玉米自交系遗传多样性与表型性状关联分析及分子ID构建
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摘要
研究玉米自交系遗传多样性、划分玉米杂种优势类群及杂种优势模式,应用于玉米自交系的选育、改良、创新及优异种质资源挖掘、利用,可提高组配玉米杂交种的效率、缩短育种年限,是玉米育种工作中的重要基础性研究;玉米重要性状QTL定位是利用分子标记辅助选择的前提条件,对于提高育种准确性和预见性有重要意义。
本论文研究目的是以我国部分重要玉米自交系、大面积推广杂交种的亲本、自选自交系等为研究对象,基于遗传距离划分优良玉米种质杂种优势群,探讨杂种优势模式。利用257份玉米自交系构成的自然群体为供试材料,选取64对核心SSR标记进行遗传多样性分析,应用STRUCTRE软件对257份玉米自交系构成的自然群体进行基于数学模型的类群划分,并计算材料相应的Q值(第i材料其基因组变异源于第k群体的概率),利用TASSEL软件中的GLM(General Linear Mode)程序,各个体的Q值作为协变量,将农艺性状表型数据对标记逐一进行回归分析,确认各性状关联位点并计算位点对表型的解释率,采用资源特征分析软件(ID Analysis1.0,软件登记号:2007SR11870)建立品种分子ID,以最少的引物来区分品种。本论文主要结论如下:
(1)目前,选育高产玉米自交系应以尽量延长生育期为主,增加穗长及百粒重,以其他性状适中为选择目标。选育品质优良的玉米自交系应定向选择穗粗与株高较佳的种质,其次为生育期、行粒数与穗粗,平衡其他农艺性状的选择,根据品质性状及与其相关性状之间的关系选用适宜的种质,协调所有影响因素,加快选育优良玉米自交系的进程。
(2)基于SSR标记的遗传距离,将供试材料划分为PA、BSSS、旅大红骨、塘四平头、Lancaster、PB六个类群。聚类结果与自交系系谱关系基本一致。本研究中Lancaster可定为A群,塘四平头与旅大红骨可归为B群,PA、PB可归为C群即PN类群,而BSSS类群由于与其他类群差异较大归为D类群,这样可将供试材料归为4大类群,共6个亚群。我国玉米种质资源遗传基础相对狭窄,因此简化类群划分数目,有利于探索杂种优势模式。核心SSR标记对于划分杂种优势群有显著的鉴别能力。
(3)64个SSR位点中,有43个标记位点与24个性状相关联,其中19个位点的变异分别与14个表型性状在P<0.01水平上呈极显著相关,其它标记与各性状均显著相关(P<0.05);与农艺、品质性状相关联的位点(次)累计有80个,与品质性状相关联的位点(次)累计有7个,关联位点数目上品质性状远少于其他农艺性状;关联分析发现的主要性状SSR位点数目远多于相同大小家系群体定位所得位点数,同一性状的关联位点在连锁群上分布较集中,如与生育期关联的位点多分布在Ch7群体上,这可能与此连锁群上使用标记较多有关;同一个位点与多个性状关联很普遍,这些性状一般为同一类性状,如umc1741与株高、全株叶数、穗位及穗长关联,Ch7连锁群上的umc2160、umc1545与生育期关联同时也与穗行数相关联,这说明玉米表型性状相关有其内在遗传因素。综合分析表明:与株高、穗位、穗长、穗行数等性状相关联的标记较多,与脂肪含量、蛋白质含量等性状相关联的标记较少,应该在试验群体内增加品质性状变异程度较大的试验材料,同一条染色体上存在一个或多个与农艺性状关联的标记位点,不同染色体上存在与同一性状关联的标记,这与玉米数量性状的QTL定位大体一致。
(4)通过SSR分子标记技术,采用资源特征分析软件(ID Analysis 1.0)对257份玉米自交系进行分析表明,仅需10对引物可将供试品种完全区分开,用这10对引物共同扩增同一品种,经电泳检测、照相可以得出品种ID照片,进而快速、准确地检测、区分玉米自交系。
Analyzing the genetic diversity of maize inbred lines and classifying the heterotic group and the model of heterosis which using to select the parents and improve, innovate, excavate and employ the good germplasm cloud improve the efficiency of selecting the parents of hybrid and shorting the duration of breeding, which was the important basis of maize breeding.QTL location of important maize traits is the basis of marker-assisted selection, which has a very important significance to improve the accuracy and foresight of breeding. The materials of this study were part of important inbred lines in our country, varieties which popularizing in large areas and inbred lines selected by ourselves. Genetic distance obtain by SSR markers were used to classify the heterotic group of good maize germplasm and analyze and discuss the model of heterosis, Association analysis was an approach to identify the relationship of molecular markers or candidate genes with traits in a given population based on linkage disequilibrium (LD). In this study, 64 pair SSR markers were used for genotyping of 257 maize inbred lines. LD of pairwise loci and population structure were analyzed, then association analysis between SSR and nine phenotypic traits such as plant height, ear height, growth duration, ear length, row number of ear, 100-seeds weight, oil content, protein content, and starch content were performed using TASSEL GLM and their explained phenotypic variation were detected, too. Molecule ID could be build up by the germplasm character analysis software (ID Analysis1.0) which developed by ourselves, and classify the materials by least primers.
