Genetic Diversity and Association Analysis of Fiber Quality Traits with SSR Markers in Germplasm Resources of Early Maturity Upland Cotton in Xinjiang

[Objective] We explored the genetic diversity of early maturity upland cotton germplasm resources in Xinjiang, China. We discovered specific germplasm resources and elite allelic variation related to fiber quality. [Method] We used 219 early maturity upland cotton accessions in our experiments and investigated 15 agronomic traits in three environments. We used a total of 128 pairs of simple sequence repeat (SSR) primers to scan for polymorphism. We then used NTsys-pc2.1 software to analyze genetic diversity, and Structure2.3.1 and Tassle5.0 software to perform association analysis of fiber quality traits based on phenotypic effect values to identify elite allele variation and conventional materials. [Result] A total of 244 loci were amplified by 128 marker pairs in 219 samples with an average of 1.91 loci per marker. The polymorphic information content ranged from 0.13 to 0.86 with an average of 0.63. The distribution range of the genetic similarity coefficient between materials in this population was 0.42-0.99 with an average of 0.61. The genetic similarity coefficient was between 0.5 and 0.7 and accounted for 90.19%. Through association analysis, we detected 11 markers that were significantly (P<0.01) associated with fiber quality traits, and discovered seven typical materials with specific allele. [Conclusion] In total, 219 Xinjiang early-maturing upland cotton germplasm resources have low genetic diversity. Based on SSR association analysis, we discovered some specific allele variations and conventional materials related to fiber quality traits.     

黄麻核心种质的遴选

遴选黄麻核心种质可为黄麻种质创新及新品种选育奠定基础。本研究以300份黄麻种质资源为基础,基于SSR分子标记和农艺性状考察,结合地理来源构建核心种质。结果表明,11个农艺性状变异系数变幅在13.06%～84.87%,表现出丰富的遗传多样性。按农艺性状聚类分析可划分为8个类群,按分子标记聚类可划分为10个类群。结合2个聚类分析、地理位置并按比例取样,建立一个由108份品种(系)组成的预选核心种质。采用44对SSR引物对其进行遗传差异分析,在遗传相似系数为0.65时,可把108份品种(系)分成圆果种和长果种两大类。根据遗传差异分析,剔除遗传相似系数大于或等于0.85的遗传冗余,获得84份品种(系)的核心种质,其中圆果种60份和长果种24份。比较84份核心种质与300份种质的农艺性状变异系数及Shannon-Wiener指数发现,两者之间相差不大,表明遴选的84份核心种质可以最大限度代表300份黄麻种质资源的遗传多样性加以利用和保存。     

Association Analysis of Agronomic and Yield-Related Traits in an Upland Cotton (Gossypium hirsutum L.) Germplasm Population Based on SSR Markers

[Objective] Molecular markers associated with agronomic and yield-related traits could provide useful references for improving the efficiency of Gossypium hirsutum breeding. [Method] In 147 upland cotton materials, 237 Simple Sequence Repeat(SSR) markers were assayed for polymorphisms. The population structure was analyzed using Structure 2.2. An association analysis between SSR markers and the agronomic and yield-related traits was performed using the Tassel 2.1 General Linear Model program. [Result] The average coefficients of variation for the five indicators varied from 7.28% to 21.06%, with an average of 13.47%, which indicated that the test population had a rich phenotypic diversity. The 237 SSR markers included 690 alleles belonging to 281 loci, with an average of 2.455 0 alleles per SSR marker (ranging from 2 to 6), and the average polymorphism information content value was 0.216 5. The test group could be divided into seven subgroups using Structure software. A genome-wide association analysis was conducted on five agronomic and yield-related traits from 2013 to 2015. There were 45 SSR loci and 1 locus significantly (P＜0.01) associated with boll weight and lint percentage, which were detected in 3 or more environments, and they explained 19.30% and 11.58%, respectively, of the phenotypic variation in the different environments (P＜0.01). Nine loci were significantly (P＜0.01) associated with agronomic traits at two or more bowing times, and one, four and four markers were associated with fruit branch number, boll number and plant height, explaining 9.96%, 7.00% and 5.57%, respectively, of the phenotypic variation in different environments. Sites significantly associated with boll weight, fruit branch number and plant height were simultaneously detected near BNL2448. [Conclusion] Many loci associated with the yield and other important agronomic traits of upland cotton were repeatedly detected in different environments using an association analysis. The results provide useful reference information for cotton molecular marker-assisted selection-based breeding.     

适于海岛棉指纹图谱构建的SNP核心位点筛选与评价

海岛棉具有纤维品质好、抗病性强等优异性状,不仅为纺织工业提供优质棉纤维原料,也是陆地棉相关性状改良的重要供体材料。然而,与陆地棉相比,开展海岛棉的遗传多样性和基因分型研究相对较少。本研究基于CottonSNP80K芯片对282份不同来源的海岛棉品种/材料进行基因组SNP分型研究,以选择高效鉴别海岛棉材料的核心位点组合。按照检出率大于95%、具多态性、最小等位频率(MAF)大于0.01、杂合率小于0.05、无冗余位点等条件筛选,获得2594个高质量SNP位点。对上述位点设置不同数目梯度筛选,确定最优核心位点数。随着位点个数的增多,位点组合对海岛棉材料的识别率逐渐增加。当位点数为200时,识别率为89%;位点数提高到1500时,识别率可达99%。进一步增加位点数,识别率无显著变化。利用中选的1500个SNP位点检测供试材料,其平均MAF值0.14,平均杂合率0.007,平均多态信息含量0.21。SNP位点的聚丙烯凝胶电泳验证表明,SNP-PCR与芯片分型结果一致性达98.3%。本研究提供了适于海岛棉指纹图谱构建、含1500位点的一套核心SNP位点组合,可用于海岛棉遗传多样性分析和品种身份鉴定。     

Association analysis of fibre traits in Gossypium arboreum accessions

Advances in the use of diploid Asiatic species in cotton breeding require an understanding of the relatedness and ancestry of diploid cotton accessions, and identification of simple sequence repeat (SSR) markers associated with agronomically important phenotypic traits, for example, fibre quality. Fifty‐six Gossypium arboreum germplasm accessions introduced from nine regions of Africa, Asia and Europe were evaluated for eight fibre characters (lint percentage, lint colour, elongation, micronaire, strength, 50% span length, 2.5% span length and maturity%) and genotyped with 98 SSR markers. When viewed across all accessions most of the SSR markers were polymorphic. Population structure analysis identified six main clusters for the accessions which corresponded to different geographic regions, indicating agreement between genetic and predefined populations. The general linear model method was used to disclose marker–trait associations. Marker–trait associations were investigated by fitting single marker regression models for phenotypic traits on marker band intensities with correction for population structure. This paper illustrates the potential of association mapping in diploid cotton, because existing phenotypic data, a modest number of SSR markers, and a pioneering statistical analysis, identified interesting associations.     

陆地棉SSR标记遗传多样性及其与农艺性状的关联分析

分析陆地棉栽培种遗传多样性,通过关联分析寻找与棉花农艺性状相关联的分子标记,为分子标记辅助选择育种和提高棉花育种效率奠定基础。本文采用74个Simple sequence repeat(SSR)标记对172份陆地棉栽培种的基因组变异进行扫描,使用NTSYS-pc 2.20进行聚类,分析该群体遗传多样性;利用Structure 2.3.4软件分析群体结构,在此基础上结合田间表型数据,采用Tassel 2.1的一般线性模型(General linear model,GLM)进行关联分析,定位与农艺性状相关的QTLs。74个标记共检测到148个多态性位点,涉及246个等位变异,变异范围2~7个,平均等位变异数为3.32;引物的多态性信息含量(PIC)为0.0281~0.3733,平均值为0.2370;遗传相似系数变异在0.2816~1,平均值为0.5369,平均遗传相似系数为0.5369,表明我国陆地棉遗传基础狭窄,尽管国外及西北内陆棉区部分材料具有较丰富的遗传变异。聚类分析将该群体划分为12个亚群,不同棉区的材料交叉分布,且聚类结果基本与系谱吻合。群体结构分析却将172份供试材料划分为3个亚群;通过关联分析,发现30个位点与铃重、衣分、黄萎病抗性显著相关(P0.05),各位点对表型变异贡献率为2.24%~5.27%。     

