發(fā)布時(shí)間：2018-01-12 15:30

  本文關(guān)鍵詞：梅花高密度遺傳圖譜構(gòu)建及部分觀賞性狀QTL分析　出處：《北京林業(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： 梅花 SLAF-seq 遺傳連鎖圖譜 QTL分析 垂枝 精細(xì)定位

【摘要】：梅花(Prunus mume Sieb. et Zucc.)是中國傳統(tǒng)名花,具有重要的文化及經(jīng)濟(jì)價(jià)值。培育集合多種優(yōu)良觀賞性狀的新品種是梅花產(chǎn)業(yè)發(fā)展的關(guān)鍵,目前梅花新品種育種工作主要采用傳統(tǒng)雜交育種途徑,然而木本植物童期較長,此外觀賞性狀遺傳機(jī)理復(fù)雜,直接找到控制這些性狀的關(guān)鍵基因十分困難。通過構(gòu)建高密度遺傳連鎖圖譜及QTL定位分析,是找到控制復(fù)雜性狀的關(guān)鍵遺傳因子和緊密連鎖分子標(biāo)記的重要方法,但其在梅花中研究較少。為了挖掘控制梅花花色、花型、花瓣數(shù)、垂枝等性狀的遺傳因子及候選基因,本研究利用性狀分離良好的‘六瓣’ב粉臺(tái)垂枝’F1群體為試驗(yàn)材料,通過SLAF-seq技術(shù)進(jìn)行大規(guī)模分子標(biāo)記開發(fā)并構(gòu)建了梅花高密度遺傳連鎖圖譜,然后對F1群體這些觀賞性狀進(jìn)行QTL分析及候選基因的挖掘。由于垂枝性狀為木本植物特有性狀,后續(xù)重點(diǎn)對其進(jìn)行了精細(xì)定位及候選基因的挖掘。主要結(jié)論如下：(1)通過梅花雜交試驗(yàn)獲得了在株型、花色、花型均有明顯分離的F1作圖群體‘六瓣’ב粉臺(tái)垂枝’,該群體大小為387。在此基礎(chǔ)上,利用SLAF-seq技術(shù)對梅花進(jìn)行全基因組范圍分子標(biāo)記的開發(fā),構(gòu)建了梅花標(biāo)記密度最大的一張遺傳連鎖圖譜,共有8條連鎖群,包含遺傳標(biāo)記8,007個(gè),標(biāo)記間平均遺傳距離為0.195 cM。其中2號(hào)連鎖群上標(biāo)記數(shù)量最多為1,722個(gè),遺傳距離為263.84cM,6號(hào)連鎖群上標(biāo)記數(shù)量為698個(gè),遺傳距離為142.48cM�？倛D距為1,550.62 cM,覆蓋梅花基因組64.31%,最終圖譜上的SLAF標(biāo)記在F1作圖群體中的平均完整度為96%。(2)基于構(gòu)建的梅花高密度遺傳連鎖圖譜,采用復(fù)合區(qū)間作圖法對梅花‘六瓣’ב粉臺(tái)垂枝’F1群體15個(gè)生長、株型、花部相關(guān)重要性狀進(jìn)行了QTL定位分析。共檢測到66個(gè)QTLs位點(diǎn),利用梅花基因組注釋信息篩選出58個(gè)可能的候選基因。(3)對垂枝梅與直枝梅枝條不同部位及發(fā)育階段細(xì)胞結(jié)構(gòu)、木材成分以及激素水平3個(gè)生理層面的差異進(jìn)行了分析。發(fā)現(xiàn)垂枝梅與直枝梅在枝條近遠(yuǎn)軸面細(xì)胞排列、木質(zhì)化程度均存在差異,且在垂枝梅枝條木質(zhì)部細(xì)胞中存在淀粉體的缺失；木質(zhì)素含量測定結(jié)果表明直枝梅枝條近軸面木質(zhì)素含量比遠(yuǎn)軸面低,垂枝枝條近軸面木質(zhì)素含量比遠(yuǎn)軸面高,田間觀察發(fā)現(xiàn)梅花枝條生長初期并未表現(xiàn)出明顯的垂枝表型,而在木質(zhì)化過程中垂枝表型越來越明顯,推測垂枝性狀形成可能與枝條木質(zhì)化過程的差異有關(guān)；采用液相質(zhì)譜的方法對5個(gè)發(fā)育階段垂枝梅和直枝梅枝條不同部位激素水平進(jìn)行比較分析,推測GA3可能與垂枝表型形成有關(guān)。(4)根據(jù)垂枝性狀遺傳分析結(jié)果推測梅花垂枝性狀可能由一個(gè)主效基因和一個(gè)或者多個(gè)微效基因共同控制。在此基礎(chǔ)上采用三種統(tǒng)計(jì)分析方法將垂枝性狀定位到梅花7號(hào)染色體10.54Mb-11.68Mb區(qū)域,同時(shí)10個(gè)SLAF分子標(biāo)記被認(rèn)為與梅花垂枝性狀緊密連鎖。在前期生理試驗(yàn)結(jié)果的基礎(chǔ)上對垂枝候選區(qū)域進(jìn)行候選基因篩選。最終,9個(gè)可能與枝條木質(zhì)化相關(guān),參與細(xì)胞壁及纖維素合成與降解的結(jié)構(gòu)基因/酶類,以及9個(gè)被預(yù)測參與基因的轉(zhuǎn)錄調(diào)控的基因被推測與梅花垂枝性狀形成相關(guān)。本研究構(gòu)建的梅花高密度遺傳連鎖圖譜將為后續(xù)重要性狀的定位奠定基礎(chǔ),15個(gè)梅花重要性狀QTL分析對指導(dǎo)這些復(fù)雜性狀的分子育種具有重要意義,而垂枝性狀的精細(xì)定位將為后續(xù)找到控制垂枝性狀的基因奠定基礎(chǔ),同時(shí)為木本植物其他重要性狀的定位提供一種研究策略。
[Abstract]:Plum (Prunus mume Sieb. et Zucc.) is China traditional flowers, has an important cultural and economic value. To cultivate new varieties of various sets of excellent ornamental traits is the key of plum industry development, the plum breeding of new varieties by traditional breeding work, but woody plant children over a long period of time, the ornamental characters of complex genetic mechanism direct, find the key genes controlling these traits is very difficult. The high density genetic linkage map and QTL mapping analysis of the construction, is an important method to find the key genetic factors controlling complex traits and molecular markers closely linked, but less research. In order to control mining in plum flower color, flower type, flower number, genetic factor and candidate gene pendula traits, this study uses characters of good separation "six flap '*' Pink Taiwan branches' F1 group as the experimental material, by SLAF-seq The technology for large-scale development of molecular markers and genetic linkage map of plum high density was established, and then the F1 group of these ornamental characters of mining and QTL analysis of the candidate genes. Because of the weeping traits of woody plant traits, the follow-up key has carried on the fine mapping and candidate gene mining. The main conclusions are as follows: (1) the plum hybridization