本文關(guān)鍵詞：煙草鉀含量遺傳變異的全基因組關(guān)聯(lián)分析　出處：《中國農(nóng)業(yè)科學(xué)院》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 煙草 鉀 SSR標(biāo)記 關(guān)聯(lián)分析 優(yōu)異等位變異

【摘要】：煙草(Nicotiana.tabacum)是我國重要的葉用性經(jīng)濟(jì)作物,煙葉鉀含量的高低嚴(yán)重影響著煙葉經(jīng)濟(jì)價值的大小。與國外優(yōu)質(zhì)煙葉相比,我國煙葉鉀含量普遍偏低,嚴(yán)重制約了我國煙草產(chǎn)值的提高,也減緩了偏遠(yuǎn)山區(qū)農(nóng)民脫貧致富的步伐。針對我國煙葉鉀含量偏低問題,傳統(tǒng)的栽培施肥措施效果并不理想。因此,通過發(fā)掘煙草中與鉀吸收轉(zhuǎn)運相關(guān)基因,開發(fā)功能標(biāo)記,進(jìn)行分子標(biāo)記輔助選擇育種的辦法,達(dá)到提高煙葉鉀含量的目的。本研究在煙草核心種質(zhì)的基礎(chǔ)上篩選了96份烤煙材料構(gòu)建了試驗群體,簡稱群體Ⅰ。同時構(gòu)建了130份煙草材料的驗證群體,簡稱群體Ⅱ。利用均勻分布于全基因組上的454個SSR標(biāo)記對群體Ⅰ進(jìn)行遺傳多樣性分析和群體結(jié)構(gòu)分析,并基于標(biāo)記-性狀關(guān)聯(lián)分析的結(jié)果,篩選與鉀吸收轉(zhuǎn)運相關(guān)的SSR標(biāo)記和候選基因,構(gòu)建了一套高鉀煙草種質(zhì)的分子標(biāo)記輔助篩選體系。主要結(jié)果如下:(1)對不同環(huán)境下煙葉鉀含量的分析發(fā)現(xiàn),在相同年份中,西昌煙葉鉀含量顯著高于諸城。與此相反,西昌煙葉鉀含量的變異系數(shù)卻明顯小于諸城煙葉鉀含量的變異系數(shù)。這從側(cè)面上說明,西昌比諸城更具有生產(chǎn)優(yōu)質(zhì)烤煙的潛力。此外,在西昌和諸城兩地,國內(nèi)種質(zhì)與國外種質(zhì)在煙葉鉀含量上的表現(xiàn)無明顯差異。(2)利用均勻分布于24條連鎖群的454個SSR標(biāo)記對96份烤煙進(jìn)行全基因組掃描。對檢測到的等位變異頻率按5%的比例進(jìn)行稀有等位過濾。過濾后具有多態(tài)性的標(biāo)記為417個。檢測到的等位變異共1038個,平均為2.5個,變幅為2-6個。平均PIC值為0.299,變幅為0.044-0.698。(3)基于Nei氏遺傳距離進(jìn)行的NJ聚類分析表明,96份的烤煙群體可以分為三類。此外利用Structure軟件進(jìn)行的群體結(jié)構(gòu)分析同樣表明,該96份烤煙群體存在3個亞群。這進(jìn)一步說明該烤煙群體存在著三種遺傳背景。(4)采用關(guān)聯(lián)分析的方法,共發(fā)掘到-log P值≥2.00的標(biāo)記16個,-log P值最高的標(biāo)記為PT61367,達(dá)到4.46。其中PT51364、PT53362、PT55319這三個標(biāo)記的重復(fù)性較好,同時關(guān)聯(lián)到了兩個環(huán)境的表型。此外,利用16個標(biāo)記在煙草基因組數(shù)據(jù)庫中的定位結(jié)果,在標(biāo)記周圍共篩選到4個與煙草鉀吸收轉(zhuǎn)運相關(guān)的候選基因,分別為Ntab0898590、Ntab0163850、Ntab0171930、Ntab0562640。其中前兩個候選基因功能類似于鉀通道基因,后兩個候選基因功能類似于鉀轉(zhuǎn)運體基因。(5)對關(guān)聯(lián)分析得到的-log P值≥2.50的11個關(guān)聯(lián)標(biāo)記進(jìn)行高鉀含量優(yōu)異等位分析。通過對各標(biāo)記各等位位點間表型差異的顯著性檢驗和年點分布規(guī)律的統(tǒng)計,最終篩選出了5個SSR標(biāo)記用于構(gòu)建煙草高鉀品種分子標(biāo)記輔助篩選體系。5個SSR標(biāo)記為PT61367、PT51364、PT30132、PT55319、PT53362,其優(yōu)異等位分別為101、102、103、101103、101。此外在供試群體Ⅰ中發(fā)現(xiàn),供試材料含有這些標(biāo)記中優(yōu)異等位的數(shù)量越多,其鉀含量越高。用群體Ⅱ進(jìn)行驗證,得到了相同結(jié)果。說明這5個標(biāo)記可應(yīng)用于高鉀煙草種質(zhì)的分子標(biāo)記輔助選擇。
[Abstract]:Tobacco (Nicotiana.tabacum) is an important economic crop leaf, the potassium content in tobacco leaves seriously affects the economic value. Compared with foreign high-quality tobacco, China's tobacco leaf potassium content is generally low, which seriously restricts the improvement of tobacco output, also slowed the pace of poverty alleviation in the remote mountainous areas aiming at the problem of our country. The low potassium content in tobacco leaves, the effect of the traditional measures of cultivation and fertilization is not ideal. Therefore, absorption and transport related genes and potassium in tobacco by exploring the development, functional marker, molecular marker assisted selection breeding method to improve the potassium content of tobacco leaf. This research is based on the selection of core tobacco germplasm 96 copies of the test material in the construction of tobacco group, referred to as group 1. Also established 130 tobacco materials verification group, referred to as group II. Using a uniform distribution throughout the genome of the 454 A SSR marker of group I analyzed genetic diversity and population structure analysis, and based