薄皮甜瓜种质资源遗传多样性的RAPD分析

RAPD分子标记对来源不同的41份薄皮甜瓜种质资源进行了遗传多样性分析。11个随机引物共扩增出78条带,平均每个引物扩增的多态性带数为5.3条,多态性带百分率为74.4%。利用NTSYS-pc软件计算种质间的遗传距离,用UPGMA法进行聚类分析,将供试的41份种质可分为3个类群8组,用EIGEN方法进行主坐标分析,将其分为4个类群12组。两种分类方法结果基本一致,后者能更为直观地反映群体之间的关系。     

基于RAPD标记的芥蓝种质资源遗传多样性分析

调查了44份芥蓝种质的植物学性状,并利用RAPD分子标记分析了其遗传多样性。结果从150条随机引物中筛选出20个引物,20个引物共扩增出177条谱带,其中多态谱带105条,平均每个引物扩增出8.9条谱带和5.3条多态性谱,多态性比率为59.32%。基于RAPD标记,利用NTSYS-pc2.11构建了聚类树状图谱,遗传距离为0.70时,44份芥蓝资源可聚成六大类群。芥蓝种质存在着一定的遗传多样性,但原产华南而且主要产区也在华南,遗传多样性要小于芸薹属其他蔬菜。     

82份瓠瓜种质资源遗传多样性的SSR分析

为了明确不同瓠瓜种质资源之间的遗传关系,本研究采用SSR分子标记对82份瓠瓜种质材料的遗传多样性进行了分析。结果表明,21对SSR引物共扩增出73条多态性位点,多态性比率为93.59%,说明引物的多态性丰富;等位基因数(Na)、有效等位基因数(Ne)、多态信息含量(PIC)的均值分别为3.71、3.09、0.63,表明份瓠瓜种质资源的遗传多样性十分丰富;通过聚类分析,当遗传距离在0.66处时,可将82份瓠瓜材料分为三大类群,并且这些材料在分子水平上的分类结果与农艺性状(瓜型)、地理分布存在一定相关性。研究结果可为瓠瓜种质资源的创新利用提供依据。     

茶树种质资源遗传多样性的RAPD分析

用随机扩增多态DNA（RAPD）标记对来自全国产茶省的31份种质资源、福建地区的10份种质资源以及湖南安化云台山种有性后代居群的30个单株的遗传多样性进行了检测。不同生态条件下的31份不同种质资源中,21个RAPD引物扩增出188条谱带,多态性比率为90.43%;福建10份种质资源,21个RAPD引物共扩增出154条谱带,多态性比率为81.81     

利用RAPD分子标记分析玉米种质遗传多样性

以RAPD分子标记技术对玉米的10个品种的全基因组DNA进行PCR扩增,再用Popgene32软件和SPSS 13.0软件分析扩增结果,研究10个玉米的种质资源遗传关系。结果表明：（1）所选20个引物可扩增出214条RAPD条带;（2）所选引物扩增条带的多态性比率为86.4%,RAPD分子标记扩增10个玉米品种间的相似性系数分别在0.316-0.654之间;（3）用RAPD-PCR扩增条带的分析结果建立了遗传相关系数矩阵、构建了分子树状图、可将10个玉米品种分为3个类群;（4）RAPD分子标记适合于构建玉米的DNA指纹图谱,进行品种鉴定和遗传分析。     

8份剑麻种质亲缘关系的ISSR和RAPD分析

为了揭示剑麻栽培品种的遗传多样性,利用ISSR和RAPD分子标记技术对8份剑麻种质的亲缘关系进行分析。结果表明,筛选后选用的8条ISSR引物和8条RAPD引物,分别产生了53条和66条扩增条带,其中多态性条带分别为44条和61条,多态性条带百分率分别为83.02%和92.42%。根据2种标记的扩增结果,用UPGMA法对8份剑麻种质进行聚类分析,供试材料之间具有较高的遗传多样性,其品种间遗传相似系数分别为0.59~0.80和0.52~0.76。2个标记的聚类结果基本一致,但有点差异,可将供试的8份剑麻种质划分为2类群,而且2个标记聚类结果呈显著相关性,相关系数为0.70。可见,剑麻种质资源的遗传多样性丰富。     

