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In vitro clonal propagation of Mucuna pruriens var. utilis and its evaluation of genetic stability through RAPD markers


N Sathyanarayana
TNB kumar
PB Vikas
R Rajesha

Abstract

The Mucuna pruriens var. utilis is an important legume cover crop. Almost all the parts of the plant are reported to contain L-3,4-dihydroxy phenylalanine (L-Dopa). Here we report a rapid and reliable method
for high fidelity micro-propagation. Auxiliary bud explants from 14-day-old seedlings were cultured on Murashige and Skoog’s (MS) medium supplemented with different concentrations of cytokinins. During the first culture on 3.5 M 6-benzylamino purine (BAP) maximum of 6.70 ± 1.15 shoots with an average shoot-length of 1.07 ± 0.21 cm were produced. The number of shoots increased up to 16.33 ± 0.58 recording average length of 1.16 ± 0.29 cm, when the intact shoots were subjected to re-culturing on the same hormonal medium. The shoots exhibited adequate elongation of 4.00 cm on 2.89 M gibberellic acid (GA3). The elongated shoots produced a maximum of 16.67 ± 2.89 roots on half-strength MS liquid medium supplemented with 16.20 ìM -naphthalene acetic acid (NAA). The plantlets were acclimatized by transferring them first to peat moss: compost (1:1) mixture followed by sand: soil (1:1) mixture, recording 95% survival. The genetic fidelity of the regenerated shoots was confirmed using randomly amplified polymorphic DNA (RAPD) analysis employing 15 operon primers. This system provides high fidelity micro-propagation system for efficient and rapid micro-propagation of this important green
manure cover crop with medicinal properties.

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eISSN: 1684-5315