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, 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

Abstract

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.

Keywords

Hypocotyl Leguminous tree Malabar kino Embryo germination Seedlings Embryogenic callus 

Abbreviations

ABA

Abscisic acid

AC

Activated charcoal

BA

6-Benzyladenine

2,4-D

2,4-Dichlorophenoxyacetic acid

MS

Murashige and Skoog medium

NAA

α-Naphthalene acetic acid

PG

1,2,3-Trihydroxy benzene

PGRs

Plant growth regulators

IBA

Indole-3-butyric acid

2iP

2-Isopentenyladenine

SE

Somatic embryogenesis

Notes

Acknowledgments

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.

References

  1. Anis M, Husain MK, Shahzad A (2005) In vitro plantlet regeneration of Pterocarpus marsupium Roxb., an endangered leguminous tree. Curr Sci 88:861–863Google Scholar
  2. Anonymous (2003) The wealth of india. A dictionary of Indian raw material and industrial products, vol. VIII. Publication and Information Directorate, CSIR, New Delhi, pp 302–305Google Scholar
  3. Ashok Kumar HG, Murthy HN, Paek KY (2002) Somatic embryogenesis and plant regeneration in Gymnema sylvestre. Plant Cell Tissue Organ Cult 71:85–88CrossRefGoogle Scholar
  4. Chand S, Singh AK (2004) In vitro shoot regeneration from cotyledonary node explants of a multipurpose leguminous tree, Pterocarpus marsupium Roxb. In Vitro Cell Dev Biol Plant 40:464–466CrossRefGoogle Scholar
  5. Chaudhuri AB, Sarkar DD (2002) Biodiversity endangered: India’s threatened wildlife and medicinal plants. Scientific Publishers, Jodhpur, pp 169–172Google Scholar
  6. Chaudhury RR (2004) Herbal remedy for diabetes finds no takers. Times of India, 9 OctoberGoogle Scholar
  7. Husain MK (2007) Tissue culture studies on the propagation of two multipurpose tree species, Pterocarpus marsupium Roxb. and Melia azedarach L. PhD thesis, Aligarh Muslim University, AligarhGoogle Scholar
  8. Husain MK, Anis M, Shahzad A (2007) In vitro propagation of Indian Kino (Pterocarpus marsupium Roxb.) using thidiazuron. In Vitro Cell Dev Biol Plant 43:59–64Google Scholar
  9. Husain MK, Anis M, Shahzad A (2008) In vitro propagation of a multipurpose leguminous tree (Pterocarpus marsupium Roxb.) using nodal explants. Acta Physiol Plant 30:353–359CrossRefGoogle Scholar
  10. Ipecki Z, Gozukirmizi N (2003) Direct somatic embryogenesis and synthetic seed production from Paulownia elongata. Plant Cell Rep 22:16–24CrossRefGoogle Scholar
  11. Junaid A, Mujib A, Bhat MA, Sharma MP (2006) Somatic embryo proliferation, maturation and germination in Catharanthus roseus. Plant Cell Tissue Organ Cult 84:325–332CrossRefGoogle Scholar
  12. Kalimuthu K, Lakshmanan KK (1995) Effect of different treatments on pod germination of Pterocarpus species. Indian J For 18:104–106Google Scholar
  13. Manickam M, Ramanatha M, Farboodniay Jahromi MA, Chausouria JPN, Ray AB (1997) Antihyperglycemic activity of phenolics from Pterocarpus marsupium. J Nat Prod 60:609–610CrossRefPubMedGoogle Scholar
  14. Murashige T, Skoog F (1962) A revised medium for rapid growth bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  15. Nomura K, Komamine A (1995) Physiological and biochemical aspects of somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer, Dordrecht, pp 249–266Google Scholar
  16. Sahrawat AK, Chand S (2001) Continuous somatic embryogenesis and plant regeneration from hypocotyls segments of Psoralea corylifolia L., an endangered and medicinally important fabaceae plant. Curr Sci 81:1328–1331Google Scholar
  17. Shahana S, Gupta SC (2002) Somatic embryogenesis in Sesbania sesban var bicolor: a multipurpose fabaceous woody species. Plant Cell Tissue Organ Cult 69:289–292CrossRefGoogle Scholar
  18. Sharma P, Koche V, Quraishi A, Mishra SK (2005) Somatic embryogenesis in Buchanania lanzan Spreg. In Vitro Cell Dev Biol Plant 41:645–647CrossRefGoogle Scholar
  19. Singh AK, Chand S (2003) Somatic embryogenesis and plant regeneration from cotyledon explants of a timber-yielding leguminous tree Dalbergia sissoo Roxb. J Plant Physiol 160:415–421CrossRefPubMedGoogle Scholar
  20. Sunnichan VG, Shivanna KR, Mohan Ram HY (1998) Micropropagation of gum karaya (Sterculia urens) by adventitious shoot formation and somatic embryogenesis. Plant Cell Rep 17:951–956CrossRefGoogle Scholar
  21. Vats V, Grover JK, Rathi SS (2002) Evaluation of antihyperglycemic effect of Trigonella foenum-graecum L., Ocimum sanctum L. and Pterocarpus marsupium, in normal and alloxanized diabetic rats. J Ethnopharmacol 79:95–110CrossRefPubMedGoogle Scholar
  22. Vlasinova H, Havel L (1999) Continuous somatic embryogenesis in Japanese maple (Acer palmatum Thumb.). J Plant Physiol 154:212–218Google Scholar
  23. Williams EG, Maheshwaran G (1986) Somatic embryogenesis: factors influencing coordinated behaviour of cells as embryonic group. Ann Bot 57:443–462Google Scholar
  24. Zimmerman JL (1993) Somatic embryogenesis: a model for early development in higher plants. Plant Cell 5:1411–1423CrossRefPubMedGoogle Scholar

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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