Micropropagation of Vitex negundo L. and Random Amplified Polymorphic DNA Analysis of Micropropagated Plants |
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| Authors: | A Yusuf S Nikhilesh P S Rao |
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| Institution: | 1. Department of Botany , University of Calicut , Kerala , India akkara.yusuf@lycos.com;3. Department of Biochemistry , University of Nebraska-Lincoln , Lincoln , Nebraska , USA;4. Department of Biotechnology , Dayananda Sagar Institutions , Kumaraswamy Layout, Bangalore , Karnataka , India |
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| Abstract: | A micropropagation protocol was established for a medicinal plant Vitex negundo. Genetic stability of micropropagated plants was investigated. Multiple shoots were induced from nodal explants cultured on Murashige and Skoog (MS) medium with 0.53 μM naphthalene acetic acid (NAA) and 11.0 μM benzyl aminopurine (BAP) along with additives (ascorbic acid, 283.9 μM; citric acid, 130.1 μM; and arginine, 143.6 μM). Shoots were further multiplied by repeated transfer of the mother explant. The shoots were further multiplied on MS medium + 0.57 μM indole-3-acetic acid (IAA) and 6.6 μM BAP. The micropropagated shoots were pulse treated with 122.5 μM indole-3-butyric acid (IBA), in liquid MS medium and then transferred to autoclaved soilrite. These rooted ex vitro. Shoots were also rooted in vitro on a half-strength MS medium + 2.45 μM IBA. The survival rate of in vitro rooted plantlets was poor during hardening compared to ex vitro rooted plantlets. About 95% of the ex vitro rooted, hardened plantlets survived in the field. Genetic stability of micropropagated plants was tested by using 25 random amplified polymorphic DNA primers. The cloned plants exhibited no variation in banding pattern in comparison with the mother plant. |
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| Keywords: | nodal explants Vitex negundo shoot clumps ex vitro rooting random amplified polymorphic DNA |
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