caa a22 4500 445322586 CHVBK 20180317142709.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10681-011-0501-3 doi (NATIONALLICENCE)springer-10.1007/s10681-011-0501-3 Pyramiding of modified cry1Ab and cry1Ac genes of Bacillus thuringiensis in transgenic chickpea ( Cicer arietinum L.) for improved resistance to pod borer insect Helicoverpa armigera [Elektronische Daten] [Meenakshi Mehrotra, Aditya Singh, Indraneel Sanyal, Illimar Altosaar, D. Amla] The modified cry1Ab and cry1Ac insecticidal genes of Bacillus thuringiensis (Bt) under the control of two different constitutive promoters have been introduced into chickpea (Cicer arietinum L.) by Agrobacterium-mediated transformation of pre-conditioned cotyledonary nodes. 118 stable transformed T0 plants as independent transformation events were obtained expressing individual cry1Ab, cry1Ac or both pyramided genes for their co-expression driven by either cauliflower mosaic virus 35S promoter with duplicated enhancer (CaMV35S) or synthetic constitutive promoter (Pcec) and their combinations. Integration and inheritance of transgenes in T0 and T1 population of transgenic chickpea plants were determined by PCR, RT-PCR and Southern hybridization. Results of Southern hybridization showed single copy integration of cry1Ab or cry1Ac genes in most of the transgenic plants developed with either single or pyramided genes and reflected Mendelian inheritance of transgenes in T1 progeny. Real time PCR of pyramided transgenic plants clearly showed differential expression of transcripts for both the genes driven by CaMV35S and Pcec promoters. Quantitative assessment of Bt Cry toxins by ELISA of T0 transgenic chickpea plants showed expression of toxin ranging from 5 to 40ngmg−1 of total soluble protein (TSP) in leaves of transgenic plants. Insect bioassay performed with transgenic plants showed relatively higher toxicity for plants expressing Cry1Ac protein as compared to Cry1Ab to Helicoverpa armigera. Pyramided transgenic plants with moderate expression levels (15-20ngmg−1 of TSP) showed high-level of resistance and protection against pod borer larvae of H. armigera as compared to high level expression of a single toxin. These results have shown the significance of pyramiding and co-expression of two Cry toxins for efficient protection against lepidopteran pests of chickpea. Springer Science+Business Media B.V., 2011 Bacillus thuringiensis nationallicence Transgenic chickpea plants nationallicence Genetic transformation nationallicence Gene pyramiding nationallicence Helicoverpa armigera nationallicence Bt-Cry toxin nationallicence Mehrotra Meenakshi Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut Singh Aditya Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut Sanyal Indraneel Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut Altosaar Illimar Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 451 Smyth Road, Ottawa, ON, Canada aut Amla D. Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut Euphytica Springer Netherlands 182/1(2011-11-01), 87-102 0014-2336 182:1<87 2011 182 10681 https://doi.org/10.1007/s10681-011-0501-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10681-011-0501-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mehrotra Meenakshi Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut NATIONALLICENCE 700 1- Singh Aditya Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut NATIONALLICENCE 700 1- Sanyal Indraneel Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut NATIONALLICENCE 700 1- Altosaar Illimar Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 451 Smyth Road, Ottawa, ON, Canada aut NATIONALLICENCE 700 1- Amla D. Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, 226 001, Lucknow, India aut NATIONALLICENCE 773 0- Euphytica Springer Netherlands 182/1(2011-11-01), 87-102 0014-2336 182:1<87 2011 182 10681 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer