珍稀濒危植物天目木兰（Magnolia amoena）遗传多样性的RAPD分析

运用随机扩增多态DNA(RAPD)技术,对天目木兰(Magnolia amoena)居群的遗传多样性进行了研究．从40个10-mer随机引物中筛选出14个能得到清晰、稳定扩增带的引物进行扩增,14个引物共检测了94个位点,其中多态性位点为23,占24．4％,计算了12个居群之间的遗传相似度和遗传距离,并运用UPGMA法进行了聚类分析,结果显示相同严地个体间(居群内)的遗传距离较小,遗传多样性水平很低;不同产地个体间(居群间)遗传距离较大,遗传多样性水平较前者高,即天目木兰个体间遗传多样性水平与它的地理分布有关,天目木兰总体较低的遗传多样性是导致它濒危的原因之一。     

RAPD分析花椰花不同品种的遗传变异

利用随机扩增多态性DNA（RAPD）方法,对13个花椰花品种的基因组DNA进行多态性分析。选用20个10bp随机引物,共扩增出175条DNA片段,其中多态性片段118例,占67．4％,结果表明,花椰花品种间具有丰富的遗传多样性。依据扩增结果进行遗传相似系数分析,构建聚类分析树状图。初步探讨了各品种间的遗传变异关系及RAPD技术在花椰花种质资源分类鉴定和育种工作中的应用前景。     

65份野生油茶种质遗传多样性的ISSR和RAPD标记分析

利用ISSR和RAPD标记,对名邛台地野生油茶种质进行遗传多样性分析。从60条简单重复序列引物中筛选出16条引物,在65份样品中共扩增出213条带,其中多态位点为203个,多态位点百分率为95.31%;从30条寡居核苷酸引物中筛选出8条引物,共扩增出105条带,其中多态性位点94个,多态位点百分率为89.52%。结果表明:名邛台地野生油茶种质具有较丰富的遗传多样性,ISSR和RAPD标记可以应用于油茶种质遗传多样性分析。     

辣椒种质遗传多样性的RAPD和ISSR及其表型数据分析

用RAPDI、SSR分子标记及28个表型性状数据对辣椒属5个栽培种的13份材料进行了分析,结果表明:23条RAPD引物共扩增出209条带,平均每个引物扩增出9.09条,多态性位点比率为83.73%;16条ISSR引物共扩增出94条带,平均每个引物扩增出5.88条,多态性位点比率为79.79%.与RAPD相比,ISSR标记检测到的有效等位基因数(Ne)及Shannon多样性指数(I)、遗传离散度(Ht)和遗传分化系数(Gst)等遗传多样性参数都较大,多态性位点比例在亲缘关系较近的一年生辣椒(Capsicum annuum)种内较高,说明ISSR有更高的多态性检测效率,并且适合亲缘关系较近的种群间遗传多样性分析.基于RAPDI、SSR的聚类与基于表型数据的聚类之间存在极显著正相关,且都能将C.annuum与其它栽培种区分开来.     

乌塌菜种质资源遗传多样性的ISSR分析

利用ISSR技术对48份乌塌菜种质资源进行遗传多样性分析。从60条随机引物中筛选出稳定性强、条带清晰且多态性丰富的9条引物进行PCR扩增,共扩增出103条谱带,平均每个引物扩增出11.4条带,其中多态性带85条,多态性位点百分率为82.68%。不同乌塌菜种质间遗传相似系数变幅为0.59~0.97,说明ISSR标记能够揭示材料间较高的遗传多样性。利用UPGMA聚类分析,ISSR标记能将48份乌塌菜品种完全区分开,48份乌塌菜种质被划分为4个类群,聚类结果与叶片颜色相关,为乌塌菜品种资源的研究利用提供参考。     

华东竹黄菌不同居群遗传分化的RAPD分析

为了解竹黄地理居群间的遗传分化,本研究采用随机引物扩增多态性DNA分子标记技术对江苏、安徽和浙江3省的8个居群共32个竹黄样本进行了遗传多样性分析.从50个RAPD引物中筛选得到了5个随机引物,对供试材料的DNA进行扩增,共检测出77个位点,其中多态性位点52个,多态性位点比率为67.53%.UPGMA聚类分析结果表明,这8个居群分为三类:安吉居群、临安居群、宜兴居群、广德居群和泾县居群聚为一类;宁国居群和休宁居群聚为一类;淳安居群单独为一类.遗传多样性分析表明8个竹黄居群中,淳安居群的遗传多样性水平最高,安吉居群的遗传多样性水平最低.Nei's基因多样性指数和Shannon信息指数均表明竹黄物种水平的遗传多样性高于居群水平.     

