本文關(guān)鍵詞：茶樹(shù)遺傳圖譜與品種指紋圖譜構(gòu)建及幾個(gè)重要性狀的QTL定位　出處：《四川農(nóng)業(yè)大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 茶樹(shù) SSR標(biāo)記 遺傳圖譜 數(shù)量性狀位點(diǎn) 品種指紋圖譜

【摘要】：茶是世界范圍內(nèi)最受歡迎的飲料之一,越來(lái)越多的證據(jù)表明飲茶可以改善人類健康。從20世紀(jì)60年代以來(lái),世界茶葉的消費(fèi)量一直保持高速增長(zhǎng),茶產(chǎn)品類型日益豐富,茶樹(shù)的栽培面積和茶葉產(chǎn)量也隨之大幅增長(zhǎng)。優(yōu)良的茶樹(shù)品種是茶葉生產(chǎn)中最關(guān)鍵的物質(zhì)資料,21世紀(jì)多元化的市場(chǎng)需求為茶樹(shù)良種選育工作帶來(lái)了新的挑戰(zhàn)。但由于其基因組復(fù)雜、育種周期長(zhǎng)、多數(shù)經(jīng)濟(jì)性狀為多基因控制的數(shù)量性狀,茶樹(shù)的遺傳研究和優(yōu)質(zhì)品種選育均較為困難�；贒NA分子標(biāo)記的遺傳圖譜是研究數(shù)量性狀的有效工具,在此基礎(chǔ)上發(fā)展的標(biāo)記輔助選擇(marker assisted selection, MAS)育種技術(shù)可以大幅提高多年生作物的育種效率。本論文首先通過(guò)開(kāi)發(fā)篩選大量簡(jiǎn)單序列重復(fù)(simple sequence repeat,SSR)標(biāo)記,基于170株‘龍井43’ב白毫早’雜交一代(F1)作圖群體,構(gòu)建起一張高密度的茶樹(shù)遺傳圖譜。接著,連續(xù)兩年觀測(cè)或測(cè)定該作圖群體多個(gè)重要性狀的表型數(shù)據(jù),利用所構(gòu)建的遺傳圖譜進(jìn)行數(shù)量性狀位點(diǎn)定位分析(quantitative trait loci mapping, QTL mapping),得到了40余個(gè)QTL。另外,本論文還從大量已定位到遺傳圖譜的標(biāo)記中篩選出八個(gè)核心SSR標(biāo)記用于構(gòu)建中國(guó)主要無(wú)性系茶樹(shù)品種的DNA分子指紋圖譜。取得的主要研究結(jié)果如下：1.為獲得足夠的SSR分子標(biāo)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(gè)SSR標(biāo)記,其中460個(gè)是根據(jù)茶樹(shù)轉(zhuǎn)錄組序列新開(kāi)發(fā)而成,另外251個(gè)是從其他兩張茶樹(shù)遺傳圖譜上轉(zhuǎn)移而來(lái)。從這些標(biāo)記中共獲得342個(gè)(48.1%)在‘龍井43’ב白毫早’作圖群體中有多態(tài)性的SSR標(biāo)記,其中206個(gè)為本研究新開(kāi)發(fā)。2.用上述342個(gè)多態(tài)性標(biāo)記對(duì)‘龍井43’ב白毫早’作圖群體的170單株代進(jìn)行基因型分析。得到的基因型數(shù)據(jù)和已有的155個(gè)SSR標(biāo)記的數(shù)據(jù)合并,用于構(gòu)建遺傳圖譜。新構(gòu)建的雙親整合圖譜含有483個(gè)SSR標(biāo)記,分布在15條連鎖群上,總遺傳圖距為1226.2 cM,相鄰標(biāo)記間平均間距為2.5 cM�；凇半p假側(cè)交”作圖理論分別構(gòu)建了兩個(gè)雜交親本各自的遺傳圖譜。母本圖譜(‘龍井43’)含有323個(gè)標(biāo)記,總圖距為1032.2 cM；父本圖譜(‘白毫早’)含有303個(gè)標(biāo)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(diǎn)。3.新建的‘龍井43’ב白毫早’雙親整合圖譜上有126個(gè)SSR標(biāo)記是與其他兩張茶樹(shù)圖譜共享的標(biāo)記位點(diǎn)�；谶@些共享位點(diǎn),建立起了三張茶樹(shù)遺傳圖譜15條連鎖群的對(duì)應(yīng)關(guān)系。來(lái)自不同圖譜對(duì)應(yīng)連鎖群的遺傳圖距、共享標(biāo)記的位置基本一致,為比較、驗(yàn)證不同作圖群體中得到的QTL位點(diǎn)奠定了基礎(chǔ)。4.連續(xù)兩年(2014和2015)觀測(cè)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(zhǎng)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(shí)用高效液相色譜測(cè)定了該作圖群體兩年夏季新梢中七個(gè)兒茶素單體(catechines)和咖啡堿(caffeine, CAF)的含量。用上述表型數(shù)據(jù)結(jié)合所構(gòu)建的遺傳圖譜定位到43個(gè)QTL,其中2個(gè)與TBF相關(guān)、5個(gè)與YSC相關(guān)、5個(gè)與MLL相關(guān)、3個(gè)與LSI相關(guān)、2個(gè)與咖啡堿含量相關(guān)、20個(gè)分別與七個(gè)兒茶素組分含量相關(guān),還有6個(gè)與酯型兒茶素與非酯型兒茶素比值相關(guān)。共有14個(gè)QTL的LOD值達(dá)到全基因組極顯著水平(P0.01),且在兩年的數(shù)據(jù)中均檢測(cè)到。5.為了將SSR標(biāo)記應(yīng)用于茶樹(shù)無(wú)性系品種鑒定,從已定位到遺傳圖譜的483個(gè)SSR標(biāo)記中篩選出30個(gè)高質(zhì)量、相互獨(dú)立的SSR標(biāo)記,結(jié)合高分辨率毛細(xì)管電泳檢測(cè)技術(shù),分析了128個(gè)中國(guó)主栽的無(wú)性系茶樹(shù)品種。結(jié)果表明這些標(biāo)記具有很高的多態(tài)性,平均每個(gè)位點(diǎn)可以檢測(cè)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(jì)顯示這些標(biāo)記也具有很強(qiáng)的品種辨別能力,根據(jù)四個(gè)位點(diǎn)的基因型即可完全區(qū)分這128個(gè)茶樹(shù)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(jìn)一步篩選出八個(gè)核心SSR標(biāo)記用于構(gòu)建茶樹(shù)無(wú)性系品種DNA指紋圖譜。6.基于上述30個(gè)SSR位點(diǎn)的基因型數(shù)據(jù),分析了128個(gè)無(wú)性系茶樹(shù)品種間可能存在的親子關(guān)系。結(jié)果共鑒定出47對(duì)可能的親子關(guān)系,其中33對(duì)與育種記錄相符,另外14對(duì)為首次發(fā)現(xiàn)。本論文所得到的SSR標(biāo)記、遺傳圖譜和QTL位點(diǎn)為茶樹(shù)功能基因鑒定、MAS育種和其他性狀的QTL定位奠定了基礎(chǔ)。篩選出來(lái)的八個(gè)核心SSR標(biāo)記和構(gòu)建的無(wú)性系品種DNA指紋圖譜數(shù)據(jù)庫(kù)為茶樹(shù)品種鑒別提供了可靠的技術(shù)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(hào)：1360525