桃蛀螟雄蛾对雌蛾体表烯烃的感受机制研究

发布时间：2018-02-03 23:28

本文关键词： 桃蛀螟性信息素体表烯烃感受机制　出处：《西南大学》2015年硕士论文　论文类型：学位论文

【摘要】：桃蛀螟(Conegethes punctiferalis),属鳞翅目草螟科(Lepidoptera:Crambidae)。其分布在东亚、南亚、澳大利亚等地区,而在我国多个地区也有分布,以幼虫钻蛀为害多种作物。目前桃蛀螟的防治方法包括农业防治、生物防治和化学防治等,其中化学防治是主要的防治方法,但该方法有诸多不足,如农药残留,农药抗性以及伤害天敌等。近年来,基于昆虫信息素具有绿色环保的特点,其已广泛用于害虫的预测预报与防治实践。桃蛀螟性信息素主要成分是反-10-十六碳烯醛和顺-10-十六碳烯醛的混合物(以下简称烯醛),但研究表明以该合成品为诱芯的诱捕器对雄蛾的引诱效果偏低。近年来有研究发现桃蛀螟雌蛾体表的两种烯烃：顺-9-二十七烯(以下简称二十七烯)和顺,顺,顺-3,6,9-二十三碳三烯(以下简称二十三碳三烯)与烯醛混合后,能够显著增强雄蛾在风洞中的行为反应(近距离滞空飞行和敲打信息素源),然而,雄蛾对这两种信息素烯烃的感受机制并不明确,影响其性诱剂技术的完善。为了明确桃蛀螟雄蛾对信息素烯烃的感受机制,本研究开展了触角电生理试验(EAG)、气相色谱触角电位仪连用(GC-EAD)检测试验、风洞行为试验,并对雄蛾触角进行了转录组测序。以下为本研究获得的主要实验结果：1.在EAG试验中,用不同浓度的烯醛对雄蛾左右触角进行测试,结果表明左右触角对结果影响并无显著性差异,为在后续试验中随机选择触角提供依据。在4种梯度浓度下(0.01；0.1；1.0和10头当量浓度),雄蛾触角分别对两种烯烃的EAG反应相对值之问,以及与正己烷之间均无差异,且均显著低于烯醛。将两种烯烃与烯醛混合后,其EAG反应相对值则与烯醛之间无显著性差异。2. GC-EAD实验中,单独测试时,雄蛾触角对二十三碳三烯和二十七烯均无触角电生理信号。将烯醛烯烃混合后进行测试时,也只测到了对烯醛的EAD反应。3.风洞行为试验方面,设置铁丝网阻隔,避免雄蛾直接接触滤纸(性信息素源)后发现,在行为指标—滞空飞行时间上,烯醛烯烃混合物的效果与性信息素粗提物相当,并且显著高于烯醛。进一步的选择性试验发现,在无阻隔的情况下,将烯醛与烯烃分别点滴在两张滤纸上,并将滤纸分别悬挂在风洞中不同的空间位置(风洞轴向10 cm间隔和径向2 cm间隔)时,雄蛾仅对烯醛有反应,对烯烃没有反应,而且对烯醛的反应强度与烯醛单独测试时相当。4.最后通过转录组技术进行了雄蛾触角转录组测序,寻找与触角嗅觉相关的气味结合蛋白(OBP)、普通气味结合蛋白(GOBP)、信息素结合蛋白(PBP),并与家蚕、棉铃虫的相关序列进行进化树比对。通过转录组数据分析,桃蛀螟雄蛾触角中鉴定出28个OBP基因,其中包括5个GOBP基因和5个PBP基因。综合EAG和风洞试验结果,桃蛀螟雄蛾能够在近距离通过嗅觉感受雌蛾体表的信息素烯烃,但是相对于烯醛的嗅觉反应而言,对烯烃的嗅觉反应相对微弱,不易通过触角电生理方法进行检测,其反应值可能低于触角电位仪的检测限。同时,从行为反应来看,只有当烯烃和烯醛同时从同一气味点释放时,雄蛾才会表现出对烯烃的行为反应。总的来说,桃蛀螟雄蛾对雌蛾体表信息素烯烃的嗅觉反应仍需进一步检验,并且嗅觉感受和行为反应之间的关系也需做深入研究。
[Abstract]:Dichocrocis punctiferalis (Conegethes punctiferalis), Lepidoptera Crambidae (Lepidoptera:Crambidae). Its distribution in East Asia, South Asia, Australia and other regions, and can also be found in many areas in China, by the larvae of borers were various crops. The control methods of dichocrocispunctiferalis including agricultural control, biological control and chemical control so, the chemical control is the main control method, but this method has many disadvantages, such as pesticide residues, pesticide resistance and damage to natural enemies. In recent years, the characteristics of insect pheromone with the green environmental protection based on the forecast and prevention of the practice has been widely used in pest. The peach borer is the main component of sex pheromone the mixture of -10- sixteen and -10- sixteen carbon carbon olefine aldehyde aldehyde (hereinafter referred to as enals), but research shows that the synthetic product is low attractant effect trapping device of male moths. In recent years, studies have found that female Dichocrocis punctiferalis Two: CIS -9- olefin moth body twenty-seven ene (hereinafter referred to as the twenty-seven ene) He Shun, CIS, CIS -3,6,9- twenty-three carbon three ene (hereinafter referred to as the twenty-three carbon three ene) and acrolein after mixing, can significantly enhance the behavior in the wind tunnel (close male moth reaction inhover flight and beat the pheromone source). However, the feelings of the two kinds of mechanism of male moth pheromone olefin is not clear, perfect the effect of sex attractant technology. In order to clear the peach borer male moths to feel the prime olefins information mechanism, this study carried out electrophysiological