利用小麦SSR标记分析鸭茅种质资源的遗传多样性

使用小麦(Triticum aestivum)SSR引物和扩增程序,以中国春小麦(T. aestivum)品种为对照,利用SSR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.结果表明,20对引物扩增出295个条带,多态性条带为187条,多态性条带比率为61.15%,鸭茅种质资源的遗传相似系数范围为0.7848～0.9513.聚类分析和主成分分析将供试材料分为6大类,聚类结果不仅能反映鸭茅生态适应性特征与其生长发育状况及生产性能相关,还显示出国产鸭茅品种遗传基础较为狭窄.研究表明将小麦SSR引物用于检测鸭茅遗传多样性行之有效.     

16份石榴RAPD扩增产物的两种电泳方法检测及其序列特征

本实验以16个石榴品种为实验材料,筛选出10个重复性及多态性均较好的引物进行RAPD分析。分别采用琼脂糖凝胶以及聚丙烯酰胺凝胶(PAGE)电泳检测方法对PCR扩增结果进行检测并对其结果进行比较,结果显示,两种电泳方式均能得到较为清晰的扩增条带,且两种电泳方式获得的条带总数及多态性条带数均有所不同,琼脂糖凝胶电泳方法共检测出76条带,其中有43条为多态性谱带,多态性比率为56.4%;而在PAGE电泳方法共检测出123条谱带,多态性谱带数为87条,多态性比率为70.95%。PAGE电泳方法检测出的条带数约为琼脂糖凝胶电泳方法检测出条带数的1.5倍。基于两种电泳方法所得RAPD标记的多态性位点,利用NYSYS软件计算遗传相似系数,并构建遗传关系聚类图,分析结果显示,石榴遗传多样性丰富,两种电泳方法所得聚类结果大致相同,可以利用RAPD分子标记及两种电泳检测方法对不同数量的石榴进行分子水平的品种鉴定和遗传多样性的分析。同时通过对来自几个引物随机挑选的17个片段进行克隆,测序结果显示17个片段都是对应引物的RAPD扩增产物,其中有3条是编码蛋白的基因片段,表明了RAPD不仅扩增基因组上的非编码蛋白序列,同时也可以扩增编...     

小麦叶锈菌毒性及分子多态性分析

以来自河北、江苏两地的22株小麦叶锈菌作供试菌株，用27个小麦抗叶锈近等基因系品系作为鉴别寄主进行毒性测定。毒性多态性分析结果为：22个菌株通过聚类被分为两组，第一组17个菌株主醚自河北，第二组5个菌株全部来自江苏，表明毒性与地理来源密切相关；菌株间遗传相似系数较高并且差异不大，为0.6296-0.9259之间，说明两地的小麦叶锈菌所含的毒性基因差异不大，应用RAPD技术，从65个随机引物中筛选了12个扩增多态性较好的随机引物，用于分子多态性分析，共扩增出DNA条带102条，其中多态性条带54条，RAPD分析结果为：22个菌株被分为两组，第一组菌株主要来自河北，第二组菌株主要来自江苏，说明DNA多态性与地理来源间具有一定的相关性；菌系间遗传相似系数相差较大，为0.3889-0.9074之间，说明两地小麦叶锈菌群体遗传结构丰富而复杂，比较22株小麦叶锈菌在27个鉴别品系上的毒性特征和54个RAPD标记建立的聚类分析树状图，发现以RAPD标记为基础的分子多态性与毒性多态性相关性不强。     

基于RAPD标记的福建省稻曲病菌遗传多样性分析

为了解福建省稻曲病菌群体的遗传多样性和遗传组成,应用随机扩增多态性RAPD (random amplified polymorphic DNA)技术分析了来自福建省不同水稻种植区的102个稻曲病菌(Ustilaginoidea virens)的遗传多样性.从100条随机引物中筛选出10条扩增带清晰、重复性好的引物进行稻曲病菌多态性扩增,共扩增出157条带,多态性条带比率为82.17％,遗传距离变化范围为0.02～0.67.遗传多样性分析表明,福建省稻曲病菌具有丰富的遗传多样性,相对于其它地区而言,福建闽西地区分离的菌株遗传多样性水平最高PPB=76.43,H=0.2212,I=0.3383),晚稻分离的菌株群体遗传多样性(PPB=91.08,H=0.2402,I=0.3655)高于早稻群体(PPB=63.06,H=0.1892,I=0.2870).聚类分析显示,在遗传距离0.349水平上,供试的所有菌株可被划分成7个遗传聚类组(R1～R7),聚类组R1为优势聚类组,包含有80个菌株,其内又存有一些亚组.这些菌株的聚类与菌株的地理来源及水稻品种无明显相关性.但是在遗传距离0.330水平上,来自宁化的10个菌株可被明显划分成早稻群体和晚稻群体.初步分析认为菌株的地理来源、水稻品种及其生长季节是影响福建省稻曲病菌遗传多样性的主要因素,在稻曲病菌的遗传变异以及该病的发生和流行中可能起重要的作用.     