The results of this study follow as:
(1) At present, the purpose of maize inbred lines breeding with high yield should focus on prolonging the growth duration and increasing the ear length and 100-seed weight, according with other triait. Breeding of maize inbred lines with high quality should directional select the germplasms with ear diameter and plant height, next with growth duration, row number per ear and ear diameter, according with other traits to correspond the relational factors and pick up the process of breeding.
(2) The materials in this study could be classified into six groups of PA, BSSS, Lvdahonggu, Tangsipingtou, Lancaster and PB. The result of clustering analysis was according with pedigree. In this study, Lancaster can be classified into group A, Tangsipingtou and Lvdahonggu in group B, PA and PB in group C (PN), and BSSS in the group D because of different from other groups. The materials in this research could be classified into 4 groups (6 subgroups). Since the genetic base of maize germplasms was narrow in our country, simplifying the number of groups could beneficial to explore the model of heterosis. Core SSR markers could classify the heterotic group.
(3) In the 64 SSR loci, there are 43 markers associated with 24 traits, in which 19 loci associated with 14 phenotypic traits at the level of 0.01 and other markers at the level of 0.05. There are 80 markers associated with agronomic traits and quality traits, in which only seven associated with quality traits, which highly less than that associated with agronomic traits. The number of SSR loci found by associating analysis were more than the number which fixed by population with the same size, the markers associated with one traits were focus in the linkage group, such as most of the markers associated with growth duration were in the linkage group 7, which may to do with there more markers in this linkage group. It is ubiquity that one marker associated with more than one trait which in one category, such as umc1741 associated with plant height, ear height and ear length; umc2160 and umc1545 in linkage group 7 associated with not only growth duration but also, which indicating that there are inner genetic factor associated with phenotypic traits of maize. The results indicated that: the markers which associated with plant height, ear height, ear length and row number per ear were more than that with oil and protein content. More materials with variation in quality should add to the research group. There are several markers associated with one or several agronomy traits in one chromosome and different markers associated with one trait in different chromosomes, most markers were according with the mapping of QTL.
(4) Resource characteristic analysis of 257 maize inbred lines by SSR indicated that: only ten pair of primers can differentiate the varieties in this study. The inbred lines could be identified quickly and exactly by amplificating one variety with ten primers and examining by electrophoresis, which acquiring the photographs of molecule ID.
本论文研究目的是以我国部分重要玉米自交系、大面积推广杂交种的亲本、自选自交系等为研究对象,基于遗传距离划分优良玉米种质杂种优势群,探讨杂种优势模式。利用257份玉米自交系构成的自然群体为供试材料,选取64对核心SSR标记进行遗传多样性分析,应用STRUCTRE软件对257份玉米自交系构成的自然群体进行基于数学模型的类群划分,并计算材料相应的Q值(第i材料其基因组变异源于第k群体的概率),利用TASSEL软件中的GLM(General Linear Mode)程序,各个体的Q值作为协变量,将农艺性状表型数据对标记逐一进行回归分析,确认各性状关联位点并计算位点对表型的解释率,采用资源特征分析软件(ID Analysis1.0,软件登记号:2007SR11870)建立品种分子ID,以最少的引物来区分品种。本论文主要结论如下:
(1)目前,选育高产玉米自交系应以尽量延长生育期为主,增加穗长及百粒重,以其他性状适中为选择目标。选育品质优良的玉米自交系应定向选择穗粗与株高较佳的种质,其次为生育期、行粒数与穗粗,平衡其他农艺性状的选择,根据品质性状及与其相关性状之间的关系选用适宜的种质,协调所有影响因素,加快选育优良玉米自交系的进程。
(2)基于SSR标记的遗传距离,将供试材料划分为PA、BSSS、旅大红骨、塘四平头、Lancaster、PB六个类群。聚类结果与自交系系谱关系基本一致。本研究中Lancaster可定为A群,塘四平头与旅大红骨可归为B群,PA、PB可归为C群即PN类群,而BSSS类群由于与其他类群差异较大归为D类群,这样可将供试材料归为4大类群,共6个亚群。我国玉米种质资源遗传基础相对狭窄,因此简化类群划分数目,有利于探索杂种优势模式。核心SSR标记对于划分杂种优势群有显著的鉴别能力。
(3)64个SSR位点中,有43个标记位点与24个性状相关联,其中19个位点的变异分别与14个表型性状在P<0.01水平上呈极显著相关,其它标记与各性状均显著相关(P<0.05);与农艺、品质性状相关联的位点(次)累计有80个,与品质性状相关联的位点(次)累计有7个,关联位点数目上品质性状远少于其他农艺性状;关联分析发现的主要性状SSR位点数目远多于相同大小家系群体定位所得位点数,同一性状的关联位点在连锁群上分布较集中,如与生育期关联的位点多分布在Ch7群体上,这可能与此连锁群上使用标记较多有关;同一个位点与多个性状关联很普遍,这些性状一般为同一类性状,如umc1741与株高、全株叶数、穗位及穗长关联,Ch7连锁群上的umc2160、umc1545与生育期关联同时也与穗行数相关联,这说明玉米表型性状相关有其内在遗传因素。综合分析表明:与株高、穗位、穗长、穗行数等性状相关联的标记较多,与脂肪含量、蛋白质含量等性状相关联的标记较少,应该在试验群体内增加品质性状变异程度较大的试验材料,同一条染色体上存在一个或多个与农艺性状关联的标记位点,不同染色体上存在与同一性状关联的标记,这与玉米数量性状的QTL定位大体一致。
(4)通过SSR分子标记技术,采用资源特征分析软件(ID Analysis 1.0)对257份玉米自交系进行分析表明,仅需10对引物可将供试品种完全区分开,用这10对引物共同扩增同一品种,经电泳检测、照相可以得出品种ID照片,进而快速、准确地检测、区分玉米自交系。
