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Development of Insect-Resistant Transgenic Cotton with Chimeric TVip3A Accumulating in Chloroplasts

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Transgenic Cotton

Part of the book series: Methods in Molecular Biology ((MIMB,volume 958))

Abstract

An optimized vip3A gene, designated as vip3A* was chemically synthesized and a thi1 gene chloroplast transit peptide coding sequence was attached to its 5′ end to produce the tvip3A*. vip3A* and tvip3A* genes were transformed into Gossypium hirsutum cv. Zhongmiansuo35 mediated by Agrobacterium tumefaciens. Four independent transgenic T1 lines with single-copy insertions and unchanged phenotypes (CTV1 and CTV2 for tvip3A*, and CV1 and CV2 for vip3A*) were selected by Polymerase chain reaction (PCR), Reverse transcription (RT)-PCR, Southern blotting, enzyme-linked immunosorbent assay (ELISA), and insect bioassay. As expected, the Vip3A* protein of CTV1 and CTV2 were transported to the chloroplasts, where they accumulated. Our results suggest that the two tvip3A* transgenic lines (CTV1 and CTV2) can be used to develop insect-resistant cultivars and could be used as a resource for raising multi-toxins-expressing transgenic cotton.

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Authors and Affiliations

  1. State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China

    Jiahe Wu & Yingchuan Tian

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  1. Jiahe Wu
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  2. Yingchuan Tian
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Correspondence to Jiahe Wu .

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Editors and Affiliations

  1. , Department of Biology, East Carolina University, Howell Science Complex S112, Greenville, 27858, North Carolina, USA

    Baohong Zhang

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Wu, J., Tian, Y. (2013). Development of Insect-Resistant Transgenic Cotton with Chimeric TVip3A Accumulating in Chloroplasts. In: Zhang, B. (eds) Transgenic Cotton. Methods in Molecular Biology, vol 958. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-212-4_20

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  • DOI: https://doi.org/10.1007/978-1-62703-212-4_20

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-211-7

  • Online ISBN: 978-1-62703-212-4

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