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Functional characterization of a pheromone-binding protein from rice leaffolder Cnaphalocrocis medinalis in detecting pheromones and host plant volatiles

  • X. Sun (a1) (a2), Z.-F. Zhao (a1), F.-F. Zeng (a1), A. Zhang (a3), Z.-X. Lu (a4) and M.-Q. Wang (a1)...


Pheromone-binding proteins (PBPs) are believed to be involved in the recognition of semiochemicals. In the present study, western blot analysis, fluorescence-binding characteristics and immunolocalization of CmedPBP4 from the rice leaffolder, Cnaphalocrocis medinalis, were investigated. Western blot analysis revealed that CmedPBP4 showed obvious antenna-specific expression patterns in female and male antenna, and made a clearly different sex-biased expression. Immunocytochemical labeling revealed that CmedPBP4 showed specific expression in the trichoid sensilla. Competitive fluorescence binding assays indicated that CmedPBP4 could selectively recognize three sex pheromone components (Z13-18:Ac, Z11-16:Al and Z13-18:OH) and eleven rice plant volatiles, including cyclohexanol, nerolidol, cedrol, dodecanal, ionone, (−)-α-cedrene, (Z)-farnesene, β-myrcene, R-(+)-limonene, (−)-limonene, and (+)-3-carene. Meanwhile the CmedPBP4 detection of sex pheromones and host odorants was pH-dependent. Our results, for the first time, provide further evidence that trichoid sensilla might be play an important role in detecting sex pheromones and host plant volatiles in the C. medinalis moth. Our systematic studies provided further detailed evidence for the function of trichoid sensilla in insect semiochemical perception.


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Abraham, D., Löfstedt, C. & Picimbon, J.F. (2005) Molecular characterization and evolution of pheromone binding protein genes in Agrotis moths. Insect Biochemistry and Molecular Biology 35, 11001111.
Allen, J.E. & Wanner, K.W. (2011) Asian corn borer pheromone binding protein 3, a candidate for evolving specificity to the 12-tetradecenyl acetate sex pheromone. Insect Biochemistry and Molecular Biology 41, 141149.
Campanacci, V., Krieger, J., Bette, S., Sturgis, J.N., Lartigue, A., Cambillau, C., Breer, H. & Tegoni, M. (2001) Revisiting the specificity of Mamestra brassicae and Antheraea polyphemus pheromone-binding proteins with a fluorescence binding assay. Journal of Biological Chemistry 276, 2007820084.
Cho, J.R., Choi, K.S., Park, H.H., Lee, S., Yum, K.H., Jung, J.K., Seo, B.Y. & Lee, M. (2013) Electroantennogram and field responses of Korean population of the rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Crambidae), to sex attractant candidates. Journal of Asia-Pacific Entomology 16, 6166.
Forstner, M., Breer, H. & Krieger, J. (2009) A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmoth Antheraea polyphemus . International Journal of Biological Sciences 5, 745757.
Gong, D.P., Zhang, H.J., Zhao, P., Xia, Q.Y. & Xiang, Z.H. (2009 a) The odorant binding protein gene family from the genome of silkworm, Bombyx mori . BMC Genomics 10, 332.
Gong, Z.J., Zhou, W.W., Yu, H.Z., Mao, C.G., Zhang, C.X., Cheng, J.A. & Zhu, Z.R. (2009 b) Cloning, expression and functional analysis of a general odorant-binding protein 2 gene of the rice striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae). Insect Molecular Biology 18, 405417.
Grater, F., Xu, W., Leal, W. & Grubmuller, H. (2006) Pheromone discrimination by the pheromone-binding protein of Bombyx mori . Structure 14, 15771586.
Grobe-Wilde, E., Gohl, T., Bouché, E., Breer, H. & Krieger, J. (2007) Candidate pheromone receptors provide the basis for the response of distinct antennal neurons to pheromonal compounds. European Journal of Neuroscience 25, 23642373.
