本文選題：水稻 + 紋枯病��； 參考：《揚州大學》2017年碩士論文

【摘要】：由立枯絲核菌引起的水稻紋枯病是世界性水稻病害。水稻對該病的抗性為部分抗性,受多基因或數量性狀基因座(quantitative trait loci/QTLs)控制,迄今未發(fā)現完全免疫的水稻種質和高抗的抗病基因。至目前,已先后有300多個抗紋枯病QTLs被報道分布于水稻全部12條染色體上。然而,由于采用的定位親本、群體類型、遺傳標記、鑒定方法以及環(huán)境條件等的不同,使得多數QTLs的位置不能直接比較,且置信區(qū)間較大,影響了育種應用。近年提出的QTLs元分析策略,其可利用已知QTLs的相關信息,通過數據整合再分析,推斷出可靠性更高、位置區(qū)間更精確的一致性QTLs(Meta-QTLs)。本研究利用該方法,對抗紋枯病QTLs以及影響紋枯病的抗性的株高和生育期性狀QTLs進行了元分析,并利用染色體片段代換系和高代回交群體對部分主效QTL進行驗證及進一步定位;同時構建了高抗紋枯病水稻新種質YSBR1在江蘇近年主推品種泰粳394(TJ394)背景下的全基因組染色體片段代換系。主要結果如下:1.收集前人有關水稻紋枯病抗性QTLs的研究報道,對符合元分析條件的188個抗紋枯病QTLs進行了元分析,獲得34個抗紋枯病一致性QTLs,命名為MQSBR(Meta QTLs ofSheathblightresistance)。這些 MQSBRs 主要分布在水稻第 1、2、3、5、7、8、9、11和12染色體上,物理區(qū)間在0.2-4.2Mb范圍內,其中近半數在1Mb區(qū)間內,明顯小于前人報道的初級定位的置信區(qū)間。通過圖譜比較,發(fā)現8個MQSBRs與MQHD(Meta QTLs of Heading Date)、MQPH(Meta QTLs of Plant Height)間連鎖或區(qū)間重疊,余下 26 個MQSBRs可能與株高及生育期性狀沒有連鎖關系。2.結合前人報道的可靠性較高的5個抗紋枯病QTLs(qSB-9TQ、qSB-11LE、qSB-11HJX74、qSB-11Tetep、qSB-12YSBR1、位置信息,發(fā)現MQSBR-25位于qSB-9TQ區(qū)間內,qSB-11LE位于MQSBR-27區(qū)間內,MQSBR-33和MQSBR-34位于qSB-12YSBR1區(qū)間內,這些結果證明元分析結果可靠性較好。本研究發(fā)現YSBR1/Lemont染色體片段代換系(Lemont背景)中有2個系的田間抗性顯著強于對照Lemont,分別攜帶MQSBR-7、MQSBR-8和MQSBR-33、MQSBR-34,為了分析這2個系中是否還攜帶其它MQSBRs,通過236個分布于水稻12條染色體的多態(tài)性分子標記,檢測發(fā)現這2個系的遺傳背景中分別攜帶有7個和2個MQSBRs。3.為了發(fā)掘更多有利用價值的抗紋枯病主效QTLs,同時創(chuàng)建抗病育種中間材料,本研究構建了 YSBR1/TJ394全基因組染色體片段代換系。累計篩選到親本間多態(tài)性分子標記266對,相對均勻的分布于水稻12條染色體上。累計獲得了 111個以T394為背景的YSBR1染色體片段代換系,其中BC6F1世代代換系14份,BC5Fi世代代換系49份,BC4F1世代代換系37份,BC3F1世代代換系11份。代換系代換片段累計覆蓋95%的YSBR1基因組信息。本研究利用覆蓋qSB-12YSBR1的BC4F2回交分離群體(YSBR1/TJ394),通過復合區(qū)間作圖法,發(fā)現一個抗紋枯病QTL,LOD值高達11.2,置信區(qū)間內包括MQSBR-34、MQPH-6和MQSBR-33的絕大部分區(qū)域。4.水稻第9染色體長臂中下端是不同研究學者多次檢測到抗紋枯病QTLs的區(qū)間,在該區(qū)域共檢測到5個MQSBRs,命名為MQSBR-22-MQSBR-26。結合紋枯病菌誘導差異表達基因信息,在MQSBR-24和MQSBR-25區(qū)間內各發(fā)現一個紋枯病菌顯著誘導表達的基因 OsPIP(Plasma membrane intrinsic protein 2c)和 OsEXPR(Expansin-related protein 2 precursor)。通過RNA干擾,我們獲得了 YSBR1背景下各基因轉錄表達顯著下調的轉基因株系。抗性鑒定結果顯示,各基因的RNAi株系的病級數據與對照YSBR1間沒有顯著差異,表明它們可能不是MQSBR-24和MQSBR-25的候選基因。為了分析兩個基因是否為qSB-9TQ的候選基因,我們利用各基因的RNA干擾系(OsPIP-Ri、OsEXPR-Ri/YSBR1背景)分別與Lemont背景下的qSB-9TQ導入系(LE-qSB-9TQ)雜交獲得大量F1,同時配制對照F1(YSBR1×LE-qSB-9TQ)種子。對三套F1雜交種植株進行紋枯病菌接種鑒定,結果顯示,不同材料間的病情差異不顯著,暗示兩個基因可能也不是qSB-9TQ的候選基因。以上結果為進一步克隆水稻抗紋枯病QTLs并開展育種應用奠定了材料基礎,具有重要的理論和實際價值。
[Abstract]:Rice sheath blight caused by Rhizoctonia Rhizoctonia is a worldwide rice disease. The resistance of rice to this disease is partial resistance, controlled by multiple gene or quantitative trait loci (quantitative trait loci/QTLs). Up to now, no fully immune rice germplasm and high resistance genes have been found. Up to now, there have been more than 300 anti sheath blight QTLs. The reports are distributed on all 12 chromosomes of rice. However, the location of the parents, population types, genetic markers, identification methods and environmental conditions are different, which make the position of most QTLs can not be directly compared, and the confidence interval is large, which affects the breeding application. The QTLs meta analysis strategy proposed in recent years can make use of the known QTLs. Relevant information, through data integration and reanalysis, infers higher reliability and more accurate consistency QTLs (Meta-QTLs) in the position interval. This study uses this method to analyze the plant height of sheath blight and the resistance to sheath blight and the growth period character QTLs, and use the chromosome fragment substitution line and the high generation backcross population to the part of the QTLs. The main effect of QTL was verified and further located, and the genome segment substitution line of YSBR1, a new rice germplasm with high resistance to sheath blight, in Jiangsu, under the background of Thai japonica 394 (TJ394) in recent years, was constructed. The main results were as follows: 1. the study of the previous studies on the resistance of rice sheath blight to the rice sheath blight resistance was reported, and 188 of the conditions were conformed to the conditions of the analysis of the rice sheath