Agrobacterium-mediated transformation of brinjal (Solanum melongena L.) using fungal resistant gene
The investigation standardized the protocol for Agrobacterium-mediated transformation system in Brinjal (Solanum melongena L), cultivar ‘Arka Neelkanth’ . Initially the regeneration system was standardized with regeneration medium (MS basal medium supplemented with 8.88 µM BAP + 1.14 µM IAA) by using 10-12 days old cotyledon explants. Transformation of the hevein, a gene responsible for fungal resistance. into cotyledon explants was carried out with Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pGPTV/hevein (Plasmid Gus Plant Transformation Vector) with marker gene neomycin phosphotransferase (npt-II). Transformation efficiency showed a higher percentage in one day pre-cultured explants, infected with bacterial suspension of optical density 0.4 at OD600 nm with 1:1 dilution for 10min and 48hrs co-cultivation on regeneration medium in the dark. Transformed explants were cultured on selection medium. The PCR molecular confirmation of amplified npt-II gene from the genomic DNA of the transformed plant indicates the integration of the hevein gene in the brinjal plant.
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