南方红豆杉紫杉烷高含量植株系RAPD初步研究

南方红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓｖａｒ． ｍａｉｒｅｉ（Ｌｅｍｅｅ ｅｔＬｅｖｌ．） Ｃｈｅｎｇ ｅｔＬ．Ｋ．Ｆｕ） 是我国提取抗癌药紫杉醇的原料植物之一,但分布于不同地区的南方红豆杉其紫杉醇含量差异很大。应用ＲＡＰＤ技术,用２１ 个引物共扩增２５６ 条条带,其中有２２４ 条有多态性（８７．８ ％） 。结合紫杉烷及紫杉醇含量测定和植物形态分类,初步探讨了紫杉烷高含量植株的分子特征、形态和生境的相互关系。在供试的１２ 个地区２４ 个样品中,提出３ 个地区存在紫杉烷高含量植株系的可能性,为资源利用和保护以及优良品种的培育提供基     

Microsatellites of <Emphasis Type="BoldItalic">Laminaria digitata</Emphasis> tested in <Emphasis Type="BoldItalic">Lessonia nigrescens</Emphasis>: Evaluation and improvement of cross amplification between kelps of two different families

The use of primers designed originally to amplify DNA for one species in a different one can save time and resources, particularly for microsatellite loci. Microsatellite amplification improvements across two kelp families are reported, where loci originally described in Laminaria digitata (Laminariaceae) were tested in Lessonia nigrescens) was observed in two localities affected by massive mortality events. Nei’s distances among five populations presented similar patterns to those of 30 multilocus dominant loci (RAPD) evaluated in the same localities. Although some success might be achieved in cross-species microsatellite amplifications, the strong mutations detected between these two Laminarian families suggests that better results of cross-amplifications should be expected at much lower taxonomic levels. Thus, although more expensive, construction of new gene libraries is strongly recommended.     

Direct evidence for biased gene diversity estimates from dominant random amplified polymorphic DNA (RAPD) fingerprints

The relevance of using dominant random amplified polymorphic DNA (RAPD) fingerprints for estimating population differentiation was investigated when typically small population sample sizes were used. Haploid sexual tissues were first used to determine genotypes at RAPD loci for 75 eastern white pines ( Pinus strobus L.) representing five populations. Dominant RAPD fingerprints were then inferred from genotypic data for each individual at each locus, and gene diversity estimates from both sources of data were compared. Genotypic information at RAPD loci indicated little or no differentiation among populations, similar to allozyme loci. However, estimates of population differentiation derived from dominant RAPD fingerprints according to various common methods of analysis were generally inflated, especially when all fragments were considered. Simulations showed that an increase in loci sampling and population sample sizes did not significantly alleviate the biases observed.     

Microevolutionary Patterns and Molecular Markers: The Genetics of Geographic Variation in Ascaris suum

Molecular markers have been used only rarely to characterize the population genetic structure of nematodes. Published studies have suggested that different taxa may show distinct genetic architectures. Isoenzyme and RAPD markers have been used to investigate geographic variation of Ascaris suum at the level of infrapopulations (nematodes within individual hosts), within localities, and among geographic regions. Independent estimates of genetic differentiation among population samples based on isoenzyme and RAPD data showed similar patterns and substantial correlation. Heterozygote deficiencies within infrapopulations and large values for inbreeding coefficients among infrapopulations suggested that the composition of these populations was not consistent with a model of random recruitment from a large panmictic pool of life-cycle stages. Both isoenzyme and RAPD markers revealed moderate levels of genetic differentiation among samples representing infrapopulations and localities. Of total gene diversity, 9.4% (isoenzyme) and 9.2% (RAPD) was partitioned among infrapopulations. Geographic localities accounted for 7.8% (isoenzyme) and 6.2% (RAPD) of total diversity. Only infrapopulations from the same farm had low levels of differentiation.     

