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Cloning and characterization of three chemosensory proteins from Spodoptera exigua and effects of gene silencing on female survival and reproduction

Published online by Cambridge University Press:  05 April 2012

L. Gong ,
Q. Luo ,
M. Rizwan-ul-Haq  and
M.-Y. Hu
L. Gong
Affiliation:
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, P.R. China, College of Natural Resources and Environment, South China Agricultural University, Guangzhou, 510642, Guangdong Province, China
Q. Luo
Affiliation:
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, P.R. China, College of Natural Resources and Environment, South China Agricultural University, Guangzhou, 510642, Guangdong Province, China
M. Rizwan-ul-Haq
Affiliation:
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, P.R. China, College of Natural Resources and Environment, South China Agricultural University, Guangzhou, 510642, Guangdong Province, China
M.-Y. Hu*
Affiliation:
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, P.R. China, College of Natural Resources and Environment, South China Agricultural University, Guangzhou, 510642, Guangdong Province, China
*
*Author for correspondence Fax: +86 02085280292 E-mail: [email protected]
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Abstract

Insect chemosensory proteins (CSPs) are supposed to transport hydrophobic chemicals to receptors on sensory neurons. However, CSPs are broadly expressed in various insect tissues, suggesting their involvement in the physiological processes beyond chemoreception. So, the exact physiological roles of CSPs in insects still need to be unraveled. In this study, three full-length of CSP genes from Spodoptera exigua have been cloned and characterized. The deduced amino acid sequences of SexiCSP1, SexiCSP2 and SexiCSP3 revealed open reading frames of 128, 128 and 126 amino acids, respectively, with four conserved cysteine residues. The expression patterns of the three SexiCSPs were further investigated by real-time PCR. Three SexiCSPs were expressed in antennae, heads, legs, wings, thoraxes, abdomens, testes and ovaries, with the highest expression level in female and male antennae. Furthermore, all three SexiCSPs mRNA were distributed extensively in the tested development stages with the highest expression level in pupae. RNAi-based gene silencing study resulted in a dramatic reduction of corresponding mRNA in female S. exigua after injection with dsRNA of all three SexiCSPs. Consequentially, 42.5% of mortalities, 68.3% (compare to DEPC water injected control) and 71.4% (compare to uninjected control) oviposition inhibition, and significantly effected egg hatching were observed in the female S. exigua injected with dsSexiCSP3 as compared to control treatments. On the other hand, dsSexiCSP1 and dsSexiCSP2 injected female adults did not show effects on survival and reproduction. Our study confirms the utility of RNAi approach to functional characterization of CSP genes in S. exigua and provides a starting point for further studies on female survival and reproduction in this insect. It also reveals the potential pest controlling method, as insect behavior regulation agent that disrupts the expression of chemosensory proteins.

Keywords

chemosensory proteinsSpodoptera exiguaRNA interferencefemale survivalreproduction
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 102 , Issue 5 , October 2012 , pp. 600 - 609
Copyright
Copyright © Cambridge University Press 2012