8份广西产绞股蓝种质的RAPD引物筛选及其遗传多样性分析

本研究的目的在于筛选合适的RAPD随机引物,应用RAPD技术对药用植物绞股蓝进行遗传多样性分析,并构建DNA指纹图谱。研究结果表明,我们利用生物信息学方法挑选出的20条引物中有19条引物的扩增条带清晰且多态性好;在清晰稳定出现的354条带中,294条具有多态性;其中有3条引物的扩增条带可清楚区分绞股蓝与混淆品种乌蔹莓,可建立其DNA指纹图谱。按UPGMA法进行聚类分析,计算其遗传相似系数,结果显示,8份绞股蓝供试材料聚为两类,聚类结果与其地理区域远近和生长环境一致。本研究中筛选出的19条引物适用于绞股蓝遗传多样性分析,且获得的DNA指纹图谱可用于鉴别绞股蓝。     

甜菜品种（系）的ISSR标记数字指纹图谱构建及聚类分析

摘要：本文针对来源于荷兰的4个引进甜菜品种和国内的6个甜菜品系（其中2个为一年生野生甜菜）进行了ISSR指纹图谱构建和聚类分析研究。筛选出稳定性高且多态性好的6个引物用于试验。利用筛选的6条引物ISSR-PCR 共扩增出51个条带, 其中多态性条带百分率为86.3%. 利用该6条引物ISSR-PCR建立的指纹图谱能将试验中的全部甜菜品种都鉴定区分开。只利用2条引物L1和UBC846 扩增的8个多态性条带构建了10个甜菜品种（系）的数字指纹识别码,该数字指纹图谱能完全区分10个甜菜品种（系）,结果显示ISSR 指纹图谱能非常有效的鉴定不同的甜菜品种。利用生物软件NTSYS-pc针对10个试验甜菜品种（系）的ISSR 扩增条带进行遗传相似性聚类分析,结果显示10个甜菜品种（系）的相似系数为0.43与0.83之间,平均为0.62。利用非加权组平均法（UPGMA）进行聚类分析,结果显示10个甜菜品种（系）聚类为2个组和3个亚组。UPGMA 聚类分析能清楚的显示10个甜菜群体间的遗传关系并且聚类结果与10个甜菜群体的特性一致, 说明ISSR标记能用于甜菜不同群体间遗传距离的评估。     

泰国产方斑东风螺养殖群体遗传多样性的RAPD分析

用随机扩增多态性DNA(RAPD)技术对泰国产方斑东风螺养殖群体的遗传多样性进行检测,从100个随机引物中筛选出21个引物对方斑东风螺的DNA进行扩增,结果表明:21个引物共检测到222条清晰且重复性好的条带,每个引物可扩增出4~16条带,分子量在200~2200bp之间,其中多态位点为156个,占70.27%;群体的Shannon多样性指数为0.2818,Nei基因多样性指数为0.2491;个体间最大遗传距离为0.291,最小遗传距离为0.066。通过与其他贝类遗传多样性的研究结果比较,可初步判断泰国产方斑东风螺养殖群体的遗传多样性比较丰富。     

广西地方食用木薯种质资源遗传多样性分析

为研究广西地方食用木薯材料的遗传多样性,以48份广西地方食用木薯为试验材料,分析其变异系数、遗传多样指数、聚类分析、多态性和遗传相似性系数。结果表明,收集的木薯材料的平均表型变异系数为37.9%,平均遗传多样性指数为0.807,其中主茎分叉角度的变异系数最大,为86.7%,块根内皮颜色的遗传多样性指数最大,为1.842。13对SSR引物共扩增出118个条带,其中多态性条带为106个,多态性比率为86.23%;分子标记聚类结果发现,遗传相似性系数在0.415~1.000之间;通过对比发现表型聚类和分子聚类结果不一致,其中表型聚类发现类群划分与地理来源之间没有关联,但与株型性状有一定的关联;在遗传相似系数为0.62时,SSR分子标记聚类为两类材料,第一类材料地理分布无规律,第二类材料大多分布在桂南。本研究结果表明,广西地方食用木薯资源遗传多样性具有一定的丰富度,可为创制优异食用木薯种质资源和新品种选育奠定基础。     

