茶树遗传图谱与品种指纹图谱构建及几个重要性状的QTL定位

发布时间：2017-12-31 16:20

  本文关键词：茶树遗传图谱与品种指纹图谱构建及几个重要性状的QTL定位　出处：《四川农业大学》2016年博士论文　论文类型：学位论文

  更多相关文章： 茶树 SSR标记 遗传图谱 数量性状位点 品种指纹图谱

【摘要】：茶是世界范围内最受欢迎的饮料之一,越来越多的证据表明饮茶可以改善人类健康。从20世纪60年代以来,世界茶叶的消费量一直保持高速增长,茶产品类型日益丰富,茶树的栽培面积和茶叶产量也随之大幅增长。优良的茶树品种是茶叶生产中最关键的物质资料,21世纪多元化的市场需求为茶树良种选育工作带来了新的挑战。但由于其基因组复杂、育种周期长、多数经济性状为多基因控制的数量性状,茶树的遗传研究和优质品种选育均较为困难。基于DNA分子标记的遗传图谱是研究数量性状的有效工具,在此基础上发展的标记辅助选择(marker assisted selection, MAS)育种技术可以大幅提高多年生作物的育种效率。本论文首先通过开发筛选大量简单序列重复(simple sequence repeat,SSR)标记,基于170株‘龙井43’ב白毫早’杂交一代(F1)作图群体,构建起一张高密度的茶树遗传图谱。接着,连续两年观测或测定该作图群体多个重要性状的表型数据,利用所构建的遗传图谱进行数量性状位点定位分析(quantitative trait loci mapping, QTL mapping),得到了40余个QTL。另外,本论文还从大量已定位到遗传图谱的标记中筛选出八个核心SSR标记用于构建中国主要无性系茶树品种的DNA分子指纹图谱。取得的主要研究结果如下：1.为获得足够的SSR分子标记用于遗传作图,以‘龙井43’、‘白毫早’以及它们的六株杂交子代的基因组DNA为PCR模板筛选了711个SSR标记,其中460个是根据茶树转录组序列新开发而成,另外251个是从其他两张茶树遗传图谱上转移而来。从这些标记中共获得342个(48.1%)在‘龙井43’ב白毫早’作图群体中有多态性的SSR标记,其中206个为本研究新开发。2.用上述342个多态性标记对‘龙井43’ב白毫早’作图群体的170单株代进行基因型分析。得到的基因型数据和已有的155个SSR标记的数据合并,用于构建遗传图谱。新构建的双亲整合图谱含有483个SSR标记,分布在15条连锁群上,总遗传图距为1226.2 cM,相邻标记间平均间距为2.5 cM。基于“双假侧交”作图理论分别构建了两个杂交亲本各自的遗传图谱。母本图谱(‘龙井43’)含有323个标记,总图距为1032.2 cM；父本图谱(‘白毫早’)含有303个标记,总图距为1322.5 cM。与父本相比,母本图谱上有更多的偏分离位点。3.新建的‘龙井43’ב白毫早’双亲整合图谱上有126个SSR标记是与其他两张茶树图谱共享的标记位点。基于这些共享位点,建立起了三张茶树遗传图谱15条连锁群的对应关系。来自不同图谱对应连锁群的遗传图距、共享标记的位置基本一致,为比较、验证不同作图群体中得到的QTL位点奠定了基础。4.连续两年(2014和2015)观测了‘龙井43’ב白毫早’作图群体的春季发芽期(timing of spring bud flush, TBF)、新梢颜色(young shoot color, YSC)、成熟叶片长度(mature leaf length, MLL)、宽度(mature leaf width, MLW)和叶形指数(leaf shape index, LSI)等性状。同时用高效液相色谱测定了该作图群体两年夏季新梢中七个儿茶素单体(catechines)和咖啡碱(caffeine, CAF)的含量。用上述表型数据结合所构建的遗传图谱定位到43个QTL,其中2个与TBF相关、5个与YSC相关、5个与MLL相关、3个与LSI相关、2个与咖啡碱含量相关、20个分别与七个儿茶素组分含量相关,还有6个与酯型儿茶素与非酯型儿茶素比值相关。共有14个QTL的LOD值达到全基因组极显著水平(P0.01),且在两年的数据中均检测到。5.为了将SSR标记应用于茶树无性系品种鉴定,从已定位到遗传图谱的483个SSR标记中筛选出30个高质量、相互独立的SSR标记,结合高分辨率毛细管电泳检测技术,分析了128个中国主栽的无性系茶树品种。结果表明这些标记具有很高的多态性,平均每个位点可以检测到10.4条等位基因,平均多态性信息含量(polymorphic information content, PIC)为0.704。统计显示这些标记也具有很强的品种辨别能力,根据四个位点的基因型即可完全区分这128个茶树品种。基于等位基因的分布频率和不同等位基因条带大小差异,进一步筛选出八个核心SSR标记用于构建茶树无性系品种DNA指纹图谱。6.基于上述30个SSR位点的基因型数据,分析了128个无性系茶树品种间可能存在的亲子关系。结果共鉴定出47对可能的亲子关系,其中33对与育种记录相符,另外14对为首次发现。本论文所得到的SSR标记、遗传图谱和QTL位点为茶树功能基因鉴定、MAS育种和其他性状的QTL定位奠定了基础。筛选出来的八个核心SSR标记和构建的无性系品种DNA指纹图谱数据库为茶树品种鉴别提供了可靠的技术支持。
[Abstract]:Tea is one of the world's most popular drink, more and more evidence that drinking tea can improve human health. Since 1960s, world consumption of tea has maintained rapid growth, tea products increasingly rich, tea cultivation area and yield of tea is also substantial growth. Excellent tea varieties is the key to the tea in the production of material, has brought new challenges in twenty-first Century diversified market demand for tea breeding work. But because of its complex genome, long breeding cycle, most economic traits were quantitative traits controlled by multiple genes, genetic research and breeding of high quality varieties of tea are more difficult. A genetic map based on DNA markers are effective tools Study on quantitative