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Odorant Binding Characteristics of Three Recombinant Odorant Binding Proteins in Microplitis mediator (Hymenoptera: Braconidae)

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Abstract

Odorant binding proteins (OBPs) are believed to be important for transporting semiochemicals through the aqueous sensillar lymph to the olfactory receptor cells within the insect antennal sensilla. In this study, three new putative OBP genes, MmedOBP8-10, were identified from a Microplitis mediator (Hymenoptera: Braconidae) antennal cDNA library. Quantitative real-time PCR (qRT-PCR) analysis revealed that all three of the OBP genes were expressed mainly in the antennae of adult wasps. The three OBPs were recombinantly expressed in Escherichia coli and purified by Ni ion affinity chromatography. Fluorescence competitive binding assays were performed using N-phenyl-naphthylamine as a fluorescent probe and 45 small organic compounds as competitors. These assays demonstrated that the three M. mediator OBPs can bind a broad range of odorant molecules with different binding affinities. They can bind the following ligands: nonane, farnesol, nerolidol, nonanal, β-ionone, acetic ether, and farnesene. In a Y-tube assay with these ligands as odor stimuli and paraffin oil as a control, all ligands, except nerolidol and acetic ether, were able to elicit behavioral responses in adult M. mediator. The wasps were significantly attracted to β-ionone, nonanal, and farnesene and repelled by nonane and farnesol. The results of this work provide insight into the chemosensory functions of the OBPs in M. mediator.

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Acknowledgments

This work was supported by the China National Basic Research Program (2012CB114104) and the National Natural Science Foundation of China (31171858,31272048, and 31321004). YJ Zhang and JJ Zhou acknowledge financial support from the international joint project between China and the UK (31111130203; JP100849). This manuscript has been edited by native English-speaking experts of Elsevier Language Editing Services.

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Authors and Affiliations

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Keming Li, Shanning Wang, Liyan Ren, Abid Ali, Yongjun Zhang & Yuyuan Guo

  2. Institute of Banana and Plantain, Chinese Academy of Tropical Agricultural Sciences, Haikou, 570102, China

    Keming Li

  3. Hanzhong Agricultural Technology Extension Center, Shanxi, 723000, China

    Kang Zhang

  4. Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, AL5 2JQ, UK

    Jingjiang Zhou

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  1. Keming Li
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  2. Shanning Wang
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  3. Kang Zhang
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  4. Liyan Ren
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  5. Abid Ali
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  6. Yongjun Zhang
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  7. Jingjiang Zhou
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  8. Yuyuan Guo
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Correspondence to Yongjun Zhang.

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Fig. S1

Expression and purification of MmedOBP8-10 proteins. SDS-PAGE analyses of the recombinant OBP proteins: MmedOBP8, MmedOBP9 and MmedOBP10. Non-induced protein (nip), induced protein (ip), inclusion bodies (ib), supernatant (s), purified protein with tag (ppwt), protein molecular weight marker (m), purified protein without tag (ppwot) and low concentration of supernatant (lcos) (GIF 786 kb)

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Li, K., Wang, S., Zhang, K. et al. Odorant Binding Characteristics of Three Recombinant Odorant Binding Proteins in Microplitis mediator (Hymenoptera: Braconidae). J Chem Ecol 40, 541–548 (2014). https://doi.org/10.1007/s10886-014-0458-5

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  • DOI: https://doi.org/10.1007/s10886-014-0458-5

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