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Functional Characterizations of Chemosensory Proteins of the Alfalfa Plant Bug Adelphocoris lineolatus Indicate Their Involvement in Host Recognition

机译：苜蓿盲蝽的化学感受蛋白的功能表征苜蓿盲蝽表明他们在主机识别参与

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Insect chemosensory proteins (CSPs) have been proposed to capture and transport hydrophobic chemicals from air to olfactory receptors in the lymph of antennal chemosensilla. They may represent a new class of soluble carrier protein involved in insect chemoreception. However, their specific functional roles in insect chemoreception have not been fully elucidated. In this study, we report for the first time three novel CSP genes (AlinCSP1-3) of the alfalfa plant bug Adelphocoris lineolatus (Goeze) by screening the antennal cDNA library. The qRT-PCR examinations of the transcript levels revealed that all three genes (AlinCSP1-3) are mainly expressed in the antennae. Interestingly, these CSP genes AlinCSP1-3 are also highly expressed in the 5th instar nymphs, suggesting a proposed function of these CSP proteins (AlinCSP1-3) in the olfactory reception and in maintaining particular life activities into the adult stage. Using bacterial expression system, the three CSP proteins were expressed and purified. For the first time we characterized the types of sensilla in the antennae of the plant bug using scanning electron microscopy (SEM). Immunocytochemistry analysis indicated that the CSP proteins were expressed in the pheromone-sensitive sensilla trichodea and general odorant-sensitive sensilla basiconica, providing further evidence of their involvement in chemoreception. The antennal activity of 55 host-related semiochemicals and sex pheromone compounds in the host location and mate selection behavior of A. lineolatus was investigated using electroantennogram (EAG), and the binding affinities of these chemicals to the three CSPs (AlinCSP1-3) were measured using fluorescent binding assays. The results showed several host-related semiochemicals, (Z)-3-hexen-1-ol, (E)-2-hexen-1-al and valeraldehyde, have a high binding affinity with AlinCSP1-3 and can elicit significant high EAG responses of A. lineolatus antennae. Our studies indicate the three antennae-biased CSPs may mediate host recognition in the alfalfa plant bug A. lineolatus.
机译：已经提出了昆虫化学传感蛋白（CSP），以将疏水性化学物质从空气中捕获并运输到触角化学感受器淋巴中的嗅觉受体。它们可能代表了与昆虫化学感受有关的一类新的可溶性载体蛋白。但是，尚未完全阐明它们在昆虫化学感受中的特定功能。在这项研究中，我们通过筛选触角cDNA文库，首次报告了苜蓿植物臭虫Adelphocoris lineolatus（Goeze）的三个新CSP基因（AlinCSP1-3）。转录水平的qRT-PCR检查显示，所有三个基因（AlinCSP1-3）主要在触角中表达。有趣的是，这些CSP基因AlinCSP1-3在第5龄若虫中也高表达，表明这些CSP蛋白（AlinCSP1-3）在嗅觉接收中的功能以及维持特定生命活动的功能。成人阶段。使用细菌表达系统，表达并纯化了三种CSP蛋白。我们首次使用扫描电子显微镜（SEM）对植物臭虫触角中的感官类型进行了表征。免疫细胞化学分析表明，CSP蛋白在信息素敏感的Trichodea敏感型和普通气味敏感的基础型敏感型Sonica中表达，为它们参与化学感受提供了进一步的证据。使用电伏安图（EAG）研究了55种与宿主相关的化学信息素和性信息素化合物在拟南芥宿主位置和配偶选择行为中的触角活性，并且这些化学物质与三种CSP（AlinCSP1-3）的结合亲和力为使用荧光结合测定法测定。结果表明，几种与宿主相关的化学信息素（Z）-3-hexen-1-ol，（E）-2-hexen-1-al和戊醛具有与AlinCSP1-3的高结合亲和力，并且可以引发显着的高EAG线虫触角的反应。我们的研究表明，三种偏向天线的CSP可能介导苜蓿植物臭虫A. lineolatus的宿主识别。

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