Agrobacterium-mediated tomato transformation and regeneration of transgenic lines expressing Tomato leaf curl virus coat protein gene for resistance against TLCV infection

Abstract

Coat protein (CP) gene of Tomato leaf curl virus (TLCV) was cloned into an expression vector and mobilized to Agrobacterium tumefaciens through triparental mating. Cotyledon leaf explants of Pusa Ruby tomato were transformed by co-cultivation with Agrobacterium containing TLCV-CP construct Kanamycin-resistant transformants were regenerated and established in glasshouse. TO-generation putative transgenic plants obtained were screened by PCR, Southern and Northern hybridization tests and Western blot assay, which confirmed the incorporation and expression of the CP gene. CP expressing transgenic plants were self-pollinated. T1-generation transgenic plants were challenged by TLCV through whiteflies which showed variable degrees of disease resistance/tolerance compared to the untransformed control.