Development and characterization of transgenic chickpea (Cicer arietinum L.) plants with cry1Ac gene using tissue culture independent protocol

Tissue culture independent Agrobacterium-mediated transformation with cry1Ac gene following the non-selective/PCR detection system using direct plant PCR screening indicated the putative transgenic nature of plants and represented transformation frequency of 13.4% and 41% in cvs. C-235 and HC-1, respectively. The putative transgenic chickpea plants were analyzed adopting multiple evaluation strategies, such as PCR, ELISA and Southern blotting, for selection of plants for further advancement. Quantitative assessment of Bt Cry toxin by ELISA in leaves of transgenic chickpea plants showed variation in expression of Cry1Ac toxin (106 to 364 ng g-1 FW), but high expressing events (> 200 ng g-1 FW) were found to exhibit phenotypic abnormalities. Results obtained from Southern blotting using gene specific probe confirmed the single copy integration of the cry1Ac gene into the chickpea genome. Evaluation of the T2 generation progenies of few promising T1 transgenic plants for entomocidal activity showed retarded larval growth but significant mortality was not observed. The present study offers a suitable approach for development of chickpea plants with novel traits in with high efficiency and short duration, with the possibility of developing marker free transgenic events, allows stacking of multiple genes and can be applicable across different genotypes/cultivars of chickpea.