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| Molecular Characterization, Expression Patterns and Binding Properties of Two Pheromone-Binding Proteins from the Oriental Fruit Moth, Grapholita molesta (Busck) |
| SONG Yue-qin, DONG Jun-feng, QIAO Hui-li , WU Jun-xiang |
1、State Key Laboratory of Crop Stress Biology for Arid Areas/Key Laboratory of Plant Protection Resources and Pest Management, Ministry of
Education/College of Plant Protection, Northwest A&F University, Yangling 712100, P.R.China
2、College of Forestry, Henan University of Science and Technology, Luoyang 471003, P.R.China
3、China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang 473061, P.R.China |
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摘要 Insect pheromone-binding proteins (PBPs) play important roles in transporting hydrophobic pheromone components across the sensillum lymph to the surface of olfactory receptors (ORs). However, the PBPs of the oriental fruit moth, Grapholita molesta, an important destructive pest of stone fruits worldwide, are not well characterized. In this study, two new putative PBP genes, GmolPBP2 and GmolPBP3, were identified from G. molesta antennae. The deduced amino-acid sequences of these two putative PBP genes are characteristic of the odorant binding protein family, containing six conserved cysteine residues. The genomic DNA sequence of each gene contained two introns. However, the lengths and positions of the introns differed. RT-PCR analyses revealed that the two GmolPBP genes are only expressed in the antennae of female and male moths. Quantitative real-time PCR indicated that the transcription levels of GmolPBP2 are far greater than those of GmolPBP3 in both female and male antennae. GmolPBP3 showed higher transcription levels in female antennae than in male antennae, while GmolPBP2 showed similar transcription levels in both female and male antennae. The transcript levels of both genes were significantly different in premating and post-coitum individuals, implying that mating affects the process of sex pheromone reception. To better understand the functions, two GmolPBPs were expressed in Escherichia coli, and the ligand binding assays were conducted. Results showed that GmolPBP2 has strong binding affinities to two sex pheromone components, E8-12:Ac and Z8-12:Ac, as well as weaker binding affinities to Z8-12:OH and 12:OH. GmolPBP2 also bound some ordinary odor molecules. However, the affinity of GmolPBP3 to both sex pheromones and ordinary odor molecules was very weak. These results show that GmolPBP2 plays the main role in pheromone discrimination and recognition in the oriental fruit moth.
Abstract Insect pheromone-binding proteins (PBPs) play important roles in transporting hydrophobic pheromone components across the sensillum lymph to the surface of olfactory receptors (ORs). However, the PBPs of the oriental fruit moth, Grapholita molesta, an important destructive pest of stone fruits worldwide, are not well characterized. In this study, two new putative PBP genes, GmolPBP2 and GmolPBP3, were identified from G. molesta antennae. The deduced amino-acid sequences of these two putative PBP genes are characteristic of the odorant binding protein family, containing six conserved cysteine residues. The genomic DNA sequence of each gene contained two introns. However, the lengths and positions of the introns differed. RT-PCR analyses revealed that the two GmolPBP genes are only expressed in the antennae of female and male moths. Quantitative real-time PCR indicated that the transcription levels of GmolPBP2 are far greater than those of GmolPBP3 in both female and male antennae. GmolPBP3 showed higher transcription levels in female antennae than in male antennae, while GmolPBP2 showed similar transcription levels in both female and male antennae. The transcript levels of both genes were significantly different in premating and post-coitum individuals, implying that mating affects the process of sex pheromone reception. To better understand the functions, two GmolPBPs were expressed in Escherichia coli, and the ligand binding assays were conducted. Results showed that GmolPBP2 has strong binding affinities to two sex pheromone components, E8-12:Ac and Z8-12:Ac, as well as weaker binding affinities to Z8-12:OH and 12:OH. GmolPBP2 also bound some ordinary odor molecules. However, the affinity of GmolPBP3 to both sex pheromones and ordinary odor molecules was very weak. These results show that GmolPBP2 plays the main role in pheromone discrimination and recognition in the oriental fruit moth.
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Received: 07 October 2013
Accepted:
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| Fund: This work was supported by the National Natural Science Foundation of China (31272043) and the the Special Fund for Agro-Scientific Research in the Public Interest, China (201103024). |
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Corresponding Authors:
WU Jun-xiang, Tel: +86-29-87092401, E-mail: junxw@nwsuaf.edu.cn
E-mail: junxw@nwsuaf.edu.cn
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| About author: SONG Yue-qin, Mobile: 15139929764, E-mail: songyueqin6@163.com; |
Cite this article:
SONG Yue-qin, DONG Jun-feng, QIAO Hui-li , WU Jun-xiang.
