SSR标记对不同黄子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,每对引物在不同素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.     

青藏高原早熟甘蓝型春油菜遗传资源研究

利用SSR和SRAP 2种分子标记研究了69份试验材料的遗传差异及其亲缘关系.29对SSR标记共扩增出118条多态性带,多态性位点占总扩增位点的97.5%,27对SRAP引物扩增出123条多态性带,多态性比率为70.3%.两种标记聚类结果表明.在相似系数0.566处所有材料可以分为A、B 2个大类群;B类在相似系数0.620处又可分为7个亚类,10个天然双低早熟甘蓝型品系、2个甘蓝型亲本和4个新型品系聚在第1亚类中,其余的51个新型甘蓝型油菜品系分别聚在其他6个亚类中.对55份新型品系进行遗传成分分析,结果表明,每个品系都合有4种带型,各品系所舍不同带所占比率不同.对各品系中含有白菜型亲本带所占比率分别与其对应的两亲本之间的遗传距离进行相关分析,结果表明新型甘蓝型油菜品系中白菜型亲本带所占比率与白菜型素本间的遗传距离为负相关(-0.52),且达到极显著水平;与甘蓝型亲本间的遗传距离为正相关(0.31),且达到显著水平.对试验材料之间的遗传距离及其来源进行分析(除与2个白菜型亲本间),遗传距离排名前20位的都来自新型品系之间或天然品系与新型之间,最大为0.544.     

青藏高原早熟甘蓝型春油菜遗传资源创新研究

摘要：利用SSR和SRAP两种分子标记研究了71份实验材料的遗传差异及其亲缘关系。结果表明：(1) 29对SSR标记共扩增出121条清晰地带,多态性位点分别占总扩增位点的97.5%,而27对SRAP引物扩增出175条清晰地带,多态性比率为70.3%。（2）两种标记混合聚类结果表明,在相似系数0.566处所有材料可以分为A、B两个大类群。B类在相似系数0.620处又可分为六个亚类,10个天然双低早熟甘蓝型品系、两个甘蓝型亲本和四个新型品系聚在第Ⅰ亚类中,其余的51个新型甘蓝型油菜品系分别聚在其他五个亚类中。（3）对55份新型品系进行遗传成分分析,结果表明,每个品系都含有四种带型,不同品系中不同带型所占总带数的比率不同;对新型甘蓝型品系中含有白菜型亲本特有的带所占比率分别与其和对应的甘蓝型和白菜型亲本之间的遗传距离进行相关分析,结果表明新型甘蓝型品系中白菜型亲本特有的带所占比率与其和白菜型亲本之间的遗传距离为负相关（-0.52）,达到极显著水平;与其和甘蓝型亲本间的遗传距离为正相关（0.31）,达到显著水平。（4）对实验材料之间的遗传距离及其来源进行分析（除与两个白菜型亲本间）,遗传距离排前20位的都来自新型品系之间或天然品系与新型之间,且最大的达到0.544。     

加工番茄种质资源的SSR分析

为探明加工番茄种质资源间的亲缘关系,利用SSR标记对20份加工番茄品种及育种材料进行了多样性分析。结果表明:从49对SSR引物中筛选出12对扩增稳定、条带清晰且多态性丰富的引物进行分析,共获得43个位点,多态性位点为37个,多态位点比率86%;供试材料间的遗传相似系数介于0.419~1.000之间,说明加工番茄种质资源间存在一定的遗传差异;通过UPGMA法聚类分析,将20份材料分为3大类群,其中亲缘关系较远的不同类群间及亚类间的种质资源可作为杂交育种的亲本。     

基于遗传和表型特征的海岛棉遗传多样性分析

依据海岛棉种质资源的遗传及表型特征对其遗传多样性进行研究非常重要,可为海岛棉杂交育种选配亲本、引进新的种质资源拓宽优异基因范围以及培育海岛棉新品种提供理论根据。本研究通过利用125对SSR分子标记鉴定结果和13个农艺性状2年田间表型调查结果,对94份海岛棉种质资源进行遗传多样性分析,将这些材料按照遗传和表型特征进行类群的划分和比较,结果表明:(1)通过标记分析共检测到420个位点,其中249个为多态性位点,并应用Nei-Li相似系数法对94份材料间的遗传相似系数进行评估,发现94份海岛棉资源材料的遗传相似系数在0.46~0.95之间浮动,同时利用SSR分子标记将94份海岛棉资源材料划分为4大类群,分类结果与系谱分析基本吻合。(2)对94份海岛棉资源材料进行13个农艺性状及品质性状2年调查分析,发现供试材料品质性状的变异范围较广,而产量性状的变异范围相对较小。其中,各品种中马克隆值的多样性指数最高,最小的是单铃重。聚类分析发现,第Ⅰ类群产量性状的平均值较高;第Ⅱ类群虽在产量性状上不占优势,但其品质性状的表现较其他组别优异;第Ⅲ类群多为低产低质品种;第Ⅳ类群品种品质较优,产量变化范围较大。(3)利用SSR标记和农艺性状分析2种方法对94份海岛棉资源材料进行分析,结果均显示出较丰富的遗传多样性;2种方法的聚类结果基本一致,聚类结果与地理分布有明显的联系。     