on the marker trait association analysis, screening and potassium uptake transporter related SSR markers and candidate genes, construct a set of high potassium of Tobacco Germplasm by molecular marker assisted selection system. The main results are as follows: (1) analysis of tobacco the content of potassium in different environment, in the same year, Xichang tobacco potassium content was significantly higher than that in Zhucheng. On the contrary, the coefficient of variation of Xichang tobacco potassium content was significantly less than Zhucheng tobacco leaf potassium content. From the side of that, Xichang has more than Zhucheng flue-cured tobacco production potential. In addition, in both Xichang and Zhucheng, no significant difference between the performance of domestic and foreign germplasm Germplasm in the potassium content in tobacco leaves on. (2) using 454 SSR markers evenly distributed in 24 linkage groups of 96 samples of Flue-cured Tobacco by whole genome scanning. The detected allele frequency of rare alleles were filtered according to the proportion of 5%. After filtration markers with polymorphism is 417. The detected alleles were 1038, the average is 2.5, the range was 2-6. The average PIC value was 0.299, ranged from 0.044-0.698. (3) analysis showed that NJ Nei's genetic distance clustering based on 96 groups of flue-cured tobacco can be divided into three categories. The population structure analysis using Structure software also shows that the 96 flue-cured tobacco group has 3 subgroups. This further indicates that the flue-cured tobacco group there are three kinds of genetic background. (4) mining method by correlation analysis the total excavation to the -log P value is more than 2 of the 16 markers, -log P is the highest marker for PT61367, 4.46. PT51364 PT53362, which reached PT55319, repeatability of these three markers is good, also related to the phenotype of the two environments. In addition, the use of 16 gene markers in tobacco The positioning results group in the database, the markings around were screened and 4 candidate genes related to tobacco potassium uptake, respectively Ntab0898590, Ntab0163850, Ntab0171930, Ntab0562640.. Two of the candidate genes similar to potassium channel gene, two candidate gene function similar to potassium transporter gene (5) on. Correlation analysis of obtained -log P for high potassium content of excellent analysis value of the 11 markers associated with = 2.50. Statistical significant test for differences in phenotype by each marker alleles between the years and point distribution, finally we selected 5 SSR markers were used to construct tobacco high potassium varieties of molecular marker assisted selection.5 SSR marked PT61367, PT51364, PT30132, PT55319, PT53362, the superior allele was 101102103101103101 in the test group I found in the tested materials containing these markers in the superior allele The higher the number, the higher the potassium content. The same results were obtained by group II. These 5 markers can be applied to marker assisted selection of high potassium tobacco germplasm.

【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S572

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