利用SSR标记分析藜麦品种的遗传多样性

为了解藜麦种质资源的多样性,本研究利用SSR引物对所搜集的41个藜麦种质的多态性及其亲缘关系进行了分析。结果表明,从54对SSR引物中筛选出了16对能明显扩增出稳定的多态性条带的引物,共检测出139个等位基因条带,每一对引物的等位基因个数为3~13,平均为8.7;16对引物的多态信息含量(PIC)变幅为0.208~0.432,平均为0.366。UPGMA聚类分析显示,41份材料的遗传相似系数(GS)在0.374~0.906之间,平均相似系数为0.626。在阀值(GS)约为0.665时,41份材料可分为4大类。其中614929与B.B.Quinoa浙Ⅰ间的遗传相似系数最小,为0.374,表明来源于不同地区的遗传距离较远,遗传基础较广泛。藜麦品种资源间的亲缘关系的揭示为藜麦资源保存和新品种选育提供了理论依据。     

甘蓝SSR标记在近缘种青花菜的通用性及其应用

本研究对50条甘蓝SSR引物在其近缘种青花菜中的通用性进行了分析,结果表明,38对引物在青花菜上可有效扩增,扩增产物分子量在100～1500bp,有效扩增比率为76%,其中18%具有较好的多态性,揭示了两种作物基因组间存在一定的相似性。同时利用获得的多态性较好的9对引物对青花菜进行基因型鉴定和遗传多样性分析,20份青花菜基因型中共检测到51个基因位点,平均每个引物组合可扩增出5.67个条带,多态性为66.7%。多引物组合可鉴别出所有青花菜基因型。聚类分析结果表明同一来源的基因型间具有相近的遗传基础,且聚类与熟性具有一定的相关性。本研究为今后青花菜种质资源的收集、利用及分子标记辅助育种提供了新的SSR标记和参考。     

RAPD and SSR markers for characterization and identification of ancient cultivars of <Emphasis Type="Italic">Olea europaea</Emphasis> L. in the Emilia region,Northern Italy

The Emilia region (Northern Italy) is characterised by the occurrence of microclimates that permit olive growing. The presence of the species, albeit sporadic, in these territories for several centuries as a fruit crop is well documented, by both archaeological and written testimony, and by a large number of plants well over a century old, located in particular sites, favourable for growth and development of the tree. Olive genetic diversity was studied using RAPD and SSR techniques, on plants growing in the Emilia territory (Reggio Emilia and Parma provinces). For genotype identification comparisons were made with 8 cultivars, some of which from Central Italy. Screening was obtained analysing patterns produced by 20 RAPD primers and 3 SSR primers, developed by other authors; the primers and we were able to discriminate olive cultivars with a sufficient degree of reliability. The dendrograms obtained from the analysis show the genetic relationship among accessions present in the Parma-Reggio Emilia district. Our results demonstrated the reliability of RAPDs and SSRs to identify all studied olive cultivars and to reveal the degree of their relatedness to each other. The analysis also reveals the presence of an interesting amount of genetic diversity among the studied individuals.     

Analysis of genetic diversity among European and Asian fig varieties (<Emphasis Type="Italic">Ficus carica</Emphasis> L.) using ISSR,RAPD, and SSR markers

Nineteen fig varieties and lines from Europe and Asia have been fingerprinted by ISSR, RAPD, and SSR markers, respectively, using 13, 19, and 13 primer combinations. All primers produced 258 loci, with the highest number of loci (119) generated by RAPD (R _p: 48.42). Clustering analysis was applied to the three marker datasets to elucidate the genetic structure and relationships among these varieties. Mean genetic similarities were 0.787, 0.717, and 0.749, respectively, as determined using ISSR, RAPD, and SSR. Each marker system produced incompletely separated clusters, although a weak binding group based on race type appeared in the combined dataset. Comparisons of coefficients revealed no correlation between different similarity matrices; congruence was observed between similarity matrices and co-phenetic matrices in all markers. Analysis of molecular variance (AMOVA) showed that most of the total polymorphism was attributable to within-group variance (ISSRs + RAPDs, 97.41%; SSRs, 90.18%). These results suggest that the genetic diversity of this fig population is low and that multiple marker utilization is critical to estimate the relatedness of figs at the variety level. Additionally, it was presumed that ‘Houraihi’, the oldest variety in Japan, was disseminated independently of other foreign varieties in the 17th century or before then.     