First comparative phenetic studies of Argentinean species of Acacia (Fabaceae), using morphometric, isozymal, and RAPD approaches

Morphological and genetic diversity among Acacia aroma, A. macracantha, A. caven, and A. furcatispina were studied with morphometric, isozymal, and RAPD approaches. The analysis of seven isozyme systems revealed 21 loci, and RAPD analysis showed 34 loci. Most of these loci allowed us to differentiate the species, with the exception of A. aroma and A. macracantha, the two most similar species. The levels of genetic variability estimated by isozymes were higher than those obtained from RAPD analyses. Morphometric characters showed highly significant differences among the species, although A. aroma and A. macracantha are differentiated only by thorn length. The phenogram obtained from isozyme data is consistent with morphological data. The RAPD phenogram based on allelic frequencies showed agreement with morphological and isozymal approaches only at the intraspecific levels, while the RAPD phenogram based on Nei and Li's similarity measures agreed with the phenograms constructed from isozyme and morphological data. High similarities and high indirect gene flow were found between A. aroma and A. macracantha, results that call the relationship between them into question.     

Evaluation of Genetic Variation in the Clown Knifefish, Chitala chitala, Using Allozymes, RAPD, and Microsatellites

Twenty-seven enzyme systems, six random amplified polymorphic DNA (RAPD) primers, and two microsatellite loci were tested to determine intraspecific divergence in the natural population of the endangered Indian featherback fish, Chitala chitala, for the first time. The 262 samples of C. chitala were collected from six riverine locations in India: the Satluj, Ganga (Ghagra, Bhagirathi, and Brahmaputra), Mahanadi, and Narmada river systems. The analysis revealed population subdivisions, with an F_ST value from 0.1235 (95% confidence 0.0868–0.1621) for RAPD and a combined F_ST of 0.0344 (95% confidence 0.0340–0.0350) for microsatellite loci. An analysis of 38 allozyme loci did not reveal any polymorphism in the samples from any of the riverine localities; a possible explanation for this could be that the ancestors of Chitala could have faced a population reduction in prehistoric periods, as low allozyme variation is also reported for other species of Chitala from south Asia.     

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.     

Genetic diversity in populations of Dalbulus maidis (DeLong and Wolcott) (Hemiptera: Cicadellidae) from distant localities in Brazil assessed by RAPD-PCR markers

Populations of Dalbulus maidis (DeLong and Wolcott) from the northeastern and central-southern regions of Brazil differ morphologically, suggesting that they could be genetically isolated. Here we used the random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) technique to estimate genetic structuring of this leafhopper species among five geographically distant localities across those regions and to estimate gene flow between populations. Ten specimens were sampled per population and genotyped with RAPD markers generated from amplification with nine oligonucleotides. The percentage of polymorphic loci was 78% in relation to the total number of amplified loci, and genetic similarity either between or within populations was higher than 0.72. Cluster analysis grouped specimens from the northeastern population (Mossoró/RN) into a single group, whereas central-southern specimens were not grouped in relation to their places of origin. Overall, the genetic subdivision index (Fst) was low (or= 0.192 and Nm 相似文献   

草鱼基因组DNA一些RAPD位点的遗传分析及分子标记筛选

RAPD技术的实验结果很容易因实验条件和反应参数的不同而造成差异。为了建立能通用的草鱼基因组多态性分析RAPD分子标记体系,需要利用RAPD位点按照孟德尔共显性规律遗传的特点,用不同遗传背景的材料对多态性的RAPD位点进行统计遗传学比较分析以判断其真实性。为此,本实验选择具有遗传多态性的湘江流域草鱼群体和经连续两代人工诱导雌核发育获得的雌核发育草鱼品系,对一些可能作为草鱼基因组DNA分子标记的RAPD位点进行了遗传学比较分析。所用的10条多态性随机引物共检测到30个多态性RAPD位点。两个不同遗传背景群体的遗传统计对比分析结果表明:这30个多态位性位点在湘江流域草鱼群体和雌核发育草鱼群体中的分布符合孟德尔遗传规律,可以作为草鱼基因组DNA分析的可靠分子标记。本实验的观察结果还表明:在人工诱导雌核发育过程中,存在RAPD位点的快速丢失现象,两次人工诱导雌核发育过程中共丢失了17个多态性位点。因此,加强对自然水体中草鱼种质资源多样性的保护和利用各种现代生物学技术纯化、筛选和组合优良性状基因,是草鱼遗传育种中同样重要和不可或缺的两个方面。       

Low isozyme diversity in Nordic and central European populations of Vicia pisiformis and V. dumetorum (Fabaceae)

