發(fā)布時(shí)間：2018-01-02 08:35

  本文關(guān)鍵詞：稀有變異揭示TCP基因調(diào)控黃瓜卷須發(fā)育　出處：《中國農(nóng)業(yè)科學(xué)院》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 稀有變異 TCP轉(zhuǎn)錄因子 黃瓜卷須 同源器官

【摘要】：卷須是葫蘆科瓜類作物株型研究最重要的方面之一,也是研究植物向觸性運(yùn)動(dòng)的重要的植物模式器官。黃瓜的基因組、轉(zhuǎn)錄組和變異組的測(cè)序工作的完成,為系統(tǒng)研究黃瓜提供了大數(shù)據(jù)平臺(tái)支持。本課題搜集了世界各地3342份黃瓜種質(zhì)資源,發(fā)現(xiàn)唯一的一份無卷須的黃瓜資源CG9192,該種質(zhì)的卷須被側(cè)枝所替代,完全失去了攀援能力。我們推測(cè)該稀有表型是由稀有的遺傳變異引起的。通過基因組學(xué)、變異組學(xué)和轉(zhuǎn)錄組學(xué)的綜合分析,并應(yīng)用稀有遺傳變異的分析策略,我們發(fā)現(xiàn)了控制黃瓜卷須發(fā)育的身份基因CsTEN,并揭示了黃瓜卷須的同源器官是側(cè)枝—這一曾經(jīng)困擾達(dá)爾文的植物發(fā)育學(xué)謎團(tuán)。本論文的主要結(jié)果如下：1.通過對(duì)變態(tài)卷須種質(zhì)CG9192和正常卷須種質(zhì)404的表型觀察和掃描電鏡分析,發(fā)現(xiàn)側(cè)枝是黃瓜卷須的同源器官,遺傳學(xué)研究表明變態(tài)卷須性狀由一個(gè)隱性單基因控制(CsTEN)。2.稀有SNP在黃瓜四個(gè)亞群的分布不均勻,印度亞群擁有的稀有SNP數(shù)目最高,占所有稀有SNP數(shù)目的81%,印證了印度是黃瓜的起源中心。應(yīng)用稀有變異分析策略,結(jié)合轉(zhuǎn)錄組數(shù)據(jù),發(fā)現(xiàn)了變態(tài)卷須種質(zhì)CG9192的一個(gè)稀有SNP (rSNPl)對(duì)應(yīng)著一個(gè)在黃瓜卷須里特異表達(dá)的基因Csa5G644520,推測(cè)該基因可能是卷須的身份基因。3.構(gòu)建遺傳大群體,應(yīng)用圖位克隆的方法將候選基因CsTEN定位區(qū)間縮小至16Kb(區(qū)間內(nèi)只有一個(gè)基因),并通過與核心種質(zhì)的基因組重測(cè)序序列比對(duì),發(fā)現(xiàn)只有發(fā)現(xiàn)的rSNP1可以解釋變態(tài)卷須表型。針對(duì)rSNP1設(shè)計(jì)dCAPS標(biāo)記,檢測(cè)了世界范圍內(nèi)的、有卷須的1386份黃瓜種質(zhì),結(jié)果顯示該稀有變異僅存在于黃瓜種質(zhì)CG9192中,從而從遺傳學(xué)上充分證明rSNP1所對(duì)應(yīng)的基因Csa5G644520就是控制黃瓜卷須發(fā)育的基因CsTEN。4. CsTEN屬于TCP家族的CYC/TB1亞類,CsTEN上的稀有SNP發(fā)生在葫蘆科特有的保守結(jié)構(gòu)域,該結(jié)構(gòu)域不存在于已知的TCP蛋白中,該結(jié)構(gòu)域是進(jìn)化上新產(chǎn)生的一個(gè)結(jié)構(gòu)域；煙草和酵母中的實(shí)驗(yàn)表明,該稀有SNP顯著影響了CsTEN的轉(zhuǎn)錄激活活性。原位雜交分析進(jìn)一步表明該CsTEN在黃瓜卷須里特異表達(dá),是控制黃瓜卷須發(fā)育的身份基因。5.噴瓜是葫蘆科植物中唯一沒有卷須的植物,通過檢測(cè)葫蘆科卷須的身份基因TEN,發(fā)現(xiàn)噴瓜的TEN基因存在至少兩個(gè)以上的提前終止變異,使得噴瓜的TEN基因完全失去了功能。6.構(gòu)建了正常卷須和變態(tài)卷須的近等基因系,并將這對(duì)近等基因系進(jìn)行轉(zhuǎn)錄組測(cè)序,發(fā)現(xiàn)CsTEN抑制葉和花等的側(cè)生器官相關(guān)基因的表達(dá),同時(shí)促進(jìn)向觸性運(yùn)動(dòng)相關(guān)基因的表達(dá)。
[Abstract]:Tendril is one of the most important aspects of plant type research in cucurbitaceae, and it is also an important plant model organ to study the thixotropic movement of plants, and the complete sequencing of cucumber genome, transcriptome and variation group. This paper collected 3342 cucumber germplasm resources from all over the world and found the only cucumber CG9192 without tendrils. The tendrils of the germplasm were replaced by lateral branches and lost the ability to climb completely. We speculate that the rare phenotype is caused by rare genetic variation. Using the analysis strategy of rare genetic variation, we found the identity gene CsTEN that controls the development of tendrils in cucumber. It also reveals that the homologous organ of cucumber tendrils is lateral branches-a phytodevelopmental mystery that once troubled Darwin. The main results of this paper are as follows:. 