本文選題：二化螟 + 抗藥性�。� 參考：《南京農(nóng)業(yè)大學》2016年碩士論文

【摘要】：隨著測序技術的飛速發(fā)展,測序通量和質(zhì)量越來越高,周期越來越短,測序成本急劇下降。數(shù)據(jù)量的空前增長帶來了生物信息學的繁榮發(fā)展,生物信息學成為當今生物學中重要的研究工具。昆蟲種類繁多、歷史悠久,和人類的生產(chǎn)、生活、生態(tài)等息息相關�；蚪M時代的昆蟲學研究已經(jīng)越來越多地依賴于基因組和轉(zhuǎn)錄組等大數(shù)據(jù),從基因組水平能夠更好地理解昆蟲生物學特性。在大數(shù)據(jù)基礎上,結(jié)合基因組編輯技術和RNAi技術,可以引領害蟲防治和益蟲利用發(fā)生革命性的變化。二化螟是重要的水稻害蟲,危害范圍廣,長期以來主要以化學防治為主。由于過度依賴化學農(nóng)藥和不恰當?shù)氖褂?造成二化螟日趨嚴重的抗藥性問題,給糧食生產(chǎn)、食品安全、生態(tài)環(huán)境帶來嚴重威脅。本文是在獲得了二化螟基因組的基礎上,從大數(shù)據(jù)和生物信息學的角度開展了研究,主要結(jié)果如下:1、二化螟抗藥性相關基因家族分析二化螟基因組數(shù)據(jù)的獲得,從基因組水平研究二化螟的抗性相關基因家族成為可能。本文選擇了家蠶、帝王蝶、小菜蛾、詩神袖蝶等四種鱗翅目昆蟲的基因組數(shù)據(jù)用于比較分析,這四個物種涉及經(jīng)濟昆蟲、觀賞昆蟲、蔬菜害蟲、水稻害蟲,其面臨的殺蟲劑選擇壓力明顯不同。通過基因組搜索和家族分析,分別獲得了 12個羧酸酯酶、108個細胞色素p450、16個谷胱甘肽S-轉(zhuǎn)移酶、32個乙酰膽堿受體、10個鈉離子通道、6個γ-氨基丁酸、27個魚尼丁受體蛋白基因,合計殺蟲劑解毒代謝或靶標基因共211個。二化螟Bt受體基因家族包含堿性磷酸酶、氨肽酶N、鈣粘蛋白、糖基轉(zhuǎn)移酶等,在基因組上分別找到5、14、33、14個基因,合計66個基因。與其他四個物種的基因組進行比較,通過進化分析,發(fā)現(xiàn)二化螟魚尼丁受體基因和氨肽酶N基因家族呈現(xiàn)明顯的擴張,顯示其對氯蟲苯甲酰胺類殺蟲劑和Bt毒素蛋白較容易產(chǎn)生靶標抗性。解毒代謝毒細胞色素P450基因家族也存在明顯的擴張,表明二化螟容易產(chǎn)生解毒代謝抗性。2、二化螟中腸抗藥性相關基因的表達量分析實驗室早期測定了二化螟中腸轉(zhuǎn)錄組,中腸是二化螟接觸和吸收農(nóng)藥的主要組織。為了研究抗藥性相關基因在中腸中表達水平,本研究選取了卵、幼蟲、蛹、成蟲組成的混合樣進行轉(zhuǎn)錄組測序作為參照,通過2個轉(zhuǎn)錄組之間的基因差異表達分析,共發(fā)現(xiàn)中腸上調(diào)基因21個,下調(diào)基因192個。其中抗藥性相關基因上調(diào)1個,下調(diào)基因13個。其中抗藥性上調(diào)基因為谷胱甘肽S-轉(zhuǎn)移酶基因,19個抗性下調(diào)基因分別為5個細胞色數(shù)P450、4個乙酰膽堿受體和4個魚尼丁受體基因。未發(fā)現(xiàn)中腸中有差異表達的Bt抗性相關基因。由于中腸轉(zhuǎn)錄組使用的是普通種群二化螟,這一結(jié)果表明,抗性相關的基因,尤其是解毒代謝酶基因,在通常情況下為正常表達,在農(nóng)藥外界因素刺激下可能導致表達增強,為誘導型表達增強型。3、二化螟基因組與轉(zhuǎn)錄組數(shù)據(jù)庫ChiloDB的構(gòu)建實驗室早期進行了基因組和多個轉(zhuǎn)錄組測序,積累了大量的二化螟基因數(shù)據(jù),為了提高數(shù)據(jù)使用效率,有利于更深層次的數(shù)據(jù)挖掘,本研究構(gòu)建了二化螟數(shù)據(jù)庫ChiloDB。該數(shù)據(jù)庫整合了二化螟基因組、轉(zhuǎn)錄組、miRNA、piRNA等數(shù)據(jù),提供序列檢索、基因查詢、基因結(jié)構(gòu)可視化、序列比對、數(shù)據(jù)下載等功能,界面交互友好,操作簡單,收錄了二化螟第一版基因組數(shù)據(jù),包含80,479條Scaffolds序列、107,037條轉(zhuǎn)錄本、10,221個蛋白質(zhì)編碼基因、262個miRNA、82,639條piRNA序列。數(shù)據(jù)庫自2014年8月上線,至今已有70個國家的研究者訪問使用,為二化螟研究提供了重要的數(shù)據(jù)平臺。
[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.

【學位授予單位】：南京農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S435.112.1

【參考文獻】

相關期刊論文 前7條

1 張傳溪;;中國農(nóng)業(yè)昆蟲基因組學研究概況與展望[J];中國農(nóng)業(yè)科學;2015年17期

2 易曉莉;馬曉;劉先方;侯成香;李木旺;;昆蟲對Bt毒素抗性研究進展[J];江蘇農(nóng)業(yè)科學;2014年07期

3 呼高偉;陳兆國;程天印;許娟;;MicroRNA及其研究現(xiàn)狀[J];中國人獸共患病學報;2012年02期

4 張贊;劉金定;黃水清;李飛;;生物信息學在昆蟲學研究中的應用[J];應用昆蟲學報;2012年01期

5 常曉麗;吳青君;王少麗;徐寶云;張友軍;;昆蟲氨肽酶N的研究進展[J];農(nóng)藥學學報;2011年03期

6 梁革梅,王桂榮,徐廣,吳孔明,郭予元;昆蟲Bt毒素受體蛋白的研究進展[J];昆蟲學報;2003年03期

7 王世貴,葉恭銀,胡萃;昆蟲對轉(zhuǎn)Bt殺蟲蛋白基因植物的抗性及其對策[J];生物技術;2000年05期

相關博士學位論文 前2條

1 姜衛(wèi)華;二化螟的抗藥性及綜合防治研究[D];南京農(nóng)業(yè)大學;2011年

2 韓招久;二化螟對殺蟲單和甲胺磷的抗性機理及神經(jīng)靶標乙酰膽堿受體的基因克隆[D];南京農(nóng)業(yè)大學;2002年

相關碩士學位論文 前3條

1 王興云;二化螟中腸Bt毒素受體—氨肽酶N基因的克隆及其功能分析[D];中國農(nóng)業(yè)科學院;2013年

2 夏艷;水稻比較基因組和進化生物學數(shù)據(jù)庫的構(gòu)建研究[D];中國科學院北京基因組研究所;2013年

3 張藝群;昆蟲基因數(shù)據(jù)庫平臺的構(gòu)建及其關鍵技術的研究[D];南京農(nóng)業(yè)大學;2012年

，

本文編號：1776241