本文關(guān)鍵詞：綠盲蝽氣味結(jié)合蛋白OBPs基因的鑒定及AlucOBP13、AlucOBP17的氣味結(jié)合特性　出處：《吉林農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 綠盲蝽 轉(zhuǎn)錄組測序 氣味結(jié)合蛋白 系統(tǒng)發(fā)育樹 組織表達譜 氣味結(jié)合特性

【摘要】：綠盲蝽(Apolygus lucorum)是我國一種重要的農(nóng)業(yè)害蟲,具有靈活機動、寄主廣泛和隱蔽為害等特點�；瘜W(xué)感受行為在綠盲蝽許多生命階段中發(fā)揮著至關(guān)重要的作用,如求偶、寄主定位和寄主轉(zhuǎn)移過程中性信息素及花香類氣味的探測。氣味結(jié)合蛋白(odorant binding proteins,OBPs)被認為參與化學(xué)信息素感知過程中的初始生化識別階段。隨著盲蝽性信息素組分的鑒定以及盲蝽性誘劑在田間應(yīng)用中的發(fā)展,干擾昆蟲的化學(xué)信息通訊已經(jīng)成為一種害蟲防治的新策略。本研究從分子水平深入探索綠盲蝽對性信息素的識別機制,為嗅覺基因的干擾或沉默及優(yōu)化性誘劑組分提供理論參考。本試驗主要研究內(nèi)容如下:1.通過綠盲蝽觸角轉(zhuǎn)錄組測序共獲得38個OBPs基因,包括26個“classic”O(jiān)BPs和12“Plus-C”O(jiān)BPs基因。系統(tǒng)發(fā)育樹結(jié)果表明綠盲蝽OBPs蛋白與半翅目中其它盲蝽的OBPs蛋白在進化樹中聚集在一起,親緣關(guān)系表現(xiàn)得更為接近。熒光定量qRT-PCR結(jié)果表明在新鑒定的23個OBPs基因中,有11個OBPs基因在綠盲蝽雌雄成蟲觸角中的表達水平顯著高于其他組織,其中有3個在雄蟲觸角表達水平顯著高于雌蟲觸角,有6個在雌蟲觸角表達水平顯著高于雄蟲觸角,暗示著它們可能在探測性信息素或寄主植物揮發(fā)物中發(fā)揮著作用;有3個OBPs基因在口器表達水平顯著高于其他組織;有4個OBPs基因在頭部表達水平顯著高于其他組織;有2個OBPs基因在足部表達水平顯著高于其他組織;有1個OBP基因在腹部表達水平顯著高于其他組織;有1個OBP基因在翅部表達水平顯著高于其他組織;有2個OBPs基因在主要組織如觸角、口器、頭部、足和翅中都有表達并且差異不顯著。大多數(shù)具有直系同源基因的綠盲蝽OBPs基因與它們的直系同源基因有著相似的表達譜,強烈地表明這些同源基因在嗅覺或味覺中具有相似的功能。2.篩選出兩個綠盲蝽male-biased觸角氣味結(jié)合蛋白AlucOBP13和AlucOBP17作為潛在的性信息素結(jié)合蛋白(PBPs),這兩個OBPs可能與綠盲蝽識別性信息素過程息息相關(guān)。對AlucOBP13和AlucOBP17進行克隆、表達和純化,通過熒光競爭結(jié)合試驗和微量熱泳動試驗對它們與9種潛在的盲蝽性信息素物質(zhì),以及59種綠盲蝽偏好寄主植物揮發(fā)物、綠葉揮發(fā)物、萜烯類物質(zhì)和引起綠盲蝽較強EAG反應(yīng)的氣味物質(zhì)的結(jié)合能力進行測定。結(jié)果表明AlucOBP13結(jié)合譜較窄,只能結(jié)合β-紫羅酮,我們推測AlucOBP13可能是普通氣味分子結(jié)合蛋白(GOBP),在綠盲蝽寄主定位和寄主轉(zhuǎn)移過程中發(fā)揮著重要的作用;AlucOBP17能與盲蝽性信息素類似物丁酸辛酯、己酸己酯以及棉花揮發(fā)物十二醛、花香類物質(zhì)β-紫羅酮有效的結(jié)合,我們推測AlucOBP17可能是潛在的PBP,并在綠盲蝽識別性信息素、寄主定位和寄主轉(zhuǎn)移過程中發(fā)揮著重要的作用。
[Abstract]:A.lucorum (Apolygus lucorum) is a kind of important agricultural pests, has the characteristics of flexible, wide host and hidden damage. Chemosensory behavior plays a vital role in many a.lucorum stages of life such as mating, host location and host transfer detection process neutral pheromone and floral scent the odorant binding protein (odorant binding, proteins, OBPs) is considered the initial biochemical identification stage in chemical pheromone perception process. With the blind sex pheromone component identification and blind sex pheromone in the field of development, chemical information and communication interference in insects has become a new strategy for pest control this study. From the in-depth exploration of the molecular level l.lucorum of sex pheromone recognition mechanism for olfactory gene interference or silencing and optimization of sex pheromone components and provide a theoretical reference. This experiment mainly study The contents are as follows: 1. by a.lucorum antennal transcriptome sequencing of 38 OBPs genes were obtained, including 26 "classic" OBPs and 12 "Plus-C" OBPs gene. The phylogenetic tree showed that other Lygus lucorum OBPs protein and OBPs protein in Hemiptera in evolutionary tree together, relationship performance get closer. Fluorescence quantitative qRT-PCR results showed that the 23 OBPs gene identification, expression level of OBPs gene in 11 lucorum antennae of male and female adults was significantly higher than that of other organizations, 3 of them expressed in male antenna were significantly higher than that of female antennae, there are 6 levels of expression in female antennae was significantly higher than that in male antenna, suggesting that they may be in the detection of sex pheromone and host plant volatiles play a role in the play; with the level of expression of 3 OBPs genes in the mouthparts was significantly higher than that in other tissues; 4 OBPs gene expression level in the head There is significantly higher than that of other organizations; the expression level of OBPs gene in 2 foot was significantly higher than that in other tissues; the expression level of OBP gene is 1 in the abdomen was significantly higher than that of other organizations; with the level of expression of 1 OBP genes in the wing was significantly higher than that of other organizations; there are 2 OBPs genes in main tissues such as antennae, mouthparts, head, have and there was no significant difference between the expression of both foot and wings. Most of the orthologous orthologs lucorum OBPs gene and they have a similar expression profile strongly suggests that these homologous genes with similar function as.2. screened two lucorum male-biased antenna odorant binding protein AlucOBP13 and AlucOBP17 as potential pheromone binding protein in smell or taste in (PBPs), the two OBPs may be closely related to a.lucorum recognition of sex pheromone. Cloning of AlucOBP13 and AlucOBP17, expression and purification, By fluorescence binding assay and microcalorimetry swimming test for them and 9 potential bug sex pheromone substances, as well as 59 kinds of a.lucorum preference of host plant volatiles, green leaf volatiles, combined with the ability of terpenes and caused strong reaction of a.lucorum EAG odorants were determined. The results showed that AlucOBP13 combined with narrow spectrum, only with betalonone, we speculate that AlucOBP13 may be a general odorant binding protein (GOBP), plays an important role in the transfer of a.lucorum host location and host in the process; AlucOBP17 and fleahopper sex pheromone analogs octyl butyrate, Hexyl Hexanoate and cotton volatiles twelve aldehyde, floral material with betalonone effectively, we speculate that AlucOBP17 may be a potential PBP, and in lucorum recognition of sex pheromone, host location and host transfer process plays an important role.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S433

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