Abstract：
The orientation of the oligophagous cone-feeding moth Dioryctria abietella (Lepidoptera: Pyralidae) to host plants primarily relies on olfactory-related proteins, particularly those candidates highly expressed in antennae. Here, through a combination of expression profile, ligand-binding assay, molecular docking and site-directed mutagenesis strategies, we characterized the chemosensory protein (CSP) gene family in D. abietella. Quantitative real-time PCR (qPCR) analyses revealed the detectable expression of all 22 DabiCSPs in the antennae, of which seven genes were significantly enriched in this tissue. In addition, the majority of the genes (19/22 relatives) had the expression in at least one reproductive tissue. In the interactions of four antenna-dominant DabiCSPs and different chemical classes, DabiCSP1 was broadly tuned to 27 plant-derived odors, three man-made insecticides and one herbicide with high affinities (Ki < 6.60 μM). By contrast, three other DabiCSPs (DabiCSP4, CSP6 and CSP17) exhibited a narrow odor binding spectrum, in response to six compounds for each protein. Our mutation analyses combined with molecular docking simulations and binding assays further identified four key residues (Tyr25, Thr26, Ile65 and Val69) in the interactions of DabiCSP1 and ligands, of which binding abilities of this protein to 12, 15, 16 and three compounds were significantly decreased compared to the wildtype protein, respectively. Our study reveals different odor binding spectra of four DabiCSPs enriched in antennae and identifies key residues responsible for the binding of DabiCSP1 and potentially active compounds for the control of this pest.
摘要：
寡食锥食蛾飞蛾（鳞翅目：Pyralidae）对寄主植物的定位主要依赖于嗅觉相关蛋白，特别是那些在触角中高度表达的候选物。这里,通过表达谱的组合，配体结合测定，分子对接和定点诱变策略，我们鉴定了D.abietella的化学感应蛋白（CSP）基因家族。定量实时PCR（qPCR）分析显示触角中所有22个DabiCSP的可检测表达，其中7个基因在该组织中显著富集。此外,大多数基因（19/22的亲属）在至少一个生殖组织中表达。在四个天线主导的DabiCSP和不同化学类别的相互作用中，DabiCSP1被广泛调整为27种植物来源的气味，三种人造杀虫剂和一种除草剂具有高亲和力(Ki<6.60μM)。相比之下,其他三个DabiCSP（DabiCSP4，CSP6和CSP17）表现出狭窄的气味结合谱，响应每种蛋白质的六种化合物。我们的突变分析结合分子对接模拟和结合测定进一步确定了DabiCS1和配体相互作用中的四个关键残基（Tyr25，Thr26，Ile65和Val69）。与野生型蛋白相比，该蛋白对12、15、16和3种化合物的结合能力显着降低，分别。我们的研究揭示了四个富含触角的DabiCSP的不同气味结合谱，并确定了负责DabiCSP1结合的关键残基以及用于控制该害虫的潜在活性化合物。
