Sun L, Xiao H-J, Gu S-H, Zhou J-J, Guo Y-Y, Liu Z-W, Zhang Y-J
Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing, China; State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
Insect Mol Biol. 2014 Aug;23(4):417-34. doi: 10.1111/imb.12089. Epub 2014 Feb 28.
Odorant-binding proteins (OBPs) are crucial in the olfactory pathway of insects. In the present study, the antenna-enriched OBP AlinOBP13 was investigated because of its potential contribution to the peripheral olfactory perception in the alfalfa plant bug Adelphocoris lineolatus. The results of quantitative reverse transcriptase-PCR showed that the transcript level of AlinOBP13 was higher in the adult stage than in the nymph stages. The transcript levels of AlinOBP13 in the male and female antennae significantly increased after 4 and 8 h of starvation, respectively. Fine ultrastructures of different types of chemosensilla in both female and male antennae were investigated using transmission electron microscopy and immunocytochemical labelling. The results revealed that the anti-AlinOBP13 antiserum strongly and specifically labelled short basiconic sensilla; this antiserum was restricted to the inner lumen and the cavities below the sensillum base of the sensilla. By contrast, multiporous sensilla trichodea, medium long sensilla basiconica, and aporous sensilla chaetica were not labelled. The present study is the first to report an OBP showing specific expression in the short basiconic sensilla of a member of the Hemipteran species. The results of a fluorescence displacement binding assay indicated that recombinant AlinOBP13 showed a more specific binding preference to terpenoids than to sex pheromones and other classes of chemicals. This binding ability was dramatically affected by pH; higher binding affinities were displayed at pH 10.0 than at pH 7.4 and 5.0. In addition, the results of dose-dependent electroantennogram recordings from the antennae showed that both female and male adult bugs responded to the terpenoids tested, suggesting an apparent physiological relevance of AlinOBP13 in A. lineolatus chemoreception. The results of this study suggest that AlinOBP13 functions as a specific carrier of terpenoids and provide insights into the mechanism of A. lineolatus in response to green volatiles.
气味结合蛋白(OBPs)在昆虫的嗅觉通路中至关重要。在本研究中,对在触角中高度富集的OBP即苜蓿盲蝽AlinOBP13进行了研究,因为其可能对苜蓿盲蝽的外周嗅觉感知有贡献。定量逆转录PCR结果显示,AlinOBP13的转录水平在成虫期高于若虫期。饥饿4小时和8小时后,雄性和雌性触角中AlinOBP13的转录水平分别显著升高。使用透射电子显微镜和免疫细胞化学标记研究了雄性和雌性触角中不同类型化学感受器的精细超微结构。结果显示,抗AlinOBP13抗血清强烈且特异性地标记了短锥形感受器;该抗血清局限于感受器的内腔和感受器基部下方的腔室。相比之下,多孔毛形感受器、中长锥形感受器和无孔刺形感受器未被标记。本研究首次报道了一种在半翅目物种成员的短锥形感受器中特异性表达的OBP。荧光位移结合试验结果表明,重组AlinOBP13对萜类化合物的结合偏好比对性信息素和其他类化学物质更具特异性。这种结合能力受pH值的显著影响;在pH 10.0时显示出比pH 7.4和5.0更高的结合亲和力。此外,对触角进行剂量依赖性触角电位记录的结果表明,成年雌雄虫均对所测试的萜类化合物有反应,这表明AlinOBP13在苜蓿盲蝽化学感受中具有明显的生理相关性。本研究结果表明,AlinOBP13作为萜类化合物的特异性载体发挥作用,并为苜蓿盲蝽对绿色挥发物的反应机制提供了见解。