本文選題：白背飛虱　切入點：南方水稻黑條矮縮病毒　出處：《中國科學技術大學》2017年博士論文　論文類型：學位論文

【摘要】：白背飛虱是一種重要的農(nóng)業(yè)害蟲,主要通過取食植物韌皮部汁液和傳播病毒兩種方式危害水稻。白背飛虱是目前已知傳播南方水稻黑條矮縮病毒(Southern rice black-streaked dwarf virus,SRBSDV)的特異介體昆蟲,SRBSDV在其體內(nèi)以持久增殖方式擴增。由于缺少白背飛虱的基因組等信息,白背飛虱的相關生物學研究進展緩慢,包括白背飛虱與SRBSDV互作機制的研究。本課題針對這種研究現(xiàn)狀,首先對白背飛虱的基因組測序,獲得了高質(zhì)量的基因組序列;系統(tǒng)鑒定了白背飛虱的免疫相關基因;對感染SRBSDV的白背飛虱轉(zhuǎn)錄組進行詳細的比較分析。結(jié)果如下:我們首先對每代白背飛虱進行同胞配對傳代,獲得基因組雜合度不斷降低的近交系。提取F6代個體的基因組DNA,構建一系列插入片段180 bp～40 kb的DNA文庫進行高通量測序,獲得了總量達到241.3 Gb測序深度為330X的原始序列。經(jīng)組裝和拼接獲得約720 Mb的白背飛虱基因組序列,其Contig N50為70.7 kb,Scaffold N50為1.18 Mb。生物信息學分析白背飛虱基因組含有21,254個蛋白編碼基因。利用64個白背飛虱F1代和F2代個體,構建了23個RAD-seq文庫,鑒定發(fā)現(xiàn)2,386個遺傳標記,構建了15個遺傳連鎖群。這些連鎖群包括529個Scaffolds,占基因組總長度的71%;含13,626個基因,占基因總數(shù)的64%�；诎妆筹w虱基因組和轉(zhuǎn)錄組信息,我們通過同源比對分析鑒定了348個免疫相關基因,其中231個基因可以歸類至28個免疫基因家族和4個功能組中。為了研究免疫基因在白背飛虱抗SRBSDV過程中的作用以及SRBSDV侵染對白背飛虱產(chǎn)生的影響,我們分析了SRBSDV侵染的白背飛虱轉(zhuǎn)錄本。傳毒后不同個體SRBSDV S9-2表達水平在第12天和15天有10∧6級別的差異。按照白背飛虱個體SRBSDV表達水平分組,提取高病毒滴度組(High viral titers,HVT)、中病毒滴度組(Median viral titers,MVT)和非侵染組(Non-viruliferous,NVF)的白背飛虱RNA進行轉(zhuǎn)錄組測序。分析發(fā)現(xiàn)SRBSDV侵染對于介體白背飛虱基因表達具有廣泛影響。和健康對照組相比,我們在HVT,MVT和NVF組鑒定了278個共同上調(diào)表達基因和406個共同下調(diào)表達基因(差異倍數(shù)=2),這些基因參與了很多種生物學過程,在初級代謝通路和氧化還原反應中有一定的富集。鑒于NVF,MVT,HVT組白背飛虱含有SRBSDV漸次增加,分析發(fā)現(xiàn)1,906個單向遞增基因(Monotonically increasing exprssion genes,MIEGs)和1,467個單向遞減基因(Monotonically decreasing exprssion genes,MDEGs),這些基因表現(xiàn)出一定程度的病毒滴度依賴。RNAi通路基因在MIEGs中顯著富集,表明RNAi是白背飛虱抗SRBSDV病毒的一種主要免疫反應。對小RNA分析同樣證明白背飛虱RNAi通路在細胞和成蟲水平上參與抗SRBSDV反應。綜上所述,通過本課題的研究,獲得了高質(zhì)量的白背飛虱基因組序列,明確了白背飛虱的免疫系統(tǒng)組成,認識了白背飛虱在感染SRBSDV后復雜的基因差異表達情況。這些數(shù)據(jù)和發(fā)現(xiàn)為深入研究SRBSDV-白背飛虱-水稻三者之間的互作機制提供了研究基礎。
[Abstract]:The white backed planthopper is one of the most important agricultural pests, harm of rice mainly through feeding plant phloem sap and the spread of the virus in two ways. Sogatellafurcifera is now known to spread southern rice black streaked dwarf virus (Southern rice black-streaked dwarf virus, SRBSDV) the specific insect, SRBSDV in the body with persistent proliferation due to the lack of amplification. Sogatellafurcifera genome information, research progress of related biological sogatellafurcifera slow, including sogatellafurcifera and SRBSDV to study the interaction mechanism. This paper based on this situation, the first genome sequencing of WBPH, obtained the genome sequences of high quality; system identification of immune related genes in white backedplanthopper; detailed comparative analysis on sogatellafurcifera transcriptome of SRBSDV infection. The results are as follows: firstly we each generation of WBPH were sib pairs were obtained Inbred to genomic heterozygosity decreased. The genomic DNA of individual extraction F6, construct a series of insert 180 BP ~ 40 KB DNA library for high-throughput sequencing, the total reached 241.3 Gb sequencing depth of the original sequence of 330X. By assembling and splicing sogatellafurcifera genomic sequences obtained about 720 Mb the Contig N50 70.7 KB Scaffold N50 1.18 Mb. bioinformatics analysis of sogatellafurcifera genome contains 21254 protein encoding gene. Using 64 sogatellafurcifera F1 and F2 generation individuals, 23 RAD-seq libraries were constructed, identified 2386 genetic markers, 15 linkage groups were constructed these linkage groups including 529 Scaffolds, accounting for 71% of the total length of the genome; containing 13626 genes, accounting for 64%. of the total gene sogatellafurcifera genome and transcriptome information based on the homologous analysis we identified 348 immune related Genes, including 231 genes can be classified into 28 immune gene family and 4 functional groups. In order to study the influence of immune genes in sogatellafurcifera anti SRBSDV process and the effect of SRBSDV infection on WBPH, we analyzed SRBSDV infection sogatellafurcifera transcripts. Expression of different individual SRBSDV S9-2 virus after the level difference at twelfth and 15 days, 10 a 6 level. In accordance with sogatellafurcifera individual expression of SRBSDV group, extraction of high virus titer group (High viral, titers, HVT), virus titer group (Median viral titers, MVT) and non infection group (Non-viruliferous, NVF) of sogatellafurcifera the RNA transcriptome sequencing. The result showed that SRBSDV infection has broad implications for the expression of mediator sogatellafurcifera gene. Compared with healthy control group, we at HVT, MVT and NVF group identified 278 common up-regulated genes and 406 down regulated table As gene (fold difference =2), these genes are involved in many biological processes in the primary metabolic pathways and redox reactions in certain concentration. In view of NVF, MVT, HVT group of sogatellafurcifera containing SRBSDV gradually increase, the analysis found that 1906 single gene (Monotonically increasing exprssion genes by MIEGs, and 1467) one gene (Monotonically decreasing exprssion genes., MDEGs), these genes showed a certain degree of virus titer dependent.RNAi pathway genes were significantly enriched in MIEGs, indicating that RNAi is a kind of white back lice primary immune responses against SRBSDV virus. The RNA analysis also showed that the small white backed planthopper RNAi pathway is involved in anti SRBSDV reaction in cells and the adult level. To sum up, through the research, obtained the WBPH genome sequence of high quality, the components of the immune system white back lice, We have known the complex gene expression of white backed planthopper after infection with SRBSDV. These data and findings provide a basis for further research on the interaction mechanism between SRBSDV-, the three white planthopper rice.

