Isolation and Culture of Protoplasts from Different Explants of Cotton (Gossypium hirsutum L.)

Abstract

Protoplasts were isolated from callus, cotyledons, hypocotyls and leaves of cotton (Gossypium hirsutum L. var. Coker 310) using different digestion media. Protoplast yield per gram of fresh mass of leaves, callus cotyledons and hypocotyls was 22.2, 5.5, 1.7 and 1.1 million respectively. Protoplasts were most viable in basic medium in presence of thidiazuro 0.663 gm/l and 2, 4, 5-trichlorophenoxy acetic acid 7 mg/l. Hypocotyl and leaf-derived protoplasts formed microcolonies. Addition of CaCl₂ and glutamine to the basic medium improved protoplast survival, and division of hypocotyl and leaf-derived protoplasts. The growth conditions, such as the light and carbon source of donor plant were the important factors for protoplast isolation and culture.