test antenna (EAG), gas chromatography electroantennography (GC-EAD). The wind tunnel test, behavior test, and the moth of transcriptome sequencing. The following is the main results obtained in this study: 1. in the EAG test, to test the male moth antennae around with different concentrations of acrolein, results show that the impact on the results and tentacles around No significant difference, and provide a basis for the random selection of antennae in subsequent experiments. In the 4 concentration gradient (0.01; 0.1; 1 and 10 equivalent concentration), the antennae of male EAG response of two kinds of relative value of olefin and n-hexane and asked, there were no differences, and were significantly lower than that of acrolein. Two olefins mixed with acrolein, the relative value of EAG reaction has no significant difference between.2. and acrolein in the GC-EAD experiment, a separate test, there were no antennae of male EAG signals in twenty-three and twenty-seven. Three carbon ene ene aldehydes will olefins after mixing test also, only detected EAD.3. wind tunnel test reaction behavior of aldehydes, set up barbed wire barrier, avoid direct contact with the filter paper (male moth sex pheromone source) found in the behavior index - inhover flight time, the effect of aldehyde olefin mixture and the sex pheromone extracts information, and Was significantly higher than that of acrolein selectivity test further. It is found that, in the case of no barrier, the aldehydes and alkenes were dropping in two on the filter paper, and the filter paper were suspended to different location in the wind tunnel (tunnel axial 10 cM interval and 2 radial cM interval), male moths react only to ene no reaction of alkenes, aldehydes, and the reaction intensity and acrolein aldehyde to separate test quite.4. finally through the transcriptome of the male moth antennal transcriptome sequencing, find related antennal olfactory odorant binding protein (OBP), egg white (GOBP), general odorant binding pheromone binding protein (PBP), and silkworm, related sequences of cotton bollworm phylogenetic comparison. Through transcriptome analysis identified 28 OBP genes of dichocrocispunctiferalis male antennae, including 5 GOBP genes and 5 PBP genes. The comprehensive EAG and wind tunnel test results, the peach borer moth can Through close enough in the olfactory receptor surface of the female moth pheromone olefin, but compared to the olfactory response of aldehyde, olfactory response of olefins is relatively weak, not easily by electrophysiological methods for detection, the detection response value may be less than electroantennography limit. At the same time, from the behavioral perspective, only when olefin and acrolein released simultaneously from the same smell when male moths will exhibit behavioral responses to olefins. In general, the olfactory response of dichocrocispunctiferalis male moth to female moth pheromone on olefin still needs further testing, and the relationship between sensory and behavioral responses also need to do in-depth research.

【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.4

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