Groot, A., Gemeno, C., Brownie, C., Gould, F. & Schal, C. (2005) Male and female antennal responses in Heliothis virescens and H. subflexa to conspecific and heterospecific sex pheromone compounds. Environmental Entomology 34, 256263.
Grosse-Wilde, E., Svatos, A. & Krieger, J. (2006) A pheromone-binding protein mediates the bombykol-induced activation of a pheromone receptor in vitro . Chemical Senses 31, 547555.
Gu, S.H., Wang, S.P., Zhang, X.Y., Wu, K.M., Guo, Y.Y., Zhou, J.J. & Zhang, Y.J. (2011) Identification and tissue distribution of odorant binding protein genes in the lucerne plant bug Adelphocoris lineolatus (Goeze). Insect Biochemistry and Molecular Biology 41, 254263.
Gu, S.H., Zhou, J.J., Wang, G.R., Zhang, Y.J. & Guo, Y.Y. (2013) Sex pheromone recognition and immunolocalization of three pheromone binding proteins in the black cutworm moth Agrotis ipsilon . Insect Biochemistry and Molecular Biology 43, 237251.
Guo, H., Huang, L.Q., Pelosi, P. & Wang, C.Z. (2012) Three pheromone-binding proteins help segregation between two Helicoverpa species utilizing the same pheromone components. Insect Biochemistry and Molecular Biology 42, 708716.
Hillier, N. & Vickers, N. (2011) Hairpencil volatiles influence interspecific courtship and mating between two related moth species. Journal of Chemical Ecology 37, 11271136.
Hooper, A.M., Dufour, S., He, X., Muck, A., Zhou, J.J., Almeida, R., Field, L.M., Svatoš, A. & Pickett, J.A. (2009) High-throughput ESI-MS analysis of binding between the Bombyx mori pheromone-binding protein BmorPBP1, its pheromone components and some analogues. Chemical Communications 38, 57255727.
Horst, R., Damberger, F., Luginbuhl, P., Guntert, P., Peng, G., Nikonova, L., Leal, W.S. & Wuthrich, K. (2001) NMR structure reveals intramolecular regulation mechanism for pheromone binding and release. Proceedings of the National Academy of Sciences of the United States of America 98, 1437414379.
Inoue, H., Kamiwada, H. & Fukamachi, S. (2004) Seasonal changes in adult density and female mating status of the rice leaf roller Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) in paddy fields of Southern Kyushu, Japan. Applied Entomology and Zoology 48, 177183.
Jin, J.Y., Li, Z.Q., Zhang, Y.N., Liu, N.Y. & Dong, S.L. (2014) Different roles suggested by sex-biased expression and pheromone binding affinity among three pheromone binding proteins in the pink rice borer, Sesamia inferens (Walker) (Lepidoptera: Noctuidae). Journal of Insect Physiology 66, 7179.
Jin, X., Brandazza, A., Navarrini, A., Ban, L., Zhang, S., Steinbrecht, R.A., Zhang, L. & Pelosi, P. (2005) Expression and immunolocalisation of odorant-binding and chemosensory proteins in locusts. Cellular and Molecular Life Sciences 62, 11561166.
Kawazu, K. & Tatsuki, S. (2002) Diel rhythms of calling behavior and temporal change in pheromone production of the rice leaffolder moth, Cnaphalocrocis medinalis (Lepidoptera: Crambidae). Applied Entomology and Zoology 37, 219224.
Kawazu, K., Hasegawa, J., Honda, H., Ishikawa, Y., Wakamura, S., Sugie, H., Kamiwada, H., Kamimuro, T., Yoshiyasu, Y. & Tatsuki, S. (2000) Geographical variation in female sex pheromones of the rice leaf folder moth, Cnaphalocrocis medinalis: identification of pheromone components in Japan. Entomologia Experimentalis et Applicata 96, 103109.
Kawazu, K., Setokuchi, O., Kohno, K., Takahashi, K., Yoshiyasu, Y. & Tatsuki, S. (2001) Sex pheromone of the rice leaffolder moth. Cnaphalocrocis medinalis (Lepidoptera: Crambidae): synthetic Indian and Philippine blends are not attractive to male C. medinalis, but are attractive to C. pilosa in the South-Western islands in Japan. Applied Entomology and Zoology 36, 471474.
