In planta transformation strategy to generate transgenic plants in chickpea: proof of concept with a cry gene.

摘要

The paper presents a non-tissue culture-based transformation of chickpea using cry1AcF gene with 5'UTR. The protocol involves raising of plant transformants (TO plants) directly from Agrobacterium-infected young seedlings. The apical meristem of the seedling axes were targeted for transformation. The resulting chimeric plants were allowed to grow in the greenhouse and the transgenics were analysed in the T1 generation. The T1 generation plants were raised in the greenhouse and initial screening was carried out in 109 plants using ELISA for the expression of the cry1AcF protein. On the basis of this, the plants were grouped as non-transformants and transformants, expressing low and high level of the cry protein. The plants expressing the cry1AcF protein in the range of 2.06-9.70 micro g/g fr wt were selected for further analysis. Bioefficacy of these 44 plants against Helicoverpa armigera allowed identification of 14 plants that not only accumulated good amount of protein but were also effective against Helicoverpa. Molecular analysis by PCR for the amplification of both the cry1AcF and nptII genes confirmed the transgenic nature of the selected plants. The protocol ensured generation of transgenic chickpea plants with considerable ease in a short time and might be applicable across different genotypes/cultivars of the crop and offers immense potential as a supplemental or an alternate protocol for generating transgenic plants of difficult-to-regenerate crops.