2014.
Molecular Characterization, Expression Patterns and Binding Properties of Two Pheromone-Binding Proteins from the Oriental Fruit Moth, Grapholita molesta (Busck). Journal of Integrative Agriculture, 13(12): 2709-2720.
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| Abraham D, Löfstedt C, Picimbon J F. 2005. Molecularcharacterization and evolution of pheromone bindingprotein genes in Agrotis moths. Insect Biochemistry andMolecular Biology, 35, 1100-1111Anton S, Dufour M C, Gadenne C. 2007. Plasticity of olfactoryguidedbehaviour and its neurobiological basis: Lessonsfrom moths and locusts. Entomologia Experimentalis etApplicata, 123, 1-11Baker T C, Cardé R T. 1979. Analysis of pheromone-mediatedbehaviors in male Grapholita molesta, oriental fruit moth.Environmental Entomology, 8, 956-968Cardé A M, Baker T C, Cardé R T. 1979. Identification of afour-component sex pheromone of the female oriental fruitmoth, Grapholitha molesta (Lepidoptera: Tortricidae).Journal of Chemical Ecology, 5, 423-427Du J, Wang Y R, Wu J X. 2010. Effect of four differentartificial diets on development and reproduction ofGrapholita molesta (Lepidoptera: Tortricidae). Journal ofShanxi Agricultural University, 30, 229-231 (in Chinese)Foret S, Maleszka R. 2006. Function and evolution of a genefamily encoding odorant binding-like proteins in a socialinsect, the honey bee (Apis mellifera). Genome Research,16, 1404-1413Forstner M, Gohl T, Breer H, Krieger J. 2006. Candidatepheromone binding proteins of the silkmoth Bombyx mori.Invertebrate Neuroscience, 6, 177-187Gräter F, Xu W, Leal W, Grubmüller H. 2006. Pheromonediscrimination by the pheromone-binding protein ofBombyx mori. Structure, 14, 1577-1586Groβe-Wilde E, Gohl T, Bouché E, Breer H, Krieger J. 2007.Candidate pheromone receptors provide the basis forthe response of distinct antennal neurons to pheromonalcompounds. European Journal of Neuroscience, 25,2364-2373Guo H, Huang L Q, Pelosi P, Wang C Z. 2012. Threepheromone-binding proteins help segregation betweentwo Helicoverpa species utilizing the same pheromonecomponents. Insect Biochemistry and Molecular Biology,42, 708-716Gu S H, Zhou J J, Wang G R, Zhang Y J, Guo Y Y. 2013.Sex pheromone recognition and immunolocalization ofthree pheromone binding proteins in the black cutwormmoth Agrotis ipsilon. Insect Biochemistry and MolecularBiology, 43, 237-251Hallem E A, Carlson J R. 2004. The odor coding system ofDrosophila. Trends in Genetics, 20, 453-459Han K S, Jung J K, Choi K H, Lee S W, Boo K S. 2001. Sexpheromone composition and male trapping of the orientalfruit moth, Grapholita molesta (Lepidoptera: Tortricidae)in Korea. Journal of Asia-Pacific Entomology, 4, 31-35Hooper A M, Dufour S, He X, Muck A, Zhou J J, AlmeidaR, Field L M, Svatoš A, Pickett J A. 2009. HighthroughputESI-MS analysis of binding between theBombyx mori pheromone-binding protein BmorPBP1, itspheromone components and some analogues. ChemicalCommunications, 38, 5725-5727Kaissling K E, Priesner E. 1970. Die Riechschwelle desSeidenspinners. Naturwissenschaften, 57, 23-28Laughlin J D, Ha T S, Jones D N M, Smith D P. 2008.Activation of pheromone-sensitive neurons is mediated byconformational activation of pheromone-binding protein.Cell, 133, 1255-1265Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2-ΔΔCT method. Methods, 25, 402-408Liu N Y, Liu C C, Dong S L. 2013. Functional differentiationof pheromone-binding proteins in the common cutwormSpodoptera litura. Comperative Biochemistry andPhysiology (Part A), 165, 254-262Maida R, Ziegelberger G, Kaissling K E. 2003. Ligandbinding to six recombinant pheromone-binding proteins ofAntheraea polyphemus and Antheraea pernyi. Journal ofComparative Physiology (B: Biochemical, Systemic, andEnvironmental