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Expression of Insecticidal Toxin Coded by Modified Full-Length and Truncated Bt-cry1Ac Gene in Transgenic Tomato for Assessment of Their Stability and Efficacy against Target Insects.

Bhupendra Koul, Devendra V. Amla, Indraneel Sanyal

Abstract


The work is structured into four different parts. The first part is dedicated to computational analysis of six variants of Bt-cry1A genes to generate an idea of their integrity and stability. The second part includes optimization of Agrobacterium-mediated tomato transformation procedure using vegetative leaf disc-explants with p35SGUS-INT and validation of the procedure using five different genes of diverse origin. The third part deals with a comparative analysis of constitutive promoter DECaMV35S and synthetic promoter Pcec (complete expression cassette) with defined regulatory sequences to enhance the expression and efficacy of Bt-cry1Ac gene against Helicoverpa armigera in transgenic tomato. Finally, the fourth part discusses molecular characterization and comparative expression analysis of transgenic tomato transformed with modified full-length and truncated version of cry1Ac genes. It also includes the performance and efficacy of truncated cry1Ab gene against Helicoverpa armigera and Spodoptera litura, in transgenic tomato. The higher level of Cry1Ab toxin (≈ 0.47 ± 0.01% TSP) did not show any detrimental effect on in vitro regeneration, plant development and efficacy against target pests in transgenic line Ab25E, contrary to the  earlier reports with Cry1Ac toxin. The full-length cry1Ac gene can be redesigned for higher expression and performance in dicots.


Keywords


Transgenic tomato, Vegetative leaf, Transformation efficiency, Expression analysis, Helicoverpa armigera

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References


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