SRAP标记在落花生属种质资源遗传多样性上的利用

利用15对SRAP标记引物分析落花生属17份材料间的遗传变异情况。结果表明17份材料共检测出119个位点,其中多态性位点为100个,多态性百分率为89.00%,平均每对引物检测出6.67个多态性位点,扩增范围为5~10。材料间的遗传相似性范围为0.310~0.967,平均为0.666。基于遗传相似,利用UPGMA聚类分析表明,17份落花生可聚为3个类群,主成分分析析结果与聚类分析结果一致。研究结果证实了落花生种质资源遗传多样性丰富,为落花生育种工作奠定了基础。     

用SSR和AFLP技术分析花生抗青枯病种质遗传多样性的比较

由Ralstonia solanacearum E.F.Smith引起的青枯病是若干亚洲和非洲国家花生生产的重要限制因子,利用抗病品种是防治这一病害最好的措施。虽然一大批抗青枯病花生种质资源材料已被鉴定出来,但对其遗传多样性没有足够的研究,限制了在育种中的有效利用。本研究以31份对青枯病具有不同抗性的栽培种花生种质为材料,通过简单序列重复(SSR)和扩增片段长度多态性(AFLP)技术分析了它们的遗传多样性。通过78对SSR引物和126对AFLP引物的鉴定,筛选出能显示抗青枯病种质多态性的SSR引物29对和AFLP引物32对。所选用的29对多态性SSR引物共扩增91条多态性带,平均每对引物扩增3.14条多态性带;32对多态性AFLP引物共扩增72条多态性带,平均扩增2.25条多态性带。在所筛选引物中,4对SSR引物(14H06,7G02,3A8,16C6)和1对AFLP引物(P1M62)检测花生多态性的效果优于其他引物。SSR分析获得的31个花生种质的遗传距离为0.12-0.94,平均为0.53,而AFLP分析获得的遗传距离为0.06～0.57,平均为0.25,基于SSR分析的遗传距离大于基于AFLP分析的遗传距离,疏枝亚种组的遗传分化相对大于密枝亚种组。基于两种分析方法所获得的聚类结果基本一致,但SSR数据聚类结果与栽培种花生的形态分类系统更为吻合。根据分析结果,对构建青枯病抗性遗传图谱群体的核心亲本和抗性育种策略提出了建议。     

应用SSR标记分析鹰嘴豆种质资源遗传多样性

种质资源的遗传多样性是育种工作的基础,本研究利用SSR标记对鹰嘴豆资源进行遗传多样性分析,旨在发掘鹰嘴豆资源中丰富的遗传变异。从48对鹰嘴豆SSR引物中筛选出18对核心多态性引物,对96份不同来源的鹰嘴豆种质资源进行SSR标记的遗传多样性分析。结果表明,18对SSR引物共检测到115个等位变异,占总检测位点的52.99%,每对SSR引物可检测出3~10个等位变异,平均6.39个;平均每个位点多态信息量(PIC)为0.8107,变化范围为0.6661~0.8984。Shannon's信息指数平均为1.4769,变化范围为0.0607~1.9584。PGMA聚类结果表明,在遗传相似系数0.59处,可将96份鹰嘴豆资源分为6个类群,类群Ⅰ包含9份资源,类群Ⅱ包含2份资源,类群Ⅲ包含28份资源,类群Ⅳ包含4份资源,类群Ⅴ包含40份资源,类群Ⅵ包含13份资源。本研究对我国鹰嘴豆种质资源的评价与利用、优异基因的挖掘、育种亲本材料的选择等提供科学依据。     