Genetic diversity,population structure and key phenotypic traits driving variation within soyabean (Glycine max) collection in Ghana

Soybean [Glycine max (L.) Merrill] is an important oilseed crop worldwide and it has recently become the crop of interest in Ghana. In this study, 142 soybean accessions were genotyped with 34 SSR markers and concurrently evaluated for five quantitative and two qualitative phenotypic traits. Twenty‐nine of the SSR markers were polymorphic with mean allele number of 5.3, polymorphic information content (PIC) of 0.51 and gene diversity of 0.55. Molecular analysis based on unweighted paired group arithmetic mean (UPGMA) clustering and principal coordinate analysis (PCoA) was similar in explaining the extent of diversity within the accessions. Structure analysis placed most of the accessions into two subpopulations with 18 (12.7%) as admixtures. Principal component analysis (PCA) based on phenotypic traits revealed two clusters. Both UPGMA clustering‐based SSR data and PCA from phenotypic data showed similar results. The assembled germplasm is genetically diverse with high variation in flowering and maturity period, and key yield components which could be exploited in developing superior varieties well adapted to Ghana and West Africa.     

谷子核心种质表型遗传多样性分析及综合评价

通过遗传种质的多样性评估可以指导深入研究资源和育种中优异互补亲本的选择进而提高优异基因的交流累加和新品种培育的效率。本研究选用了来自世界各地的份谷子核心种质通过个表型性状的综合鉴定评估遗传多样性和筛选优异种质资源结果表明我国谷子资源的表型遗传多样性丰富单穗粒重、穗长、穗粗、株高、茎节数和生育期均表现了丰富的变异谷子育成品种遗传多样性相比农家品种下降明显育种的遗传增益主要体现在株高和穗长的适度减低,以及茎粗、茎节数、穗粗、单穗粒重、单穗重及生育期的适度增加系统聚类分析将谷子资源分成类第类以来源为东北欧国家的品种为主第类以北美和非洲的品种为主第类以东亚、南亚的品种为主我国种质可划分为春播型、春夏兼播型和南方型类型采用主成分分析法和逐步回归分析法综合评判表明叶鞘色、刚毛长度、粒色、米色、株高、穗长、茎粗和单穗粒重个性状可作为谷子表型鉴定的主要指标。     

海岛棉纤维品质与其种子短绒多寡的相关性分析

【目的】为探讨海岛棉纤维品质与其种子短绒多寡之间的相关关系,提高海岛棉品质育种效率。【方法】以218份海岛棉种质资源为供试材料,通过2年纤维品质指标的检测及短绒多寡的调查与分级,分析短绒各分级间纤维品质指标的差异性及纤维品质与短绒多寡的相关性。【结果】方差分析表明,各级间除马克隆值差异不大外,其他纤维品质指标随着短绒的增加而增高,且差异呈极显著或显著(P0.05或P0.01)水平。Spearman相关系数表明,纤维上半部平均长度、断裂比强度、长度整齐度指数和断裂伸长率分别与短绒级别间呈极显著(P0.01)正相关,马克隆值与级别间呈显著(P0.05)负相关。【结论】海岛棉纤维品质与其种子短绒多寡间有统计学意义的正相关性,短绒多寡可作为海岛棉纤维品质优劣的参照之一。     

首批海岛棉基因组来源的微卫星标记的分离、评价和定位

海岛棉(Gossypium barbadense)是世界上最重要的栽培棉种之一。海岛棉纤维品质优良,是优质棉的重要产源。为了研究海岛棉的遗传多样性,为海岛棉育种提供参考依据,从海岛棉遗传标准系中分离基因组来源的微卫星标记用于海岛棉遗传评价。采用两种方法分离微卫星标记,一是用ISSR (inter simple sequence repeat) 引物扩增Pima3-79,克隆测序后从中开发微卫星标记;二是利用简并引物扩增Pima3-79,克隆测序后从中开发微卫星标记。共挑选1 447个克隆,筛选出239个独立克隆。测序后得到214个单一序列,其中包含微卫星并可用于引物设计的序列70个,获得86对引物。86对引物用于扩增56个海岛棉材料和4个陆地棉材料,16对引物没有扩增,43对引物在所有材料中没有多态性;27对引物在海岛棉和陆地棉之间有多态性,19对引物在海岛棉中表现多态性。利用Jaccard相似系数和UPGMA方法进行聚类分析可以明显区分陆地棉和海岛棉,并且将海岛棉分为4类。14对引物在BC₁群体中表现多态性,产生14个位点。9个位点整合到BC₁连锁图的7个染色体上,4个位于A亚基因组,5个位于D亚基因组。海岛棉微卫星标记扩展了棉花微卫星标记,有助于海岛棉遗传多样性的研究,有利于棉花遗传图谱的进一步丰富。     

大豆核心种质和微核心种质的构建、验证与研究进展

我国作物种质资源长期库中保存大豆资源2.3万余份,数量居世界之首。然而,在大豆新品种培育中的利用率仅为1%左右,导致大豆育成品种的遗传基础趋于狭窄。主要原因是缺少对其重要经济性状的鉴定,尤其是缺少多年多点的评价,难以有目的选择有重要价值的育种亲本。为了加速大豆种质资源的评价并促进其利用,在国家基础研究项目(973)的连续资助下,开展了“大豆核心种质构建(1998-2003)”和“大豆微核心种质基因多样性(2004-2009)”研究,目的是浓缩大豆资源的遗传多样性,强化其表型和基因型鉴定,为发掘和利用大豆资源中的优异基因提供指导。本文在研究构建不同比例(占总体2%~5%)大豆核心种质和大豆微核心种质(占总体1%)的同时,介绍了核心种质补充和完善的研究进展。为了验证核心种质的代表性,在构建方法方面,从SSR位点、样本组成、取样比例、低频率等位变异4个方面对代表性进行了分析,并用随机抽样方法对核心种质代表性进行了检测和验证。文中还介绍了利用核心种质和微核心种质在新基因发掘、种质创新和育种利用方面的研究进展,尤其介绍了与育种单位密切合作,建立基于核心种质的种质创新与利用体系的研究成效。围绕遗传多样性、核心种质利用方式进行了讨论,指出大豆核心种质为性状鉴定、新基因发掘、新种质创造和新品种培育等理论研究和实际应用提供材料基础,具有潜在的应用前景。实践证明,大豆种质资源的系统研究与利用,将促进我国大豆种质资源由数量保存型向研究应用型转变。     

棉花适宜机采相关性状的SSR标记关联分析及优异等位基因挖掘

棉花机械采收对品种的生育期、株型及对脱叶剂敏感度有较高的要求。本研究利用覆盖全基因组有多态性的214对SSR标记对118份含有一个或多个机采性状的种质资源的株高、始节高、始节位、第一果枝平均长度、生育期及脱叶率6个机采相关性状进行关联分析。利用Structure 2.3.1软件进行群体结构分析,并结合2年2点12个重复的田间表型数据,采用Tassel 5.0软件的混合线性模型MLM关联定位。结果检测到460个等位基因,涉及905个基因型,基因多样性指数平均为0.5151,PIC值平均为0.4587,基因多样性指数和PIC值都大于平均数的标记有99个,占总标记数的46.3%,说明该批SSR标记具有较多的等位变异数和较高的遗传多样性。群体结构分析将118份供试材料划分为4个亚群,结果显示各类群中材料与地理来源无对应关系。关联分析结果显示4种环境中,在显著条件下(P0.05),共检测到124个与6个机采相关性状相关的位点,对表型变异解释率范围为2.23%~14.15%;其中在极显著条件下(P0.01),共检测到20个与机采相关性状相关的位点,对表型变异解释率范围为4.84%~14.15%。基于本研究的结果,鉴定出典型的载体材料11份,分别为系7、金垦9号、Y11、豫棉18、AY-4、K2、朝阳棉2号、DZ22、中棉所43、C2和关农长早B14。以上发掘出的控制棉花适宜机采性状的优异等位基因及优异亲本资源,可为机采棉的分子辅助选择育种提供理论依据。     

海岛棉遗传多样性的SRAP标记分析

利用SRAP标记, 选用132对带型清晰的多态性引物, 对我国引入海岛棉以来培育的36个国内品种及20个国外品种进行遗传多样性分析。共检测到419个多态性位点, 每组合的多态性条带数从2~9不等, 平均为3.17。利用NTSYS-pc 2.10e 软件采用Jaccard’s相似系数和UPGMA方法进行聚类分析。结果表明, 大部分具有亲缘关系的品种聚在同类中, 说明其结果与系谱具有一定的相符性; 56个品种的平均遗传相似系数为0.497, 变化范围在0.312~0.876之间, 说明我国海岛棉品种在分子水平上存在较大差异; 我国3个育种时期育成品种的平均相似系数依次为0.501、0.507和0.548, 表明我国现在育成的品种相对于早期品种遗传多样性在逐渐降低。这些结果为我国海岛棉育种提供了有益的参考。     

中棉所70分离群体农艺性状相关QTL定位

【目的】定位棉花产量相关性状的数量性状基因座(Quantitative trait locus,QTL)。【方法】以中棉所70的F_2分离群体为遗传作图群体,利用从14 820对简单序列重复(Simple sequence repeat,SSR)引物中筛选出的267对两亲本间的多态性引物检测F_2群体250个单株的标记基因型,利用Joinmap 4.0进行连锁分析,并通过WinQTLCart 2.5复合区间作图法对F_(2:3)群体的株高、单株结铃数和单株果枝数性状进行QTL定位。【结果】在F_2群体中共获得342个SSR标记位点,并构建了包括312个标记、35个连锁群,总长1 929.9 cM的遗传连锁图谱(标记间平均距离为9.2 cM,覆盖棉花基因组的43.4%)。经QTL定位,共检测到19个QTL,其中涉及株高的7个、单株果枝数4个、单株结铃数8个,这些QTL分布在8条染色体上,解释0.25%~11.28%的表型变异。【结论】这些与农艺性状相关的QTL有助于棉花产量分子标记辅助选择。     

95份南瓜种质资源亲缘关系的SSR分析

为研究95份南瓜资源的遗传关系,提高南瓜育种效率,应用150对简单重复序列(SSR)标记及聚类分析方法对95份核心资源进行遗传多样性分析。结果显示,有32对引物能够在不同南瓜种质间扩增出清晰而稳定的多态性标记。利用MEGA 4.0统计软件中UPGMA聚类分析,95份南瓜的平均遗传距离为0.583,最大为0.9412,最小为0;分支总长度SBL=10.3384。在分支距离为0.39时,32对SSR引物可以将95份资源可以分为3个类群。第Ⅰ类群,主要为印度南瓜,第Ⅱ类群为中国南瓜,第Ⅲ类群71号为美洲南瓜。本研究为种质资源的鉴定评价,优良种质的挖掘以及品种选育提供了参考。     

Development of a core set of SSR markers for the characterization of <Emphasis Type="Italic">Gossypium</Emphasis> germplasm

Molecular markers such as simple sequence repeats (SSR) are a useful tool for characterizing genetic diversity of Gossypium germplasm. Genetic profiles by DNA fingerprinting of cotton accessions can only be compared among different collections if a common set of molecular markers are used by different laboratories and/or research projects. Herein, we propose and report a core set of 105 SSR markers with wide genome coverage of at least four evenly distributed markers per chromosome for the 26 tetraploid cotton chromosomes. The core marker set represents the efforts of ten research groups involved in marker development, and have been systematically evaluated for DNA polymorphism on the 12 genotypes belonging to six Gossypium species [known collectively as the cotton marker database (CMD) panel]. A total of 35 marker bins in triplex sets were arranged from the 105 markers that were each labeled with one of the three fluorescent dyes (FAM, HEX, and NED). Results from this study indicated that the core marker set was robust in revealing DNA polymorphism either between and within species. Average value of polymorphism information content (PIC) among the CMD panel was 0.65, and that within the cultivated cotton species Gossypium hirsutum was 0.29. Based on the similarity matrix and phylogenetic analysis of the CMD panel, the core marker set appeared to be sufficient in characterizing the diversity within G. hirsutum and other Gossypium species. The portability of this core marker set would facilitate the systematic characterization and the simultaneous comparison among various research efforts involved in genetic diversity analysis and germplasm resource preservation.     

