Establishment of cell suspension lines of indica rice.

Abstract

Cell suspension cultures were initiated from 2-3 months old calli of eight cultivars of indica rice (Oryza sativa L.). The calli were originated from mature embryos, immature embryos and immature inflorescences. Seven to ten days old root segments of aseptically grown 8 rice cultivars were also employed to develop cell suspensions. Several liquid media i.e., MS, MS-T and Amino acid (AA) with a range of auxins (0.5 to 8.0 mg/l 2,4-dichlorophenoxy acetic acid) were used. After repeated selection, AA medium with 2.0 mg/l 2,4-D was found to be optimum for the growth and maintenance of cell lines. Five fast growing suspension lines were established from 3 cultivars (DM-25; B-370 and B. Kashmir). The cell lines of other cultivars are being developed. Most of the suspensions developed roots on MS or N6 regeneration media. Sporadic somatic embryogenesis were obtained in two cell lines of B-370 and B. Kashmir. The doubling time of DM-25-Inf and B-370-S in AA medium was found to be 3 and 4 days respectively. Maximum protoplast yield of newly established cell line (DM-25-Inf) was obtained with enzyme mixture of 150 mg cellulose R-10, 15 mg Pectolyase and 50 mg macerozyme in 10 ml CPW at pH 6.0 and 0.6 M mannitol as osmoticum.