基于SRAP标记的大花蕙兰种质资源遗传多样性分析

采用SRAP技术分析了来源于不同国家和地区的42个大花蕙兰品种及4个国兰原生种之间的遗传亲缘关系。34对引物组合中筛选出29对带型稳定、多态性较好的引物组合,共扩增出398条谱带,其中387条为多态带,多态性比率97.2%,平均每个引物组合扩增多态性带13.3条。46份种质之间的相似系数变化范围为0.60～0.99,平均为0.76。基于29个SRAP标记的扩增结果,利用NTSYS 2.10e软件计算Dice遗传相似系数,并建立UPGMA聚类图,结果表明:在遗传相似系数0.69处将供试材料分为4类,较好地揭示了大花蕙兰品种及国兰原生种间的遗传多样性与亲缘关系,可为大花蕙兰种质资源利用及杂交育种中亲本选择提供科学依据。     

基于SSR标记的132份甘薯种质指纹图谱的构建及遗传多样性分析

利用SSR分子标记技术,构建132份甘薯种质的DNA指纹图谱,并进行遗传多样性分析,旨在为甘薯种质资源亲缘关系鉴定、分类提供理论依据。利用筛选的核心引物进行PCR扩增,通过聚丙烯酰胺凝胶电泳检测显示,19对引物共扩增出232个条带,其中多态性条带165条,多态性比率为71.1%,平均每对引物扩增出8.68个条带,多态性信息含量变化范围在0.6706~0.9331之间,平均为0.8158;其中引物SSR9和引物C33可将132份种质完全区分开,并构建供试材料的DNA指纹图谱,供试材料遗传距离在0.0363~0.5939之间,平均为0.4087,表明种质资源间遗传多样性丰富。基于SSR标记对供试材料进行聚类分析,将供试材料分为2个类群,第Ⅰ类群分为两个亚类,第Ⅰ-1亚类包括济薯25和3份日本引进品种日本金千贯、安納芋、日本薯;第Ⅰ-2亚类包括济徐薯23、苏丹、济薯09281。第Ⅱ类群分为两个亚类,第Ⅱ-1亚类由S07甘薯品系和与其亲缘关系较近的20份甘薯种质组成;第Ⅱ-2亚类由剩余的70份甘薯种质组成,为甘薯分子辅助育种中亲本的选择提供理论依据。     

鸭茅种质资源遗传多样性的SRAP研究

利用SRAP(Sequence-related amplified polymorphism)标记对来自国内外的45份鸭茅（Dactylis glomerata L.）种质资源进行遗传多样性研究。21对引物扩增出438个条带,多态性条带为363条,多态性条带比率为82.08%,每对引物组合的多态性带数平均为17.29条。GS值范围在0.6248-0.9686间,平均GS值为0.7958,显示来源广泛的鸭茅种质资源间存在着丰富的遗传变异。聚类分析及主成分分析能将所有材料聚为4类,能较准确的反映材料的来源分布情况及供试材料的染色体倍性差异,表明鸭茅的遗传多样性与染色体倍性及地理分布密切相关。同时清楚的揭示出国产鸭茅品种遗传基础较为狭窄。本研究为育种和探讨鸭茅种质资源遗传变异奠定了较好的理论基础。     

观赏羽衣甘蓝SSR标记分型与亲缘关系研究

观赏羽衣甘蓝凭借优良的观赏特性和抗逆性已经成为重要的冷季观赏植物。国内观赏羽衣甘蓝育种起步较晚,并且缺乏对种质资源遗传背景的系统研究。本研究应用SSR标记对不同类型的观赏羽衣甘蓝材料进行标记分型和亲缘关系分析。从99对均匀分布于甘蓝基因组的SSR引物中筛选出46对多态性好的引物,对27份不同类型的观赏羽衣甘蓝材料进行标记分型,共扩增出210个多态性位点,平均PIC值为0.58。进一步利用标记分型结果进行STRUCTURE群体结构、UPGMA聚类和聚类热图分析,结果显示3种分析结果基本一致,可以将27份材料分为圆叶、羽叶和皱叶3种类型,其中圆叶和羽叶类型的亲缘关系更近,与皱叶类型的亲缘关系较远;STRUCTURE分析还可以将双亲为不同类型的杂交种材料进行区分;聚类热图分析可以将标记分型结果形象的展示出来。本研究为进一步建立观赏羽衣甘蓝分子指纹图谱,明确种质资源的遗传背景,建立观赏羽衣甘蓝分子标记辅助选择育种体系,培育具有自主知识产权的新品种奠定基础。     

