發(fā)布時(shí)間：2018-05-19 04:05

  本文選題：煙草 + 青枯病�。� 參考：《廣州大學(xué)》2017年碩士論文

【摘要】：煙草青枯病是一種在我國(guó)南方各大煙區(qū)嚴(yán)重影響煙草生長(zhǎng)導(dǎo)致產(chǎn)量和品質(zhì)下降的主要病害,煙草青枯病抗性與一些農(nóng)藝性狀是考察和評(píng)價(jià)煙草生長(zhǎng)、產(chǎn)量和品質(zhì)的重要指標(biāo)。眾所周知,煙草青枯病抗性和多數(shù)農(nóng)藝性狀均屬于數(shù)量性狀遺傳,受多基因共同控制,且易受到環(huán)境影響;而找出與青枯病抗性及一些農(nóng)藝性狀密切相關(guān)的分子標(biāo)記,開展分子標(biāo)記輔助選擇(Marker-assisted selection,MAS),可加速煙草優(yōu)良品種的選育進(jìn)程。因此,本研究利用2個(gè)煙草親本(父本“118-3”,母本“粵煙98”)建立的F7代重組自交系(Recombinant inbred line,RIL)群體為材料,采用SSR標(biāo)記構(gòu)建煙草遺傳圖譜;在此基礎(chǔ)上,定位出煙草青枯病抗性和一些農(nóng)藝性狀的數(shù)量性狀基因座(Quantitative trait locus,QTL)。主要研究結(jié)果如下:1、建立了一種簡(jiǎn)捷快速檢測(cè)簡(jiǎn)單重復(fù)序列(Simple repeated sequence,SSR)標(biāo)記的聚丙烯酰胺凝膠銀染法。聚丙烯酰胺銀染法是一種具有高靈敏度檢測(cè)具有幾個(gè)堿基差異的SSR標(biāo)記的方法,針對(duì)傳統(tǒng)銀染方法中操作繁瑣、費(fèi)時(shí)和所需試劑種類較多等不足,在前人所建立的銀染方法的基礎(chǔ)上,本研究建立了只有染色和顯色兩個(gè)主要步驟,只需硝酸銀、甲醛和氫氧化鈉三種試劑,整個(gè)銀染過(guò)程僅需時(shí)6~7分鐘的一種簡(jiǎn)捷快速檢測(cè)SSR的聚丙烯酰胺凝膠銀染法;此外,該銀染法檢測(cè)SSR標(biāo)記擴(kuò)增產(chǎn)物的DNA條帶還具有顯色強(qiáng)度高、對(duì)比度大、分辨力高和重現(xiàn)性好等特點(diǎn),適用于大規(guī)模SSR標(biāo)記的檢測(cè),且已應(yīng)用于本研究煙草SSR標(biāo)記的檢測(cè)工作中。2、利用“118-3”和“粵煙98”為親本的RIL群體作材料,構(gòu)建了煙草遺傳圖譜。從1,405對(duì)SSR標(biāo)記中篩選出214對(duì)在兩親本材料中具有多態(tài)性SSR標(biāo)記,并利用這些標(biāo)記對(duì)RIL群體進(jìn)行基因分型。根據(jù)基因分型數(shù)據(jù),運(yùn)用Map Maker 2.0軟件構(gòu)建出一張具有24個(gè)連鎖群的煙草遺傳圖譜。圖譜總長(zhǎng)度達(dá)到3,126.10 cM,各連鎖群長(zhǎng)度在48.51~292.04 cM之間,平均為130.25 cM;該圖譜含有426個(gè)SSR等位變異位點(diǎn),各連鎖群上的等位變異位點(diǎn)數(shù)在6~38個(gè)之間,平均圖距為7.34 cM。此遺傳連鎖圖譜可用于煙草青枯病抗性和農(nóng)藝性狀的QTL定位。3、定位出煙草青枯病抗性和一些農(nóng)藝性狀的QTLs。測(cè)定了種植在廣東省煙草南雄科學(xué)研究所試驗(yàn)田RIL群體的表型性狀,這些表型性狀為青枯病抗性、莖圍、節(jié)距、SPAD值、最大上部葉葉長(zhǎng)、最大上部葉葉寬、最大上部葉葉重、最大中部葉葉長(zhǎng)、最大中部葉葉寬、最大中部葉葉重、最大下部葉葉長(zhǎng)、最大下部葉葉寬和最大下部葉葉重。根據(jù)RIL群體各個(gè)表型性狀和基因分型數(shù)據(jù),利用MapQTL 5.0軟件的多QTL模型作圖法(Multiple QTL mapping,MQM)對(duì)13個(gè)性狀進(jìn)行QTL定位,共探測(cè)到14個(gè)QTLs;其中2個(gè)QTLs與煙草青枯病抗性相關(guān),5個(gè)QTLs與莖圍相關(guān),與最大中部葉葉重相關(guān)的QTLs 2個(gè),與最大中部葉葉長(zhǎng)、最大下部葉葉重、最大下部葉葉寬、最大上部葉葉長(zhǎng)以及節(jié)距相關(guān)的QTL各1個(gè)。另外,遺傳圖譜中一些區(qū)域存在不同性狀QTLs的重疊現(xiàn)象。
[Abstract]:Tobacco blight is a major disease which seriously affects the yield and quality of tobacco growth in the large tobacco areas of the south of China. The resistance of tobacco blight and some agronomic traits are important indexes to investigate and evaluate the growth, yield and quality of tobacco. It is known that the resistance of tobacco blight and most agronomic traits are quantitative traits. Heredity, controlled by multiple genes and easy to be affected by the environment, and identifying molecular markers that are closely related to the resistance of bacterial wilt and some agronomic traits and carrying out molecular marker assisted selection (Marker-assisted selection, MAS) can accelerate the breeding process of fine tobacco varieties. Therefore, this study uses 2 tobacco parents (male parent "118-3", mother). The F7 generation Recombinant inbred line (RIL) group established by "Guangdong Tobacco 98" was used as the material to construct the genetic map of tobacco using SSR markers. On this basis, the quantitative trait loci (Quantitative trait locus, QTL) of tobacco bacterial wilt resistance and some agronomic traits were located. The main results were as follows: 1, a simple method was established. Polyacrylamide gel silver staining with simple repeat sequence (Simple