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journal1 ›› 2018, Vol. 34 ›› Issue (6): 838-847.DOI: 10.16409/j.cnki.2095-039x.2018.06.006

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Fusion Expression and Insecticidal Activity of Two Spider Toxin Peptides with Cry1Ac Protein from Bacillus thuringiensis

CHEN Junjun1, LIU Fang2, LIAO Xianqing2, ZHANG Zhigang2, MIN Yong2, RAO Ben2, YANG Ziwen2, ZHOU Ronghua2, LIU Xiaoyan2   

  1. 1. College of Life Science, Wuhan University, Wuhan 430072, China;
    2. Biopesticide Branch, Hubei Innovation Centre of Agricultural Science and Technology/Hubei Biopesticide Engineering Research Centre/Hubei Academy of Agricultural Sciences, Wuhan 430064, China
  • Received:2018-09-03 Online:2018-12-08 Published:2018-12-08

Abstract: Spider venom contains many insecticidal peptides, thus showing strong insecticidal effect and able to kill agricultural and forestry pests quickly. The spider toxin peptides (ω-ACTX-Hv1a and ω-ACTX-Hv2a) were isolated from the venom of Hadronyche versuta, which are toxic to insects but not to mammals. In this study, the insecticidal gene cry1Ac from Bacillus thuringiensis (Bt) wild type strain NBIC380 was fused with the two synthetic virulent peptide genes (hv1a and hv2a) and the fused gene and the individual genes were expressed in BCIC380. Seven recombinant strains were obtained, BtBM-Ve (empty plasmid control), BtBM-Ac (carrying cry1Ac gene), BtBM-1a (carrying hv1a gene), BtBM-2a (carrying hv2a gene), BtA1a (carrying fusion gene cry1Ac-hv1a), BtA2a (carrying fusion gene cry1Ac-hv2a), BtA(1+2)a (carrying fusion gene cry1Ac-hv1a-hv2a). The insecticidal activities of the recombinant strains were tested against the 2nd instar larvae of Helicoverpa armigera, Caenorhabditis elegans and Tetranychus cinnabarinus. The results showed that BtBM-Ve had no insecticidal and synergistic effects, BtBM-Ac, BtBM-1a, BtBM-2a, BtA1a, BtA2a and BtA(1+2)a had varying insecticidal and synergistic activities against different pests. BtA(1+2)a was the most effective recombinant against the three pests, with an increase in insecticidal efficiency of 93.43% against the H. armigera 2nd instars, 34.48% against C. elegans, and 13.46% against T. cinnabarinus. The genetically engineered strains constructed in this study are of significant potential in control of Lepidopteran, mite and nematode pests.

Key words: spider toxin peptide, Bacillus thuringiensis, fusion protein, insecticidal activity

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