桃蚜传播黄瓜花叶病毒机理的研究
发布时间：2019-05-29 22:38
【摘要】：桃蚜 Myzus persicae(Sulzer),属半翅目Hemiptera,蚜科Aphididae,是一种几乎遍布全球的多食性害虫。桃蚜不仅可以吸取植物汁液,还可以作为介体传播多种植物病毒,是常见的媒介昆虫,给农业生产带来了严重的损失。桃蚜主要以非持久性的方式来传播黄瓜花叶病毒(Cucumber mosaic virus,CMV),这种方式主要是指病毒在昆虫体内仅停留几秒至几分钟,病毒不会随昆虫的繁殖而传递给下一代。目前,关于桃蚜传播CMV的传毒机制尚不清楚。本研究一方面根据文献的报道,利用qPCR、酵母双杂交、RNAi等分子生物学方法,研究桃蚜表皮蛋白基因在传毒过程中的作用;另一方面利用RNA-seq和iTRAQ等组学研究技术,进一步挖掘与传毒相关的潜在传毒因子,为全面理解桃蚜的传毒机制奠定理论基础,同时也为生产中桃蚜和病毒病的防治策略提供参考依据。在研究表皮蛋白基因的作用方面,前期筛选出用于扩增MPCP1、MPCP2、MPCP4、MPCP5的引物,并优化了反应条件;用整个虫体和口针扩增的各表皮蛋白基因序列是一致的,故桃蚜整个虫体可以作为后续的研究对象。在基因水平上,CMVCP的相对表达量与各表皮蛋白基因的变化趋势相似,并且MPCP1、MPCP2和MPCP4的相对表达量明显高于MPCP5;在酵母双杂交系统里,只有基因MPCP4编码的蛋白质与CMVCP是相互作用的。在功能研究上,基因MPCP4的沉默可以降低桃蚜获得病毒的效率。综上所述,桃蚜表皮蛋白相关基因参与了传毒过程,且基因MPCP4在其中起主要作用。在挖掘与传毒相关的潜在传毒因子方面,对带毒的和不带毒的桃蚜分别进行了转录组和蛋白质组研究。在转录组测序中,一共找到20550个差异表达的基因。其中,9731个差异基因是上调表达的,10818个差异基因是下调表达的。在KEGG数据库中,差异表达的基因共注释到501个通路,其中有32个是富集分布的(p-value≤0.05),多集中在cAMP信号通路(cAMP signaling pathway)和药物代谢通路(drug metabolism)。在蛋白质组测序中,共找到744个差异表达的蛋白。其中,上调表达的差异蛋白有437个,下调表达的差异蛋白有307个。在KEGG数据库中,差异表达蛋白富集(p-value≤0.05)分布在了 14个通路里,其中包括脂肪酸代谢途径(fatty acid metabolism)、碳代谢途径(carbon metabolism)和各种氨基酸代谢途径等。最后,对桃蚜转录组和蛋白质组进行联合分析,发现两者存在一定的相关性,并且有282个蛋白质在转录组数据中可以找到联系,这其中表达模式(均上调表达或均下调表达)一致的有207个蛋白质,包括表皮蛋白、核糖体蛋白和细胞色素P450等。
[Abstract]:Aphididae, of Hemiptera, aphids of the genus Myzus persicae (Sulzer), is a kind of polyfeed pest almost all over the world. Peach aphid can not only absorb plant juice, but also spread a variety of plant viruses as a mediator. It is a common vector insect, which has brought serious losses to agricultural production. Peach aphids mainly transmit cucumber mosaic virus (Cucumber mosaic virus,CMV in a non-persistent way, which mainly means that the virus stays in insects for only a few seconds to a few minutes, and the virus is not passed on to the next generation with the reproduction of insects. At present, the mechanism of transmission of CMV by peach aphids is not clear. On the one hand, according to the reports in the literature, the role of epidermis protein gene in the transmission process of peach aphid was studied by qPCR, yeast two-hybrid, RNAi and other molecular biological methods. On the other hand, RNA-seq and iTRAQ were used to further excavate the potential transmission factors related to virus transmission, which laid a theoretical foundation for a comprehensive understanding of the transmission mechanism of peach aphid. At the same time, it also provides a reference for the control strategy of peach aphid and virus disease in production. In the study of the role of epidermis protein gene, primers for amplification of MPCP1,MPCP2,MPCP4,MPCP5 were selected in the early stage, and the reaction conditions were optimized. The sequence of epidermis protein genes amplified by the whole insect body and oral needle is the same, so the whole aphid body can be used as the follow-up research object. At the gene level, the relative expression of CMVCP was similar to that of each epidermis protein gene, and the relative expression of MPCP1,MPCP2 and MPCP4 was significantly higher than that of MPCP5;. In yeast two-hybrid system, only the protein encoded by gene MPCP4 interacts with CMVCP. In functional research, the silencing of gene MPCP4 can reduce the efficiency of peach aphid in obtaining virus. In conclusion, the epidermis protein related genes of peach aphid are involved in the transmission process, and the gene MPCP4 