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

【摘要】：隨著測(cè)序技術(shù)的飛速發(fā)展,測(cè)序通量和質(zhì)量越來(lái)越高,周期越來(lái)越短,測(cè)序成本急劇下降。數(shù)據(jù)量的空前增長(zhǎng)帶來(lái)了生物信息學(xué)的繁榮發(fā)展,生物信息學(xué)成為當(dāng)今生物學(xué)中重要的研究工具。昆蟲(chóng)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(guān)�；蚪M時(shí)代的昆蟲(chóng)學(xué)研究已經(jīng)越來(lái)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(chóng)生物學(xué)特性。在大數(shù)據(jù)基礎(chǔ)上,結(jié)合基因組編輯技術(shù)和RNAi技術(shù),可以引領(lǐng)害蟲(chóng)防治和益蟲(chóng)利用發(fā)生革命性的變化。二化螟是重要的水稻害蟲(chóng),危害范圍廣,長(zhǎng)期以來(lái)主要以化學(xué)防治為主。由于過(guò)度依賴化學(xué)農(nóng)藥和不恰當(dāng)?shù)氖褂?造成二化螟日趨嚴(yán)重的抗藥性問(wèn)題,給糧食生產(chǎn)、食品安全、生態(tài)環(huán)境帶來(lái)嚴(yán)重威脅。本文是在獲得了二化螟基因組的基礎(chǔ)上,從大數(shù)據(jù)和生物信息學(xué)的角度開(kāi)展了研究,主要結(jié)果如下:1、二化螟抗藥性相關(guān)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(guān)基因家族成為可能。本文選擇了家蠶、帝王蝶、小菜蛾、詩(shī)神袖蝶等四種鱗翅目昆蟲(chóng)的基因組數(shù)據(jù)用于比較分析,這四個(gè)物種涉及經(jīng)濟(jì)昆蟲(chóng)、觀賞昆蟲(chóng)、蔬菜害蟲(chóng)、水稻害蟲(chóng),其面臨的殺蟲(chóng)劑選擇壓力明顯不同。通過(guò)基因組搜索和家族分析,分別獲得了 12個(gè)羧酸酯酶、108個(gè)細(xì)胞色素p450、16個(gè)谷胱甘肽S-轉(zhuǎn)移酶、32個(gè)乙酰膽堿受體、10個(gè)鈉離子通道、6個(gè)γ-氨基丁酸、27個(gè)魚(yú)尼丁受體蛋白基因,合計(jì)殺蟲(chóng)劑解毒代謝或靶標(biāo)基因共211個(gè)。二化螟B(niǎo)t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(gè)基因,合計(jì)66個(gè)基因。與其他四個(gè)物種的基因組進(jìn)行比較,通過(guò)進(jìn)化分析,發(fā)現(xiàn)二化螟魚(yú)尼丁受體基因和氨肽酶N基因家族呈現(xiàn)明顯的擴(kuò)張,顯示其對(duì)氯蟲(chóng)苯甲酰胺類殺蟲(chóng)劑和Bt毒素蛋白較容易產(chǎn)生靶標(biāo)抗性。解毒代謝毒細(xì)胞色素P450基因家族也存在明顯的擴(kuò)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(guān)基因的表達(dá)量分析實(shí)驗(yàn)室早期測(cè)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(guān)基因在中腸中表達(dá)水平,本研究選取了卵、幼蟲(chóng)、蛹、成蟲(chóng)組成的混合樣進(jìn)行轉(zhuǎn)錄組測(cè)序作為參照,通過(guò)2個(gè)轉(zhuǎn)錄組之間的基因差異表達(dá)分析,共發(fā)現(xiàn)中腸上調(diào)基因21個(gè),下調(diào)基因192個(gè)。其中抗藥性相關(guān)基因上調(diào)1個(gè),下調(diào)基因13個(gè)。其中抗藥性上調(diào)基因?yàn)楣入赘孰腟-轉(zhuǎn)移酶基因,19個(gè)抗性下調(diào)基因分別為5個(gè)細(xì)胞色數(shù)P450、4個(gè)乙酰膽堿受體和4個(gè)魚(yú)尼丁受體基因。未發(fā)現(xiàn)中腸中有差異表達(dá)的Bt抗性相關(guān)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(guān)的基因,尤其是解毒代謝酶基因,在通常情況下為正常表達(dá),在農(nóng)藥外界因素刺激下可能導(dǎo)致表達(dá)增強(qiáng),為誘導(dǎo)型表達(dá)增強(qiáng)型。3、二化螟基因組與轉(zhuǎn)錄組數(shù)據(jù)庫(kù)ChiloDB的構(gòu)建實(shí)驗(yàn)室早期進(jìn)行了基因組和多個(gè)轉(zhuǎn)錄組測(cè)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(kù)ChiloDB。該數(shù)據(jù)庫(kù)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(duì)、數(shù)據(jù)下載等功能,界面交互友好,操作簡(jiǎn)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(gè)蛋白質(zhì)編碼基因、262個(gè)miRNA、82,639條piRNA序列。數(shù)據(jù)庫(kù)自2014年8月上線,至今已有70個(gè)國(guó)家的研究者訪問(wèn)使用,為二化螟研究提供了重要的數(shù)據(jù)平臺(tái)。
[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(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.112.1

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本文編號(hào)：1776241