基于SSR标记的陆地棉早熟相关种质遗传多样性分析

丰富的遗传变异对于提高作物的环境适应性和遗传改良进度至关重要。为了解我国早熟陆地棉种质资源遗传多样性,本研究利用136对SSR引物对186份陆地棉材料(96份早熟陆地棉材料和90份中、晚熟陆地棉材料)进行了遗传多样性分析,共检测到等位基因变异355个,平均2.61个。在早熟棉材料中,有134对多态性SSR引物扩增出341个条带,平均2.54个;中、晚熟材料中有133对多态性SSR引物,扩增出345个条带,平均2.59个。早熟棉材料的位点多态性信息含量(PIC)、有效等位基因数(Ne)、基因型多样性(H')分别为0.684、3.994和1.361;中、晚熟棉材料的PIC、Ne、H'分别为0.668、3.852和1.343。早熟棉材料和中、晚熟棉材料的Jaccard相似性系数分别在0.349~0.935和0.270~0.907之间,平均为0.635、0.666。遗传相似性系数总体平均值接近,但早熟棉变化范围较中、晚熟棉小。用类平均法(UPGMA)进行聚类可将186份材料分成2个类群。总体上来看,供试材料遗传相似性系数较高,说明我国陆地棉早熟相关种质遗传基础狭窄。本研究结果为早熟棉育种亲本选配,早熟棉种质创新提供依据。     

乌塌菜种质资源遗传多样性的ISSR分析

利用ISSR技术对48份乌塌菜种质资源进行遗传多样性分析。从60条随机引物中筛选出稳定性强、条带清晰且多态性丰富的9条引物进行PCR扩增,共扩增出103条谱带,平均每个引物扩增出11.4条带,其中多态性带85条,多态性位点百分率为82.68%。不同乌塌菜种质间遗传相似系数变幅为0.59~0.97,说明ISSR标记能够揭示材料间较高的遗传多样性。利用UPGMA聚类分析,ISSR标记能将48份乌塌菜品种完全区分开,48份乌塌菜种质被划分为4个类群,聚类结果与叶片颜色相关,为乌塌菜品种资源的研究利用提供参考。     

甜瓜种质资源遗传多样性的SRAP分析

为研究甜瓜材料之间的亲缘关系及其分类, 更有效地利用种质资源, 为培育新品种提供依据, 文章采用 SRAP (Sequence-related amplified polymorphism)技术对61份甜瓜种质资源的遗传多样性进行研究。结果表明: 从42对引物组合中筛选出16对扩增条带清晰、多态性高的引物组合分析供试材料, 共检测出452个位点, 其中265个为多态性位点, 多态性比率达58.63％, 平均每对引物组合产生28.56个位点和16.56个多态性位点。61份材料间的相似系数为0.48～0.93, 平均为0.73。这些结果说明, 供试甜瓜材料具有较为丰富的遗传多样性。聚类分析结果表明, 61个甜瓜品种中首先可分为薄皮甜瓜与厚皮甜瓜两大类, 彼此亲缘关系最远。以遗传相似系数0.74为截值, 可把供试材料分为5个类群。在生态区域中, 新疆厚皮甜瓜的Nei’s基因多样性指数(0.2231)和Shannon’s信息指数(0.3422)最高。     

Genetic diversity analysis in sorghum germplasm as estimated by AFLP, SSR and morpho-agronomical markers