大白菜种质资源的遗传多样性分析

采用RAPD技术对64份大白菜种质资源的遗传多样性进行了研究。从OPE,OPF,OPG和OPH 4套共80条引物中选出了16个条带清晰、且具有多态性的引物分析供试材料,结果共产生100条扩增带,其中72条带具有多态性,多态性频率平均为72%。由RAPD分析结果可初步得出芜菁、小白菜和大白菜的特征谱带,这些特征谱带可分别作为这3个亚种的RAPD标记,据此可鉴定这3个亚种。将扩增出来的72条多态性片断进行统计,采用UPGMA进行聚类分析,取相似距离0.721为阈值,将大白菜种质资源分为6个类群,而小白菜(66号,苏州青)和芜菁(67号)各成一类,共8个类群。     

西藏油菜种质资源的RAPD分子标记分析

采用RAPD分子标记, 对西藏地区部分白菜型和芥菜型油菜资源的遗传多样性进行了分析, 结果显示, 22条引物在106份白菜型油菜中共扩增出236条带, 多态性位点比率为89%; 24条引物在50份芥菜型油菜中共扩增出276条带, 多态性位点比率为94%. 通过UPGMA聚类分析, 将西藏白菜型油菜分为Ⅰ和Ⅱ类群; 芥菜型油菜分为Ⅰ、Ⅱ、Ⅲ和Ⅳ类群     

龙眼种质资源的RAPD分析

利用RAPD技术对龙眼的95份种质资源和一份荔枝种质资源进行研究。从200条随机引物中筛选出18条多态性引物,共扩增出197条带,DNA扩增带的多态性为100%,这表明96份材料的遗传多样性和遗传资源的丰富性。利用SPSS13.0统计软件对RAPD表型数据矩阵进行分析。根据欧氏距离（ED）法计算各样品间的遗传距离,采用UPGMA（类平均法）,进行分析,建立亲缘关系聚类树状图。结果表明,RAPD标记能在分子水平揭示龙眼的遗传背景与亲缘关系     

新疆薰衣草种质资源遗传多样性的ISSR分析

本研究应用ISSR标记对19份薰衣草种质的遗传多样性进行分析。结果显示:从100条引物中筛选出9条多态性明显的引物,19份材料DNA共扩增出94条谱带,其中多态性谱带40条,多态性比率为42.6%,平均每个引物扩增条带数为10.4条;9条ISSR引物可将所有样品完全分开,当遗传相似阈值为0.69时,可将样品分为3个类群:第Ⅰ类群13个品种(系)可分为2个亚类,第Ⅱ类群5个品种(系)可分为2个亚类,第Ⅲ类群1个品种(系)。样品间的遗传相似性系数为0.325~0.975(平均0.737)。群体的有效等位基因数、基因多样度、Shannon信息指数分别为1.454 0、0.280 5和0.435 4。研究结果表明,19个新疆薰衣草种质资源遗传多样性水平处于较高。     

白菜类蔬菜种质资源亲缘关系的RAPD分析

利用200个随机引物对21份白菜类蔬菜种质资源材料的基因组DNA进行了扩增,研究了它们的亲缘关系。结果表明,有22个随机引物可得到稳定的RAPD谱带,扩增带总数为135条,其中多态性带为125条,多态性比例为92.6%。系统聚类分析结果显示,在遗传距离(D)=0.25处,可将供试材料分为2类10组,其中第1类包括5个甘蓝品种,另外还包括芜菁;而第2类包括白菜和芥菜2个亚类。这表明芜菁和白菜的亲缘关系较远。芜菁和散白菜的杂交后代与芜菁和其他白菜的杂交后代形态相似,不能从形态学上为李家文的杂交起源假说提供佐证。推测白菜可能起源于野生小白菜的原始类型,在向北方传播的过程中相互杂交,经过长期的自然环境选择,优胜劣汰逐渐演化才终于出现了结球结实的大白菜。     

