普通菜豆生长习性相关基因定位
发布时间：2019-06-25 19:16
【摘要】：普通菜豆生长习性是一个与驯化密切相关的重要农艺性状,易受光周期等环境影响。该性状对株高、花期、成熟期、水分利用率、产量等都有重要影响,也是决定能否适宜机械化生产的关键性状,是普通菜豆重要育种策略。因此,开展生长习性研究不仅对探索普通菜豆的起源驯化具有重要的理论意义,而且对普通菜豆育种和产业发展具有重要的实践意义。本研究选择无限蔓生品种和有限丛生品种配置杂交组合,对分离群体进行遗传分析,利用基于菜豆基因组数据开发的SSR标记,采用分组分离群体法,初步将该基因定位在第一连锁群。同时针对候选区段进一步开发SSR和In/Del标记,对目标基因进行精细定位,为克隆和阐释该基因的作用机制奠定了分子基础。获得主要结果如下:1、分离群体构建及遗传分析选取无限蔓生型育成品种“连农紫芸一号”和有限丛生地方品种“兔子腿”(F0404)构建分离群体,获得F2及F2:3群体,在北京昌平(398株)、河南南阳(338株)、黑龙江哈尔滨(295株)多地种植,对F1、F2、F2:3群体进行表型调查。F1群体表型为无限蔓生型,F2后代分离表型为无限蔓生型和有限丛生型,不同观察地点群体遗传分析均符合3:1分离比,表明该性状由单基因控制,其中无限蔓生对有限丛生为显性。2、多态性标记筛选从基于菜豆基因组数据开发的2,802个均匀分布在各染色体上的SSR标记,筛选到310个亲本间多态性标记。在初步定位基础上,针对目标区段开发55个SSR标记,筛选到19个亲本间多态性标记。进一步选择该区段内15个候选基因克隆测序,开发了3个In/Del标记In87、In89和In93用于精细定位,在B01染色体上位置依次为45,513,666 bp、45,532,820bp、45,575,103bp。3、目标基因初步定位利用310个均匀分布各染色体上多态性标记,对隐性混池和隐性亲本进行连锁分析,将目标基因定位在第一条染色体;利用第一染色体上的标记对作图群体进行遗传分析,运用IciMapping v4.0构建连锁遗传图谱,将目标基因初定位于第1条染色体p1s86和p1s91之间,命名为gh-lz,遗传距离15.32cM,根据普通菜豆参考基因组序列,确定标记间的物理距离约为2.5Mb。4、gh-lz位点精细定位针对初步定位区间开发19个多态性SSR标记,进一步将目标基因精细定位于45,453,003bp和45,588,349bp之间,长度为135,346bp。对目标区段预测基因进行克隆,开发了3个In/Del标记,利用In/Del标记检测分离群体重组单株,In87和In89检测到0个重组单株,该标记与目标基因共分离,In93检测到2个重组株。最终,将目的基因定位在SSR标记p1t52和In/Del标记In93之间,区间大小为122,100bp,预测候选区段共包含12个基因,命名为Gene1~Gene12。将预测基因在NCBI中BLAST,对12个基因进行注释,其中Gene12注释为普通菜豆基因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..
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S643.1
本文编号：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..
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S643.1
【参考文献】
相关期刊论文 前3条
1 吴星波;岳欢;郝俊杰;张晓艳;李红卫;王珍青;吕享华;;普通菜豆抗锈病基因SCAR标记鉴定[J];生物技术通报;2013年10期
2 张文慧,陈庆山,杨庆凯,李文滨,王文辉,刘春燕,陈立君,刘海燕,单继勋;大豆灰斑病1号生理小种抗性基因的SSR标记分析[J];大豆科学;2004年03期
3 张志永,陈受宜,盖钧镒;大豆花叶病毒抗性基因Rsa的分子标记[J];科学通报;1998年20期
，本文编号：2505935
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2505935.html
最近更新
教材专著