基于表型性状构建中国花生地方品种骨干种质

中国花生地方品种遗传多样性丰富,是花生新品种选育的重要亲本来源。本研究以种质库保存的2741份地方种质为材料,基于种植区划和植物学类型分组,平方根法确定取样量,组内按13个表型数据进行UPGMA聚类分析,类内随机取样,构建骨干种质。利用t检验、F测验、卡方测验、极差、表型保留比例、表型相关性等对骨干种质代表性进行检验和评价;并利用主成分分析和直方图对骨干种质进行确认。结果表明,构建了包含259份种质的中国花生地方品种骨干种质,占全部种质的9.4%,包括多粒型14份、珍珠豆型85份、龙生型42份、普通型103份、中间型15份。在P 0.05概率条件下,骨干种质13个性状的均值、方差、变异系数、香农指数与全部种质无显著差异,且保留了全部种质的分布范围、表型保留比例和表型相关性;二者的植物学类型组成和生态分布是一致的,具有相似的遗传结构和分布频率。建立的骨干种质很好地代表了全部种质的遗传变异和群体结构,可为花生种质创新和优异等位基因发掘奠定良好的基础。     

Genetic Dissection of Allelic Loci Associated with Economic Traits of Upland Cottons in Xinjiang

[Objective] We completed anassociation analysis of economic traits for upland cotton using simple sequence repeat (SSR) markers. We then explored the allelic variation sites to analyze the genetic basis of economically important traits, studied the genetic mechanism of Xinjiang upland cotton, and aimed to accelerate efficient breeding of upland cotton. [Method] We carried out polymorphic scanning on 156 upland cotton varieties in Xinjiang by screening 73 pairs of SSR markers encompassing the whole cotton genome. We constructed boxplot maps using R statistical computing software and graphics language and used TASSEL software to correlate yield or fiber quality traits with significant allelic variation loci. [Results] We obtained 10 allelic variation loci related to yield traits using the correlation analysis of Xinjiang upland cotton varieties from six different environments. The interpretation rate of phenotypic variation ranged from 6.69% to 9.88% with an average of 8.43%. Twenty-three allelic variation loci associated with fiber quality traits and phenotypic variation interpretation rates ranged from 3.73% to 13.22% with an average of 7.52%. The 22 detected quantitative trait loci were reported in previous studies and 10 showed the same associated traits as previously reported. [Conclusion] The population genetic structure of Xinjiang upland cotton varieties is simple, the linkage disequilibrium level is low, and the phenotypic traits show a stable trend under six environments. Using association analysis, we discovered unique allelic variation genes related to yield and fiber quality and diverse allele loci.     

利用陆地棉MAGIC群体定位产量、生育期和株高性状的QTL

棉花的产量、生育期和株高是重要的农艺性状, 决定着棉花的经济效益和生产方式。为了研究这些性状的遗传基础, 利用8个亲本构建的包含960个株系的陆地棉MAGIC (multi-parent advanced generation inter-cross)群体和SSR标记对这些性状进行关联分析。对产量、生育期和株高共8个相关性状进行了3年3个地点共5个环境的田间试验。经过最佳线性无偏预测(BLUP)综合多环境的表型数据, 发现MAGIC群体比亲本有更丰富的表型变异。8个性状的广义遗传力(H ²)变化范围为0.17~0.71。结合284个SSR标记基因型数据, 利用混合线性模型对产量、生育期和株高相关性状进行关联分析, 分别检测到51、27和9个显著关联的位点, 这些位点都表现出微效性, 表明该陆地棉MAGIC群体在性状位点的挖掘方面具有高效性。检测到20个标记位点或区间控制多个性状, 还发现单株有效铃数、单铃皮棉重、第一果枝节位和株高存在染色体热点区域, 对多性状综合研究或单性状深入挖掘具有重要价值。本研究为后续深入利用MAGIC群体进行遗传研究提供参考, 一些表型优良的材料和关联到的位点为育种改良奠定了基础。