二化螟抗药性相关基因家族分析及数据库的构建

发布时间：2018-04-20 04:24

本文选题：二化螟 + 抗药性　；参考：《南京农业大学》2016年硕士论文

【摘要】：随着测序技术的飞速发展,测序通量和质量越来越高,周期越来越短,测序成本急剧下降。数据量的空前增长带来了生物信息学的繁荣发展,生物信息学成为当今生物学中重要的研究工具。昆虫种类繁多、历史悠久,和人类的生产、生活、生态等息息相关。基因组时代的昆虫学研究已经越来越多地依赖于基因组和转录组等大数据,从基因组水平能够更好地理解昆虫生物学特性。在大数据基础上,结合基因组编辑技术和RNAi技术,可以引领害虫防治和益虫利用发生革命性的变化。二化螟是重要的水稻害虫,危害范围广,长期以来主要以化学防治为主。由于过度依赖化学农药和不恰当的使用,造成二化螟日趋严重的抗药性问题,给粮食生产、食品安全、生态环境带来严重威胁。本文是在获得了二化螟基因组的基础上,从大数据和生物信息学的角度开展了研究,主要结果如下:1、二化螟抗药性相关基因家族分析二化螟基因组数据的获得,从基因组水平研究二化螟的抗性相关基因家族成为可能。本文选择了家蚕、帝王蝶、小菜蛾、诗神袖蝶等四种鳞翅目昆虫的基因组数据用于比较分析,这四个物种涉及经济昆虫、观赏昆虫、蔬菜害虫、水稻害虫,其面临的杀虫剂选择压力明显不同。通过基因组搜索和家族分析,分别获得了 12个羧酸酯酶、108个细胞色素p450、16个谷胱甘肽S-转移酶、32个乙酰胆碱受体、10个钠离子通道、6个γ-氨基丁酸、27个鱼尼丁受体蛋白基因,合计杀虫剂解毒代谢或靶标基因共211个。二化螟Bt受体基因家族包含碱性磷酸酶、氨肽酶N、钙粘蛋白、糖基转移酶等,在基因组上分别找到5、14、33、14个基因,合计66个基因。与其他四个物种的基因组进行比较,通过进化分析,发现二化螟鱼尼丁受体基因和氨肽酶N基因家族呈现明显的扩张,显示其对氯虫苯甲酰胺类杀虫剂和Bt毒素蛋白较容易产生靶标抗性。解毒代谢毒细胞色素P450基因家族也存在明显的扩张,表明二化螟容易产生解毒代谢抗性。2、二化螟中肠抗药性相关基因的表达量分析实验室早期测定了二化螟中肠转录组,中肠是二化螟接触和吸收农药的主要组织。为了研究抗药性相关基因在中肠中表达水平,本研究选取了卵、幼虫、蛹、成虫组成的混合样进行转录组测序作为参照,通过2个转录组之间的基因差异表达分析,共发现中肠上调基因21个,下调基因192个。其中抗药性相关基因上调1个,下调基因13个。其中抗药性上调基因为谷胱甘肽S-转移酶基因,19个抗性下调基因分别为5个细胞色数P450、4个乙酰胆碱受体和4个鱼尼丁受体基因。未发现中肠中有差异表达的Bt抗性相关基因。由于中肠转录组使用的是普通种群二化螟,这一结果表明,抗性相关的基因,尤其是解毒代谢酶基因,在通常情况下为正常表达,在农药外界因素刺激下可能导致表达增强,为诱导型表达增强型。3、二化螟基因组与转录组数据库ChiloDB的构建实验室早期进行了基因组和多个转录组测序,积累了大量的二化螟基因数据,为了提高数据使用效率,有利于更深层次的数据挖掘,本研究构建了二化螟数据库ChiloDB。该数据库整合了二化螟基因组、转录组、miRNA、piRNA等数据,提供序列检索、基因查询、基因结构可视化、序列比对、数据下载等功能,界面交互友好,操作简单,收录了二化螟第一版基因组数据,包含80,479条Scaffolds序列、107,037条转录本、10,221个蛋白质编码基因、262个miRNA、82,639条piRNA序列。数据库自2014年8月上线,至今已有70个国家的研究者访问使用,为二化螟研究提供了重要的数据平台。
[Abstract]:With the rapid development of sequencing technology, the flux and quality of sequencing are getting higher and higher, the cycle is shorter and shorter, and the cost of sequencing has fallen sharply. The unprecedented growth of data has brought about the prosperity and development of bioinformatics. Bioinformatics has become an important research tool in biology. There are many kinds of insects, long history, and human production, life and life. The study of Entomology in the genomic era has become more and more dependent on large data such as genome and transcriptional groups, which can better understand the biological characteristics of insects from the genome level. On the basis of large data, combining genome editing and RNAi techniques, it can lead to the revolution of pest control and the utilization of beneficial insects. Change. The rice borer is an important pest of rice, which has a wide range of harm. It has been mainly based on chemical control for a long time. Due to excessive dependence of chemical pesticides and inappropriate use, the growing resistance of borer borer has been caused seriously. It has brought serious threat to grain production, food safety and ecological environment. From the perspective of large data and bioinformatics, the main results are as follows: 1, the gene family of the resistance related genes of the suppressalis suppressalis is obtained, and it is possible to study the resistance related gene family of the borer from the genome level. This article selected four species of silkworm, monarch butterfly, Plutella xylostella, and Shi Shen butterfly. Genomic data of Lepidoptera are used for comparative analysis. These four species involve economic insects, ornamental insects, vegetable pests, and rice pests. The pressure