repeated sequence, SSR) is fast detection. The polyacrylamide silver staining method is a method with high sensitivity to detect several base differences, and it has many shortcomings in the traditional silver staining method, such as tedious operation, time consuming and many kinds of reagents needed. On the basis of the silver staining method established, this study established only two main steps of dyeing and coloring, only three kinds of reagent, only silver nitrate, formaldehyde and sodium hydroxide, and a simple and rapid detection of SSR polyacrylamide gel silver staining in the whole process of silver staining with only 6~7 minutes. In addition, the silver staining method was used to detect the DNA of the amplified products of SSR. The strip also has the characteristics of high color intensity, high contrast, high resolution and good reproducibility. It is suitable for the detection of large scale SSR markers and has been applied to the detection work of tobacco SSR markers in this study. The genetic map of tobacco was constructed by using "118-3" and "Guangdong Tobacco 98" as parent, and the genetic map of tobacco was screened from 1405 SSR markers. Select 214 pairs of polymorphic SSR markers in two parent materials and use these markers to genotyping RIL populations. According to the genotyping data and using Map Maker 2 software, a tobacco genetic map with 24 linkage groups was constructed. The total length of the map was 3126.10 cM and the length of each linkage group was between 48.51~292.04 cM and average. 130.25 cM, the atlas contains 426 SSR alleles, and the number of heterotopic points on each linkage group is between 6~38, and the average distance is 7.34 cM.. The genetic linkage map can be used for QTL positioning.3 for tobacco bacterial wilt resistance and agronomic traits. The QTLs. of tobacco bacterial wilt resistance and some agronomic traits is determined to be planted in Guangdong. The phenotypic traits of the RIL population in the experimental field of the Nanxiong Institute of tobacco science are resistant to bacterial wilt, the stem circumference, the pitch, the SPAD value, the maximum upper leaf leaf length, the maximum upper leaf blade width, the maximum upper leaf leaf weight, the largest middle leaf leaf length, the largest middle leaf width, the largest central leaf leaf weight, the largest lower leaf leaf length and the largest lower Ye Yekuan. According to the phenotypic and genotyping data of the RIL population, using the multiple QTL model mapping of MapQTL 5 software (Multiple QTL mapping, MQM) to locate the 13 character QTL, and detect 14 QTLs, of which 2 QTLs are related to the resistance of Tobacco Wilt, 5 QTLs and the stem circumference, and the largest central Ye Yezhong. Related QTLs 2, with the largest central leaf leaf length, the maximum lower leaf leaf weight, the maximum lower leaf blade width, the maximum upper Ye Yechang and the pitch related QTL each. In addition, some regions of the genetic map overlap with different characters of QTLs.
【學(xué)位授予單位】：廣州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S435.72

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李海洋;李榮華;夏巖石;袁清華;張振臣;趙偉才;郭培國(guó);;基于RAD-seq數(shù)據(jù)開發(fā)煙草多態(tài)性SSR標(biāo)記[J];中國(guó)煙草科學(xué);2018年01期

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本文編號(hào)：1908625