Khan, Z.R., Barrion, A.T., Litsinger, J.A., Castilla, N.P. & Joshi, R.C. (1988) A bibliography of rice leaffolders (Lepidoptera: Pyralidae). Insect Science 9, 129174.
Krieger, J. & Breer, H. 1999. Olfactory reception in invertebrates. Science 286, 720723.
Krieger, J., von Nickisch-Rosenegk, E., Mameli, M., Pelosi, P. & Breer, H. (1996) Binding proteins from the antennae of Bombyx mori . Insect Biochemistry and Molecular Biology 26, 297307.
Leal, W.S. (2003) Proteins that make sense. pp. 447476 in Blomquist, G.J. & Vogt, R.G. (Eds) Insect Pheromone Biochemistry and Molecular Biology, the Biosynthesis and Detection of Pheromone and Plant Volatiles. London, Elsevier Academic Press.
Leal, W.S., Chen, A.M., Ishida, Y., Chiang, V.P., Erickson, M.L., Morgan, T.I. & Tsuruda, J.M. (2005) Kinetics and molecular properties of pheromone binding and release. Proceedings of the National Academy of Sciences of the United States of America 102, 53865391.
Liu, N.Y., He, P. & Dong, S.L. (2012) Binding properties of pheromone-binding protein 1 from the common cutworm Spodoptera litura . Comparative Biochemistry and Physiology – Part B: Biochemistry & Molecular Biology 161, 295302.
Lou, Y.G., Ma, B. & Cheng, J.A. (2005) Attraction of the parasitoid Anagrus nilaparvatae to rice volatiles induced by the rice brown planthopper Nilaparvata lugens . Journal of Chemical Ecology 31, 23572372.
Maida, R., Ziegelberger, G. & Kaissling, K.E. (2003) Ligand binding to six recombinant pheromone-binding proteins of Antheraea polyphemus and Antheraea pernyi . Journal of Comparative Physiology B 173, 565573.
Michel, E., Damberger, F.F., Ishida, Y., Fiorito, F., Lee, D., Leal, W.S. & Wuthrich, K. (2011) Dynamic conformational equilibria in the physiological function of the Bombyx mori pheromone-binding protein. Journal of Molecular Biology 408, 922931.
Mohl, C., Breer, H. & Krieger, J. (2002) Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus . Invertebrates Neuroscience 4, 165174.
Nathan, S.S., Chung, P.G. & Murugan, K. (2004) Effect of botanical insecticides and bacterial toxins on the gut enzyme of the rice leaffolder Cnaphalocrocis medinalis . Phytoparasitica 32, 433443.
Pelosi, P. (1994) Odorant-binding proteins. Critical Reviews in Biochemistry and Molecular Biology 29, 199228.
Pelosi, P. & Maida, R. (1990) Odorant-binding proteins in vertebrates and insects: similarities and possible common function. Chemical Senses 15, 205215.
Picimbon, J.F. & Gadenne, C. (2002) Evolution of noctuid pheromone binding proteins: identification of PBP in the black cutworm moth, Agrotis ipsilon . Insect Biochemistry and Molecular Biology 32, 839846.
Prestwich, G.D., Du, G. & LaForest, S. (1995) How is pheromone specificity encoded in proteins? Chemical Senses 20, 461469.
Roelofs, W.L. (1995) Chemistry of sex attraction. Proceedings of the National Academy of Sciences of the United States of America 92, 4449.
Schneider, D. (1964) Insect antennae. Annual Review of Entomology 9, 103122.
Steinbrecht, R.A. (1998) Odorant-binding proteins: expression and function. Annals of the New York Academy of Sciences 855, 323332.
Sun, M., Liu, Y. & Wang, G. (2013 a) Expression patterns and binding properties of three pheromone binding proteins in the diamondback moth, Plutella xyllotella . Journal of Insect Physiology 59, 4655.
