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A proteomic approach to study the mechanism of tolerance to Bt toxins in Ostrinia furnacalis larvae selected for resistance to Cry1Ab

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Abstract

A Cry1Ab-resistant population of Asian corn borer (ACB-AbR) exhibiting approximately 100 times greater resistance to activated Cry1Ab than a susceptible population (Ostrinia furnacalis; ACB-BtS), was previously shown to exhibit high levels of cross-resistance to Cry1Ah (131-fold), but no cross-resistance to Cry1Ie. It was suggested that the proposed mechanism of resistance was due to the alteration of specific receptors for Cry toxins in the midgut brush border membrane. In the present study a proteomic-based approach was used to identify proteins from brush border membrane vesicles (isolated from both resistant and susceptible Ostrinia furnacalis larvae) interacting with biotinylated Cry1Ab, Cry1Ah, and Cry1Ie. 2D-Electrophoresis in combination with ligand blots were employed and putative protein identities obtained using MALDI-ToF/ToF mass spectrometry. The V-type proton ATPase catalytic subunit A and heat shock 70 kDa proteins were identified as interacting with the Cry toxins tested in the ACB-AbR and ACB-BtS larvae. The biotinylated Cry toxins showed markedly stronger interactions with proteins in the resistant compared to the susceptible larvae, suggesting an up-regulation of the V-type proton ATPase catalytic subunit A and heat shock 70 kDa proteins in the resistant (ACB-AbR) larvae. Interestingly, Cry1Ie interactions with the V-type proton ATPase catalytic subunit A in the ACB-BtS larvae appeared to be absent.

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Acknowledgments

This research was supported in part by Genetically Modified Organisms Breeding Major Projects (2011ZX08011-003), The Royal Society (RS) and National Natural Science Foundation of China (NSFC) Joint Project (30610236) and NSFC (30571238).

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Authors and Affiliations

  1. The State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 West Yuanmingyuan Road, Beijing, 100193, People’s Republic of China

    Lina Xu, Zhenying Wang, Jie Zhang & Kanglai He

  2. Newcastle Institute for Research on Environment and Sustainability, School of Biology, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK

    Lina Xu, Natalie Ferry, Martin G. Edwards & Angharad M. R. Gatehouse

  3. Institute of Plant Protection and Agro-Products Safety, Anhui Academy of Agricultural Sciences, No. 40 South Nongke Road, Hefei, 230031, People’s Republic of China

    Lina Xu

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  1. Lina Xu
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  2. Natalie Ferry
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  3. Zhenying Wang
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  4. Jie Zhang
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Correspondence to Angharad M. R. Gatehouse or Kanglai He.

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Xu, L., Ferry, N., Wang, Z. et al. A proteomic approach to study the mechanism of tolerance to Bt toxins in Ostrinia furnacalis larvae selected for resistance to Cry1Ab. Transgenic Res 22, 1155–1166 (2013). https://doi.org/10.1007/s11248-013-9718-3

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  • DOI: https://doi.org/10.1007/s11248-013-9718-3

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