Analyzing the genetic diversity of maize inbred lines and classifying the heterotic group and the model of heterosis which using to select the parents and improve, innovate, excavate and employ the good germplasm cloud improve the efficiency of selecting the parents of hybrid and shorting the duration of breeding, which was the important basis of maize breeding.QTL location of important maize traits is the basis of marker-assisted selection, which has a very important significance to improve the accuracy and foresight of breeding. The materials of this study were part of important inbred lines in our country, varieties which popularizing in large areas and inbred lines selected by ourselves. Genetic distance obtain by SSR markers were used to classify the heterotic group of good maize germplasm and analyze and discuss the model of heterosis, Association analysis was an approach to identify the relationship of molecular markers or candidate genes with traits in a given population based on linkage disequilibrium (LD). In this study, 64 pair SSR markers were used for genotyping of 257 maize inbred lines. LD of pairwise loci and population structure were analyzed, then association analysis between SSR and nine phenotypic traits such as plant height, ear height, growth duration, ear length, row number of ear, 100-seeds weight, oil content, protein content, and starch content were performed using TASSEL GLM and their explained phenotypic variation were detected, too. Molecule ID could be build up by the germplasm character analysis software (ID Analysis1.0) which developed by ourselves, and classify the materials by least primers.
The results of this study follow as:
(1) At present, the purpose of maize inbred lines breeding with high yield should focus on prolonging the growth duration and increasing the ear length and 100-seed weight, according with other triait. Breeding of maize inbred lines with high quality should directional select the germplasms with ear diameter and plant height, next with growth duration, row number per ear and ear diameter, according with other traits to correspond the relational factors and pick up the process of breeding.
(2) The materials in this study could be classified into six groups of PA, BSSS, Lvdahonggu, Tangsipingtou, Lancaster and PB. The result of clustering analysis was according with pedigree. In this study, Lancaster can be classified into group A, Tangsipingtou and Lvdahonggu in group B, PA and PB in group C (PN), and BSSS in the group D because of different from other groups. The materials in this research could be classified into 4 groups (6 subgroups). Since the genetic base of maize germplasms was narrow in our country, simplifying the number of groups could beneficial to explore the model of heterosis. Core SSR markers could classify the heterotic group.
(3) In the 64 SSR loci, there are 43 markers associated with 24 traits, in which 19 loci associated with 14 phenotypic traits at the level of 0.01 and other markers at the level of 0.05. There are 80 markers associated with agronomic traits and quality traits, in which only seven associated with quality traits, which highly less than that associated with agronomic traits. The number of SSR loci found by associating analysis were more than the number which fixed by population with the same size, the markers associated with one traits were focus in the linkage group, such as most of the markers associated with growth duration were in the linkage group 7, which may to do with there more markers in this linkage group. It is ubiquity that one marker associated with more than one trait which in one category, such as umc1741 associated with plant height, ear height and ear length; umc2160 and umc1545 in linkage group 7 associated with not only growth duration but also, which indicating that there are inner genetic factor associated with phenotypic traits of maize. The results indicated that: the markers which associated with plant height, ear height, ear length and row number per ear were more than that with oil and protein content. More materials with variation in quality should add to the research group. There are several markers associated with one or several agronomy traits in one chromosome and different markers associated with one trait in different chromosomes, most markers were according with the mapping of QTL.
(4) Resource characteristic analysis of 257 maize inbred lines by SSR indicated that: only ten pair of primers can differentiate the varieties in this study. The inbred lines could be identified quickly and exactly by amplificating one variety with ten primers and examining by electrophoresis, which acquiring the photographs of molecule ID.
引文
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