plays a major role in it. In the aspect of excavating the potential transmission factors related to virus transmission, the transcripts and proteome of peach aphids with and without virus were studied respectively. A total of 20550 differentially expressed genes were found in the transcriptional group. Among them, 9731 differentially expressed genes and 10818 differentially expressed genes were down-regulated. In the KEGG database, the differentially expressed genes were commented to a total of 501 pathways, 32 of which were enriched and distributed (p-value 鈮，
本文编号：2488280
[Abstract]:Aphididae, of Hemiptera, aphids of the genus Myzus persicae (Sulzer), is a kind of polyfeed pest almost all over the world. Peach aphid can not only absorb plant juice, but also spread a variety of plant viruses as a mediator. It is a common vector insect, which has brought serious losses to agricultural production. Peach aphids mainly transmit cucumber mosaic virus (Cucumber mosaic virus,CMV in a non-persistent way, which mainly means that the virus stays in insects for only a few seconds to a few minutes, and the virus is not passed on to the next generation with the reproduction of insects. At present, the mechanism of transmission of CMV by peach aphids is not clear. On the one hand, according to the reports in the literature, the role of epidermis protein gene in the transmission process of peach aphid was studied by qPCR, yeast two-hybrid, RNAi and other molecular biological methods. On the other hand, RNA-seq and iTRAQ were used to further excavate the potential transmission factors related to virus transmission, which laid a theoretical foundation for a comprehensive understanding of the transmission mechanism of peach aphid. At the same time, it also provides a reference for the control strategy of peach aphid and virus disease in production. In the study of the role of epidermis protein gene, primers for amplification of MPCP1,MPCP2,MPCP4,MPCP5 were selected in the early stage, and the reaction conditions were optimized. The sequence of epidermis protein genes amplified by the whole insect body and oral needle is the same, so the whole aphid body can be used as the follow-up research object. At the gene level, the relative expression of CMVCP was similar to that of each epidermis protein gene, and the relative expression of MPCP1,MPCP2 and MPCP4 was significantly higher than that of MPCP5;. In yeast two-hybrid system, only the protein encoded by gene MPCP4 interacts with CMVCP. In functional research, the silencing of gene MPCP4 can reduce the efficiency of peach aphid in obtaining virus. In conclusion, the epidermis protein related genes of peach aphid are involved in the transmission process, and the gene MPCP4 plays a major role in it. In the aspect of excavating the potential transmission factors related to virus transmission, the transcripts and proteome of peach aphids with and without virus were studied respectively. A total of 20550 differentially expressed genes were found in the transcriptional group. Among them, 9731 differentially expressed genes and 10818 differentially expressed genes were down-regulated. In the KEGG database, the differentially expressed genes were commented to a total of 501 pathways, 32 of which were enriched and distributed (p-value 鈮，
本文编号：2488280
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