Genetic diversity analysis of hulless barley from Shangri-la region revealed by SSR and AFLP markers

For adding the hulless barley resources of Shangri-la region to the global barley resource library, a basic work was done by us to assess their genetic diversity of this region. The genetic diversity of 60 hulless barley samples collected from three counties in Shangri-la region of Yunnan Province, were studied using SSR (simple sequence repeats) and AFLP (amplified fragment length polymorphism) markers. A total of 70 alleles were detected for 19 pairs of SSR primers, and 525 band containing 464 polymorphic bands were revealed for 5 pairs of AFLP primers. The value of polymorphism information content (PIC) ranged from 0.03 to 0.86 for SSR primers. The total numbers of alleles were 51, 55, 43 in three populations and the polymorphic bands were 188, 205 and 141. The genetic distances and genetic identity among the three populations showed their close relationship. The gene diversity among populations relative to the total population diversity (Gst) was 0.13 for SSR markers and 0.02 for AFLP markers and indicated that just 13 and 2% variations were among populations, respectively. The UPGMA cluster analysis revealed that all of the samples grouped randomly rather than clustered into distinct groups corresponding to their populations, row types and spring/fall types. We concluded that there was high genetic diversity in the population of Shangri-la region and the formation of diversity was related to complex environment and inhabitants’ traditional practices.     

银杏内生真菌刺盘孢遗传多样性的初步分析

为了研究银杏内生真菌的刺盘孢菌株的遗传分化,选用了分离自江苏5个县市的14个菌株,利用RAPD技术进行遗传多样性分析。从20个随机引物中筛选出5个引物,扩增出62条DNA带,在此基础上分析并建构了遗传相关聚类图。结果表明,不同地区的刺盘孢菌株其遗传差异性不同,14个菌株可聚为两大类,其中启东、邳州的6株聚为一类,南京、南通和泰兴的8株又聚为一类。所有的14个菌株的相似性系数范围在46%~89%。     

三个野生种群马氏珠母贝遗传多样性的RAPD分析

摘要：为了解野生种群马氏珠母贝（Pimctada martensii (Dunker.)）的遗传结构背景和开展遗传改良育种,运用RAPD技术分析了海南三亚（SW）、广东大亚湾（DW）和广西北海（BW）3个野生种群的遗传多样性。采用了18个10碱基的随机引物对3个野生种群进行分析,其中6个引物能扩增出清晰的多态带谱,扩增的DNA片段大小在含0.2~3.0 kb之间。SW、DW和BW 3个种群内的遗传相似性指数分别为0.642、0.672和0.688,Shannon多样性值分别为0.266、0.211和0.174。马氏珠母贝3个野生种群的遗传相似性依次为SW< DW < BW,而遗传多样性依次是SW > DW> BW。DW和SW相对遗传距离为0.104;DW和BW相对遗传距离为0.094;SW和BW相对遗传距离为0.212。讨论了马氏珠母贝野生种群遗传多样性差异的可能原因、种群间杂交子代的杂交优势预测及人工养殖对野生种群遗传结构的可能影响。     

Microsatellites and RAPD Markers to Study Genetic Relationships Among Cowpea Breeding Lines and Local Varieties in Senegal

Genetic diversity in local cowpea varieties and breeding lines from Senegal were studied using random amplified polymorphic DNA (RAPD) and microsatellite (SSR) techniques. Among the 61 RAPD primers used, twelve show polymorphism. Fifteen of the 30 microsatellite primer pairs were polymorphic, detecting one to nine alleles per locus. The RAPD and SSR data were analyzed both separately and in combination to assess relationships among genetic lines. Although RAPD provided information on levels of genetic diversity, microsatellite markers are most effective in determining the relationship among cowpea accessions and varieties. The SSR results support the genetic diversification of cowpea in Senegal and underscore their potential in elucidating patterns of germplasm diversity of cowpea in Senegal. An erratum to this article is available at .     

