Efficient micropropagation protocol of Spilanthes acmella L. possessing strong antimalarial activity
- 435 Downloads
Micropropagation has been achieved in a promising larvicidal asteraceous taxon Spilanthes acmella L. using seedling leaf explants. The explants were reared on a variety of growth regulators, namely 2,4-dichlorophenoxyacetic acid, 1-naphthalene acetic acid, Indole-3-butyric acid, N6-benzyladenine, and kinetin either alone or in combination on Murashige and Skoog’s (MS) medium. The best green and compact callus was obtained on 1 μM NAA and 10 μM benzyladenine (BA) in 15 d. The callus on subculture to the same but fresh medium after every 30 d differentiated an average of 12.90 ± 0.32 shoot buds in 50% cultures. Elongation in shoot buds occurred only if they were transferred to NAA lacking MS+BA medium. An average number of 4.22 ± 0.83 shoots and 15 ± 0.84 shoot buds per explant were obtained in 70.3% cultures on MS + 10 μM BA in 30 d. One hundred percent excised shoots rooted in MS(1/2) + 0.1 μM IBA within 2 wk. The plants were gradually hardened and established in soil where they flowered and set viable seeds. The regenerated plants were morphologically similar to the field grown plants and showed 100% larvicidal activity against malaria and filarial vectors.
KeywordsBA Larvicidal activity Leaf explant Multiple shoots NAA Plantlets
Veena Agrawal is grateful to University Grants Commission for financial assistance and Vibha Pandey to CSIR, New Delhi for the award of JRF and SRF. Authors are also thankful to Dr. A.P. Dash and Dr. K. Raghavendra, National Institute of Malaria Research, Delhi for providing assistance in conducting the bioassay tests.
- Anonymous. World Health Organization: Instructions for determining the susceptibility or resistance of mosquito larvae to insecticides; 1981.Google Scholar
- Anonymous The Wealth of India: a dictionary of Indian raw materials and industrial products, Vol.10. CSIR, New Delhi, pp 11–12; 1989.Google Scholar
- Charlwood B. V.; Pletsch M. Manipulation of natural product accumulation in plants through genetic engineering. In: Johnson C. B.; Franz C. (eds) Breeding research on aromatic and medicinal plants. Haworth Herbal Press, NY, pp 139–151; 2002.Google Scholar
- Eklof S.; Astot C.; Blackwell J.; Moritz T.; Olsson O.; Sandberg G. Auxin–cytokinin interactions in wild-type and transgenic tobacco. Plant Cell Physiol. 38: 225–235; 1997.Google Scholar
- Finney D. J. Probit analysis. 3rdrd ed. Cambridge University Press, Cambridge, UK, pp 1–333; 1971.Google Scholar
- Haw A. B.; Keng C. L. Micropropagation of Spilanthes acmella L. a bio-insecticide plant, through proliferation of multiple shoots. J. Appl. Hortic. 5: 65–68; 2003.Google Scholar
- Jansen R. K. Systematics of Acmella (Asteraceae: Heliantheae). Syst. Bot. Mongr. 8: 115; 1985.Google Scholar
- Nordstrom A.; Tarkowski P.; Tarkowska D.; Norbaek R.; Astot C.; Dolezal K.; Sandberg G. Auxin regulation of cytokinin biosynthesis in Arabidopsis thaliana: a factor of potential importance for auxin-cytokinin regulated development. Proc. Natl. Acad. Sci. 101: 8039–8044; 2004. doi: 10.1073/pnas.0402504101.PubMedCrossRefGoogle Scholar
- Pandey V.; Agrawal V.; Raghavendra K.; Dash A. P. Strong larvicidal activity of three species of Spilanthes (Akarkara) against malaria (Anopheles stephensi Liston, Anopheles culicifacies, species C) and filaria vector (Culex quinquefasciatus Say). Parasitol. Res. 102: 171–174; 2007. doi: 10.1007/s00436-007-0763-9.PubMedCrossRefGoogle Scholar
- Pitasawat B.; Choochote W.; Kanjanapothi D.; Panthong A.; Jitpakdi A.; Chaithong U. Screening for larvicidal activity of ten carminative plants. South East Asian J. Trop. Med. Public Health 29: 660–662; 1998.Google Scholar
- Ramawat K. G. Production of alkaloids. In: Ramawat K. G.; Merillon J. M. (eds) Biotechnology secondary metabolites. Oxford Publications, New Delhi, India, pp 193–218; 2002.Google Scholar
- Richard, A. Spilanthes. http://www.chatlink.com/herbseed/Spilanthes. 1996.
- Saraf D. K.; Dixit V. K. Spilanthes acmella Murr.: study on its extract spilanthol as larvicidal compound. Asian J. Exp. Sci. 16: 9–19; 2002.Google Scholar
- Skoog F.; Miller C. O. Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp. Soc. Exp. Biol. 11: 118–131; 1957.Google Scholar
- Storey C.; Salem J. I. Lay use of amazonian plants for the treatment of tuberculosis. Acta Amazonica 27: 175; 1997.Google Scholar
- Watt P. M.; Brayer-Brandwijk M. C. The medicinal and poisonous plants of Sourthern and Eastern Africa. 2nd ed. E&S Livingstone, Edinburgh1962.Google Scholar
- Willis J. C. A dictionary of the flowering plants and ferns. Cambridge University Press, Cambridge1977.Google Scholar