blight. The resistance to sheath blight QTLs was analyzed by meta analysis, and 34 conformance QTLs of resistance to sheath blight were obtained, named MQSBR (Meta QTLs ofSheathblightresistance). These MQSBRs were mainly distributed on the 1,2,3,5,7,8,9,11 and 12 chromosomes of rice. The physical interval was within the 0.2-4.2Mb range, and nearly half of them were within the 1Mb interval, obviously smaller than those of the previous reports. By comparison, 8 MQSBRs and MQHD (Meta QTLs of Heading Date), MQPH (Meta QTLs of Plant) are linked or overlapped, and the remaining 26 may have no linkage to plant height and growth stage traits. 11HJX74, qSB-11Tetep, qSB-12YSBR1, location information, found that MQSBR-25 is located in the qSB-9TQ interval, qSB-11LE is located in the MQSBR-27 interval, MQSBR-33 and MQSBR-34 are in the qSB-12YSBR1 interval. These results show that the results of the meta analysis are reliable. This study found that there are 2 fields in the YSBR1/Lemont chromophore segment substitution system (Lemont background). Compared with control Lemont, MQSBR-7, MQSBR-8 and MQSBR-33, MQSBR-34 were carried respectively. In order to analyze the other MQSBRs in the 2 lines and 236 polymorphic markers distributed on the 12 chromosomes of rice, we found that 7 and 2 MQSBRs.3. were carried in the genetic background of the 2 lines to explore more utilization. The main effect of anti sheath blight was QTLs, and the intermediate material of disease resistance breeding was created at the same time. The YSBR1/TJ394 genome fragment substitution line was constructed in this study. A total of 266 pairs of polymorphic molecular markers were screened and distributed on 12 chromosomes of rice. 111 YSBR1 chromosome fragments with T394 as the background were obtained. Substitution lines, of which 14 generations of BC6F1 generation, 49 generations of substitution lines for generation BC5Fi, 37 generations of substitution lines in BC4F1 generation and 11 generations of substitution lines for generation BC3F1. The substitution line substitution fragment covers 95% of YSBR1 genome information. This study uses BC4F2 backcross segregated populations (YSBR1/ TJ394) covering qSB-12YSBR1, and discovers an anti grain by composite interval mapping method. The value of QTL, LOD, is up to 11.2. In the confidence interval, the middle and lower end of the ninth chromosome ninth chromosome long arm of.4. rice, including MQSBR-34, MQPH-6 and MQSBR-33, is the interval between different researchers on the anti sheath blight QTLs, and 5 MQSBRs are detected in the region, and the differential expression gene is induced by the MQSBR-22-MQSBR-26. combined with the Rhizoctonia solani. We found a gene OsPIP (Plasma membrane intrinsic protein 2C) and OsEXPR (Expansin-related protein 2 precursor) in the MQSBR-24 and MQSBR-25 intervals. The results showed that there was no significant difference between the disease level data of the RNAi strains of each gene and the control YSBR1, indicating that they may not be candidate genes for MQSBR-24 and MQSBR-25. In order to analyze whether the two genes are candidate genes for qSB-9TQ, we use the RNA interference lines of each gene (OsPIP-Ri, OsEXPR-Ri/YSBR1 background) with qSB-9T in the Lemont background, respectively. A large number of F1 were obtained by Q Introduction (LE-qSB-9TQ) hybridization and control of F1 (YSBR1 * LE-qSB-9TQ) seeds. Three sets of F1 hybrid plants were inoculated with Rhizoctonia solanae. The results showed that the difference between different materials was not significant, suggesting that two genes might not be the candidate genes of qSB-9TQ. The above results were the further cloning of rice anti grain. The QTLs and the application of breeding have laid the foundation for the disease and have important theoretical and practical value.

【學位授予單位】：揚州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S435.111.42
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本文編號：1881659