甘薯杂交F_1群体材料的遗传多样性研究

甘薯(Ipomoea batatas(L.)Lam.)作为世界上一种重要的粮食、饲料、工业原料及新型能源作物,从有性生殖F1选择优良实生系进行选择育种一直是甘薯育种的重要方式。为了优化甘薯杂交育种方法,合理选配杂交组合,提高育种效率,本试验利用SSR标记研究了甘薯杂交群体基于SSR标记及13个农艺性状的遗传多样性,得到了群体内的聚类图,并且筛选出了甘薯的高产株系。群体SSR标记的聚类分析结果显示,群体材料与各亲本遗传距离比较远,被聚为3类,而亲本单独聚在另外一类。13个农艺性状的聚类将亲本与部分群体材料聚在了一起,且将群体材料和亲本材料作为一个整体时,其遗传距离的变异高达30%以上,远远高于SSR标记所获得的遗传变异系数。     

基于遗传多样性构建金针菇的核心种质群体及分子身份证

金针菇Flammulina filiformis是我国产量最高的工厂化栽培食用菌。为提高优良工厂化栽培金针菇种质的育种效率,本研究以国内外收集的105份金针菇种质为材料,开展体细胞不亲和评价,并采用SSR分子标记的方法对所有种质进行遗传多样性分析和聚类分析。20对SSR引物在105份种质中共扩增得到209个等位基因位点,所有种质间的遗传相似系数为0.71-1.00,在遗传距离0.76处可分为5个大类群。105份金针菇种质共包含67种不同的遗传背景,野生金针菇种质比栽培种质具有更丰富的遗传多样性。基于SSR的聚类分析结果和体细胞不亲和评价结果既相互印证,又可互为借鉴。本研究构建了包含44份金针菇种质的核心种质群体,占所有供试材料的41.90%,保留了100%等位基因。核心种质群体覆盖区域广泛,最大限度地保留了原始群体的遗传多样性和表型变异,可为育种的亲本选择提供参考。进一步构建了能同时反映每份金针菇种质SSR分子标记指纹图谱、收集地区、子实体颜色和栽培性状的分子身份证编码,并转换成可视二维码,为金针菇种质的高效标识和快速溯源提供了科学依据。     

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。     

甘蓝型黄籽油菜种皮色泽QTL作图

甘蓝型黄籽油菜具有低纤维、高蛋白及高含油量的优点,因而己成为广大油菜育种工作者研究的重点之一。利用甘蓝型黑籽品系油研2号作父本,计蓝型黄籽品系GH06为母本,获得132个单株的F2群体;以AFLP和SSR为主要分析方法,构建了包括164个标记的甘蓝型油菜遗传连锁图谱,其中包括125个AFLP标记、37个SSR标记及一个RAPD和一个SCAR标记,分布在19个连锁群上,覆盖油菜基因组2549．8cM,标记间平均距离15．55cM。利用多区间作图法,对种皮色泽QTL进行分析,在第5及第19连锁群上各检测到一个QTL位点,分别解释表型变异46％及30．9％。     

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。     

Fine mapping of the yellow seed locus in Brassica juncea L

The yellow mustard plant in Northern Shaanxi is a precious germplasm, and the yellow seed trait is controlled by a single recessive gene. In this report, amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) techniques were used to identify markers linked to the brown seed locus in an F(2) population consisting of 1258 plants. After screening 256 AFLP primer combinations and 456 pairs of SSR primers, we found 14 AFLP and 2 SSR markers that were closely linked to the brown seed locus. Among these markers, the SSR marker CB1022 showed codominant inheritance. By integrating markers previously found to be linked to the brown seed locus into the genetic map of the F(2) population, 23 markers were linked to the brown seed locus. The two closest markers, EA02MC08 and P03MC08, were located on either side of the brown seed locus at a distance of 0.3 and 0.5 cM, respectively. To use the markers for the breeding of yellow-seeded mustard plants, two AFLP markers (EA06MC11 and EA08MC13) were converted into sequence-characterized amplified region (SCAR) markers, SC1 and SC2, with the latter as the codominant marker. The two SSR markers were subsequently mapped to the A9/N9 linkage group of Brassica napus L. by comparing common SSR markers with the published genetic map of B. napus. A BLAST analysis indicated that the sequences of seven markers showed good colinearity with those of Arabidopsis chromosome 3 and that the homolog of the brown seed locus might exist between At3g14120 and At3g29615 on this same chromosome. To develop closer markers, we could make use of the sequence information of this region to design primers for future studies. Regardless, the close markers obtained in the present study will lay a solid foundation for cloning the yellow seed gene using a map-based cloning strategy.     

甘蓝型特高含油量油菜种质资源及主栽品种指纹图谱构建

本研究选择特高含油量资源7份,与中国各油菜主产区具有代表性的主栽品种16份,利用SSR多引物组合法开展指纹图谱构建研究。选择多态丰富、图谱清晰稳定且来自不同连锁群的引物28对,对所有材料进行指纹图谱分析,共获SSR指纹条带302条,其中多态性条带为279条,每引物所获条带6-16条,平均10.79条,平均多态率92. 38%,通过指纹图谱将所有材料有效地区别开来。用非加权类平均法（UPGAM）聚类分析显示：高油材料之间以及高油材料与主栽品种之间遗传距离均有较大差异,在遗传距离0.171处可将23份材料分成9个类群,其中7份高油材料分处4个类群,遗传距离差异显著;而其他8份主栽品种被分别聚类在另外5个类群中;所有材料间皆具有丰富的遗传多样性,其中高油材料与主栽品种间遗传差异更大。     

The development of multiplex simple sequence repeat (SSR) markers to complement distinctness,uniformity and stability testing of rape (Brassica napus L.) varieties

To assess the potential of multiplex SSR markers for testing distinctness, uniformity and stability of rape (Brassica napus L.) varieties, we developed three multiplex SSR sets composed of five markers each. These were used to measure the extent of diversity within and between a set of ten varieties using a fluorescence-based semi-automated detection technology. Also, we evaluated the significance of any correlation between SSRs, pedigree and five of the morphological characters currently used for statutory distinctness, uniformity and stability testing of rape varieties. An assignment test was allowed to identify 99% of the plants examined, with the correct variety based on the analysis of 48 individual plants for each variety. Principal coordinate analysis confirmed that a high degree of separation between varieties could be achieved. Varieties were separated in three groups corresponding to winter, spring and forage types. These results suggested that it should be possible to select a set of markers for obtaining a suitable separation. Diversity within varieties varied considerably, according to the variety and the locus examined. No significant correlation was found between SSR and morphological data. However, genetic distances measured by SSRs were correlated to pedigree. These results suggested that SSRs could be used for pre-screening or grouping of existing and candidate varieties, allowing the number of varieties that need to be grown for comparison to be reduced. Multiplex SSR sets gave high-throughput reproducible results, thus reducing the costs of SSR assessment. Multiplex SSR sets are a promising way forward for complementing the current variety testing system in B. napus.     

鉴定烟草种质资源SSR核心引物筛选和验证

核心引物对遗传多样性分析、种质资源鉴定、品种纯度和真实性检测、指纹图谱构建等研究具有重要价值。本研究利用均匀分布于烟草24条染色体的278对SSR引物,对20份亲缘关系相对较远的烟草材料进行初步筛选,筛选出32对引物。随后再加上10份亲缘关系较近的材料（共30份）对引物进行复筛,最终确定14对为核心引物。将14对引物对39份烟草材料进行进行系谱分析、品种遗传多样性分析和农艺性状分析聚类,结果表明该套SSR核心引物适用于烟草种质资源鉴定和遗传多样性分析。     

Genetic differentiation analysis of African cassava (Manihot esculenta) landraces and elite germplasm using amplified fragment length polymorphism and simple sequence repeat markers

Molecular‐marker‐aided evaluation of germplasm plays an important role in defining the genetic diversity of plant genotypes for genetic and population improvement studies. A collection of African cassava landraces and elite cultivars was analysed for genetic diversity using 20 amplified fragment length polymorphic (AFLP) DNA primer combinations and 50 simple sequence repeat (SSR) markers. Within‐population diversity estimates obtained with both markers were correlated, showing little variation in their fixation index. The amount of within‐population variation was higher for landraces as illustrated by both markers, allowing discrimination among accessions along their geographical origins, with some overlap indicating the pattern of germplasm movement between countries. Elite cultivars were grouped in most cases in agreement with their pedigree and showed a narrow genetic variation. Both SSR and AFLP markers showed some similarity in results for the landraces, although SSR provided better genetic differentiation estimates. Genetic differentiation (Fst) in the landrace population was 0.746 for SSR and 0.656 for AFLP. The molecular variance among cultivars in both populations accounted for up to 83% of the overall variation, while 17% was found within populations. Gene diversity (H_e) estimated within each population varied with an average value of 0.607 for the landraces and 0.594 for the elite lines. Analyses of SSR data using ordination techniques identified additional cluster groups not detected by AFLP and also captured maximum variation within and between both populations. Our results indicate the importance of SSR and AFLP as efficient markers for the analysis of genetic diversity and population structure in cassava. Genetic differentiation analysis of the evaluated populations provides high prospects for identifying diverse parental combinations for the development of segregating populations for genetic studies and the introgression of desirable genes from diverse sources into the existing genetic base.     

Diversity array technology markers: genetic diversity analyses and linkage map construction in rapeseed (Brassica napus L.)

We developed Diversity Array Technology (DArT) markers for application in genetic studies of Brassica napus and other Brassica species with A or C genomes. Genomic representation from 107 diverse genotypes of B. napus L. var. oleifera (rapeseed, AACC genomes) and B. rapa (AA genome) was used to develop a DArT array comprising 11 520 clones generated using PstI/BanII and PstI/BstN1 complexity reduction methods. In total, 1547 polymorphic DArT markers of high technical quality were identified and used to assess molecular diversity among 89 accessions of B. napus, B. rapa, B. juncea, and B. carinata collected from different parts of the world. Hierarchical cluster and principal component analyses based on genetic distance matrices identified distinct populations clustering mainly according to their origin/pedigrees. DArT markers were also mapped in a new doubled haploid population comprising 131 lines from a cross between spring rapeseed lines 'Lynx-037DH' and 'Monty-028DH'. Linkage groups were assigned on the basis of previously mapped simple sequence repeat (SSRs), intron polymorphism (IP), and gene-based markers. The map consisted of 437 DArT, 135 SSR, 6 IP, and 6 gene-based markers and spanned 2288 cM. Our results demonstrate that DArT markers are suitable for genetic diversity analysis and linkage map construction in rapeseed.     

Divergent patterns of allelic diversity from similar origins: the case of oilseed rape (Brassica napus L.) in China and Australia.

Oilseed rape (Brassica napus) in Australia and China have similar origins, with introductions from Europe, Canada, and Japan in the mid 20th century, and there has been some interchange of germplasm between China and Australia since that time. Allelic diversity of 72 B. napus genotypes representing contemporary germplasm in Australia and China, including samples from India, Europe, and Canada, was characterized by 55 polymorphic simple sequence repeat (SSR) markers spanning the entire B. napus genome. Hierarchical clustering and two-dimensional multidimensional scaling identified a Chinese group (China-1) that was separated from "mixed group" of Australian, Chinese (China-2), European, and Canadian lines. A small group from India was distinctly separated from all other B. napus genotypes. Chinese genotypes, especially in the China-1 group, have inherited unique alleles from interspecific crossing, primarily with B. rapa, and the China-2 group has many alleles in common with Australian genotypes. The concept of "private alleles" is introduced to describe both the greater genetic diversity and the genetic distinctiveness of Chinese germplasm, compared with Australian germplasm, after 50 years of breeding from similar origins.     

上海地区芸薹属蔬菜遗传多样性研究

利用SSR分子标记分析和农艺性状鉴定对上海地区的17份甘蓝、44份白菜,共61份芸薹属蔬菜进行遗传多样性评价。UPGMA聚类结果显示,61份材料被聚类为两大类,甘蓝类和白菜类,其中白菜型蔬菜品种的相似系数在0．96～0．77之间,甘蓝型蔬菜品种的相似系数在0．93～0．63之间。聚类分析的结果说明,上海地区的甘蓝类蔬菜遗传多样性比较丰富,而白菜类蔬菜的遗传基础比较单一,急需保护现有的白菜型蔬菜品种的种质资源,防止遗传流失。本实验还说明通过SSR分子标记技术与农艺性状鉴定相结合,综合评价芸薹属蔬菜的遗传多样性比采用单一的方法更加准确有效。     

Genetic diversity analysis of North Africa’s barley using SSR markers

It was demonstrated that some North Africa barley accessions have diverse tolerance sources for abiotic stresses and a good nutritional quality, but the studies done were incomplete since they were realized separately in each country apart.To implement a more complete analysis, 31 barley accessions originated from North Africa (Algeria, Tunisia and Egypt) were analyzed using 11 SSR markers selected from the seven barley linkage groups for studying the genetic diversity among these chosen barley accessions.Over the 11 SSR markers, a total of 478 reproducible bands were scored with an average of 2.13 alleles/primer and the average polymorphism information content of 0.5.Genetic distance analysis of the 31 accessions showed a large genetic diversity and high number of different groups. The most accessions are clustered according to their eco-geographical origin, according to their pedigree and agronomic characters or according to the caryopsis character (hulled or naked caryopsis). This high number of obtained groups is an invaluable aid in crop improvement strategies and confirms the opinion suggesting that North Africa could be a secondary center of origin of barley. The various growing conditions and the multiple uses of barley in each country may be the cause of the large variability of the barley germplasm in each region.