应用RAPD标记分析黑麦属品种的遗传多样性

四川农业大学小麦研究所侯永翠、郑有良、魏育明等研究人员对黑麦遗传多样性课题作了研究。他们采用随机扩增多态性DNA(RAPD)标记 ,对黑麦属 (SecaleL) 7个品种共 1 2份材料进行了遗传多样性检测 ,发现被检测材料间RAPD标记多态性较高 ,在 4 0个随机引物中 ,有 2 5个引物约占整个的 6 2 .5 %的扩增产物具有多态性。这 2 5个中共扩增出 1 6 7条带 ,其中 89条带约占 5 3.2 %具有多态性 ,每个引物可扩增出 1～ 1 0条多态性带 ,平均为 3～ 6条。RAPD标记遗传距离GD变异范围为 0 .1 382～ 0 .4 5 1 2 ,平均为 0 .2 71 2。通过聚类分析表…     

美国白蛾种群遗传分化的RAPD分析

应用随机扩增多态性DNA(RAPD)技术,对9个不同地理种群美国白蛾进行了遗传多样性和UPMGA聚类分析.结果表明:25个RAPD引物共扩增191个位点,片段大小在200～2000 bp,其中158个是多态位点,多态百分率为82.72%,遗传距离指数在0.2043～0.5108,遗传相似性系数在0.4892～0.7957;9个不同地理种群的美国白蛾可以分成3类:辽宁省和山东类群、北京和天津类群、美国类群,表明美国白蛾不同地理种群间的遗传分化与地理位置相关.     

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。     

云南茶树资源遗传多样性分析(英文)

利用EST标记对云南134份茶树资源遗传多样性和亲缘关系进行了分析.结果显示:(1)30对引物共检测到等位位点127个,平均每对引物产生4.23个;共检测到263个基因型,平均每对引物所扩增的基因型有8.8个,遗传多态性信息含量变异范围为0.014～0.736,平均达0.50l.(2)资源间的平均遗传距离和相似系数分别为0.413和0.597,聚类可将134份资源划分为4大组.(3)8个种群间的遗传相似系数变异范围为0.753～0.981,平均遗传相似系数为0.891.结果表明,云南茶树资源间的遗传差异比较大,遗传基础较宽,具有丰富的遗传多样性,而不同种间的遗传差异比较小.     

Analysis of Genetic Diversity in Cicer arietinum L Using Random Amplified Polymorphic DNA Markers

PCR-based random amplified polymorphic DNA (RAPD) markers were employed to assess genetic diversity in 23 chickpea genotypes. Forty of the 100 random primers screened revealed polymorphism among the genotypes. Most of the primers revealed single polymorphic band, and only 14.1 2% of the products were polymorphic. Estimates of genetic similarity based on Jaccard’s coefficient ranged from 0.92 to 0.99, indicating narrow genetic variability among the genotypes based on RAPD markers.The 23 chickpea genotypes formed two major clusters in the dendrogram.The low RAPD polymorphism among chickpea genotypes suggests that more number of polymorphic primers need to be analysed to determine genetic relationships. It was observed that RAPD analysis employing 30 polymorphic primers could provide better estimates of genetic relationships in chickpea.     

Genetic diversity of different Tunisian fig (Ficuscarica L.) collections revealed by RAPD fingerprints

The genetic diversity in Tunisian fig (Ficus carica L.) was studied using RAPD markers. Thirty-five fig cultivars originating from diverse geographical areas and belonging to three collections were analysed. Random decamer primers were screened to assess their ability to detect polymorphisms in this crop. Forty-four RAPD markers were revealed and used to survey the genetic diversity and to detect cases of mislabelling. As a result, considerable genetic diversity was detected among the studied F. carica accessions. The relationships among the 35 varieties were studied by cluster analysis. The dendrogram showed two main groups composed of cultivars with similar geographic origin. Moreover, the male accessions (caprifigs) were clustered indistinctively within the female ones, suggesting a narrow genetic diversity among these accessions. Our data proved that RAPD markers are useful for germplasm discrimination as well as for investigation of patterns of variation in fig. Since this designed procedure has permitted to establish a molecular database of the reference collections, the opportunity of this study is discussed in relation to the improvement and rational management of fig germplasm.     

