普通菜豆生長習性相關(guān)基因定位
發(fā)布時間：2019-06-25 19:16
【摘要】：普通菜豆生長習性是一個與馴化密切相關(guān)的重要農(nóng)藝性狀,易受光周期等環(huán)境影響。該性狀對株高、花期、成熟期、水分利用率、產(chǎn)量等都有重要影響,也是決定能否適宜機械化生產(chǎn)的關(guān)鍵性狀,是普通菜豆重要育種策略。因此,開展生長習性研究不僅對探索普通菜豆的起源馴化具有重要的理論意義,而且對普通菜豆育種和產(chǎn)業(yè)發(fā)展具有重要的實踐意義。本研究選擇無限蔓生品種和有限叢生品種配置雜交組合,對分離群體進行遺傳分析,利用基于菜豆基因組數(shù)據(jù)開發(fā)的SSR標記,采用分組分離群體法,初步將該基因定位在第一連鎖群。同時針對候選區(qū)段進一步開發(fā)SSR和In/Del標記,對目標基因進行精細定位,為克隆和闡釋該基因的作用機制奠定了分子基礎。獲得主要結(jié)果如下:1、分離群體構(gòu)建及遺傳分析選取無限蔓生型育成品種“連農(nóng)紫蕓一號”和有限叢生地方品種“兔子腿”(F0404)構(gòu)建分離群體,獲得F2及F2:3群體,在北京昌平(398株)、河南南陽(338株)、黑龍江哈爾濱(295株)多地種植,對F1、F2、F2:3群體進行表型調(diào)查。F1群體表型為無限蔓生型,F2后代分離表型為無限蔓生型和有限叢生型,不同觀察地點群體遺傳分析均符合3:1分離比,表明該性狀由單基因控制,其中無限蔓生對有限叢生為顯性。2、多態(tài)性標記篩選從基于菜豆基因組數(shù)據(jù)開發(fā)的2,802個均勻分布在各染色體上的SSR標記,篩選到310個親本間多態(tài)性標記。在初步定位基礎上,針對目標區(qū)段開發(fā)55個SSR標記,篩選到19個親本間多態(tài)性標記。進一步選擇該區(qū)段內(nèi)15個候選基因克隆測序,開發(fā)了3個In/Del標記In87、In89和In93用于精細定位,在B01染色體上位置依次為45,513,666 bp、45,532,820bp、45,575,103bp。3、目標基因初步定位利用310個均勻分布各染色體上多態(tài)性標記,對隱性混池和隱性親本進行連鎖分析,將目標基因定位在第一條染色體;利用第一染色體上的標記對作圖群體進行遺傳分析,運用IciMapping v4.0構(gòu)建連鎖遺傳圖譜,將目標基因初定位于第1條染色體p1s86和p1s91之間,命名為gh-lz,遺傳距離15.32cM,根據(jù)普通菜豆參考基因組序列,確定標記間的物理距離約為2.5Mb。4、gh-lz位點精細定位針對初步定位區(qū)間開發(fā)19個多態(tài)性SSR標記,進一步將目標基因精細定位于45,453,003bp和45,588,349bp之間,長度為135,346bp。對目標區(qū)段預測基因進行克隆,開發(fā)了3個In/Del標記,利用In/Del標記檢測分離群體重組單株,In87和In89檢測到0個重組單株,該標記與目標基因共分離,In93檢測到2個重組株。最終,將目的基因定位在SSR標記p1t52和In/Del標記In93之間,區(qū)間大小為122,100bp,預測候選區(qū)段共包含12個基因,命名為Gene1~Gene12。將預測基因在NCBI中BLAST,對12個基因進行注釋,其中Gene12注釋為普通菜豆基因TFL1。
[Abstract]:The growth habit of common bean is an important agronomic trait closely related to domestication, which is easy to be affected by photoperiod and other environments. This character has important effects on plant height, flowering stage, maturity, water use efficiency, yield and so on. It is also the key to determine whether it is suitable for mechanized production, and it is an important breeding strategy for common bean. Therefore, the study of growth habits is of great theoretical significance not only to explore the origin and domestication of common bean, but also to the breeding and industrial development of common bean. In this study, infinite trailing varieties and limited cluster varieties were selected for genetic analysis of isolated populations. SSR markers based on soybean genome data and group segregation method were used to locate the gene in the first linkage group. At the same time, SSR and In/Del markers were further developed for candidate regions to localize the target gene, which laid a molecular foundation for cloning and explaining the mechanism of action of the gene. The main results were as follows: 1. The segregation population construction and genetic analysis were carried out by selecting unlimited trailing type variety "Liannong Ziru No. 1" and limited cluster local variety "Rabbit leg" (F0404) to construct the isolated population. F2 and F2 populations were obtained and planted in Changping (398) in Beijing, Nanyang (338) in Henan and 295 in Harbin (Heilongjiang). The phenotypic investigation of F1 population was carried out. The phenotypic of F1 population was infinite trailing type, and the segregation phenotype of F2 progenies was infinite trailing type and finite cluster type. The genetic analysis of different observation sites was in accordance with the 3:1 segregation ratio, indicating that the trait was controlled by single gene, in which infinite trailing pair was dominant. 2. 2802 SSR markers based on soybean genome data were screened by polymorphism markers. 310 polymorphism markers between parents were screened. On the basis of preliminary localization, 55 SSR markers were developed for the target region, and 19 polymorphism markers between parents were screened. Furthermore, 15 candidate genes in this region were cloned and sequenced, and three In/Del markers In87,In89 and In93 were developed for fine mapping. 45513666 bp,45532820bp,45575103bp.3, target genes were located on B01 chromosome in turn, and the recessive mixed pool and recessive parents were analyzed by linkage analysis, and the target gene was located on the first chromosome. The genetic analysis of the mapping population was carried out by using the markers on the first chromosome, and the linkage genetic map was constructed by IciMapping v4.0. The target gene was initially mapped between the first chromosome p1s86 and p1s91, named gh-lz, genetic distance 15.32 cm. According to the reference genome sequence of common bean, the physical distance between the markers was determined to be about 2.5 Mb.4. 19 polymorphism SSR markers were developed for the preliminary mapping interval of gh-lz loci, and the target genes were further mapped between 45453003bp and 45588349bp with a length of 135346bp. Three In/Del markers were developed to clone the target region prediction gene. In/Del markers were used to detect the recombinant individual plants, In87 and In89 were used to detect 0 recombinant individual plants. The marker was co-isolated from the target gene and two recombinant strains were detected by In93. Finally, the target gene was located between SSR marker p1t52 and In/Del marker In93, and the interval was 122100bp. the predicted candidate region contained 12 genes named Gene1~Gene12.. The predicted genes were commented on 12 genes by BLAST, in NCBI, in which Gene12 was annotated as common bean gene TFL1..