of insecticide selection is significantly different. By genome search and family analysis, 12 carboxylesterases, 108 cytochrome p450,16 glutathione S- transferases, and 32 cytochrome p450,16 are obtained. Acetylcholine receptor, 10 sodium ion channels, 6 gamma aminobutyric acid, 27 fish nidin receptor protein genes, combined insecticide detoxification metabolism or target gene 211. The Bt receptor gene family of the suppressalis suppressalis contains alkaline phosphatase, aminopeptidase N, cadherin, glycosyltransferase and so on. 5,14,33,14 genes are found in the genome, the total of 66 genes. Gene. Compared with the genomes of the other four species, through evolutionary analysis, it was found that the gene and the aminopeptidase N gene family of the suppressalis suppressalis were obviously dilated, showing that they were more susceptible to the target resistance to the benzamide insecticides and the Bt toxin protein, and the detoxified metabolic cytochrome P450 gene family also existed. The dilatation showed that the suppressalis suppressalis was prone to detoxification and metabolic resistance.2, and the expression of resistance related genes in the midgut of the suppressalis suppressalis was measured early in the midgut transcriptional group. The midgut is the main tissue for the contact and absorption of pesticides. The mixed sample of larvae, pupae and adult was sequenced as a reference. Through the analysis of gene differential expression in the 2 transcriptional groups, 21 and 192 down-regulation genes were found. Among them, the resistance related genes were up regulated by 1 and down regulated by 13. The anti drug up-regulated gene was the glutathione S- transferase gene and 19 resistant genes. The genes of the down regulated genes were 5 cell chromatic number P450,4 acetylcholine receptor and 4 fish neidine receptor genes. No differentially expressed Bt resistance related genes were found in the midgut. As the midgut transcriptome used the common population of the suppressalis suppressalis, the results showed that the resistance related genes, especially the detoxification metabolizing enzyme genes, were in general condition. For normal expression, the expression enhanced by the external factors of pesticides may lead to enhanced expression, for inducible expression enhanced.3. The genome and transcriptional database ChiloDB of the stem borer genome and transcriptional database were sequenced by genome and multiple transcripts in the early laboratory, and a large number of gene data were accumulated. In order to improve the efficiency of data use, it was beneficial to the more. Deep data mining, this study constructed a database ChiloDB. of borer suppressalis, which integrates the data of the genome, transcriptional group, miRNA, piRNA and so on. It provides sequence retrieval, gene query, gene structure visualization, sequence alignment, data download and so on. The interface is friendly and easy to operate. The data, including 80479 Scaffolds sequences, 107037 transcripts, 10221 protein coding genes, 262 miRNA, and 82639 piRNA sequences. The database has been launched in August 2014 and has been accessed by researchers in 70 countries, providing an important data platform for the study of the borer.

【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.112.1

【参考文献】