The use of RAPD markers for detecting genetic diversity,relationship and molecular identification of Chinese elite tea genetic resources [<Emphasis Type="Italic">Camellia sinensis</Emphasis> (L.) O. Kuntze] preserved in a tea germplasm repository

The genetic diversity, relationship and molecular identification of 15 well known, widely planted traditional Chinese elite tea genetic resources [Camellia sinensis (L.) O. Kuntze] preserved in the China National Germplasm Hangzhou Tea Repository in the Tea Research Institute of the Chinese Academy of Agricultural Sciences located in Zhejiang province, China, were investigated using RAPD markers. A total of 1050 bands with an average of 52.5 bands per primer, 70 bands per genetic resource were generated by the 20 selected primers from the 15 tea genetic resources. In the total of 137 amplified products, 129 were polymorphic, corresponding to 94.2% genetic diversity. The relative frequency of polymorphic products was from 0.24 to 0.83, with an average of 0.47. In general, this average frequency was relatively high. The genetic distances among the genetic resources were from 0.16 to 0.62, with an average of 0.37. The 15 tea genetic resources were grouped into three groups by UPGMA cluster analysis based on RAPD data. By using the presence of 20 unique RAPD markers and the absence of 11 unique markers, all the 15 investigated tea genetic resources could be easily identified. RAPD markers provided a practical method not only to evaluate the genetic diversity and relationship, but also to identify tea genetic resources.     

Genetic variation in a wild population of the 'sleep' passion fruit (Passiflora setacea) based on molecular markers

Little is known about the molecular genetic diversity of most Passiflora species. We used RAPD markers to evaluate the genetic diversity of 24 genotypes of the 'sleep' passion fruit (Passiflora setacea). Twelve primers generated 95 markers, 88% of which were polymorphic. The genetic distance estimated by the complement of the Dice index ranged from 0.29 (among accessions Ps-G1 and Ps-G13) to 0.69 (among accessions Ps-G21 and Ps-G23). Genotype grouping based on the UPGMA algorithm showed considerable variability among genotypes. We conclude that P. setacea has a broad genetic base that could be exploited in breeding programs.     

Development of Molecular Tools for Characterization and Genetic Diversity Analysis in Tunisian Fig (Ficus carica) Cultivars

Fig, Ficus carica L., is a useful genetic resource for commercial cultivation. In this study, RAPD (60), ISSR (48), RAMPO (63), and SSR (34) markers were compared to detect polymorphism and to establish genetic relationships among Tunisian fig tree cultivars. The statistical procedures conducted on the combined data show considerable genetic diversity, and the tested markers discriminated all fig genotypes studied. The identification key established on the basis of SSR permitted the unambiguous discrimination of cultivars and confirmed the reliability of SSR for fingerprinting fig genotypes. The study findings are discussed in relation to the establishment of a national reference collection that will aid in the conservation of Tunisian fig resources.     

Comparative Analysis of RAPD and ISSR Markers for Characterization of Sesame (Sesamum indicum L) Genotypes

The determination of genetic differences among crop genotypes has become the primary need to grant patent and the protection of Plant Breeder Rights (PBR). In the present study RAPD and ISSR markers were employed for the characterization of 16 sesame genotypes. Twenty six RAPD and 17 ISSR primers that generated clear and reproducible banding patterns amplified 194 and 163 bands, respectively among 16 sesame genotypes. Both RAPD and ISSR primers showed maximum discrimination power, and produced putative variety specific bands, which could be used for the identification of all the sesame genotypes, individually. However, only AG and CA based ISSR primers were found effective in the discrimination of genotypes. A poor correlation was observed between the matrices produced by RAPD and ISSR primers, which might be due to the array of different sites of the genome. Though, there was greater similarity among sesame genotypes (0.78 for RAPD and 0.71 for ISSR), the observed genetic diversity (0.22 for RAPD and 0.29 for ISSR), was found effective for the characterization of sesame genotypes. It is suggested that putative variety specific RAPD and ISSR markers could be converted to Codominant sequence characterized amplified region/sequence tagged site (SCAR /STS) markers to develop robust variety specific markers.     

Molecular characterization of <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek genotypes based on nuclear ribosomal DNA and RAPD polymorphism

