SSR和SRAP标记揭示甘蓝型油菜遗传多样性的差异分析

分别用SSR和SRAP标记分析了来自国内外的19个甘蓝型油菜（Brassicanapus L.）品种间的遗传距离及其与选育年代的相关关系,并构建聚类图。从726个SRAP引物组合中筛选出23对多态性较好的组合扩增得到了234个多态性标记;35对SSR引物获得了138条多态性标记。结果显示,SSR标记揭示的遗传距离大于SRAP标记。SSR揭示的国内甘蓝型油菜品种间平均遗传距离大于国外品种,而SRAP标记得到的结果与SSR标记分析所得结果相反。SSR平均遗传距离与育成年代的相关性较小,SRAP平均遗传距离与育成年代存在一定的负相关,其相关系数为-0.34。研究认为,SRAP标记更适用于通过分析遗传距离以获得杂种优势;从种质资源保存的角度出发,使用SSR标记能较全面保证种质资源遗传多样性;结合SSR标记和SRAP标记即能从全基因组又能有重点的从功能基因组分析育种材料的遗传多样性,对于中长期的育种目标有较大的意义。     

甜菜品种SSR指纹图谱的构建及遗传多样性分析

筛选出20对多态性高、带型清晰、重复性好的SSR标记引物对供试甜菜品种进行PCR扩增,根据扩增产物构建SSR指纹图谱,并分析其遗传多样性。结果显示,共检测出112个等位基因,平均每对引物5.6个,利用获得的等位基因计算遗传距离,107个品种的遗传距离变化范围在0.065~0.467之间,平均遗传距离0.298。Shannon’s多样性指数变异范围0.78~2.90;PIC值介于0.08~0.83;Nei’s指数介于0.39~1.87。利用类平均法（UPGMA）进行聚类分析,可将107个甜菜品种分为2个类群。类群Ⅰ29个品种,类群Ⅱ78个品种。类群Ⅰ和Ⅱ又可分为多个亚群。结果表明,每个甜菜品种有区别于其他品种唯一的数字指纹,说明用于试验的20对SSR标记适用于甜菜品种真实性的鉴定,同时甜菜品种指纹图谱库的构建也为甜菜品种鉴定提供技术基础。     

高粱SSR和EST-SSR标记在割手密中的通用性分析

目前, 割手密中开发的分子标记有限, 为增加其分子标记数量, 本研究对高粱分子标记在割手密中的通用性进行分析。选用高粱的 29对基因组 SSR标记和 20对 EST-SSR标记在割手密种质 GSM39中进行筛选, 以评价其通用性。进一步利用 14份割手密材料评价标记的多态性和遗传多样性。结果显示： 70%的 EST-SSR引物在割手密中得到成功扩增, 可用的 EST-SSR引物中多态性比率占 50%。SSR引物在割手密中的通用性比率只有 34.5%, 但多态性比率为 70%。14对多态性引物在 14份割手密中共检测到33个等位基因, 平均观测等位基因数为2.4286, 平均有效等位基因数为1.7, 平均观测表观杂合度为0.544, 平均期望杂合度为 0.3858, Nei’ s基因多样性指数平均值为 0.3716。结果表明, EST-SSR引物的通用性高于 SSR引物, 但 SSR引物的多态性更高。这对割手密遗传多样性研究和比较基因组学研究具有重要意义。     

陕西汉中地区主栽水稻品种的SSR多态性分析

掌握陕西省汉中地区主栽水稻品种的遗传多样性,了解其遗传背景,明确各品种间的亲缘关系。从水稻12条染色体上的30个SSR标记中筛选出14对有效引物,对陕西省汉中地区20个主要种植的水稻品种的遗传多样性进行分析。结果共检测到69个等位基因,平均每个标记检测到5个等位基因,每个SSR位点的遗传多态性信息含量在0.27~0.74之间,平均值为0.48。聚类分析表明,20个水稻品种的遗传相似系数集中在0.55~0.94之间,遗传相似度较高,亲缘关系较近。笔者认为,SSR标记所作的品种聚类分析与系谱法趋势一致,汉中地区水稻种质资源遗传基础相近是其水稻产量和品质难以突破的原因之一。     

基于转录组SSR的三角梅遗传多样性分析

为揭示三角梅的遗传背景,本研究采用软件MicroSAtellite搜索其苞片转录组SSR位点,使用Primer Premier 5软件设计50对SSR引物,将筛选出的多态性引物用于55份三角梅材料的聚丙烯酰胺凝胶电泳,采用Popgene 32、PowerMarker V3.25和Structure V2.3.4等软件分析三角梅遗传多样性。结果显示,从50对SSR引物中筛选出25对条带清晰且多态性好的引物,25对SSR引物共检测到76个等位基因,平均每对引物有3个等位基因,平均有效等位基因数为2.30,引物多态性信息量(PIC)在0.01～0.66之间,均值为0.38。55个三角梅材料间的遗传距离为0.01～0.92,平均遗传距离为0.43;在遗传距离0.44处,聚类分析将55份材料分为3个群体。与印度三角梅栽培种比较而言,本实验研究的三角梅材料的遗传多样性水平不高,结果可为三角梅品种鉴定、保护及进化提供一定参考价值。     

