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Expression of a synthetic cry1EC gene for resistance against Spodoptera litura in transgenic peanut (Arachis hypogaea L.)

  • Genetic Transformation and Hybridization
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Abstract

The tobacco cutworm (Spodoptera litura) is a polyphagous foliage insect and a major pest on peanut (Arachis hypogaea L.). S. litura is susceptible to the chimeric δ-endotoxin Cry1EC reported earlier. De-embryonated cotyledon explants of peanut were transformed using Agrobacterium tumefaciens strain EHA101 harboring a synthetic cry1EC gene driven by the CaMV 35S promoter. Transgenic plants of peanut with a single copy insertion of cry1EC were selected in the T0 generation by Southern blot hybridization. Real-time PCR, Western blot and ELISA analysis indicated that expression of the cry1EC gene was higher in single copy T1 plants. Immunoassay showed expression of Cry1EC up to 0.13% of total soluble protein in T1 plants. Leaf feeding bioassay on highly expressing transgenic lines showed 100% killing of larvae at the 2nd instar stage of S. litura. This is the first report of transgenic peanut plants with resistance to S. litura.

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Abbreviations

BAP:

6-Benzylaminopurine

hptII :

Hygromycin phosphotransferase

SIM:

Shoot induction medium

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Acknowledgments

The authors are grateful to Dr. K. Chandrasekhar and S. M. H. Abidi for insect bioassays, Council of Scientific and Industrial Research, Government of India for financial support and fellowship to Siddharth Tiwari, Devesh K. Mishra and Ankit Singh.

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Authors and Affiliations

  1. Plant Molecular Biology and Genetic Engineering Division, National Botanical Research Institute, Rana Pratap Marg, Lucknow, UP, 226001, India

    Siddharth Tiwari, Devesh K. Mishra, Ankit Singh, P. K. Singh & Rakesh Tuli

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  1. Siddharth Tiwari
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  2. Devesh K. Mishra
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  3. Ankit Singh
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  4. P. K. Singh
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  5. Rakesh Tuli
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Correspondence to Rakesh Tuli.

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Communicated by P. Ozias-Akins.

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299_2008_525_MOESM3_ESM.doc

Table 1S Quantification of cry1EC transcript by real-time PCR. The amplification plots shown in Fig. 2S were used to calculate 2-DDCT values for the five transgenic lines. The results given in the last column are presented graphically in Fig. 4 in the manuscript (doc 109 kb)

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Tiwari, S., Mishra, D.K., Singh, A. et al. Expression of a synthetic cry1EC gene for resistance against Spodoptera litura in transgenic peanut (Arachis hypogaea L.). Plant Cell Rep 27, 1017–1025 (2008). https://doi.org/10.1007/s00299-008-0525-x

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  • DOI: https://doi.org/10.1007/s00299-008-0525-x

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