Physiology), 173, 565-573Mittapalli O, Wise I L, Shukle R H. 2006. Characterization of a serine carboxypeptidase in the salivary glands andfat body of the orange wheat blossom midge, Sitodiplosismosellana (Diptera:Cecidomyiidae). Insect Biochemistryand Molecular Biology, 36, 154-160Nagnan-Le Meillour P, Huet J C, Maïbèche M, Pernollet JC, Descoins C. 1996. Purification and characterization ofmultiple forms of odorant/pheromone binding proteins inthe antennae of Mamestra brassicae (Noctuidae). InsectBiochemistry and Molecular Biology, 26, 59-67Olsson S B, Hansson B S. 2012. A flux capacitor for mothpheromones. Chemical Senses, 37, 295-298Picimbon J F, Gadenne C. 2002. Evolution of noctuidpheromone binding proteins:identification of PBP in theblack cutworm moth, Agrotis ipsilon. Insect Biochemistryand Molecular Biology, 32, 839-846Robertson H M, Martos R, Sears C R, Todres E Z, WaldenK K O, Nardi J B. 1999. Diversity of odourant bindingproteins revealed by an expressed sequence tag projecton male Manduca sexta moth antennae. Insect MolecularBiology, 8, 501-518Roelofs W L, Comeau A, Selle R. 1969. Sex pheromone ofthe oriental fruit moth. Nature, 224, 723.Rospars J P, Gu Y, Grémiaux A, Lucas P. 2010. Odourtransduction in olfactory receptor neurons. Chinese Journalof Physiology, 53, 364-372Rothschild G H L, Vickers R A. 1991. The biology, ecologyand control of Oriental fruit moth. In: Van Der Geest L P S,Evenhuis H H, eds., Tortricid Pests: Their Biology, NaturalEnemies and Control. Elsevier, Amsterdam. pp. 389-412Rutzler M, Zwiebel L J. 2005. Molecular biology of insectolfaction:recent progress and conceptual models. Journalof Comparative Physiology (A-Neuroethology SensoryNeural and Behavioral Physiology), 191, 777-790Sun M J, Liu Y, Wang G R. 2013. Expression patterns andbinding properties of three pheromone binding proteinsin the diamondback moth, Plutella xylostella. Journal ofInsect Physiology, 59, 46-55Tamura K, Peterson D, Peterson N, Stecher G, Nei M, KumarS. 2011. MEGA 5: Molecular evolutionary geneticsanalysis using maximum likelihood, evolutionary distance,and maximum parsimony methods. Molecular Biology andEvolution, 28, 2731-2739Vogt R G, Prestwich G D, Lerner M R. 1991. Odorantbinding-protein subfamilies associate with distinctclasses of olfactory receptors neurons in insect. Journalof Neurobiology, 22, 74-84Vogt R G, Riddiford L M. 1981. Pheromone binding andinactivation by moth antennae. Nature, 293, 161-163Wang G R, Wu K M, Guo Y Y. 2003. Cloning, expression andimmunocytochemical localization of a general odorantbindingprotein gene from Helicoverpa armigera (Hübner).Insect Biochemistry and Molecular Biology, 33, 115-124Xiu W M, Dong S L. 2007. Molecular characterization of twopheromone binding proteins and quantitative analysis oftheir expression in the beet armyworm, Spodoptera exiguaHübner. Journal of Chemical Ecology, 33, 947-961Yang C Y, Han K S, Boo K S. 2001. Occurrence of and damageby the oriental fruit moth, Grapholita molesta (Busck)(Lepidoptera: Tortricidae) in pear orchards. Journal ofApplied Entomology, 40, 117-123Zhang T T, Gu S H, Wu K M, Zhang Y J, Guo Y Y. 2011.Construction and analysis of cDNA libraries fromthe antennae of male and female cotton bollwormsHelicoverpa armigera (Hübner) and expression analysisof putative odorant-binding protein genes. Biochemicaland Biophysical Research Communications, 407, 393-399Zhang T T, Mei X D, Feng J N, Berg B G, Zhang Y J, GuoY Y. 2012. Characterization of three pheromone-bindingproteins (PBPs) of Helicoverpa armigera (Hübner) andtheir binding properties. Journal of Insect Physiology,58, 941-948Zhou J J. 2010. Odorant-binding proteins in insects. Vitaminsand Hormones, 83, 241-272. |
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