本文選題：豌豆蚜 + 報警信息素�。� 參考：《吉林大學(xué)》2015年碩士論文

【摘要】：昆蟲依靠靈敏的嗅覺系統(tǒng)來感知環(huán)境中特異性氣味分子，進(jìn)而啟動相應(yīng)的行為反應(yīng)，例如尋找食物、尋找產(chǎn)卵或生殖場所、逃避天敵等。蚜蟲是農(nóng)業(yè)和林業(yè)上重要的害蟲，其在受到外界威脅時會分泌一種報警信息素，，這種報警信息素的主要成分是(E)-β-farnesene（EBF），它不僅可以有效防止蚜蟲聚集、控制蚜蟲數(shù)量，而且為天敵搜尋蚜蟲提供了可靠的線索。闡明蚜蟲識別報警信息素的分子機(jī)制，可以為從分子水平篩選和設(shè)計比EBF更穩(wěn)定的、不易揮發(fā)的蚜蟲報警信息素類似物。本實驗室從棉蚜和豌豆蚜基因組中鑒定了全部的氣味結(jié)合蛋白和氣味受體家族基因。本文建立了蚜蟲RNAi干擾的方法，通過比較基因組學(xué)結(jié)合RNAi和行為測定等技術(shù)初步明確了蚜蟲識別報警信息素的分子機(jī)制。主要研究結(jié)果如下： 1．明確了豌豆蚜通過嗅覺受體識別報警信息素。從豌豆蚜觸角中鑒定和克隆了非典型氣味受體Orco基因，研究了Orco基因的時空表達(dá)譜，明確了Orco基因主要在蚜蟲觸角中表達(dá)。體外合成Orco基因dsRNA，注射后72h， Orco基因相對表達(dá)量與對照相比下降了50%左右，觸角電位記錄結(jié)果表明注射了dsRNA的蚜蟲對EBF的反應(yīng)顯著減弱，Y型管行為實驗結(jié)果顯示，注射了dsRNA的蚜蟲對EBF沒有趨避行為。 2．利用RNAi技術(shù)證明OBP3和OBP7基因在豌豆蚜體內(nèi)參與了對報警信息素EBF的識別。單獨(dú)注射OBP3或OBP7的dsRNA，雖然基因表達(dá)量顯著下降，但是豌豆蚜對EBF的趨避行為和EAG反應(yīng)與對照組相比都沒有顯著性差異。同時注射OBP3和OBP7的dsRNA，處理組蚜蟲對EBF喪失了趨避行為，相應(yīng)的EAG反應(yīng)顯著減弱。 3．兩個嗅覺受體基因參與豌豆蚜對報警信息素EBF的識別。單獨(dú)注射OR5或OR20的dsRNA，基因表達(dá)量顯著下降，而且豌豆蚜對EBF的趨避行為和EAG反應(yīng)顯著減弱。利用EAG玻璃電極遮蓋豌豆蚜觸角第五第六節(jié)的EAG實驗表明，豌豆蚜觸角感受EBF的部位主要在觸角頂端兩節(jié)。實時熒光定量PCR檢測到這兩條基因在豌豆蚜觸角中高度表達(dá)。
[Abstract]:Insects rely on a sensitive olfactory system to sense the specific odour molecules in the environment, and then initiate corresponding behavioral responses, such as looking for food, finding spawning or reproductive sites, avoiding natural enemies, and so on. Aphids are important pests in agriculture and forestry. Aphids secrete an alarm pheromone when threatened by the outside world. The main component of this warning pheromone is Ean- 尾 -farneseneeEBFN, which can not only effectively prevent aphids from gathering and control the number of aphids. It also provides reliable clues for natural enemies to search for aphids. The molecular mechanism of aphids recognizing alarm pheromones can be used to screen and design more stable and non-volatile aphids warning pheromones at molecular level than EBF. All the odor binding proteins and odor receptor genes were identified from the genome of cotton aphids and pea aphids in our laboratory. In this paper, a method of RNAi interference in aphids was established. The molecular mechanism of aphids' recognition of alarm pheromones was preliminarily clarified by means of comparative genomics, RNAi and behavior determination techniques. The main results are as follows: 1. The alarm pheromone was recognized by the olfactory receptors of the pea aphids. The atypical odor receptor Orco gene was identified and cloned from the antennae of the pea aphid. The spatial and temporal expression profiles of the Orco gene were studied. The results showed that the Orco gene was mainly expressed in the antennae of the aphid. Orco gene dsRNAs were synthesized in vitro. The relative expression of Orco gene decreased by about 50% compared with the control at 72 h after injection. The results of antenna potential recording showed that the response of aphids injected with dsRNA to EBF was significantly decreased. Aphids injected with dsRNA did not avoid EBF. 2. RNAi technique was used to prove that OBP3 and OBP7 genes were involved in the recognition of alarm pheromone EBF in pea aphids. DsRNAs of OBP3 or OBP7 alone had no significant difference in EBF avoidance behavior and EAG response to EBF, although the gene expression level was significantly decreased. After dsRNAs of OBP3 and OBP7 were injected at the same time, the aphids in the treatment group lost the behavior of avoidance to EBF, and the corresponding EAG response was significantly weakened. 3. Two olfactory receptor genes were involved in the recognition of the alarm pheromone EBF in pea aphids. DsRNAs of OR5 or OR20 alone decreased gene expression, and the resistance to EBF and EAG response of aphids decreased significantly. The EAG experiment using EAG glass electrode to cover the fifth and sixth session of the antennae of the pea aphid showed that the part of the antennae of the apical pea aphid felt EBF mainly in the two segments of the top of the antennae. The two genes were highly expressed in the antennae of the pea aphid by real time fluorescence quantitative PCR.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S433

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本文編號：2010897