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

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. | R. SATHYANARAYANA, V. KUMAR, C. K. RAMESH, M. PARMESHA, M. H. M. KHAN Turk J Biol 36 (2012) 285-292 © TÜBİTAK doi: A preliminary attempt for efficient genetic transformation and regeneration of legume Mucuna pruriens L. mediated by Agrobacterium tumefaciens Raghavendra SATHYANARAYANA1, Vadlapudi KUMAR2, Chapeyil Kumaran RAMESH3, Mahadevappa PARMESHA4, Mahaboob Habeebulla Moinuddin KHAN5 1 Department of Biochemistry, College of Horticulture, University of Horticultural Sciences, Bagalkot, Karnataka - INDIA 2 Department of Biochemistry, Davangere . Centre, Shivagangothri, Davangere 577002, Karnataka - INDIA 3 Department of Biotechnology, Sahyadri Science College, Shivamogga 577203, Karnataka - INDIA 4 Department of Biotechnology, GMIT, Davangere 577001, Karnataka - INDIA 5 Department of Chemistry, Jawaharlal Nehru National College of Engineering, Shivamogga 577201 - INDIA Received: 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 .