Agrobacterium-mediated genetic transformation of groundnut (Arachis hypogaea L.): An assessment of factors affecting regeneration of transgenic plants
1998
Venkatachalam, P. (Bharathidasan Univ., Tiruchirappalli (India)) | Geetha, N. | Jayabalan-Saravanababu, N. | Sita, L.
Transgenic groundnut (Arachis hypogaea L.) plants were produced efficiently by inoculating different explants with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBM21 containing uidA (GUS) and nptll (neomycin phosphotransferase) genes. Genetic transformation frequency was found to be high with cotyledonary node explants followed by 4 d cocultivation. This method required 3 days of precultivation period before cocultivation with Agrobacterium. A concentration of 75 mg/l kanamycin sulfate was added to regeneration medium in order to select transformed shoots. Shoot regeneration occurred within 4 weeks excised shoots were rooted on MS medium containing 50 mg/l kanamycin sulfate before transferring be soil. The expression of GUS gene (uidA gene) in the regenerated plants was verified by histochemical and fluorimetric assays. The presence of uidA and nptll genes in the putative transgenic lines was confirmed by PCR analysis. Insertion of the nptll gene in the nuclear genome of transgenic plants was verified by genomic Southern hydridization analysis. Factors affecting transformation efficiency are discussed
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