Sun, X., Wang, M.Q. & Zhang, G.A. (2011) Ultrastructural observations on antennal sensilla of Cnaphalocrocis medialis (Lepidoptera: Pyralidae). Microscopy Research and Technique 74, 113121.
Sun, X., Zhou, W., Liu, H., Zhang, A.J., Ai, C.R., Zhou, S.S., Zhou, C.X. & Wang, M.Q. (2013 b) Transgenic Bt rice does not challenge host preference of the target pest of rice leaffolder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae). PLoS ONE 8, e79032.
Sun, X., Liu, Z., Zhang, A., Dong, H., Zeng, F.F., Pan, X.Y., Wang, Y. & Wang, M.Q. (2014) Electrophysiological Responses of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) to synthetic plant volatiles. Journal of Insect Science V14, 70.
Visser, J.H. (1986) Host odor perception in phytophagous insects. Annual Review of Entomology 31, 121144.
Vogt, R.G. (1987) The molecular basis of pheromone reception: its influence on behavior. pp. 385431 in Prestwich, G.D. & Blomquist, G.J. (Eds) Pheromone Biochemistry. New York, Academic Press.
Vogt, R.G. & Riddiford, L.M. (1981) Pheromone binding and inactivation by moth antennae. Nature 293, 161163.
Vogt, R.G., Riddiford, L.M. & Prestwich, G.D. (1985) Kinetic properties of a sex pheromone-degrading enzyme: the sensillar esterase of Antheraea polyphemus . Proceedings of the National Academy of Sciences of the United States of America 82, 88278831.
Vogt, R.G., Prestwich, G.D. & Lerner, M.R. (1991) Odorant-binding-protein subfamilies associate with distinct classes of olfactory receptors neurons in insect. Journal of Neurobiology 22, 7484.
Vogt, R.G., Rogers, M.E., Franco, M.D. & Sun, M. (2002) A comparative study of odorant binding protein genes: differential expression of the PBP1-GOBP2 gene cluster in Manduca sexta (Lepidoptera) and the organization of OBP genes in Drosophila melanogaster (Diptera). Journal of Experimental Biology 205, 719744.
Watanabe, H., Tabunoki, H., Miura, N., Matsui, A., Sato, R. & Ando, T. (2009) Identification of a new pheromone-binding protein in the antennae of a geometrid species and preparation of its antibody to analyze the antennal proteins of moths secreting type II sex pheromone components. Bioscience, Biotechnology & Biochemistry 73, 14431446.
Wechgama, K., Laopaiboom, L. & Laopaiboon, P. (2008) Quantitative analysis of main volatile and other compounds in traditional distilled spirits from Thai rice. Biotechnology 7, 718724.
Weng, C., Fu, Y., Jiang, H., Zhuang, S. & Li, H. (2015) Binding interaction between a queen pheromone component HOB and pheromone binding protein ASP1 of Apis cerana . International Journal of Biological Macromolecules 72, 430436.
Willett, C.S. & Harrison, R.G. (1999) Pheromone binding proteins in the European and Asian corn borers: no protein change associated with pheromone differences. Insect Biochemistry and Molecular Biology 29, 277284.
Zeng, F.F., Sun, X., Dong, H.B. & Wang, M.Q. (2013) Analysis of a cDNA library from the antenna of Cnaphalocrocis medinalis and the expression pattern of olfactory genes. Biochemical and Biophysical Research Communications 433, 463469.
Zhang, S.G., Maida, R. & Steinbrecht, R.A. (2001) Immunolocalization of odorant-binding proteins in noctuid moths (Insecta, Lepidoptera). Chemical Senses 26, 885896.
Zhang, T.T., Gu, S.H., Wu, K.M., Zhang, Y.J. & Guo, Y.Y. (2011) Construction and analysis of cDNA libraries from the antennae of male and female cotton bollworms Helicoverpa armigera (Hubner) and expression analysis of putative odorant-binding protein genes. Biochemical and Biophysical Research Communications 407, 393399.



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