Vicia pisiformis and V. dumetorum are closely related perennial species occurring in small, disjunct populations throughout Europe. In Sweden, they are classified as "vulnerable" and "care-demanding", respectively. They possess low variation in RAPD (Random Amplified Polymorphic DNA) markers but relatively high morphological variation. We estimated the amount and distribution of isozyme diversity within and among marginal (Sweden, Norway) and central (Czech Republic) populations. Vicia dumetorum was monomorphic for all 14 isozyme loci in the 22 populations investigated, whereas V. pisiformis had 2.94 % polymorphic loci and a mean of 1.02 alleles per loci. The differentiation (F_sr) of populations of V pisiformis were 0.848, but no clear geographical pattern was found. Preliminary results from artificial pollination experiments suggests self-compatibility in V. dumetorum . The low overall polymorphism in Swedish and Czech localities in both species is explained in terms of common postglacial colonization events, recent anthropogenic activity, selfing and clonal reproduction. Finally, the implications of the data for conservation genetics is discussed.     

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.     

DNA polymorphism in the living fossil Ginkgo biloba from the eastern United States.

Random amplified polymorphic DNA (RAPD) analysis is a valuable tool in studying inter- and intra-specific genetic variations, patterns of gene expression, and for the identification of specific genes using nearly isogenic variants. Here we used RAPD analysis to study the genetic variation in Ginkgo biloba grown in the eastern United States. Our results support the evidence that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern than dye-stained gels or Southern blot hybridization of RAPD blots using probes made from purified PCR products. Using these techniques, we observed a high degree of relatedness among plants grown in certain localities although significant genetic variation may exist in the species, and could be a possible explanation for the observed variations in the efficacy of medications derived from G. biloba extract.     

Strong genetic differentiation among east Atlantic populations of the sword razor shell (<Emphasis Type="Italic">Ensis siliqua</Emphasis>) assessed with mtDNA and RAPD markers

The sword razor shell Ensis siliqua (Linnaeus, 1758) is a bivalve with a high commercial value being appreciated in fresh and processed markets. However, the genetic studies carried out in populations of E. siliqua are scarce. In this work, the genetic variability and differentiation of the sword razor shell was assessed using PCR–RFLPs of a fragment of the 16S rRNA mitochondrial gene and random amplified polymorphic loci (RAPD) in nine localities from Ireland, Spain, and Portugal. In the 314 individuals examined for the mitochondrial fragment, 12 composite haplotypes were observed; meanwhile, a unique phenotype was observed for each of the 242 individuals analyzed with 61 RAPD loci. Two of the mitochondrial composite haplotypes accounted for the majority of individuals (89.81%) and showed a remarkably disjoint distribution between Irish and Iberian samples, with the exception of Aveiro which exhibited as the most frequent haplotype the same found in Ireland. The level of variability observed for each sample was generally correlated with both types of markers and the results obtained suggest the existence of a strong population differentiation between Irish and Iberian localities, except for the Portuguese sample from Aveiro which is surprisingly closer to Irish individuals, although it is probably highly differentiated.     

Population Genetic Structure of Astyanax scabripinnis (Teleostei, Characidae) from an Urban Stream

Despite the great anthropogenic interference on urban streams, information is still scarce about the genetic variability and structure of native fish populations inhabiting such streams. In the present study, random amplified polymorphic DNA (RAPD) markers were used to analyze genetic variability and structure of populations assigned to the Neotropical fish species Astyanax scabripinnis from an urban stream located in Londrina, Paraná State, southern Brazil. Thirty individuals of this species were collected from three sites throughout the upper Cambé stream. A total of 10 primers amplified 159 loci, of which 128 (80.5%) were polymorphic. Each of the three populations showed very similar proportions of polymorphic loci, which ranged from 63.5 to 64.8%. Unbiased genetic distances varied from 0.0612 to 0.0646. Theta_p-test values indicated moderate to high genetic differentiation among individuals from different localities. The number of migrants varied from 1.34 to 1.46, suggesting a low gene flow between populations. The genetic similarity among all individuals studied ranged from 0.424 to 0.848. The results suggest that populations of A. scabripinnis in Cambé stream are undergoing genetic differentiation.     

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.     