六倍体小黑麦种质资源遗传多样性分析

利用RAPD标记对10份六倍体小黑麦、1份普通小麦和1份黑麦种质资源遗传多样性进行了分析,19个RAPD引物中,有10个引物(52.63%)可扩增出清晰且具多态性的条带。10个引物共产生385条DNA片段,其中384条具有多态性,多态性比率为99.74%,不同引物扩增带数变幅为8~71条,平均为38.5条,扩增产物的片段大小为298~3 054 bp,材料间的遗传距离变化范围在0.724~0.833之间。对12份供试材料的遗传关系进行聚类分析,在平均GD值0.77水平上可将12份供试材料分为3类。研究结果为将六倍体小黑麦的优良基因导入到普通小麦中提供了理论依据。     

抗PVY马铃薯品种遗传多样性的RAPD分析

采用RAPD技术分析了国内外37种抗PVY马铃薯资源。提取马铃薯(Solanumtuberosum)新鲜叶片的总DNA,用17种RAPD引物进行随机多态性扩增,利用遗传多样性信息,进行亲源关系的分子鉴定。试验表明:平均每10个碱基引物扩增出6￣21条谱带,共获得164条特异性谱带,平均每个引物扩增获得9.8条多态性谱带,多态性比率平均为76.3%。RAPD分析表明37种抗PVY马铃薯资源之间的遗传距离介于0.06￣0.68之间,平均值为0.35,平均遗传距离介于0.27￣0.50之间,聚类分析将37种抗PVY材料划分为三个类,聚类结果同材料的血缘关系密切,并且表现出一定的地域特性。根据扩增的特异性谱带可进行抗PVY马铃薯资源的分子鉴定,为抗PVY育种亲本选配提供了依据。     

Molecular characterization of taro (Colocasia esculenta) using RAPD markers

Forty-four taro (Colocasia esculenta), two tanier (Xanthosoma species) and one Colocasia gigantea accessions were evaluated for genetic diversity using random amplified polymorphic DNA (RAPD) primers. Seventy-three of 112 primers amplified PCR DNA products used to fingerprint the accessions. Thirty-two primers were considered highly informative because they amplified more than 5 bands or amplified one or more polymorphic bands that distinguished between accessions. RAPDs showed high genetic diversity in taro accessions from Indonesia, were capable in distinguishing between Hawaiian accessions, and could separate triploid from diploid accessions. UPGMA cluster analysis of genetic similarity estimates (Jaccard's coefficient), separated the accessions into 3 main groups with C. esculenta divided into 5 subgroups. These primers will be useful for future genetic analysis and provide taro breeders with a genetic basis for selection of parents for crop improvement. Polymorphic markers identified in the DNA fingerprinting study will be useful to screen a segregating population which is being generated in our laboratory aimed at developing a taro genetic linkage map. This revised version was published online in July 2006 with corrections to the Cover Date.     

Contrastant banana accessions for resistance to the burrowing nematode,based on molecular markers RAPD