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References

Angeli, S., Ceron, F., Scaloni, A., Monti, M., Monteforti, G., Minnocci, A., Petacchi, R. & Pelosi, P. (1999) Purification, structural characterization, cloning and immunocytochemical localization of chemoreception proteins from Schistocerca gregaria. Europen Journal of Biochemistry 262(3), 745–754.CrossRefGoogle ScholarPubMed
Bohbot, J., Sobrio, F., Lucas, P. & Nagnan-Le Meillour, P. (1998) Functional characterization of a new class of odorant binding proteins in the moth Mamestra brassicae. Biochemical and Biophysical Research Communications 253, 489–494.CrossRefGoogle ScholarPubMed
Chen, J., Tang, B., Chen, H., Yao, Q., Huang, X., Chen, J., Zhang, D.W. & Zhang, W.Q. (2010) Different functions of the insect soluble and membrane-bound trehalase genes in chitin biosynthesis revealed by RNA interference. PLoS ONE 5(4), e10133.CrossRefGoogle ScholarPubMed
Chen, X., Tian, H., Zou, L., Tang, B., Hu, J. & Zhang, W. (2008) Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference. Bulletin of Entomological Research 98, 613–619.CrossRefGoogle ScholarPubMed
Emanuelsson, O., Brunak, S., Heijne, G.V. & Nielsen, H. (2007) Locating proteins in the cell using TargetP, SignalP and related tools. Nature Protocols 2, 953–971.CrossRefGoogle ScholarPubMed
Forêt, S., Kevin, W. & Wanner, M.R. (2007) Chemosensory proteins in the honey bee: Insights from the annotated genome, comparative analyses and expressional profiling. Insect Biochemistry and Molecular Biology 37, 19–28.CrossRefGoogle ScholarPubMed
Gong, D.P., Zhang, H.J., Zhao, P., Lin, Y., Xia, Q.Y. & Xiang, Z.H. (2007) Identification and expression pattern of the chemosensory protein gene family in the silkworm, Bombyx mori. Insect Biochemistry and Molecular Biology 37, 266–277.CrossRefGoogle ScholarPubMed
Gong, L., Chen, Y., Cheng, G. & Zhong, G.H. (2009) Insect chemosensory proteins. Chinese Bulletin of Entomology 46(4), 646–652.Google Scholar
Gong, L., Zhong, G.H., Hu, M.Y., Luo, Q. & Ren, Z.Z. (2010) Molecular cloning, expression profile and 5′regulatory region analysis of two chemosensory protein genes from the diamondback moth, Plutella xylostella. Journal of Insect Science 10, 143.CrossRefGoogle Scholar
Higgins, D., Thompson, J., Gibson, T. & Thompson, J.D. (1994) CLUSTALW improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673–4680.Google Scholar
Kitabayashi, A.N., Arai, T., Kubo, T. & Natori, S. (1998) Molecular cloning of cDNA for p10, a novel protein that increases in regenerating legs of Periplaneta americana (American cockroach). Insect Biochemistry and Molecular Biology 28, 785–790.CrossRefGoogle ScholarPubMed
Lartigue, A., Campanacci, V., Roussel, A., Larsson, A.M., Jones, T.A. & Tegoni, M. (2002) X-ray structure and ligand binding study of a moth chemosensory protein. Journal of Biology Chemical 277, 32094–32098.CrossRefGoogle ScholarPubMed
Leal, W.S., Nikonova, L. & Peng, G. (1999) Disulfide structure of the pheromone binding protein from the silkworm moth, Bombyx mori. FEBS Letter 464, 85–90.CrossRefGoogle ScholarPubMed
Liu, R., He, X., Lehane, S., Lehane, M., Hertz-Fowler, C., Berriman, M., Field, L.M., & Zhou, J.J. (2011) Expression of chemosensory proteins in the tsetse fly Glossina morsitans morsitans is related to female host-seeking behaviour. Insect Molecular Biology 21, 41–48.CrossRefGoogle ScholarPubMed
Livak, K.J. & Schmittgen, T.D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔC T method. Methods 25, 402–408.CrossRefGoogle Scholar
Maike, F., Heinz, B. & Jürgen, K. (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(7), 745–757.Google Scholar
Maleszka, J., Forê, S., Saint, R. & Maleszka, R. (2007) RNAi-induced phenotypes suggest a novel role for a chemosensory protein CSP5 in the development of embryonic integument in the honeybee (Apis mellifera). Development Genes and Evolution 217, 189–196.CrossRefGoogle ScholarPubMed
