Agrobacterium-mediated genetic transformation of tomato (Solanum lycopersicum L.) with Cry1Ac gene for resistance against fruit borer

Abstract

Agrobacterium-mediated genetic transformation was used to produce transgenic plants of tomato (Solanum lycopersicum L.) resistant to fruit borer. Cotyledon and leaf explants from in vitro tomato seedlings of variety ‘Punjab Upma’ were co-cultivated with A. tumefaciens strain GV 3101 containing npt II (plant selectable marker gene providing resistance to kanamycin) under the control of nopaline marker gene providing promoter (pNOS) and coding region containing a plant intron linked to the cauliflower mosaic 35 S (CaMV35S) promoter. Out of the 376 cotyledons used for infection, 18 got selected on kanamycin (30 mg L–1) showing 4.78% regeneration and of the 336 leaf explants, 11 got selected on kanamycin (30 mg L–1) showing 3.27% regeneration. Histochemical GUS assay of the kanamycin selected cotyledon explants revealed 47.82% and in case of leaf explants it was 40%. PCR analysis confirmed presence of the transgene of 1.2 kb in five individual plantlets. These results signify the successful introduction of Cry1Ac gene into tomato plants.