This work aimed to proceed molecular characterization of seven banana accessions (Borneo, Grand Naine, 1304-06, 4249-05, 0337-02, 0323-03 and 4279-06) resistance to the nematode Radopholus similis. These accessions were selected taking in account the reproduction factor (RF) among 26 banana genotypes from a working collection belonging to Embrapa Mandioca e Fruticultura Tropical. The genomic DNA of the seven accessions was extracted, and 36 decamere primers had been used to obtain RAPD markers. The resulting markers were converted into a matrix of binary data. From that matrix the genetic distance between the accessions were estimated, for further clustering and graphic dispersion analyses. From a total of 521 RAPD markers generated, 420 (81%) were polymorphic, including 140 (27%) potentially promising for application on works related to genetic mapping of the resistance to R. similis. OPE-15, OPH-17, and OPG-09 were the primers that contributed to the highest number of bands promising for genetic mapping of resistance (12, 8, and 8, respectively). The genetic distances between accessions ranged from 0.106 to 0.455, with the longest one observed between cv. Borneo and the genotype 4279-06, considered as highly susceptible and resistant, respectively, to the nematode according to the RF. The graphic dispersion distinguished three groups of accessions, and most of resistant genotypes clustered together in the same group. The most contrastant genotypes for resistance (Borneo and 4249-05) were separated by a genetic distance of 0.374, and possessed a total of 114 polymorphic bands promising for genetic mapping of resistance. In addition, the results of pathogenicity tests were congruent with those obtained by RAPD analyses.     

西南横断山区野生狗牙根遗传多样性的RAMP分析

采用RAMP标记技术,分析了西南横断山区25份野生狗牙根材料的遗传多样性。结果表明,被测材料间RAMP标记多态性较高。10个引物组合扩增出的56条带中,39条具多态性,PPB为69.64%。每个引物组合可扩增出2~6条多态性带,平均3.9条。RAMP标记遗传相似性系数(GS)变化范围为0.6667~0.9474,平均值为0.8129。RAMP标记可将所有25份狗牙根材料区分开,供试材料可以明显的划分为4大类。聚类结果与材料的地理分布有一定关系。据此认为,西南横断山区野生狗牙根种质资源在分子水平确实存在较大的遗传差异,RAMP是评价狗牙根遗传多样性的有效方法。     

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。     

利用RAPD标记研究燕麦属不同种的遗传差异

利用RAPD标记对我国8个燕麦种共21份材料的遗传差异及类群划分进行了研究。22个多态性引物在21个基因型间共扩增出114条带，其中97条带具有多态性，占85．1％。用这些条带计算了不同基因型间的遗传距离，发现各种间的遗传距离(0．053～0．510)普遍大于不同种内的遗传距离(0．022～0．087)。聚类结果显示，21份材料可划归为5大类群8个亚群，并与传统的分类结果基本一致。由此表明，用RAPD标记对燕麦进行遗传关系和分类研究是可行的。     

Estimation of between and within accession variation in selected Spanish melon germplasm using RAPD and SSR markers to assess strategies for large collection evaluation

The population structure of 15 Spanish melon (C. melo L.)accessions, mostly of Group Inodorus, was assessed by the analysis of 16individuals of each accession using 100 random amplified polymorphic DNA (RAPD) bands produced by 36 primers, and allelic variation at 12microsatellite (SSR) loci (23 alleles). A relatively high level of polymorphism (25.6%) was detected using RAPD markers, and eight SSR loci (66.7%) were useful in discriminating accessions. Cluster analysis using RAPD- and SSR-based genetic distance estimates resulted in similar and consistent groupings of most of the accessions studied. The mean genetic distance and standard error among accessions estimated by RAPD variation was 0.421 ± 0.099, and mean SSR-based genetic distance estimate was 0.285 ± 0.141. Albeit many dominant markers examined were fitted to a 3:1 test ratio, deviation from this ratio and from Hardy-Weinberg expectations for many SSR loci suggests that some populations were in genotypic disequilibrium. Moreover, a higher level of genetic variation was observed between Cassaba market classes than within accessions, suggesting that, depending upon the accession, allelic fixation has occurred in these populations. The relatively high level of heterogeneity observed (different band morphotypes and cluster grouping within a particular market class), however, indicates that the Spanish melons examined possess a relatively broad genetic background. An appraisal of accession population structure such as the one reported herein indicates that bulk sampling techniques coupled with molecular analysis techniques that employ a unique array of discriminating markers can provide information leading to effective strategies for diversity analyses of large collections. This revised version was published online in August 2006 with corrections to the Cover Date.