21种不同类型葡萄种质资源遗传多样性的SSR分析

本研究通过SSR标记解析不同类型葡萄种质遗传多样性差异和亲缘关系,为资源分类与品种鉴定提供理论依据。利用15对SSR引物对21种不同类型葡萄种质进行PCR扩增,对扩增情况、遗传距离与亲缘关系等进行分析。15对SSR引物共扩增出等位基因位点91个,其中多态性位点89个,多态性比率高达97.8%,其中13对引物扩增多态性百分率达到100%。21份供试材料间遗传相似系数变化范围在0.45~0.91之间,并在遗传相似系数0.62处被分为2个类群。研究表明SSR标记从分子水平上解析了不同类型葡萄种质的遗传差异性,揭示了材料间亲缘关系,为育种研究提供了一定参考依据。     

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。     

利用SSR荧光标记构建20个高粱品种指纹图谱

为建立高粱种子纯度及真实性鉴定技术体系,构建高粱栽培品种DNA指纹图谱数据库是必要的。利用SSR荧光标记技术筛选出36对SSR荧光标记引物,构建我国高粱杂种优势利用以来中晚熟区主推的20个品种的SSR指纹图谱。36对SSR引物共扩增出235个等位基因,平均每个等位基因检测到多态性位点7个,多态性位点变化范围为2~12个。20个高粱品种36个SSR位点的遗传多样性指数和多态性信息量的变化范围分别为0.3490~0.8813和0.3144~0.8696,平均值分别为0.6976和0.6571。从36对SSR引物中筛选到多态性丰富的2对引物作为高粱品种鉴定的特征引物,2对SSR特征引物组合可区分所有供试品种。20个高粱品种的SSR指纹图谱互不相同,可以作为各品种特定的图谱,为品种鉴别提供依据。     

利用荧光SSR和MFLP标记技术分析烟草核心种质的遗传多样性

了解烟草种质材料的遗传多样性,可为其有效利用与保护提供依据。据此,本研究采用荧光SSR和荧光MFLP技术,对来源于12个不同国家和地区94份烟草核心种质材料进行遗传多样性分析。结果显示,17对SSR和52对MFLP引物组合分别检测出34个和136个具多态性的等位变异位点,平均每对SSR和MFLP引物分别为2个和2.61个。SSR标记的多态性信息量(PIC)变幅为0.126 4~0.466 2,平均为0.281 1;MFLP标记的PIC值在0.119 5~0.803 4之间,平均为0.462 4。遗传多样性分析表明,94份烟草核心种质材料间的遗传距离在0.095 2~0.902 6之间,平均为0.453 9;遗传相似系数在0.444 4~0.905 5之间,平均为0.664 8。UPGMA聚类分析和主坐标分析均将94份材料分为6大类,且两种方法能相互对应、补充和验证。此外,依据12个不同地区烟草种质材料间的遗传一致度,可将其聚类为4个大组,其中第IV大组可以分为4个亚组。本研究结果表明94份烟草核心种质的遗传多样性较丰富,合理地利用这些种质材料,将有利于拓宽中国烟草品种的遗传基础。     

贵州马铃薯引进品种SSR分子标记及遗传多样性分析

为了鉴定马铃薯品种间的亲缘关系,采用5对SSR分子标记引物,对18个贵州马铃薯生产品种进行SSR分子标记及遗传多样性分析.结果表明:5对SSR引物共扩增出77个多态性条带,每个组合的多态性条带数为10～24不等,平均每个引物组合产生15.4个多态性条带.18个马铃薯材料之间的遗传距离范围在0.376 6～0.909 0之间,平均为0.701 1.经聚类分析,18个马铃薯材料在遗传距离0.57水平上全部聚为一类,以遗传距离0.60为基准,可明显聚为4个类群.第Ⅰ类包括5个材料;第Ⅱ类仅1个材料;第Ⅲ类包括4个材料;第Ⅳ类包括8个材料.     

