Somatic embryogenesis and plant regeneration in Pterocarpus marsupium Roxb.
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Somatic embryogenesis (SE) has been achieved from hypocotyl-derived callus culture in Pterocarpus marsupium. Ninety percent of hypocotyl explants (excised from 12-day-old in vitro germinated axenic seedlings) produced callus on Murashige and Skoog medium supplemented with 5 μM 2,4-dichlorophenoxyacetic acid and 1 μM a 6-benzyladenine (BA). Induction of SE occurred after transfer of callus clumps (200 ± 20 mg fresh mass) to MS medium supplemented with BA at 2.0 μM, where a maximum of 23.0 ± 0.88 globular stage embryos per callus clump were observed after 4 weeks of culture. Subculturing of these embryos on MS medium supplemented with 0.5 μM BA, 0.1 μM α-naphthalene acetic acid and 10 μM abscisic acid significantly enhanced the maturation of somatic embryos to early cotyledonary stage, where 21.4 ± 0.32 embryos per callus clump were recorded after 4 weeks of culture. Of 30-well developed somatic embryos, 16.6 ± 0.33 germinated and subsequently converted into plantlets on half-strength MS medium supplemented with 1.0 μM BA. The morphologically normal plantlets with well-developed roots were first transferred to 1/4-liquid MS medium for 48 h and then to pots containing autoclaved soilrite and acclimatized in a culture room. Thereafter, they were transferred to a greenhouse, where 60% of them survived.
KeywordsHypocotyl Leguminous tree Malabar kino Embryo germination Seedlings Embryogenic callus
Murashige and Skoog medium
α-Naphthalene acetic acid
Plant growth regulators
The award of Research Associateship to M. K. Husain by the Council of Scientific and Industrial Research (CSIR), Government of India, New Delhi, is greatly acknowledged. We also acknowledge the Department of Biotechnology (DBT), Government of India, New Delhi, for financial assistance and Dr. Shamim A. Ansari, Scientist, Tropical Forest Research Institute (TFRI), Jabalpur, for his helpful discussion. The authors are also thankful to Dr. K. Klimaszewska, Communicating Editor, for critical evaluation and valuable comments on the manuscript.
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