烟草遗传图谱的构建及青枯病抗性与一些农艺性状的QTL定位

发布时间：2018-05-19 04:05

本文选题：烟草 + 青枯病　；参考：《广州大学》2017年硕士论文

【摘要】：烟草青枯病是一种在我国南方各大烟区严重影响烟草生长导致产量和品质下降的主要病害,烟草青枯病抗性与一些农艺性状是考察和评价烟草生长、产量和品质的重要指标。众所周知,烟草青枯病抗性和多数农艺性状均属于数量性状遗传,受多基因共同控制,且易受到环境影响;而找出与青枯病抗性及一些农艺性状密切相关的分子标记,开展分子标记辅助选择(Marker-assisted selection,MAS),可加速烟草优良品种的选育进程。因此,本研究利用2个烟草亲本(父本“118-3”,母本“粤烟98”)建立的F7代重组自交系(Recombinant inbred line,RIL)群体为材料,采用SSR标记构建烟草遗传图谱;在此基础上,定位出烟草青枯病抗性和一些农艺性状的数量性状基因座(Quantitative trait locus,QTL)。主要研究结果如下:1、建立了一种简捷快速检测简单重复序列(Simple repeated sequence,SSR)标记的聚丙烯酰胺凝胶银染法。聚丙烯酰胺银染法是一种具有高灵敏度检测具有几个碱基差异的SSR标记的方法,针对传统银染方法中操作繁琐、费时和所需试剂种类较多等不足,在前人所建立的银染方法的基础上,本研究建立了只有染色和显色两个主要步骤,只需硝酸银、甲醛和氢氧化钠三种试剂,整个银染过程仅需时6~7分钟的一种简捷快速检测SSR的聚丙烯酰胺凝胶银染法;此外,该银染法检测SSR标记扩增产物的DNA条带还具有显色强度高、对比度大、分辨力高和重现性好等特点,适用于大规模SSR标记的检测,且已应用于本研究烟草SSR标记的检测工作中。2、利用“118-3”和“粤烟98”为亲本的RIL群体作材料,构建了烟草遗传图谱。从1,405对SSR标记中筛选出214对在两亲本材料中具有多态性SSR标记,并利用这些标记对RIL群体进行基因分型。根据基因分型数据,运用Map Maker 2.0软件构建出一张具有24个连锁群的烟草遗传图谱。图谱总长度达到3,126.10 cM,各连锁群长度在48.51~292.04 cM之间,平均为130.25 cM;该图谱含有426个SSR等位变异位点,各连锁群上的等位变异位点数在6~38个之间,平均图距为7.34 cM。此遗传连锁图谱可用于烟草青枯病抗性和农艺性状的QTL定位。3、定位出烟草青枯病抗性和一些农艺性状的QTLs。测定了种植在广东省烟草南雄科学研究所试验田RIL群体的表型性状,这些表型性状为青枯病抗性、茎围、节距、SPAD值、最大上部叶叶长、最大上部叶叶宽、最大上部叶叶重、最大中部叶叶长、最大中部叶叶宽、最大中部叶叶重、最大下部叶叶长、最大下部叶叶宽和最大下部叶叶重。根据RIL群体各个表型性状和基因分型数据,利用MapQTL 5.0软件的多QTL模型作图法(Multiple QTL mapping,MQM)对13个性状进行QTL定位,共探测到14个QTLs;其中2个QTLs与烟草青枯病抗性相关,5个QTLs与茎围相关,与最大中部叶叶重相关的QTLs 2个,与最大中部叶叶长、最大下部叶叶重、最大下部叶叶宽、最大上部叶叶长以及节距相关的QTL各1个。另外,遗传图谱中一些区域存在不同性状QTLs的重叠现象。
[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.
【学位授予单位】：广州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.72

【引证文献】