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Keywords:
Mucuna pruriens var. utilis auxiliary bud multiple shoots rapid micro-propagation fidelity RAPD.
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<i>In vitro</i> clonal propagation of <i>Mucuna pruriens var. utilis</i> and its evaluation of genetic stability through RAPD markers
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.
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.