利用SSR荧光标记技术分析烟草种质的遗传多样性

针对中国烟草栽培品种遗传基础狭窄的情况,分析烟草种资资源的遗传多样性将可为烟草新品种选育、引种和资源保护提供重要信息。据此,利用20个SSR引物组合,采用荧光SSR标记法对来自不同国家和地区的50个烟草种质材料的遗传多样性进行分析。研究结果显示,20对SSR引物在50份烟草种质中检测到81个等位变异,平均每个SSR为4.1,平均多态性位点比率为68.4%;其中引物PT20457的鉴别能力最高,它的扩增多态位点数为6个,多态位点比率为100%,PIC值为0.944。研究的UPGMA聚类分析结果在遗传相似系数约为0.65处将50份烟草种质明显分成了3个组群,组群Ⅰ中只有1个广东晒烟,组群Ⅱ包括2个白肋烟,组群Ⅲ包括来自4个国家的47个品种。主坐标分析将所有种质分成了4个组群8个亚类。2种分析方法均较好地揭示了烟草属种间或栽培种品种类型间的遗传多样性与亲缘关系,可为烟草遗传育种和遗传连锁图谱构建的杂交亲本选择提供科学依据。     

山东省近期育成小麦品种遗传多样性的SSR分析

为了揭示山东省近年来小麦品种的遗传多样性.选用分布于小麦21条染色体上的68个SSR引物,对44个山东省小麦品种进行了遗传多样性分析.共检测到248个等位基因,每对引物的等位基因数变幅为2～9个,平均为3.65个.每个SSR位点的多态性信息量(PIC)变化范围为0.087～0.837,平均为0.562.44个品种间的遗...     

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。     

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.     

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.     

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.     

DNA Fingerprint Construction and Genetic Diversity Analysis Based on SSR Markers for Upland Cotton in Xinjiang

[Objective] The aim of this study was to construct a DNA fingerprinting database of 120 upland cotton cultivars from Xinjiang and to analyze their genetic diversity based on SSR markers. [Methods] Seventy-eight evenly distributed SSR primer pairs with high polymorphism and good repeatability were successfully screened out from 586 candidates to construct the fingerprinting database. [Result] A total of 392 alleles from 120 varieties were screened using 78 pairs of core primers, 324 of which were polymorphic loci with a polymorphism rate of 82.7%. Seventeen cultivars had specific genotypes determined using 24 primer pairs and 120 upland cotton cultivars could be identified by only 12 primer combinations. Cluster analysis indicated that genetic similarity coefficient for the 120 upland cotton cultivars ranged from 0.50 to 0.96, with an average of 0.73, indicating that upland cotton resources possess high genetic similarity and have an accordingly narrow genetic basis. [Conclusion] The primer combination method is one of the most effective methods for constructing DNA fingerprinting. The 120 upland cotton varieties were divided into three types with the genetic similarity coefficient matrix; these groups were strongly consistent with their pedigrees.     

32个棉花主栽品种DNA指纹图谱构建及遗传多样性分析

 以我国2010年32个主栽品种为材料,利用SSR标记进行DNA指纹图谱的构建和遗传多样性分析。从95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40对引物,共扩增出161种多态性基因型,平均每对引物扩增出4.025种。引物多态信息量（PIC）0.2989～0.7585,平均为0.5407。11对引物在13个品种上有特征带型,最少利用5对引物就可以将32份材料完全区分开。利用NTSYS-pc V2.10软件聚类分析表明：长江流域棉区品种间遗传差异最大,黄河流域棉区次之,新疆棉区最小;常规种遗传基础较杂交种窄。     

Development of novel microsatellite markers for effective applications in Anthurium cultivar identification

Anthurium andraeanum is one of the most economically important floral crops and potted flowers marketed worldwide. Microsatellite markers are currently the preferred molecular marker owing to the many desirable attributes, including hypervariability, codominance, and amenability to high-throughput genotyping; however, there are few polymorphic molecular markers available for Anthurium. The object of this study was to develop and characterize novel microsatellite markers using the Araceae sequences in GenBank of the National Center for Biotechnology Information (NCBI) to contribute to molecular identification for cultivar protection. Using 1,579 Araceae expressed sequence tags (ESTs) and the related nucleotide sequences, 100 candidates contained simple sequence repeat (SSR) motifs that were suitable for primer design. Furthermore, 100 pairs of SSR primers were screened against a set of 28 diverse genotypes representing 24 cultivars that included four registration cultivars which were bred from the Taiwan Agricultural Research Institute (TARI) and 20 commercial cultivars, appended with three hybrid progeny and a mutant line. From the selected six polymorphic SSR loci, 52 alleles were amplified and 27 distinct genotypes were found, except for ‘Tropical’ and its mutant, with a mean number of eight alleles per locus. The polymorphism information content (PIC) ranged from 0.86 to 0.93. Based on these results, we proposed a key identification set using four microsatellite markers that is sufficient to discriminate among 24 cultivars. Because the Anthurium microsatellite markers developed in this study are primarily from expressed sequence tags or related genomic sequences, they can be used for cultivar identification and, accordingly, contribute to genetic evaluations in breeding programs.