Full Length Research Paper

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

N. Sathyanarayana*, T. N. Bharath kumar, P. B. Vikas and R. Rajesha

Department of Bio-Technology, Sir M. Visvesvaraya Institute of Technology, Bangalore – 562 157, India.

*Corresponding author. E-mail: sathya_dixit@yahoo.com. 

Abbreviations: BAP,  6-benzylamino purine; GA3,  gibberellic acid; NAA,  α-naphthalene acetic acid.

Accepted 20 February, 2008

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 μM 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 μM 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 µM α-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.

Key words: Mucuna pruriens var. utilis; auxiliary bud, multiple shoots, rapid micro-propagation, fidelity, RAPD.