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Bacillus thuringiensis Cry11Ba works synergistically with Cry4Aa but not with Cry11Aa for toxicity against mosquito Culex pipiens (Diptera: Culicidae) larvae

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Abstract

A 2,175-bp modified gene (cry11Ba-S1) encoding Cry11Ba from Bacillus thuringiensis subsp. jegathesan was designed and the recombinant protein was expressed as a fusion protein with glutathione S-transferase in Escherichia coli. The recombinant Cry11Ba was highly toxic against Culex pipiens mosquito larvae, being nine and 17 times more toxic than mosquitocidal Cry4Aa and Cry11Aa from Bacillus thuringiensis subsp. israelensis, respectively. Interestingly, a further increase in the toxicity of the recombinant Cry11Ba was achieved by mixing with Cry4Aa, but not with Cry11Aa. These findings suggested that Cry11Ba worked synergistically with Cry4Aa, but not with Cry11Aa, in exhibiting toxicity against C. pipiens larvae. On the other hand, the amount of Cry toxin bound to brush border membrane vesicles (BBMVs) did not significantly change between individual toxins and the toxin mixtures, suggesting that the increase in toxins binding to BBMVs was not a reason for the observed synergistic effect. It is generally accepted that synergism of toxins is a potentially powerful tool for enhancing insecticidal activity and managing Cry toxin resistance in mosquitoes. The mixture of Cry4Aa and Cry11Ba in order to increase toxicity would be very valuable in terms of mosquito control.

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Acknowledgments

C. pipiens eggs were kindly supplied by the Research and Development Laboratory at Dainihon Jochugiku, Osaka. This work was supported by the Japan Society for the Promotion of Science KAKENHI (Grant Nos. JP24380034 and JP26660268).

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Authors and Affiliations

  1. Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushima-Naka, Okayama, 700-8530, Japan

    Tohru Hayakawa, Naoya Yoneda, Kouji Okada, Ayuko Higaki, Mohammad Tofazzal Hossain Howlader & Toru Ide

  2. Department of Entomology, Bangladesh Agricultural University, Mymensisngh, 2202, Bangladesh

    Mohammad Tofazzal Hossain Howlader

Authors
  1. Tohru Hayakawa
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  2. Naoya Yoneda
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  3. Kouji Okada
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  4. Ayuko Higaki
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  5. Mohammad Tofazzal Hossain Howlader
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  6. Toru Ide
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Corresponding author

Correspondence to Tohru Hayakawa.

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13355_2016_454_MOESM1_ESM.pptx

Supplemental Fig. 1 Alignment of the native and the synthetic cry11Ba gene. The amino acid sequence is shown above the nucleotide sequence. The nucleotide residues modified in accordance with the codon preference of E. coli gene are indicated. Nucleotide and amino acid numbers are shown on the right. Supplemental Fig. 2 Schematic structure of Cry protoxins and the recombinant Cry toxins used in this study. Insecticidal toxin regions are shaded. Arrow heads Internal cleavage site located in the insecticidal toxin region. After ingestion by susceptible mosquito larvae, Cry toxins are processed by residential proteases in the midgut, and the protease-resistant segments which form a heterodimer as an insecticidal toxin are generated (PPTX 182 kb)

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Hayakawa, T., Yoneda, N., Okada, K. et al. Bacillus thuringiensis Cry11Ba works synergistically with Cry4Aa but not with Cry11Aa for toxicity against mosquito Culex pipiens (Diptera: Culicidae) larvae. Appl Entomol Zool 52, 61–68 (2017). https://doi.org/10.1007/s13355-016-0454-z

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  • DOI: https://doi.org/10.1007/s13355-016-0454-z

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