –Vitro culture of plant cell, tissue, organs, seeds on a defined Nutrient Media under aseptic conditions.  German Botanist Gottelieb Haberlandt (1902) was the first to culture isolated fully differentiated cells in nutrient media and is regarded as “Father of Tissue Culture”. He put forth the concept of Totipotency.

Basic Facilities For Tissue Culture Well equipped laboratory with • Washing and Storage facility. • Media preparation, sterilization & storage. • Transfer area for aseptic manipulation. • Culture room (Dark/ Light rooms) • Data collection and documentation facility. • Attached Green House facility.

Basic Technique of Tissue Culture  Selection of Explant- Stem tip, node, meristem, embryo or seed.  Sterilization and growing on Culture Media aseptically.  Undifferentiated mass of cells- callus formed, is transferred on to another media for shoot growth or callus formation or multiple shoots directly produced.  Hardening of plant, transfer into Green house and then into the field.

MICROPROPOGATION

CALLUS

TISSUE CULTURED PLANTLETS MULTIPLE SHOOT PROLIFERATION

Important factors to be monitered • Maintenance of aseptic conditions • Providing suitable environment for plant growth. • Use of specific growth media with growth harmones.

Nutrient media & disinfectants Nutrient Media MS- Murashige & Skoog`s Media LS- Linsmares & Skoogs Media. White`s media, B5, N6 media also used.  For callus formation Auxins and cytokinins are required.  Auxin is required for shoot formation/ organogenesis.

Disinfectants Sodium hypochloride(0.5-5%) Mercuric chloride (0.1- 1%)

Applications of tissue Culture • Small amount of tissue can be used to raise hundreds of plants in short period. • High yielding verities of several plants can be developed at low cost. • Several commercial products, drugs, secondary metabolites can be produced in a short duration • Eradication of virus is outstanding contribution of tissue culture. • In vitro pollination help in overcoming incompatibility barrier.

• Induction of haploidy through anther culture. • Protoplast fusion helped in introducing somatic hybrids. • Genetic transformation. • Protection of novel plants. • Conservation of plants of medicinal importance, aesthetic value and plants at verge of extinction. • Altogether conservation of germplasm.