A comparison of the different methods of the estimation of genetic diversity is important to evaluate their utility as a tool in germplasm conservation and plant breeding. Amplified fragment length polymorphism (AFLP), microsatellites or SSR and morphological traits markers were used to evaluate 45 sorghum germplasm for genetic diversity assessment and discrimination power. The mean polymorphism information content (PIC) values were 0.65 (AFLPs) and 0.46 (SSRs). The average pairwise genetic distance estimates were 0.57 (morphological traits), 0.62 (AFLPs) and 0.60 (SSRs) markers data sets. The Shannon diversity index was higher for morphological traits (0.678) than AFLP (0.487) and SSR (0.539). The correlation coefficients obtained by the Mantel matrix correspondence test, which was used to compare the cophenetic matrices for the different markers, showed that estimated values of genetic relationship given for AFLP and SSR markers, as well as for morphological and SSR markers were significantly related (p <0.001). However, morphological and AFLP data showed non-significant correlation (p >0.05). Both data sets from AFLP and SSR allowed all accessions to be uniquely identified; two accessions could not be distinguished by the morphological data. In summary, AFLP and SSR markers proved to be efficient tools in assessing the genetic variability among sorghum genotypes. The patterns of variation appeared to be consistent for the three marker systems, and they can be used for designing breeding programmes, conservation of germplasm and management of sorghum genetic resources.     

中俄茄子种质资源遗传多样性研究

利用25对SSR分子标记和24个表型性状对105份中俄茄子材料进行遗传多样性分析。表型变异分析结果表明:24个表型性状在中俄材料间均表现出了不同程度的多样性,但是同一性状在中俄材料中多样性不同;主成分分析可将24个表型性状概括为果形因子、颜色因子、果实外观因子、叶片形态因子、果萼刺和花药条纹6个指标,其中果实特征占主要成分;利用UPGMA法进行聚类,遗传相似系数在0.4~0.8之间,平均值0.6。25对多态性SSR标记,扩增出122个条带,含有等位基因82个,其中有效等位数24.8个,PIC值为0.3~0.7。分子聚类的遗传相似系数在0.5~1之间,平均值是0.7。表型聚类和分子标记聚类的结果相似,105份茄子种质资源间的类群划分与地理来源之间没有直接关系,但与茄子的果实性状有一定的相关性。     

用SSR标记分析辣椒属种质资源的遗传多样性

利用21对引物在33个辣椒材料中共检测到54个等位基因,每对SSR引物检测到2~4个等位基因变异,平均为2.6个,说明辣椒种质资源的遗传多样性相对较少。通过MVSP3.13f软件对SSR数据进行聚类分析,把33个辣椒材料分为五类,同个种的辣椒种质资源基本聚在一起,说明用SSR标记来区分辣椒种质是可行的,是研究辣椒种质资源遗传多样性的有效手段。     

我国陆地棉基础种质遗传多样性的SSR分子标记分析

利用398对BNL、JESPR、TMB等SSR引物,对不同亲本来源、不同选育时期、不同种植生态区的43份陆地棉基础种质进行了遗传多样性的SSR分子标记分析。扩增产物用8％的非变性聚丙烯酰胺凝胶检测,银染观察并照相。遗传多样性带型分析按位点多态信息量（PIC）,Shannon-weaver多样性指数（H^＋）等方法,利用NTSYSpc2．1软件计算品种间的遗传相似系数（Jaccard系数）,并用类平均法（UPGMA）进行聚类。结果表明所选择多态性引物分布在棉花基因组的第3、4、5、8、9、10、16、18、20、23号等染色体上,36对多态性引物在基础种质中扩增等位基因130个,其中多态性等位基因占80％,每个引物扩增等位基因2～8个,平均3．6个,PIC为0．278～0．865,平均0．62,基因型多样性（H^＋）为0．451～2．039,平均1．102,基础种质问SSR遗传相似系数平均为0．610,变幅为0．409～0．865,这说明所选基础种质基因组水平的多样性较丰富,变化范围大、代表性强。按品种不同选育时期来讲,第一、二、三期基础种质的SSR分子标记平均遗传相似系数分别是0．587、0．630、0．630,说明现代基础种质比早期基础种质在基因组水平的差异呈下降的趋势,可能是由于育种者偏重于使用优质高产性状的亲本品种,致使我国棉花的育种基础逐渐变窄。不同棉区基础种质SSR标记性状差异大,北部特早熟棉区基础种质间的SSR标记的多样性大于黄河、长江棉区,主要原因是长江、黄河棉区的育种过分强调高产、优质品种选育,品种间的差异变小;基础种质中的国内品种SSR相似系数（0．624）比引进品种（0.85）高,说明国内品种在遗传多样性上目前还没有超越国外品种。总之,我国棉花现代基础种质比早期基础种质的遗传多样性呈下降的趋势,黄河、长江主产棉区基础种质的遗传多样性还没有超过国外基础种质,品种间的遗传背景较为狭窄,还必须采用多种途径丰富我国棉花种质资源的遗传多样性。     

SSR标记对不同黄籽甘蓝型油菜亲本材料的遗传分析

杂交育种中,亲本选配是育种成败的关键。本研究以重庆市油菜工程技术研究中心提供的180份甘蓝型黄子油菜亲本种质为材料,应用分布于不同连锁群的60对SSR标记进行了分析,共检测出308个标记位点,每对引物在不同亲本材料之间的等位基因数在1～11个之间,平均位点为5.1个。其中多态性位点207个,多态率达67.2%。对SSR扩增结果进行UPGMA分析,在遗传距离0.566处,180个品种(系)分为3个类群,聚类结果与种质来源比较一致,本研究为甘蓝型油菜黄子杂交育种和优势组合的选配提供了理论依据。     

苦荞地方种质资源的遗传多样性分析

利用SSR分子标记技术对中国苦荞主产区陕西、云南、四川、西藏等地的82份苦荞地方种质的遗传多样性进行了分析,以揭示中国特有的作物种质--苦荞地方种质资源的遗传多样性,促进苦荞优良品种的选育.结果显示:(1)所用25个SSR引物中有13个引物在苦荞地方品种中具有多态性,且扩增条带的稳定性较好,共扩增出208条条带,其中多态性条带200条,占总数的96.2%;(2)聚类分析结果显示,82份苦荞材料的遗传相似系数(GS)分布于0.52～0.85之间,平均值为0.69,在GS值为0.722的水平上,82份材料被聚为10大类群.研究表明82份苦荞品种间遗传多样性明显,具有丰富的遗传基础.     

Transferability of microsatellite markers from Brachypodium distachyon to Miscanthus sinensis, a potential biomass crop

Miscanthus sinensis has high biomass yield and contributed two of the three genomes in M. x giganteus, a bioenergy crop widely studied in Europe and North America, and thus is a potential biomass crop and an important germplasm for Miscanthus breeding. Molecular markers are essential for germplasm evaluation, genetic analyses and new cultivar development in M. sinensis. In the present study, we reported transferability of simple sequence repeat (SSR) markers from Brachypodium distachyon to M. sinensis. A set of 57 SSR markers evenly distributed across the B. distachyon genome were deliberately designed. Out of these B. distachyon SSR markers, 86.0% are transferable to M. sinensis. The SSR loci amplified in M. sinensis were validated by re-sequencing the amplicons. The polymorphism information content (PIC) of the transferable SSR markers varied from 0.073 to 0.375 with a mean of 0.263, assessed based on 21 M. sinensis genotypes. Phylogenetic tree based on 162 alleles detected by 49 SSR markers could unambiguously distinguish B. distachyon from M. sinensis, and cluster 21 M. sinensis genotypes into three groups that are basically in coincidence with their geographical distribution and ecotype classifications. The markers developed by the comparative genomic approach could be useful for germplasm evaluation, genetic analysis, and marker-assisted breeding in Miscanthus.     

Genetic Diversity of Source Germplasm of Upland Cotton in China as Determined by SSR Marker Analysis

The genetic diversity of 43 sources of Upland cotton germplasm with different parental origins, breeding periods, and ecological growing areas in China were studied on the basis of simple sequence repeat (SSR) markers. A total of 130 gene alleles with 80% polymorphism were detected from 36 SSR primers. The number of alleles per primer ranged from two to eight with an average of 3.6. The polymorphism information content (PIC) range was 0.278-0.865, with an average of 0.62. The average genotype diversity index (H') was 1.102, the highest was 2.039 and the lowest was 0.451. The average coefficient of the genetic similarity of SSR markers among source germplasm was 0.610, ranging from 0.409 to 0.865. These indicated that the genetic diversity at the genomic level of the selected source germplasm was rich, and was representative of the diversity of the germplasms, in general. The diversity at the genome level of the base germplasm from the second and third breeding periods was decreased compared to that of the first period, indicating that the cotton genetic background in China became narrow gradually. The diversity of SSR markers among the base germplasm from early maturity cotton growing areas in the north was higher than those from the Huanghe and Yangtze growing areas. The molecular marker genetic similarity index of the domestic varieties was higher than that in the introduced varieties, which indicates that the genetic diversity in domestic cultivars was lower than that in the introduced varieties. This study gives an overview of the genetic diversity of the cotton germplasm base in China, and provides a guide for breeders to develop new cultivars efficiently.     

棉花耐盐相关种质资源遗传多样性分析

为了解我国棉花耐盐相关种质资源的遗传变异,利用88对SSR引物对23份棉花耐盐材料和24份盐敏感材料进行遗传多样性分析。88个SSR位点在47份材料中共检测出338个等位基因变异,平均每个位点有3.841个;其中耐盐材料中检测出333个,盐敏感材料中检测出312个。耐盐材料的位点多态信息含量(PIC)、每个位点的有效等位基因数(Ne)、基因型多样性(H′)分别为0.613、2.929和1.083,盐敏感材料的PIC、Ne、H′分别为0.605、2.883和1.071。耐盐材料和盐敏感材料的Jaccard相似性系数分别在0.530–0.979和0.525–0.878之间,遗传相似性系数总体平均值接近,但耐盐材料的变化幅度更大。用类平均法(UPGMA)聚类将47份材料分成3个类群。总体而言,大多数材料之间的遗传相似性系数较高,表明我国陆地棉耐盐相关种质资源遗传基础狭窄。本结果为棉花耐盐育种中亲本的选配和优势组合的预测以及耐盐资源的合理利用等提供了基础资料。     

Assessment of genetic diversity among sorghum landraces and their wild/weedy relatives in western Kenya using simple sequence repeat (SSR) markers

Understanding the extent of gene exchange between cultivated sorghum and its wild/weedy relatives and the evolutionary processes (including farmers’ practices) that act to shape the structure of genetic diversity within and between them is an important aspect for germplasm conservation strategies, biosafety risk assessment, and crop improvement programs. In this study, molecular characterization and genetic diversity analyses were conducted on wild, weedy and cultivated sorghums collected at a local-scale in a traditional farming system in the Lambwe Valley of western Kenya. Nine simple sequence repeat (SSR) markers were used to genotype 294 cultivated sorghum and 200 wild sorghum individuals. The nine SSR markers were highly polymorphic with a number of alleles that varied from 2 to 19. Overall, wild sorghums had higher genetic diversity, observed heterozygosity, total number of alleles, polymorphic information content and more genotypes per locus than the cultivated types. A Mantel test demonstrated that there was significant isolation-by-distance for wilds and cultivated materials. STRUCTURE, cluster and principal coordinate analyses consistently assigned wild and cultivated individuals to different groups but failed to place hybrids/weedy types as a single separate group from wilds. Our results provide strong evidence of significant genetic diversity retained within wilds, larger divergence between wild and cultivated materials and reduced gene flow than those previously reported in Kenya. These results demonstrate the value of the Lambwe Valley region as a genetic reservoir and the importance to conduct genetic diversity studies at the local scale to design and execute appropriate in situ conservation programs and policies.