A preliminary attempt for efficient genetic transformation and regeneration of legume Mucuna pruriens L. mediated by Agrobacterium tumefaciens

Authors: RAGHAVENDRA SATHYANARAYANA, VADLAPUDI KUMAR, CHAPEYL.KUMARAN RAMESH, MAHADEVAPPA PARESHA, MAHABOOB HABEEBULLA MOINUDDIN KHAN

Abstract: Mucuna pruriens belongs to the family Papillonaceae. The seeds and leaves of these plants contain a large amount of L-DOPA. L-DOPA (L-3,4-dihydroxyphenylalanine) is one of the most widely used drugs in the treatment of Parkinson disease. Development of an efficient gene transfer method is an absolute requirement for genetically improving Mucuna pruriens and creating plants with more desirable traits. A simple protocol was developed for the Agrobacterium-mediated stable genetic transformation of M. pruriens. Agrobacterium tumefaciens strain EHA 101, containing the vector pCAMBIA1305 and the hptII and GUS plus genes, was used for the gene transfer experiments. Putative transgenic shoots were obtained on medium supplemented with kanamycin (50 mg L^-^1) and cefotaxime (400 mg L^-^1). GUS histochemical analysis of the putative transgenic tissues further confirmed the transformation event. Genomic polymerase chain reaction (PCR) analysis was performed to verify the presence of transgenes and their stable integration. Transformation mediated by Agrobacterium in M. pruriens is reported for the first time.

Keywords: M. pruriens, genetic transformation, Agrobacterium, GUS, PCR

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