experiments obtained in plant type, color, F1 mapping population 'patterns are obviously separated with six' x 'powder Taiwan branches', the population size of 387. on the basis of this, the development of molecular markers for genome wide Meihua using SLAF-seq technology, one of the largest plum marker density genetic linkage to build a map, a total of 8 linkage groups, including 8007 genetic markers, markers between the average genetic distance of 0.195 cM. markers in 2 linkage groups, including the number of up to 1722, the genetic distance was 263. 84cM, No. 6 linkage group mark number is 698, the genetic distance of 142.48cM. total length is 1550.62 cM, 64.31% SLAF markers covering the plum genome, the final map on average in F1 mapping population in the degree of 96%. (2) to construct the plum high density genetic linkage map based on composite interval mapping method of plant type on the "six petal plum '*' powder Taiwan branches' F1 Group 15 growth, flower related traits were important for QTL positioning analysis. 66 QTLs loci were detected by plum genome annotation screened 58 candidate genes. (3) the different parts of the vertical and straight plum plum branches and the developmental stages of cell structure, the difference of wood components and hormone level 3 physiological level were analyzed. Found weeping plum and plum branches in nearly straight abaxial cell arrangement, there are differences in the degree of lignification, and vertical plum branches of wood There is lack of starch cells; the determination results of lignin content shows that the straight plum branches adaxial than abaxial surface of the lignin content is low, the weeping branches of the adaxial surface lignin content than the abaxial surface of high field observation found plum branch growth early shows no significant branches in phenotype. In lignification of pendula phenotype is more and more obvious, the difference that branches and branches may form characters of the lignification process; using the method of liquid chromatography mass spectrometry on the 5 developmental stages of vertical and straight plum plum branches in different parts of hormone levels were compared, suggesting that GA3 may from the weeping phenotype formation. (4) according to the weeping genetic analysis results suggest that characteristics of Prunus mume could be controlled by a major gene and one or more minor genes. On the basis of using three methods of statistical analysis will be weeping traits to Mei Take the 10.54Mb-11.68Mb region on chromosome 7, and 10 SLAF markers were considered closely linked with characteristics of Prunus mume. Based on the previous physiological test results on the vertical branch candidate region candidate gene screening. Finally, 9 possible branches and lignification related genes involved in cell wall structure, / and cellulose synthesis and degradation enzymes, as well as 9 are predicted to be involved in transcriptional regulation gene presumably associated with characteristics of Prunus mume. The plum high density genetic linkage map will lay the foundation for the subsequent location of important traits, 15 plum blossom shaped QTL importance to guide the analysis of molecular breeding of these complex traits has important significance, and fine mapping of weeping traits will find the control for the subsequent pendula gene lay the foundation, at the same time as other important traits of woody plants provide a research strategy.

【學(xué)位授予單位】：北京林業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S685.17
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本文編號(hào)：1414863