苎麻疫霉群体的RAPD分析

利用从126个RAPD(RandomAmplifiedPolymorphicDNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的8条引物,对采集自江苏、安徽和江西不同寄主的45个Phytophthoraboehmeriae菌株进行全基因组DNARAPD标记遗传多样性分析。选用引物共标出DNA指纹图带68条,其中多态性条带20条,多态性检测率为29.4%,表明该种内不同地区和寄主来源的菌株间变异较小。利用Popgene软件计算供试菌株间的遗传距离并绘制聚类树状图,供试菌株被划分为2个遗传聚类组。菌株间的遗传相似性与菌株的寄主来源有一定的相关性,来自江苏、江西和安徽棉花上的27个菌株被划分在同一遗传聚类组内,而分离自构树、枫杨和苎麻的18个菌株被划分在另一个遗传聚类组。结果还表明菌株间遗传相似性与其地区来源无直接相关性。     

苎麻疫霉群体的RAPD分析*

利用从126个RAPD(RandomAmplifiedPolymorphicDNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的8条引物,对采集自江苏、安徽和江西不同寄主的45个Phytophthoraboehmeriae菌株进行全基因组DNARAPD标记遗传多样性分析。选用引物共标出DNA指纹图带68条,其中多态性条带20条,多态性检测率为29.4%,表明该种内不同地区和寄主来源的菌株间变异较小。利用Popgene软件计算供试菌株间的遗传距离并绘制聚类树状图,供试菌株被划分为2个遗传聚类组。菌株间的遗传相似性与菌株的寄主来源有一定的相关性,来自江苏、江西和安徽棉花上的27个菌株被划分在同一遗传聚类组内,而分离自构树、枫杨和苎麻的18个菌株被划分在另一个遗传聚类组。结果还表明菌株间遗传相似性与其地区来源无直接相关性。     

大豆灰斑病菌生理小种的RAPD标记*

运用随机扩增多态性DNA（RandomamplifiedpolymorphicDNA,RAPD）技术对发生于中国东北的大豆发斑病菌（Cercosporidiumsojinum）的10个生理小种进行基因组DNA多态性分析。用13个10-核苷酸随机引物共计获得了111个RAPD标记,其中86.5％具有多态性,通过聚类分析确定了供试小种间的亲缘关系。试验证明,RAPD技术分析大豆灰斑病菌遗传变异可提供大量分子标记,综合分析13个随机引物的扩增谱带可将供试菌株清楚分开。RAPD技术是一项操作简单、快速和灵敏的方法,极具对病菌群体遗传分析的潜力。     

5个泥蚶群体遗传多样性的RAPD分析

采用随机扩增多态性DNA(RAPD)技术 ,对 5个泥蚶群体的遗传多样性进行了研究。在选择的 2 0个随机引物中共检测到 15 4个扩增片段 ,长度在 30 0bp～ 2 30 0bp之间 ,每个个体扩增条带在 1～ 11条不等。利用Popgen3.2和PHILIP统计软件 ,获得实验数据 ,确立了 5个群体间的亲缘关系。结果表明 :( 1)泥蚶的 5个地理群体分化不明显 ,没有形成不同的地理种群 ;( 2 )聚类分析显示 ,韩国群体与浙江群体亲缘关系最近 ,然后依次为山东群体和福建群体 ;( 3) 5个群体无论是在多态位点比例还是在平均遗传杂合度上都处于较高水平 ,说明泥蚶当前种质资源状况良好 ,遗传多样性水平较高 ,泥蚶养殖有很好的发展前景 ;( 4)与韩国野生群体相比 ,其他 4个泥蚶群体在多态位点比例和遗传平均杂合度上都有不同程度的降低 ,表明人为等因素已经开始影响到泥蚶的种质资源状况。因此 ,为持续发展泥蚶养殖业 ,必须加强保护泥蚶现有的种质资源并制定相应的渔业管理措施。     

大豆灰斑病菌生理小种的RAPD标记

运用随机扩增多态性DNA（RandomamplifiedpolymorphicDNA,RAPD）技术对发生于中国东北的大豆发斑病菌（Cercosporidiumsojinum）的10个生理小种进行基因组DNA多态性分析。用13个10-核苷酸随机引物共计获得了111个RAPD标记,其中86.5％具有多态性,通过聚类分析确定了供试小种间的亲缘关系。试验证明,RAPD技术分析大豆灰斑病菌遗传变异可提供大量分子标记,综合分析13个随机引物的扩增谱带可将供试菌株清楚分开。RAPD技术是一项操作简单、快速和灵敏的方法,极具对病菌群体遗传分析的潜力。