In Vitro Cellular & Developmental Biology - Plant

, Volume 43, Issue 6, pp 585–592 | Cite as

In vitro regeneration through somatic embryogenesis and organogenesis using cotyledons of Cassia angustifolia Vahl

Developmental Biology/Morphogenesis

Abstract

In vitro regeneration through somatic embryogenesis as well as organogenesis using cotyledon of a woody medicinal legume, Cassia angustifolia is reported. The cotyledons dissected from semi-mature seeds, if inoculated on Murashige and Skoog’s medium (MS) supplemented with auxin alone or in combination with cytokinin, produced direct and indirect somatic embryos. A maximum of 14.36 ± 2.26 somatic embryos per 20 mg of explants including callus were produced in 70% cultures on MS medium with 2.5 μM benzyladenine (BA) + 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Although the percentage of embryogenic cultures was higher (83.33%) at 10 μM 2,4-D + 1 μM BA, the average number of somatic embryos was much less (7.6 ± 0.85) at this level, whereas at 2.5 μM BA and 5 μM 2,4-D, there was a simultaneous formation of both somatic embryos and shoots. The somatic embryos, although started germinating on the same medium, developed into full plantlets only if transferred to MS basal with 2% sucrose. Cytokinins alone did not induce somatic embryogenesis, but formed multiple shoots. Five micromolar BA proved optimum for recurrently inducing shoots in the competent callus with a maximum average of 12.04 ± 2.10 shoots and shoot length of 2.26 ± 0.03 cm. Nearly 91.6% shoots (2–2.5 cm in size) organized an average of 5.12 ± 0.58 roots on half strength MS + 10 μM indole-3-butyric acid. All the plantlets have been transferred successfully to soil. Types of auxin and its interaction with cytokinin significantly influenced somatic embryogenesis.

Keywords

Benzyladenine Indole-3-butyric acid Rooting Plantlet 

References

  1. Agrawal, V.; Prakash, S.; Gupta, S.C. Somatic embryogenesis in jojoba (Simmondsia chinensis). In: Jain, S.M.; Gupta, P.K. & Newton, R.J. (eds), Somatic Embryogenesis in Woody Plants. Kluwer, Dordrecht, p. 587–604; 2000.Google Scholar
  2. Agrawal, V.; Prakash, S.; Gupta, S.C. Effective protocol for in vitro shoot production through nodal explant of Simmondsia chinensis. Biol Plant. 45: 449–453; 2002.CrossRefGoogle Scholar
  3. Agrawal, V.; Sardar, P.R. In vitro organogenesis and histomorphological investigations in Senna (Cassia angustifolia)—a medicinally valuable shrub. Physiol. Mol. Biol. Plants 9: 131–140; 2003.Google Scholar
  4. Agrawal, V.; Sardar, P.R. In vitro propagation through leaflet and cotyledon derived callus in Senna (Cassia angustifolia)—a medicinally valuable drought resistant legume. Biol. Plant. 50: 118–122; 2006.CrossRefGoogle Scholar
  5. Anonymous. The Wealth of India, Raw Materials, Vol. 3, CSIR, New Delhi, pp. 354–363; 1992.Google Scholar
  6. Arnold, S.V.; Sabala, I.; Bozhkov, P.; Dyachok, J.; Filonova, L. Developmental pathways of somatic embryogenesis. Plant Cell Tissue Org. Cult. 69: 233–249; 2002.CrossRefGoogle Scholar
  7. Bohra, N.K.; Sankhla, P.S. Senna angustifolia crop for arid region. Vaniki Sandesh 21:19–23; 1997.Google Scholar
  8. Dudits, D.; Bogre, I.A.; Gyorgyey, J. Molecular and cellular approaches to the analysis of plant embryo development from somatic cells in vitro. J. Cell Sci. 99: 475–484; 1991.Google Scholar
  9. Dure, L. Embryogenesis and gene expression during seed formation. Oxford Survey. Plant Mol. Cell. Biol. 2: 179–197; 1985.Google Scholar
  10. Gaj, M.D. Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Regul. 43: 27–47; 2004.CrossRefGoogle Scholar
  11. Gaj, M.D.; Zhang, S.; Harada, J.J. Leafy cotyledon genes are essential for induction of somatic embryogenesis of Arabidopsis. Planta 222: 977–988; 2005.PubMedCrossRefGoogle Scholar
  12. Kumar, S.; Agrawal, V.; Gupta, S.C. Somatic embryogenesis in the woody legume Calliandra tweedii. Plant Cell Tissue Organ Cult. 71: 77–80; 2002.CrossRefGoogle Scholar
  13. Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497; 1962.CrossRefGoogle Scholar
  14. Pulliah, T. Medicinal plants in India, Regency Publ., New Delhi, India, pp 137–139; 2002.Google Scholar
  15. Sharma, A.K.; Goyal, R.K.; Gupta, J.P. Senna the best choice for sandy wastelands. Indian Farming 6: 18–20; 1999.Google Scholar
  16. Sharp, W.R.; Sondahl, M.R.; Caldas, L.S.; Maraffa, S.B. The physiology of in vitro asexual embryogenesis. Hortic. Rev. 2: 268–310; 1980.Google Scholar
  17. Sterk, P.; de Vries, C. Molecular markers for plant embryos, In: K. Redenbaugh (Ed.), Synseeds, Applications of Synthetic Seeds to Crop Improvement, CRC, London, pp. 116–132; 1993.Google Scholar

Copyright information

© The Society for In Vitro Biology 2007

Authors and Affiliations

  1. 1.Department of BotanyUniversity of DelhiDelhiIndia

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