title: CLONING, EXPRESSION, AND FUNCTIONAL ANALYSIS OF THREE ODORANT-BINDING PROTEINS OF THE ORIENTAL FRUIT MOTH, Grapholita molesta (BUSCK) (LEPIDOPTERA: TORTRICIDAE).
abstract: Odorant-binding proteins (OBPs) act in insect olfactory processes. OBPs are expressed in the olfactory organs and serve in binding and transport of hydrophobic odorants through the sensillum lymph to olfactory receptor neurons within the antennal sensilla. In this study, three OBP genes were cloned from the antennal transcriptome database of Grapholita molesta via reverse-transcription PCR. Recombinant GmolOBPs (rGmolOBPs) were expressed in a prokaryotic expression system and enriched via Ni ion affinity chromatography. The binding properties of the three rGmolOBPs to four sex pheromones and 30 host-plant volatiles were investigated in fluorescence ligand-binding assays. The results demonstrated that rGmolOBP8, rGmolOBP11, and rGmolOBP15 exhibited high binding affinities with the major sex pheromone components (E)-8-dodecenyl acetate, (Z)-8-dodecenyl alcohol, and dodecanol. The volatiles emitted from peach and pear, decanal, butyl hexanoate, and α-ocimene, also showed binding affinities to rGmolOBP8 and rGmolOBP11. Hexanal, heptanal, and α-pinene showed strong binding affinities to rGmolOBP15. Results of the electrophysiological recording experiments and previous behavior bioassays indicated that adult insects had strong electroantennogram and behavioral responses toward butyl hexanoate, hexanal, and heptanal. We infer that the GmolOBP8 and GmolOBP11 have dual functions in perception and recognition of host-plant volatiles and sex pheromones, while GmolOBP15 was mainly involved in plant volatile odorants' perception.
authors.affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, China.|Key Laboratory of Applied Entomology, Northwest A&F University, Yangling, China.
authors.forename: Guang-Wei
authors.initials: GW
authors.lastname: Li
authors.affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, China.|Key Laboratory of Applied Entomology, Northwest A&F University, Yangling, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, China.|Key Laboratory of Applied Entomology, Northwest A&F University, Yangling, China.
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Li
authors.affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, China.|Key Laboratory of Applied Entomology, Northwest A&F University, Yangling, China.
authors.forename: Jun-Xiang
authors.initials: JX
authors.lastname: Wu
authors.affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, China.|Key Laboratory of Applied Entomology, Northwest A&F University, Yangling, China.
authors.forename: Xiang-Li
authors.initials: XL
authors.lastname: Xu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057506:Arthropod Antennae / Q000378:metabolism*; D001483:Base Sequence; D002846:Chromatography, Affinity; D055724:Electrophysiological Phenomena; D005260:Female; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000378:metabolism*; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012724:Sex Attractants; D013050:Spectrometry, Fluorescence; D059467:Transcriptome
chemical_list: D019476:Insect Proteins; D018035:Receptors, Odorant; D012724:Sex Attractants
keywords: Grapholita molesta (Busck); fluorescence binding assay; odorant-binding protein; olfaction