traits, marker assisted selection on the basis of the development of (marker assisted selection, MAS) can significantly improve the number of breeding technology The breeding efficiency of annual crops. Firstly, through the development of screening a large number of simple sequence repeat (simple sequence, repeat, SSR) markers based on 170 strains of "Longjing 43" X "baihaozao" hybrid (F1) mapping population, build a high density genetic map of tea. Next, two consecutive years of observation or determination of the phenotype data of several important traits in the mapping population, for QTL analysis using genetic map constructed (quantitative trait loci mapping, QTL mapping), has been more than 40 QTL. in addition, this paper also from large amount have been mapped on the genetic map markers screened eight core SSR markers construction of DNA molecular fingerprinting system mainly China tea varieties asexual. The main results are as follows: 1. for SSR markers for genetic mapping with enough, "Longjing 43", and "tea" The six strains of hybrids genome DNA screened 711 SSR markers as PCR template, 460 of which are based on the transcriptome sequence and the development of new tea, the other 251 are transferred from the other two piece tea genetic map to get 342 from these markers of the Communist Party of China (48.1%) SSR markers in the "Longjing 43" X "baihaozao 'linkage group, of which 206 are the new research development of.2. by the 342 polymorphic markers of" Longjing 43 "x 170" baihaozao "mapping population per plant generation genotype was analyzed. 155 SSR markers from genotype data and the with the data, for the construction of genetic mapping. Parents integration newly constructed containing 483 SSR markers distributed in 15 linkage groups, the total genetic distance of 1226.2 cM, the average distance between adjacent markers was 2.5 cM. based on the" double false side "mapping theory to construct Two hybrid parents their genetic map. The female map ("Longjing 43") containing 323 markers, a total distance of 1032.2 cM; the male map ("baihaozao") containing 303 markers, the total distance is 1322.5 cM. compared with the male, female were more segregation points of a new.3. "Longjing 43 'x' baihaozao 'parents integrated map with 126 SSR markers are shared with other sites of two Zhang Chashu map. These shared sites based on the established three piece tea genetic map of 15 linkage groups corresponding relationship. Genetic map from different linkage groups from the corresponding map, mark the location of the basic shared consistent, for comparison,.4. provides a basis for two consecutive years to verify the QTL sites of different mapping populations (2014 and 2015) were observed in' Longjing 43 '*' baihaozao 'mapping population in spring (timing of spring bud in germination period flush, TB F),鏂版ⅱ棰滆壊(young shoot color, YSC),鎴愮啛鍙剁墖闀垮害(mature leaf length, MLL),瀹藉害(mature leaf width, MLW)鍜屽彾褰㈡寚鏁，

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