caa a22 4500 469028076 CHVBK 20180323132735.0 cr unu---uuuuu 170328e19920901xx s 000 0 eng 10.1007/BF00236266 doi (NATIONALLICENCE)springer-10.1007/BF00236266 Agrobacterium -mediated genetic transformation of oilseed Brassica campestris [Elektronische Daten] Transformation frequency is strongly influenced by the mode of shoot regeneration [A. Mukhopadhyay, N. Arumugam, P. Nandakumar, A. Pradhan, V. Gupta, D. Pental] Summary: Protocols were developed for efficient shoot regeneration from hypocotyl and cotyledon explants of oilseed Brassica campestris (brown sarson) cv. ‘Pusa Kalyani'. These were used for genetic transformation by an Agrobacterium based binary vector carrying neomycin phosphotransferase (npt) gene and β-glucuronidase (gus)-intron gene for plant cell specific expression. Transformed plants were recovered from hypocotyl explants at a frequency of 7-13%. Addition of silver nitrate markedly enhanced shoot regeneration in hypocotyl explants under non-selection conditions and was found to be an absolute requirement under selection conditions. Cotyledon explants, inspite of being more regenerative, proved to be highly refractory to transformation. Only two chimeric transformed shoots were obtained from more than 10,000 cotyledons treated with Agrobacterium. In hypocotyl explants, shoot regeneration occurred from the vascular parenchyma both with and without the intervention of callus phase. Only the shoot buds differentiating from callus tissue were positive for GUS activity. In cotyledons, shoot buds originated only directly from the vascular parenchyma, generally at a distance of about 450-625 μ from the cut surface. Such shoots were negative for GUS activity. Springer-Verlag, 1992 Genetic transformation nationallicence Oilseed Brassica campestris nationallicence Agrobacterium nationallicence Shoot differentiation nationallicence Mukhopadhyay A. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Arumugam N. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Nandakumar P. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Pradhan A. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Gupta V. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Pental D. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut Plant Cell Reports Springer-Verlag 11/10(1992-09-01), 506-513 0721-7714 11:10<506 1992 11 299 https://doi.org/10.1007/BF00236266 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00236266 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mukhopadhyay A. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 700 1- Arumugam N. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 700 1- Nandakumar P. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 700 1- Pradhan A. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 700 1- Gupta V. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 700 1- Pental D. Tata Energy Research Institute, 90 Jor Bagh, 110 003, New Delhi, India aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 11/10(1992-09-01), 506-513 0721-7714 11:10<506 1992 11 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer