本文關鍵詞：茶樹遺傳圖譜與品種指紋圖譜構建及幾個重要性狀的QTL定位　出處：《四川農業(yè)大學》2016年博士論文　論文類型：學位論文

  更多相關文章： 茶樹 SSR標記 遺傳圖譜 數(shù)量性狀位點 品種指紋圖譜

【摘要】：茶是世界范圍內最受歡迎的飲料之一,越來越多的證據表明飲茶可以改善人類健康。從20世紀60年代以來,世界茶葉的消費量一直保持高速增長,茶產品類型日益豐富,茶樹的栽培面積和茶葉產量也隨之大幅增長。優(yōu)良的茶樹品種是茶葉生產中最關鍵的物質資料,21世紀多元化的市場需求為茶樹良種選育工作帶來了新的挑戰(zhàn)。但由于其基因組復雜、育種周期長、多數(shù)經濟性狀為多基因控制的數(shù)量性狀,茶樹的遺傳研究和優(yōu)質品種選育均較為困難。基于DNA分子標記的遺傳圖譜是研究數(shù)量性狀的有效工具,在此基礎上發(fā)展的標記輔助選擇(marker assisted selection, MAS)育種技術可以大幅提高多年生作物的育種效率。本論文首先通過開發(fā)篩選大量簡單序列重復(simple sequence repeat,SSR)標記,基于170株‘龍井43’ב白毫早’雜交一代(F1)作圖群體,構建起一張高密度的茶樹遺傳圖譜。接著,連續(xù)兩年觀測或測定該作圖群體多個重要性狀的表型數(shù)據,利用所構建的遺傳圖譜進行數(shù)量性狀位點定位分析(quantitative trait loci mapping, QTL mapping),得到了40余個QTL。另外,本論文還從大量已定位到遺傳圖譜的標記中篩選出八個核心SSR標記用于構建中國主要無性系茶樹品種的DNA分子指紋圖譜。取得的主要研究結果如下：1.為獲得足夠的SSR分子標記用于遺傳作圖,以‘龍井43’、‘白毫早’以及它們的六株雜交子代的基因組DNA為PCR模板篩選了711個SSR標記,其中460個是根據茶樹轉錄組序列新開發(fā)而成,另外251個是從其他兩張茶樹遺傳圖譜上轉移而來。從這些標記中共獲得342個(48.1%)在‘龍井43’ב白毫早’作圖群體中有多態(tài)性的SSR標記,其中206個為本研究新開發(fā)。2.用上述342個多態(tài)性標記對‘龍井43’ב白毫早’作圖群體的170單株代進行基因型分析。得到的基因型數(shù)據和已有的155個SSR標記的數(shù)據合并,用于構建遺傳圖譜。新構建的雙親整合圖譜含有483個SSR標記,分布在15條連鎖群上,總遺傳圖距為1226.2 cM,相鄰標記間平均間距為2.5 cM�；凇半p假側交”作圖理論分別構建了兩個雜交親本各自的遺傳圖譜。母本圖譜(‘龍井43’)含有323個標記,總圖距為1032.2 cM；父本圖譜(‘白毫早’)含有303個標記,總圖距為1322.5 cM。與父本相比,母本圖譜上有更多的偏分離位點。3.新建的‘龍井43’ב白毫早’雙親整合圖譜上有126個SSR標記是與其他兩張茶樹圖譜共享的標記位點�；谶@些共享位點,建立起了三張茶樹遺傳圖譜15條連鎖群的對應關系。來自不同圖譜對應連鎖群的遺傳圖距、共享標記的位置基本一致,為比較、驗證不同作圖群體中得到的QTL位點奠定了基礎。4.連續(xù)兩年(2014和2015)觀測了‘龍井43’ב白毫早’作圖群體的春季發(fā)芽期(timing of spring bud flush, TBF)、新梢顏色(young shoot color, YSC)、成熟葉片長度(mature leaf length, MLL)、寬度(mature leaf width, MLW)和葉形指數(shù)(leaf shape index, LSI)等性狀。同時用高效液相色譜測定了該作圖群體兩年夏季新梢中七個兒茶素單體(catechines)和咖啡堿(caffeine, CAF)的含量。用上述表型數(shù)據結合所構建的遺傳圖譜定位到43個QTL,其中2個與TBF相關、5個與YSC相關、5個與MLL相關、3個與LSI相關、2個與咖啡堿含量相關、20個分別與七個兒茶素組分含量相關,還有6個與酯型兒茶素與非酯型兒茶素比值相關。共有14個QTL的LOD值達到全基因組極顯著水平(P0.01),且在兩年的數(shù)據中均檢測到。5.為了將SSR標記應用于茶樹無性系品種鑒定,從已定位到遺傳圖譜的483個SSR標記中篩選出30個高質量、相互獨立的SSR標記,結合高分辨率毛細管電泳檢測技術,分析了128個中國主栽的無性系茶樹品種。結果表明這些標記具有很高的多態(tài)性,平均每個位點可以檢測到10.4條等位基因,平均多態(tài)性信息含量(polymorphic information content, PIC)為0.704。統(tǒng)計顯示這些標記也具有很強的品種辨別能力,根據四個位點的基因型即可完全區(qū)分這128個茶樹品種�；诘任换虻姆植碱l率和不同等位基因條帶大小差異,進一步篩選出八個核心SSR標記用于構建茶樹無性系品種DNA指紋圖譜。6.基于上述30個SSR位點的基因型數(shù)據,分析了128個無性系茶樹品種間可能存在的親子關系。結果共鑒定出47對可能的親子關系,其中33對與育種記錄相符,另外14對為首次發(fā)現(xiàn)。本論文所得到的SSR標記、遺傳圖譜和QTL位點為茶樹功能基因鑒定、MAS育種和其他性狀的QTL定位奠定了基礎。篩選出來的八個核心SSR標記和構建的無性系品種DNA指紋圖譜數(shù)據庫為茶樹品種鑒別提供了可靠的技術支持。
[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，

本文編號：1360525