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Transgenic rice expressing the cry2AX1 gene confers resistance to multiple lepidopteran pests

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Abstract

A chimeric Bacillus thuringiensis toxin (Bt) gene, cry2AX1was cloned in a bi-selectable marker free binary vector construct. The cry2AX1 gene, driven by the Chrysanthemum rbcS1 promoter, was introduced into JK1044R, the restorer line (Oryza sativa L. ssp. Indica) of a notified commercially grown rice hybrid in India, by Agrobacterium-mediated transformation. Its effect against two major lepidopteran insect pests viz., yellow stem borer (YSB) Scirpophaga incertulas, rice leaf folder (RLF) Cnaphalocrocis medinalis and one minor insect pest, oriental army worm (OAW) Mythimna separata was demonstrated through bioassays of transgenic rice plants under laboratory and greenhouse conditions. The rbcS1 promoter with chloroplast signal peptide was used to avoid Cry2AX1 protein expression in rice seed endosperm tissue. A total of 37 independent transformants were generated, of which after preliminary molecular characterization and YSB bioassay screening, five events were selected for their protein expression and bioefficacy against all three rice insect. One elite transgenic rice line, BtE15, was identified with Cry2AX1 expression ranging from 0.68 to 1.34 µg g−1 leaf fresh weight and with 80–92 % levels of resistance against rice pests at the vegetative and reproductive stages. Increase in Cry2AX1 protein concentration was also observed with crop maturity. The Cry2AX1protein concentration in the de-husked seeds was negligible (as low as 2.7–3.6 ng g−1). These results indicate the potential application of cry2AX1 gene in rice for protection against YSB, RLF and OAW.

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Acknowledgments

We are thankful to Mr. Sanjay Gupta, Director J.K Agri. Genetics Ltd. and we acknowledge all type of inputs towards this research from all co-workers in J. K Biotech Lab. This work is supported and Funded by the Department of Biotechnology, Government of India (BIRAC Grant No. BT/BIPP/0320/07/10). The authors wish to thank the anonymous referees for their constructive suggestion.

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  1. Debashis Rana

    Present address: Bayer CropScience-Seeds, Bayer (South East Asia) Pte Ltd, Singapore, Singapore

Authors and Affiliations

  1. Biotechnology Division, J.K Agri. Genetics Pvt. Ltd., Hyderabad, 500016, India

    M. Chakraborty, P. Sairam Reddy, G. Mustafa, G. Rajesh & Debashis Rana

  2. Department of Biotechnology, Jawaharlal Nehru Technological University (JNTU), Hyderabad, 500072, India

    M. Chakraborty & V. M. Laxmi Narasu

  3. Department of Plant Molecular Biology and Biotechnology, Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, 641 003, India

    V. Udayasuriyan

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Chakraborty, M., Reddy, P.S., Mustafa, G. et al. Transgenic rice expressing the cry2AX1 gene confers resistance to multiple lepidopteran pests. Transgenic Res 25, 665–678 (2016). https://doi.org/10.1007/s11248-016-9954-4

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