Council of Scientific and Industrial Research

Madhu Sharma

Paramvir Singh Ahuja

Amit A Bhattacharya

Indra Sandal

P S Ahuja

Pratap Kumar Pati

The present invention relates to novel method for micropropagation of tea plants from explants obtained from completely folded, half opened or fully expanded leaves via callus phase, by culturing the explants on different media.

Method for microproduction of tea plants from leaf explants

The invention a novel and efficient method for protoplast culture comprising the steps of isolating the protoplasts from a cell suspension, mixing the protoplasts with equal volumes of alginate solution, placing 40-50 &mgr;l of CaCl solution on a glass microslide, placing a mixture of protoplasts and alginate solution on the glass microslide and immediately covering by a glass coverglass, adding CaCl solution from the sides of coverglass, sliding down the coverglass towards one side and placing it in a petridish containing protoplast culture medium, sealing the petridishes, incubating it and transferring the extra thin alginate layer with celled colonies to regeneration medium for development of culture.

Efficient method of protoplast culture

The invention provides a one step method for micropropagation of tea plants from leaf explants via callus phase, wherein the explants are maintained in basal Murashige and Skoog medium (Murashige T. and Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497; 1962) supplemented with vitamins like thiamine-HCl (0.05 to 2.0 mg/l), pyridoxine-HCl (0.25 to 1.5 mg/l) and nicotinic acid (0.25 to 1.5 mg/l) together with glycine (1.0 to 3.0 mg/l) and either 2.5, 5.0 or 7.5 mg/l 2,4-Dichlorophenoxy acetic acid for a period of 14-16 weeks, from the stage of induction of callus to rhizogenesis and shoot formation.

Growing community of inventors

Himachal Pradesh, India

Madhu Sharma