Martine, M.C., Christine, M., François, M.C., Isabelle, Q., Patrick, P. & Emmanuelle, J.J. (2004) Putative odorant-degrading esterase cDNA from the moth Mamestra brassicae: cloning and expression patterns in male and female antennae. Chemical Senses 29, 381–390.Google Scholar
Mosbah, A., Campanacci, V., Lartigue, A., Tegoni, M., Cambillau, C. & Darbon, H. (2003) Solution structure of a chemosensory protein from the moth Mamestra brassicae. Biochemical Journal 369, 39–44.CrossRefGoogle ScholarPubMed
Nagnan-Le Meillour, P., Cain, A.H., Jacquin-Joly, E., Francois, M.C., Ramachandran, S., Maida, R. & Steinbrecht, R.A. (2000) Chemosensory proteins from the proboscis of Mamestra brassicae. Chemical Senses 25, 541–553.CrossRefGoogle ScholarPubMed
Ozaki, K., Utoguchi, A., Yamada, A. & Yoshikawa, H. (2008) Identification and genomic structure of chemosensory proteins (CSP) and odorant binding proteins (OBP) genes expressed in foreleg tarsi of the swallowtail butterfly Papilio xuthus. Insect Biochemistry and Molecular Biology 38, 969–976.CrossRefGoogle ScholarPubMed
Pelletier, J. & Leal, W.S. (2011) Characterization of olfactory genes in the antennae of the Southern house mosquito, Culex quinquefasciatus. Journal of Insect Physiology 57, 915–929.CrossRefGoogle ScholarPubMed
Pelosi, P. (1996) Perireceptor events in olfaction. Journal of Neurobiology 30, 3–19.3.0.CO;2-A>CrossRefGoogle ScholarPubMed
Pelosi, P., Zhou, J.J., Ban, L.P. & Calvello, M. (2006) Soluble proteins in insect chemical communication. Cell and Molecular Life Science 63, 1658–1676.CrossRefGoogle ScholarPubMed
Picimbon, J.F., Dietrich, K., Krieger, J. & Breer, H. (2001) Identity and expression pattern of chemosensory proteins in Heliothis virescens (Lepidoptera, Noctuidae). Insect Biochemistry and Molecular Biology 31, 1173–1181.CrossRefGoogle ScholarPubMed
Sabatier, L., Jouanguy, E., Dostert, C., Zachary, D., Dimarcq, J.L., Bulet, P. & Imler, J.L. (2003) Pherokine-2 and -3. Two Drosophila molecules related to pheromone/odor-binding proteins induced by viral and bacterial infection. European Journal Biochemistry 270, 3398–3407.CrossRefGoogle Scholar
Sanchez-Gracia, A., Vieira, F.G. & Rozas, J. (2009) Molecular evolution of the major chemosensory gene families in insects. Heredity 103, 208–216.CrossRefGoogle ScholarPubMed
SAS Institute, Inc. (2000) SAS User's Guide: Statistics. SAS Institute, Cary, New York, USA.Google Scholar
Sengul, M.S. & Tu, Z. (2010) Identification and characterization of odorant-binding protein 1 gene from the Asian malaria mosquito, Anopheles stephensi. Insect Molecular Biology 19(1), 49–60.CrossRefGoogle ScholarPubMed
Stathopoulos, A., Van Drenth, M., Erives, A., Markstein, M. & Levine, M. (2002) Whole genome analysis of dorsal-ventral patterning in the Drosophila embryo. Cell 111, 687–701.CrossRefGoogle ScholarPubMed
Tamura, K., Dudley, J., Nei, M. & Kumar, S. (2007) Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24, 1596–1599.CrossRefGoogle ScholarPubMed
Tang, B., Wang, S.G. & Zhang, F. (2010) Two storage hexamerins from the beet armyworm Spodoptera exigua: Cloning, characterization and the effect of gene silencing on survival. BMC Molecular Biology 11, 65.CrossRefGoogle ScholarPubMed
Wanner, K.W., Willis, L.G., Theilmann, D.A., Isman, M.B., Feng, Q. & Plettner, E. (2004) Analysis of the insect os-d-like gene family. Journal Chemical Ecology 30, 889–911.CrossRefGoogle ScholarPubMed
Wanner, K.W., Isman, M.B., Feng, Q., Plettner, E. & Theilmann, D.A. (2005) Developmental expression patterns of four chemosensory protein genes from the Eastern spruce budworm, Chroistoneura fumiferana. Insect Molecular Biology 14(3), 289–300.CrossRefGoogle ScholarPubMed
Zhou, D.S., Wanga, C.Z. & van Loon, J.J.A. (2009) Chemosensory basis of behavioural plasticity in response to deterrent plant chemicals in the larva of the Small Cabbage White butterfly Pieris rapae. Journal of Insect Physiology 792(55), 788–792.CrossRefGoogle Scholar
Zhao, Y.Y., Liu, F., Yang, G. & You, M.S. (2011) PsOr1, a potential target for RNA interference-based pest management. Insect Molecular Biology 20(1), 97–104.CrossRefGoogle ScholarPubMed
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