Direct somatic embryogenesis, plant regeneration and in vitro flowering in rapid-cycling Brassica napus |
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Authors: | W. L. Koh C. S. Loh |
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Affiliation: | (1) Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543 e-mail: dbslohcs@nus.edu.sg Fax: +65-779-5671, SG |
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Abstract: | A simple method to induce somatic embryogenesis from seeds of rapid-cycling Brassica napus is described. Seedlings cultured on Murashige and Skoog (MS) basal medium produced somatic embryos directly on hypocotyls and cotyledons after 2 to 3 subcultures onto the same medium. A low pH of the medium (3.5–5) was more conducive to somatic embryogenesis than a higher pH (6 and 7). Embryogenic potential of the seeds was inversely correlated to seed age: about 41–68% of immature seeds between the ages of 14 and 28 days after pollination (DAP) formed somatic embryos compared to 0–11% of the seeds obtained 29–37 DAP. About 54% of the somatic embryos produced secondary embryos after subculturing onto the same medium. The embryogenic potential of the cultures has been maintained on MS basal medium for 2 years (12 generations) without diminution. Up to 75% of the secondary embryos developed into plantlets on MS medium enriched with 10–6 M zeatin, and 40% of these produced flowers when transferred to an optimised flower-induction medium. Viable seeds were produced in self-pollinated in vitro flowers. Received: 15 February 2000 / Revision received: 18 July 2000 / Accepted: 19 July 2000 |
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Keywords: | Rapid-cycling Brassica napus Somatic embryogenesis Secondary embryogenesis Regeneration In vitro Howering |
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