Agrobacterium-mediated Genetic Transformation of Groundnut (Arachis hypogaea L.) using Epicotyl Explants
Yogranjan, S. Eapen
Nuclear Agriculture and Biotechnology Division, Plant Biotechnology and Secondary Product Section, BARC, Mumbai-400 094, India
S. Tiwari
Plant Tissue Culture and Genetic Transformation Division, Biotechnology Centre, J. N. Agricultural University, Jabalpur-482 004, India
M. K. Tripathi *
Plant Tissue Culture and Genetic Transformation Division, Biotechnology Centre, J. N. Agricultural University, Jabalpur-482 004, India
*Author to whom correspondence should be addressed.
Abstract
Transformation of groundnut (Arachis hypogaea L.) was achieved using Agrobacterium tumefaciens strain LBA4404 harboring binary vectors pHS102 (containing uidA, nptII and Cry1Ac genes) pBI121 (containing uidA (GUS) and nptII genes) pBinAR (possesses a fusion chimeric gene Cry1Ac-F gene). The transformation frequency varied from 4% (with pHS102) to 12% (with pBI121). A strong GUS expression was detected in the putative transformed shoots by the histochemical assay. Transformation was confirmed by the presence of 0.70 kb amplified fragments for nptII and 1.40 kb for uidA genes by PCR analysis. Shoot differentiation via callusing was attained on MS medium supplemented with 2 mg.l-1 BA, 0.1 mg.l-1 IAA, and 150 mg.l-1 adenine sulphate, whereas direct shoot differentiation was obtained on medium containing 0.1 mg.l-1 TDZ.
Keywords: Arachis hypogaea, Agrobacterium, Epicotyl, Transformation, Transgenic plants