caa a22 4500 469028033 CHVBK 20180323132735.0 cr unu---uuuuu 170328e19920401xx s 000 0 eng 10.1007/BF00232171 doi (NATIONALLICENCE)springer-10.1007/BF00232171 Rapid and efficient plant regeneration from hypocotyl protoplasts of Brassica carinata [Elektronische Daten] [S. Narasimhulu, P. Kirti, S. Prakash, V. Chopra] Protoplasts were isolated from hypocotyls of 7-d-old seedlings of three genotypes of Brassica carinata after enzymatic digestion in cellulase R-10 (0.5%) and pectolyase Y-23 (0.025%). The protoplasts were stabilized with 0.4 M mannitol used as osmoticum, and were cultured in darkness in Kao's liquid medium containing 0.4 M glucose and the growth regulators 2,4-D (1.0 mg/l), NAA (0.1 mg/l) and zeatin riboside (0.5 mg/l). Protoplasts were transferred to 16 h photoperiod conditions after 3 d of dark culture, and the medium was diluted to reduce the osmoticum on the seventh and tenth days of culture. Microcolonies were thus obtained which, upon transfer to MS agarose medium with 2,4-D (0.1 mg/l), BAP (1 mg/l) and 0.1 M sucrose, proliferated further to produce callus clumps. The plating efficiency of the three genotypes varied from 1 to 2%. Calli 2-3 mm in diameter were transferred to MS agarose plates with zeatin (2 mg/l) where they produced shoot buds and shoots with frequencies ranging from 22.5 to 74.2% for the three genotypes. The shoots were rooted in medium with IBA (1 mg/l) and were then established in soil. The time required for protoplast to plant development was 8 to 10 weeks. Springer-Verlag, 1992 Narasimhulu S. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut Kirti P. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut Prakash S. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut Chopra V. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut Plant Cell Reports Springer-Verlag 11/3(1992-04-01), 159-162 0721-7714 11:3<159 1992 11 299 https://doi.org/10.1007/BF00232171 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00232171 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Narasimhulu S. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut NATIONALLICENCE 700 1- Kirti P. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut NATIONALLICENCE 700 1- Prakash S. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut NATIONALLICENCE 700 1- Chopra V. Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 11/3(1992-04-01), 159-162 0721-7714 11:3<159 1992 11 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer