Genetic diversity and population structure of Indian soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.] revealed by simple sequence repeat markers

Genetic diversity in 90 Indian soybean cultivars was assessed using 45 SSR markers distributed on 20 soybean chromosomes. Forty-five SSR markers generated 232 alleles with an average of five alleles/locus. The observed frequencies of the 232 alleles ranged from 0.01 to 0.94 with an average of 0.19. The polymorphic information content (PIC) value of the SSR markers varied from 0.10 to 0.83 with an average of 0.61 and about 71% markers have a PIC value of >0.5. In this study, 54 rare alleles including 19 genotype specific alleles were also identified. The observed hetrozygosity for SSR markers ranged from 0 to 0.11 with a mean of 0.10. Cluster analysis grouped the 90 soybean cultivars into three major clusters and principal coordinates analysis (PCoA) results were similar to those of the cluster analysis. A combination of eight SSR markers successfully differentiated all 90 soybean cultivars. The population structure analysis distributed the 90 soybean genotypes into two populations with mean alpha (α) value of 0.1873. In AMOVA analysis, proportion of variation within population was high (88%), whereas only 12% occurred among populations. In cluster and structure analyses, most of the genotypes with similar pedigree were grouped together. Soybean cultivars DS228, MACS-13, LSb-1, Hardee, Improved Pelican, and Pusa-24 were the six most genetically distinct cultivars identified. The study reported a moderate genetic diversity in Indian soybean cultivars and findings would be useful to the soybean breeders in selecting genetically distinct parents for a soybean improvement program.     

Assessment of genetic diversity in cabbage cultivars using RAPD and SSR markers

Genetic relationship and diversity among seven cabbage cultivars were analyzed using RAPD and SSR markers. These cultivars are of great commercial value in India and are confirmed for their reaction to black rot caused by Xanthomonas campestris pv. campestris. However, so far the extent of genetic diversity and relatedness has not been studied in these cultivars. A total of 17 selected RAPD primers generated 90 bands, 76 of which were polymorphic (84.44%). In addition, 27 selected SSR primers generated 67 amplified bands with 59 of which were polymorphic (87.6%). Though both the marker techniques were able to discriminate the cultivars effectively, analysis of combined data of markers (RAPD and SSR) resulted in better distinction of cultivars. By combining both the markers, a total of 157 bands were detected of which 135 bands (85.98%) were polymorphic, i.e. an average of 5.95 bands per primer. High level of polymorphism (> 85%) recorded with two different marker systems indicated a high level of genetic variation existing among the cultivars. Genetic relationship estimated using similarity co-efficient (Jaccard’s) values between different pairs of cultivars varied from 0.21 to 0.77 in RAPD, 0.42 to 0.82 in SSR, and 0.43 to 0.89 with combined markers. A high correspondence had been recorded between the values of genetic variations generated by UPGMA, clustering, and scatter plot diagrams. The cultivars ‘January King Sel. Improved’ and ‘Golden Acre’ are highly divergent cultivars as demonstrated by both the marker systems.     

2012年北方春大豆国家区试大豆品种纯度鉴定、分子ID构建及遗传多样性分析

为了鉴定北方春大豆组国家区域试验大豆品种的纯度、构建参试大豆品种的分子ID及品种间遗传关系分析,从而有效地指导中国北方春大豆新品种的选育和推广。通过对参加2012年北方春大豆国家区试的94份大豆材料用分布在大豆基因组8个连锁群的13对SSR标记进行分析。结果表明：94份参试大豆品种纯度分布范围为53.85%~100%,平均纯度为99.02%;利用Satt231、Satt288、Satt160、Satt193和Sat-092这5对引物可以将94份参试大豆品种区分开,并获得唯一的分子ID;94个参试大豆品种间遗传相似系数为0~0.846,平均相似系数为0.2783,说明参试大豆品种间遗传差异性较大。通过SSR标记分析,可以有效地鉴定参试大豆品种的纯度、建立参试大豆品种的分子ID及实现品种间遗传关系分析。     

大豆品种成熟期基因型推测的研究

选择不同来源中国大豆品种23份和国外引进的成熟期近等基因系35份进行SSR分析,目的是鉴定与成熟期基因型紧密连锁的标记,进而推测中国大豆品种的成熟期基因型。结果表明,（1）210对SSR标记中125对在成熟期近等基因型中具有多态性,推测与成熟期有关的标记有8个;（2）在Clark近等基因系中,筛选出成熟期基因E3/e3特异性标记Satt229,E4/e4的特异SSR标记Sct_010、Satt294、Satt247、Satt452和Satt156;在Clark和Harosoy近等基因系中,筛选出E7/e7的特异性SSR标记为Satt071、Satt178;（3）根据8个与成熟期相关的标记的分子数据,构建了国外大豆近等基因系的UPGMA聚类图,共聚为4类,背景来源相同或相似的材料被聚为一类,明显分为Clark近等基因系和Harosoy近等基因系。（4）与近等基因系成熟期基因(E7)分子标记比对,推测出25份中国大豆品种的成熟期基因。     

Molecular characterisation of cultivars of apple (Malus × domestica Borkh.) using microsatellite (SSR and ISSR) markers

In this study, two microsatellite-based methodologies (SSR and ISSR) were evaluated for potential use in fingerprinting and determination of the similarity degree between 41 commercial cultivars of apple previously characterised using RAPD and AFLP markers. A total of 13 SSR primer sets was used and 84 polymorphic alleles were amplified. Seven ISSR primers yielded a total of 252 bands, of which 176 (89.1%) were polymorphic. Except for cultivars obtained from somatic mutations, all cultivars were easily distinguishable employing both methods. The similarity coefficient between cultivars ranged from 0.20 to 0.87 for SSR analysis and from 0.71 to 0.92 using the ISSR methodology. Dendrograms constructed using UPGMA cluster analysis revealed a phenetic classification that emphasises the existence of a narrow genetic base among the cultivars used, with the Portuguese cultivars revealing higher diversity. This study indicates that the results obtained based on the RAPD, AFLP, SSR and ISSR techniques are significantly correlated. The marker index, based on the effective multiplex ratio and expected heterozygosity, was calculated for both analyses (MI = 1.7 for SSR and MI = 8.4 for ISSR assays) and the results obtained were directly compared with previous RAPD and AFLP data from the same material. The SSR and ISSR markers were found to be useful for cultivar identification and assessment of phenetic relationships, revealing advantages, due to higher reproducibility, over other commonly employed PCR-based methods, namely RAPD and AFLP. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

黑龙江省主栽大豆品种遗传多样性和群体结构分析

利用187对SSR标记对近25年（1992-2017）在黑龙江省栽培的202个大豆品种进行遗传多样性和群体结构分析。结果表明,从试验材料的基因组DNA中扩增出多态性位点808个,平均每对引物扩增出多态性位点4.42个;多态性位点最多的引物是satt703和satt311,均为10个;等位变异频率最高的引物是satt417和satt575,等位变异频率均为99.5%。供试品种间的遗传相似系数为0.283~0.930,平均值为0.519。同一个育种单位育成的部分品种具有较高的遗传相似性。群体结构、主坐标分析和NJ聚类将202个品种划分的结果是一致的,均为3个类群。类群中的部分材料血缘不是独立的,而是相互渗透的。     

新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析

以新疆截止2012年审定的23份新彩棉品种为材料,利用SSR标记进行DNA指纹图谱的构建和遗传多样性分析。从5000对SSR引物中,挑选出多态性高、稳定性好、均匀分布在棉花26条染色体上的52对引物,在23份新彩棉品种中筛选出核心引物47对,SSR扩增检测到多态性基因型位点数共计162个,每个标记检测到的基因型位点数在2~7之间,平均为3.45个;引物多态信息量(PIC)值介于0.4537~0.8686之间,平均值为0.7096。结果显示:在23份新彩棉品种中,14份品种采用特异或特征引物可以一次性区分开,其余9份品种需要采用引物组合来实现区别该品种与其他品种。最少选用18对特异引物及组合引物就可以完全区分开新彩棉1~23号品种。利用18对SSR标记构建了新彩棉1号至23号品种的指纹图谱。利用NTSYS-pcV2.10软件聚类分析表明:23个新彩棉品种遗传相似系数变化范围是0.3781~0.9298,平均为0.5511,表明新彩棉品种之间存在着丰富的遗传多样性。     

Random amplified polymorphic DNA variation among German and Dutch asparagus cultivars

DNA polymorphism among nine cultivars of Asparagus officinalis L. was measured using random amplified polymorphic DNA (RAPD). Of 69 reproducible amplification products from 12 arbitrary decamer primers, 49 RAPD markers were polymorphic and could be used to distinguish six German and three Dutch asparagus cultivars. Even with very small sample sizes, genetic similarity measurements based on the RAPD data allowed accurate grouping of the nine cultivars into distinct clusters, with the exception of two individuals which clustered to closely related varieties. Two German cultivars showed high genetic similarity and were distinct from the remaining German varieties. The German and Dutch cultivars were clearly separated by a relatively large genetic distance.     

Genetic diversity in soybean as determined by RAPD and microsatellite analysis

The random amplified polymorphic DNA (RAPD) and microsatellite techniques were used to evaluate the genetic diversity among 18 soybean genotypes selected for a breeding programme to increase the protein content of varieties adapted for central European growing conditions. Out of 33 random primers used in RAPD reactions, only 12 showed polymorphism useful for characterization of these genotypes. In contrast, all 12 microsatellite primer pairs used in this study detected polymorphism with 2–6 alleles per locus. Similarity measures and cluster analysis were made using RAPD and simple sequence repeat (SSR) data, separately and together. The resulting dendrograms were compared with each other and with the available pedigree information as a control. The dendrogram derived from RAPD data showed some divergence from the pedigree information available for the lines. The dendrograms based on SSR data and SSR data combined with RAPD gave very good agreement with pedigree information. It can be concluded that the combined use of a limited number of RAPD and SSR markers is a useful and reliable means of evaluating genetic relationships of genotypes in the absence of pedigree data.     

利用与大豆灰斑病抗性基因连锁的SSR标记构建品种(系)的分子身份证

以黑龙江省29个大豆育种单位的103份已鉴定大豆灰斑病3个生理小种抗性的大豆品种(系)为材料,选择与大豆灰斑病抗病基因连锁的19个SSR标记检测,获得等位变异数86个,每个标记检测到的等位变异数分布在2~6个之间,平均为4.42个。应用遗传统计软件(genetics statistics 3.0)分析表明, 标记的多样性指数介于0.198~0.751之间,平均多样性指数为0.606。品种(系)特异指数差异较大,介于46.592~481.541之间,平均为87.415。根据标记的等位基因数,使用ID Analysis 1.0软件分析表明,利用与大豆抗灰斑病基因连锁的7个SSR标记(Satt565、Satt547、Satt431、Sct_186、SOYGPATR、Satt244、Sat_151)就能有效区分各品种(系),因此利用这7个标记构建了供试品种(系)的分子身份证。     

青稞遗传多样性及其农艺性状与SSR标记的关联分析

利用92个SSR标记对108份青稞亲本材料进行多态性扫描,分析其遗传多样性,旨在寻找与农艺性状相关联的分子标记,为青稞杂交组合的配制及分子标记辅助育种提供依据。挑选48个多态性标记进行群体遗传结构分析,在此基础上采用Tassel 2.1 GLM (general linear model)和MLM (mixed linear model)方法进行标记与农艺性状的关联分析。共检测出156个等位变异,每个位点2~6个等位变异。供试群体的Shannon指数为0.6727~1.1368,材料间遗传相似系数为0.2250~1.0000,平均0.7585。通过群体遗传结构分析将供试材料划分成4个亚群。以GLM分析,发现12个与株高、穗长、穗粒数和分蘖数相关联的标记,对表型变异的解释率分别为11.5%~17.6%、19.4%~45.4%、15.4%~22.1%和29.2%;以MLM分析,发现8个与株高、分蘖数和小穗数相关的标记,各标记对表型变异的解释率分别为31.7%~49.8%、28.1%~37.2%、22.7%~32.7%。关联标记分布在基因组全部6个连锁群上。     

应用SSR和ISSR标记分析栽培香稻品种的遗传多样性

本研究利用24对SSR引物和36个ISSR引物,分析33份来源于亚洲10个国家的香稻品种的遗传多样性。分别获得93条和181条多态性片段,每个SSR座位可检测3～8个等位基因,平均为4．23个;每个ISSR引物可检测3～8个多态性位点,平均为5．03个。根据SSR和ISSR标记计算的品种间遗传相似系数分别在0．294～0．884之间和0．595～0．867之间。聚类分析表明,利用两种标记所得的聚类结果基本上一致,与品种所处的3种气候类型变化基本相符。进一步证实SSR和ISSR标记是研究水稻种质资源分类有效的工具。     

应用RAPD分析四川马铃薯地方品种与部分栽培品种间遗传关系

为探讨四川省马铃薯地方品种与栽培品种间遗传关系,评价其在遗传研究与育种利用价值,指导杂交亲本选配,对21份地方品种和10个栽培品种进行了RAPD分析。选取8份差异较大的材料对170条RAPD引物进行筛选,获得20条多态性高、条带清晰、稳定的RAPD引物。20条RAPD引物共扩增出191个位点,其中多态性位点172个;平均多态性位点比率90.1%,平均多态性信息量(PIC)为0.822;栽培品种、地方品种平均PIC分别为0.823和0.814。遗传距离与聚类分析表明：31份材料间平均遗传距离0.4274,变幅为0.1481~0.5852,地方品种间平均遗传距离和变幅均高于栽培品种间,地方品种与栽培品种间平均遗传距离为0.4496;表明地方品种中可能存在更广泛的遗传变异,具有较高的育种利用价值。大部分地方品种出乎意料与栽培品种聚在不同的类中,与供试栽培品种间亲缘关系较远,可能具有较高的杂交组配潜力;而N5-33、N5-38和N6-22可能分别源自于大西洋、疫不加与南湖塔(或米拉),值得深入对比研究。     

RAPD genetic diversity of Albanian olive germplasm and its relationships with other Mediterranean countries

RAPD markers were used for the study of 19Albanian olive cultivars and two wild olives (oleasters). A total of 76polymorphic bands (4.8 polymorphic markers per primer) out of 107 reproducible were obtained using 16 primers. The number of bands per primer ranged from 4 to 10,whereas the number of polymorphic bands ranged from 1 to 9, corresponding to 71%of the total amplification products. All the accessions could be identified by the combination of four primers: OPA-19;OPA-02; OPK-16 and OPP-19. The dendrogram,based on Jaccard's index, included three major groups according to their origin: 1)most of the cultivars from the area of Berat (South of Albania) 2) cultivars from the Centre and Centre-North of Albania and3) cultivars from the Centre and North-West of Albania along with the oleaster from Elbasan. In order to evaluate the origin of Albanian cultivars they were compared to those diffused in other countries like Greece, Italy and Turkey, due to geographical and historical affinity among these countries, by using a one way AMOVA. Although most of the genetic diversity was attributable to differences among cultivars within each country (91.47%) significantφ-values among countries(φ_st = 0.085; p < 0.001)suggested the existence of RAPD phenotypic differentiation. Significant φ-values in all pairs formed by Albania with the other countries were observed. These results are consistent with the autochthonous origin of Albanian cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.     

新疆早熟棉品种SSR指纹图谱构建与品种鉴别

以新疆自1978年以来审定命名的42个早熟棉品种为材料,利用SSR分子标记对新疆早熟棉品种进行研究。从2300对SSR引物中筛选出了52对具有稳定多态性的引物,每对引物可检测到3～24个多态性片段,共检测到506个,平均9.7个,片段大小介于100～2000bp之间。对所得到的52个SSR指纹图谱进行组合和综合分析,用其中2对就可将所有供试品种区分开来。同时,分别对42个早熟棉品种进行指纹分析,获得各个品种的特异性指纹,可为今后棉花种子真伪快速鉴别奠定基础。     

Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.     

Molecular Marker based Genetic Diversity Analysis in Rice (Oryza sativa L.) using RAPD and SSR markers

The availability of an array of molecular marker systems allowed comparing the efficiency of two of these marker systems to estimate the relationships among various taxa. The objective of this study was to assess the genetic diversity among 40 cultivated varieties and five wild relatives of rice, Oryza sativa L. involving simple sequence repeat (SSR) randomly amplified polymorphic DNA (RAPD) markers. The accessions were evaluated for polymorphisms after amplification with 36 decamer primers and 38 SSR primer pairs. A total of 499 RAPD markers were produced among the 40 cultivated varieties and five wild relatives with a polymorphism percentage of 90.0. Out of 38 SSR primer pairs used, only one locus viz., RM115 was monomorphic. The average Polymorphism Information Content (PIC) value was 0.578 and it ranged from a low of zero (RM 115) to a high of 0.890 (RM 202). The Mantel matrix correspondence test was used to compare the similarity matrices and the correlation coefficient was 0. 582. The test indicated that clusters produced based on RAPD and SSR markers were not conserved since matrix correlation value was 0.582 as against the minimum required value of 0.800. The two marker systems contrasted most notably in pair-by-pair comparisons of relationships. SSR analysis resulted in a more definitive separation of clusters of genotypes indicating a higher level of efficiency of SSR markers for the accurate determination of relationships between accessions that are too close to be accurately differentiated by RAPD markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and identification of cymbidium cultivars as measured by random amplified polymorphic DNA (RAPD) markers

DNA from thirty-six cymbidium cultivars was examined using polymerase chain reaction (PCR) to determine the efficiency of randomly amplified polymorphic DNA (RAPD) markers in identifying cultivars and determining levels of genetic variability. A total of 132 RAPD markers, 78% of which were polymorphic, were produced from 15 10mer arbitrary primers. All the cultivars were distinguishable when a number of primers was considered. One cultivar, Blue Smoke ‘Green Meadow’ could be distinguished from all the rest based only on lack of the OPA5-370 fragment. Genetic distances among the cultivars were estimated based on the amount of band sharing and ranged from 0.08–0.50 with an average of 0.29. Cluster analysis of genetic distance estimates grouped siblings together with each other and parents with offsprings, thereby agreeing with known parentage information and corroborating isozyme data obtained from a separate study. The possible application of the observed polymorphism and variation to cymbidium breeding is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.