1. Phenotypic observation and scanning electron microscope analysis of metamorphosed tendrils (CG9192) and normal tendrils (404). It was found that lateral branches were the homologous organs of cucumber tendrils. Genetic studies showed that metamorphosis of tendrils was controlled by a single recessive gene, CsTEN. 2. The distribution of rare SNP in four subpopulations of cucumber was not uniform. India subgroup has the highest number of rare SNP, accounting for 81% of all rare SNP, which confirms that India is the origin center of cucumber. A rare SNP rSNP of CG9192, a metamorphosis tendrils, was found to correspond to a gene Csa5G644520 specifically expressed in cucumber tendrils. It is speculated that this gene may be an identity gene of tendrils. A large genetic population was constructed, and the mapping method was used to narrow the mapping interval of candidate gene CsTEN to 16kb (there was only one gene in the region). The results showed that only the rSNP1 could explain the phenotype of abnormal tendrils. DCAPS markers were designed for rSNP1 and detected in the world. There were 1386 cucumber germplasms with tendrils. The results showed that the rare variation existed only in the cucumber germplasm CG9192. Therefore, it is fully proved from genetics that the gene Csa5G644520 corresponding to rSNP1 is the gene CsTEN.4which controls the development of tendrils in cucumber. CsTEN belongs to the CYC/TB1 subclass of the TCP family. Rare SNP on CsTEN occurs in the conserved domain of Cucurbitaceae, which does not exist in known TCP proteins. Experiments in tobacco and yeast showed that the rare SNP significantly affected the transcriptional activation activity of CsTEN. In situ hybridization analysis showed that the CsTEN was specifically expressed in the tendrils of cucumber. It is the identity gene that controls the development of cucumber tendrils. 5. Spray melon is the only plant without tendrils in Cucurbitaceae. The identity gene TEN of tendrils of Cucurbitaceae is detected. It was found that there were at least two early termination mutations in the TEN gene of Spoux melon, which completely lost the function of TEN gene. 6. The near-isogenic lines of normal tendrils and metamorphosed tendrils were constructed. CsTEN inhibited the expression of genes related to lateral organs in leaves and flowers, and promoted the expression of genes associated with thixotropism.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S642.2

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