【學位授予單位】：中國科學技術大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S435.11

【參考文獻】

相關期刊論文 前10條

1 張\~;谷少華;李顯春;;昆蟲學研究進入基因組學時代[J];中國科學:生命科學;2016年10期

2 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

3 洪健;周雪平;;ICTV第九次報告以來的植物病毒分類系統(tǒng)[J];植物病理學報;2014年06期

4 劉萬才;陸明紅;翟保平;王建強;高小麗;Hoang Anh Tuan;Ho Dang Cu;;越南水稻生產(chǎn)及其遷飛性害蟲發(fā)生情況[J];中國植保導刊;2014年10期

5 劉萬才;陸明紅;黃沖;周益軍;周國輝;;南方水稻黑條矮縮病大區(qū)流行規(guī)律初探[J];中國植保導刊;2014年04期

6 黃俊;杜堯東;王華;黃珍珠;;廣東省稻飛虱遷入軌跡及蟲源地分析[J];中國農(nóng)學通報;2013年24期

7 曹楊;潘峰;周倩;李冠華;劉雙清;黃志農(nóng);李有志;;南方水稻黑條矮縮病毒介體昆蟲白背飛虱的傳毒特性[J];應用昆蟲學報;2011年05期

8 鐘天潤;劉宇;劉萬才;;2010年我國南方水稻黑條矮縮病發(fā)生原因及趨勢初析[J];中國植保導刊;2011年04期

9 姜玉英;郭榮;劉宇;馮曉東;;越南的水稻病毒病發(fā)生和防治概況[J];中國植保導刊;2010年08期

10 郭榮;周國輝;張曙光;;水稻南方黑條矮縮病發(fā)生規(guī)律及防控對策初探[J];中國植保導刊;2010年08期

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