Mungbean germplasm characterization, evaluation and improvement are fundamentally based on morpho-agronomic traits. The lack of break-through in mungbean production has been due to non-availability of genetic variability for high yield potential. Forty-four genotypes of mungbean [Vigna radiata (L.)Wilczek] were subjected to random amplified polymorphic DNA (RAPD) analysis to assess the genetic diversity and relationships among the genotypes. Multilocus genotyping by twelve RAPD primers generated 166 markers and detected an average of intraspecific variation amounting to 82% polymorphism in banding patterns. Dendrogram obtained from cluster analysis delineated all the 44 genotypes into six clusters. Higher values of Nei’s gene diversity (h) and Shannon information index (i) and genetic distance analysis validate existence of wide genetic diversity among mungbean genotypes tested. Besides internal transcribed spacer (ITS) length variations, single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELS) were detected at number of sites in nuclear rDNA region and the sequences of representatives of each sub-cluster and all distinct genotypes have been submitted to NCBI database and assigned Gen accession numbers HQ 148136-148147. Multiple sequence alignment revealed further lineages of distinct genotypes with main RAPD clusters. The measures of relative genetic distances among the genotypes of mungbean did not completely correlate the geographical places of their development. The homogeneous phenotypic markers proved insufficient in exhibiting genetic divergence among mungbean genotypes studied. RMG-62, RMG-976, and NDM-56 have been identified as potential source of parents for crop improvement. RAPD primers, OPA-9 and OPA-2 as polymorphic genetic markers and number of pods/plant and number of seeds/plant as dependable phenotypic markers have been identified for improving yield potentials. This genetic diversity will be of significance in developing intraspecific crosses in mungbean crop improvement programme.     

Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes

Ten snap bean (Phaseolus vulgaris) genotypes were screened for polymorphism with 400 RAPD (random amplified polymorphic DNA) primers. Polymorphic RAPDs were scored and classified into three categories based on ethidium bromide staining intensity. An average of 5.19 RAPD bands were scored per primer for the 364 primers that gave scorable amplification products. An average of 2.15 polymorphic RAPDs were detected per primer. The results show that primer screening may reduce the number of RAPD reactions required for the analysis of genetic relationships among snap-bean genotypes by over 60%. Based on the analysis of the distribution of RAPD amplification, the same number of polymorphic RAPDs were amplified from different genotypes for all RAPD band intensity levels. A comparison of RAPD band amplification frequency among genotypes for the three categories of bands classified by amplification strength revealed a measurable difference in the frequencies of RAPDs classified as faint (weakly amplifying) compared to RAPD bands classified as bold (strongly amplifying) indicating a possible scoring error due to the underscoring of faint bands. Correlation analysis showed that RAPD bands amplified by the same primer are not more closely correlated then RAPD bands amplified by different primers but are more highly correlated then expected by chance. Pairwise comparisons of RAPD bands indicate that the distribution of RAPD amplification among genotypes will be a useful criterion for establishing RAPD band identity. For the average pairwise comparison of genotypes, 50% of primers tested and 15.8% of all scored RAPDs detected polymorphism. Based on RAPD data Nei's average gene diversity at a locus was 0.158 based on all scorable RAPD bands and 0.388 if only polymorphic RAPD loci were considered. RAPD-derived 1 relationships among genotypes are reported for the ten genotypes included in this study. The data presented here demonstrate that many informative, polymorphic RAPDs can be found among snap bean cultivars. These RAPDs may be useful for the unique identification of bean varieties, the organization of bean germplasm, and applications of molecular markers to bean breeding.     

萝卜种质资源亲缘关系的RAPD分析

利用筛选出的12个随机引物,对来源于不同国家和地区的有代表性的56份萝卜种质资源遗传多样性进行了RAPD分析,共扩增出109条带,其中多态性带为72条,多态性带百分率为62.9%,种质平均期望杂合度为0.289.系统聚类分析将供试材料分为2大类、9组,主坐标分析将其分为3大类、7组,两种分类方法结果基本一致,后者能更为直观和清楚地反映群体之间的关系.     

Estimation of genetic distance based on RAPDs between 11 cotton accessions varying in heat tolerance

The genetic distance of 11 cotton genotypes varying in heat tolerance was studied using RAPD markers. Fifty-three random decamer primers were used for the estimation of genetic distance. Among the 53 RAPD primers, which were custom synthesized by GeneLink Inc., UK, 32 were polymorphic and 21 were monomorphic. The 32 polymorphic primers produced 273 fragments, with a mean of 8.3 fragments per primer. The number of polymorphic bands produced in the 11 cotton accessions ranged from 1 to 31. Primer GLC-20 produced 31 polymorphic bands, while two primers, GLB-5 and GLC-12, produced one polymorphic band each. A range of 88.89 to 42.48% genetic similarity was observed among the 11 cotton accessions. The highest genetic similarity was observed between FH-945 and BH-160 (88.89%), whereas the lowest value was found between NIAB-801/2 and FH-945 (42.48%). Unique amplification profiles were produced by most of the cultivars; the differences were sufficient to distinguish them from other genotypes. This confirms the efficacy of RAPD markers for the identification of plant genotypes. An accumulative analysis of amplified products generated by RAPDs was sufficient to assess the genetic diversity among the genotypes. This information should be helpful for formulating breeding and genome mapping programs.