HIGH-FREQUENCY SOMATIC EMBRYOGENESIS AND PLANTLET REGENERATION FROM HYPOCOTYL PROTOPLAST CULTURES OF BRASSICA-NAPUS

A simple protocol has been developed for high frequency protoplast regeneration via somatic embryogenesis in B. napus. Protoplasts isolated from hypocotyl tissue of 8-12 day old seedlings of Brassica napus ISN706 (AACC) when cultured in KM(A) medium resulted in divisions with a, frequency ranging from 30-35%. Regeneration of plantlets was possible by both organogenesis and embryogenesis. Nearly 80% of the call transferred on to MS medium supplemented with 5.0 mg 1(-1) 2iP, 0.1 mg 1(-1) NAA, 0.001 mg 1(-1) GA(3), 0.5 g 1(-1) PVP and 0.5 g 1(-1) MES displayed somatic embryogenesis. The somatic embryos developed into normal plantlets, and also displayed secondary, repetitive embryogenesis.