, Volume 24, Issue 4, pp 781–787 | Cite as

Somatic embryogenesis and plant regeneration in Pterocarpus marsupium Roxb.

  • Mohd Kashif HusainEmail author
  • Mohammad Anis
  • Anwar Shahzad
Original Paper


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.


Hypocotyl Leguminous tree Malabar kino Embryo germination Seedlings Embryogenic callus 



Abscisic acid


Activated charcoal




2,4-Dichlorophenoxyacetic acid


Murashige and Skoog medium


α-Naphthalene acetic acid


1,2,3-Trihydroxy benzene


Plant growth regulators


Indole-3-butyric acid




Somatic embryogenesis



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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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Mohd Kashif Husain
    • 1
    Email author
  • Mohammad Anis
    • 2
    • 3
  • Anwar Shahzad
    • 2
  1. 1.Biotechnology DivisionIndian Institute of Integrative Medicine (IIIM), Council of Scientific and Industrial Research (CSIR)Jammu-TawiIndia
  2. 2.Plant Biotechnology Laboratory, Department of BotanyAligarh Muslim UniversityAligarhIndia
  3. 3.Department of Plant Production, College of Food and Agricultural ScienceKing Saud UniversityRiyadhSaudi Arabia

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