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S643.1
本文編號：2505935
[Abstract]:The growth habit of common bean is an important agronomic trait closely related to domestication, which is easy to be affected by photoperiod and other environments. This character has important effects on plant height, flowering stage, maturity, water use efficiency, yield and so on. It is also the key to determine whether it is suitable for mechanized production, and it is an important breeding strategy for common bean. Therefore, the study of growth habits is of great theoretical significance not only to explore the origin and domestication of common bean, but also to the breeding and industrial development of common bean. In this study, infinite trailing varieties and limited cluster varieties were selected for genetic analysis of isolated populations. SSR markers based on soybean genome data and group segregation method were used to locate the gene in the first linkage group. At the same time, SSR and In/Del markers were further developed for candidate regions to localize the target gene, which laid a molecular foundation for cloning and explaining the mechanism of action of the gene. The main results were as follows: 1. The segregation population construction and genetic analysis were carried out by selecting unlimited trailing type variety "Liannong Ziru No. 1" and limited cluster local variety "Rabbit leg" (F0404) to construct the isolated population. F2 and F2 populations were obtained and planted in Changping (398) in Beijing, Nanyang (338) in Henan and 295 in Harbin (Heilongjiang). The phenotypic investigation of F1 population was carried out. The phenotypic of F1 population was infinite trailing type, and the segregation phenotype of F2 progenies was infinite trailing type and finite cluster type. The genetic analysis of different observation sites was in accordance with the 3:1 segregation ratio, indicating that the trait was controlled by single gene, in which infinite trailing pair was dominant. 2. 2802 SSR markers based on soybean genome data were screened by polymorphism markers. 310 polymorphism markers between parents were screened. On the basis of preliminary localization, 55 SSR markers were developed for the target region, and 19 polymorphism markers between parents were screened. Furthermore, 15 candidate genes in this region were cloned and sequenced, and three In/Del markers In87,In89 and In93 were developed for fine mapping. 45513666 bp,45532820bp,45575103bp.3, target genes were located on B01 chromosome in turn, and the recessive mixed pool and recessive parents were analyzed by linkage analysis, and the target gene was located on the first chromosome. The genetic analysis of the mapping population was carried out by using the markers on the first chromosome, and the linkage genetic map was constructed by IciMapping v4.0. The target gene was initially mapped between the first chromosome p1s86 and p1s91, named gh-lz, genetic distance 15.32 cm. According to the reference genome sequence of common bean, the physical distance between the markers was determined to be about 2.5 Mb.4. 19 polymorphism SSR markers were developed for the preliminary mapping interval of gh-lz loci, and the target genes were further mapped between 45453003bp and 45588349bp with a length of 135346bp. Three In/Del markers were developed to clone the target region prediction gene. In/Del markers were used to detect the recombinant individual plants, In87 and In89 were used to detect 0 recombinant individual plants. The marker was co-isolated from the target gene and two recombinant strains were detected by In93. Finally, the target gene was located between SSR marker p1t52 and In/Del marker In93, and the interval was 122100bp. the predicted candidate region contained 12 genes named Gene1~Gene12.. The predicted genes were commented on 12 genes by BLAST, in NCBI, in which Gene12 was annotated as common bean gene TFL1..
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S643.1
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