Genetic diversity and differentiation in Ethiopian populations of Phytolacca dodecandra as revealed by AFLP and RAPD analyses

Amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) analyses were performed on six populations (a total of 89 individuals) of Phytolacca dodecandra (endod) collected in Ethiopia. The populations were selected based on our previous investigation to represent two altitude groups: lowland/central-highland (1600–2500 m) and highland (2501–3000 m). A total of 197 AFLP and 68 RAPD markers were scored from 5 primer pairs and 12 random primers, respectively. The overall patterns obtained for AFLPs and RAPDs from diversity, cluster and principal component analyses were very comparable. However, the moderate correlation (r = 0.56) between AFLP and RAPD similarity matrices as well as the discrepancies in diversity estimates between the two techniques in some populations and in the lowland/central-highland plants could be due to differences in sensitivity of reaction conditions, bias in scoring of bands, number of markers used for analyses, and/or parts of the genome surveyed. For both AFLP and RAPD, the lowland/central-highland populations showed higher polymorphism and Shannon information measure (H) than the highlands. Cluster and principal component analyses performed for both marker types have also clearly demonstrated the differentiation of all the lowland/central highland plants from those of the highlands, in agreement with our previous conclusion. Markers scored from any of the five AFLP primer pairs were sufficient to clearly distinguish the two altitude groups; with RAPD, selection of about 8 informative markers produced by seven random primers was needed for the same purpose.     

Relationships between an Italian Strawberry Ecotype and its Ancestor using RAPD Markers

Random-amplified polymorphic DNA (RAPD) markers were used to evaluate genetic variability among populations of an Italian strawberry ecotype, and to determinate genetic relationships between genotypes and their putative ancestor. A total of 65 selections and one cultivar ‘Madame Moutot’ (MM), were analysed to evaluate genetic variability present in Etna mountain area and to confirm as MM was one of the cultivars that originated the ecotype. A total of 222 RAPD markers was obtained using 16 decamer primers and 6 longer primers, 90.8% of the markers obtained by selected primers resulted polymorphic at least within analysed genotypes. RAPDs were used to calculate genetic similarity coefficients and to generate dendrograms representing genetic relationships among genotypes analysed. Cluster analysis displays as RAPD polymorphisms were able to characterize the genotype variability among closely related groups. The data show as MM could be considered the ancestral genotypes introduced in that area. The results obtained confirm that RAPD markers could be used as reliable markers to perform phylogenetic studies in Fragaria×ananassa Duch. ex Rozier. Giuseppe Bertino and Piero Spada - Coauthor involved in genotype selection and field management     

Characterization of Some Italian Common Bean (<Emphasis Type="Italic">Phaseolus Vulgaris</Emphasis> L.) Landraces by RAPD,Semi-random and ISSR Molecular Markers

Randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and a semi-random PCR system were used to analyze the genetic diversity of 16 Italian common bean landraces and their relationship to four commercial cultivars. Of the primers tested, 8 ISSR, 6 RAPD and 7 semi-random primers produced polymorphic and reproducible DNA fragments. A higher proportion of polymorphic bands were observed using ISSR (85%) and semi-random (90%) primers than RAPD (69%) method. The combination of any two semi-random markers allowed the identification of all 20 bean genotypes. In contrast ISSR (except for primer (CAC)₃GC) and RAPD markers appeared to be less informative as more than two markers were necessary to achieve the same diagnostic level. Moreover, 7 ISSR, 2 RAPD and 8 semi-random exclusive bands were identified as putative population-specific markers. Semi-random and ISSR derived dendrograms showed similar tendencies in terms of genetic relatedness, whereas clustering of genotypes within groups was not similar when compared with the RAPD technique. Despite the different ability to resolve genetic variation among the investigated landraces, two major clusters with less than 60% (ISSR) and 40% (RAPD and semi-random) genetic similarity were formed with all three marker systems. The two groups were correlated with the phaseolin patterns and seed size of the landraces. The analysis showed that the cultivar ȁ8Lingua di Fuocoȁ9 and most of the landraces (13 out of 16) collected in Italy belong to the Andean gene pool, whereas only the three populations from Pratomagno belong to the Middle American gene pool.