Rapid and stable in vitro regeneration of plants through callus morphogenesis in two varieties of Mucuna pruriens L. – an anti Parkinson’s drug yielding plant

摘要

Mucuna pruriens L. is an important medicinal plant reported to contain L-3,4-dihydroxy phenylalanine (L-DOPA). The present report describes a simple protocol for plant regeneration through callus morphogenesis in four strains belonging to two different varieties of Mucuna pruriens. Friable, soft and nodular callus was induced from nodal and internodal segments of in vitro-grown seedlings on modified Murashige and Skoog’s (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). For regeneration of shoots, the callus tissues were transferred to medium supplemented with various levels of cytokinin. Highest shoot regeneration from the callus tissue was achieved in modified MS medium fortified with BAP at 1.33 μM level. The regenerated shoots were rooted in vitro in half-strength of liquid MS medium supplemented with various levels of NAA. The regenerates were acclimatized for 2–3 weeks and showed about 80 % survival rate after transferring to the field. Cytological analysis revealed chromosome number stability of all the regenerates (2n = 22) with complete absence of aneuploidy. Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) analyses of total genomic DNA supported stability at the molecular level among all the clones of these strains when compared with their mother plants.