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。     

Study of genetic diversity in safflower genotypes using agro-morphological traits and RAPD markers

This research was conducted to study the genetic diversity in safflower (Carthamus tinctorius L.) using agro-morphological traits and RAPD markers. Sixteen selected lines derived from landraces growing in various agro-climatic regions of Iran along with four exotic genotypes were evaluated in a randomized complete block design with three replications under field conditions. Days to emergence, days to initial flowering, days to flowering, days to maturity, plant height, branches per plant, capitula per plant, seeds per capitulum, 1,000-seed weight, seed yield per plant, seed yield, and reaction to powdery mildew (Leveillula taurica Arnaud) were evaluated in this study. Genetic diversity of the genotypes was assessed by RAPD markers. The results indicated significant differences among genotypes for the agro-morphological traits and clustering based on these traits classified the genotypes into five groups. Analysis of the RAPD markers revealed 15 polymorphic primers out of 50 used primers. Based on RAPD data, the highest genetic similarity was observed between the cultivars of “AC Sunset,” “AC Sterling” from Canada and the lowest relatedness observed between a local breeding line “E₂₄₂₈” and genotype “GE₆₂₉₂₃” from Germany. Cluster analysis based on RAPD markers and 54% coefficient of similarity divided the genotypes into five distinct groups. Comparing the clusters based on agro-morphological traits with those from molecular markers showed slight similarities. The finding of high genetic variation for agro-morphological traits and polymorphism at DNA level reveal that agronomic traits can be improved by selection programs.     

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。     

RAPD variability in rice (Oryza sativa L.) plants derived from desiccation-tolerant calli

Genetic modification from selfed progenies of 18 rice (Oryza sativa L.) plants regenerated from callus tissues which survived desiccation, were investigated at the DNA level using the random amplified polymorphic DNA (RAPD) method. Twelve 10-mer random primers were used to amplify DNA of progenies from the regenerated plants, and a total of 228 PCR products and 1780 DNA fragments were obtained by primers, generating between four to thirteen major bands. The size of the amplified fragments ranged from 0.2 to 2.55 kb. The results showed that 10 out of 12 primers produced polymorphic bands, two primers (RA31 and RA185) showed no polymorphism among plants tested. A dendrogram of the genetic distance was constructed based on their polymorphism, demonstrating that somaclonal variation exists in rice plants regenerated from callus which survived the desiccation treatment. Part of this variation can be useful in rice breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic diversity of hexaploid wheat cultivars estimated by RAPD markers, morphological traits and coefficients of parentage

Estimates of genetic diversity can be based on different types of data. The aim of this research were to study genetic diversity among Croatian wheat cultivars by random amplified polymorphic DNA (RAPD) markers, morphological traits and pedigree records; to analyse differences between wheat cultivars from two breeding centres; and to evaluate usability of RAPD markers for estimation of genetic diversity among wheat cultivars in comparison with morphological traits and pedigree record data. Studies were conducted on 14 wheat cultivars and breeding lines from two breeding centres in Croatia. For the RAPD analysis 36 primers were screened and the 14 most polymorphic ones yielded 341 polymorphic bands. Twelve morphological traits were used for morphological analysis. Pedigrees were composed of seven generations of ancestors. RAPD markers showed a high level of polymorphism among the cultivars examined and the breeding lines. No significant correlations were observed among the methods tested.     

胡萝卜种质资源遗传多样性的RAPD分析

利用RAPD技术对34份胡萝卜种质资源进行遗传多样性研究,以探明各材料间的亲缘关系、遗传多样性及其分类,为进一步利用和创新品种提供参考信息。结果表明,20条随机引物共扩增出139个DNA片段,其中99条带表现出多态性,占总带数的76.74%,对其进行数据化处理后聚类,34份胡萝卜材料可划分为5个类群。材料的归组与根形、根长有一定的相关性。     

Purity control of F1-hybrid tomato cultivars by RAPD markers

Randomly amplified polymorphic DNA (RAPD) markers were applied in purity control of hybrid seed production of tomato (Lycopersicon esculentum Mill.). DNA from three commercial F₁-hybrid cultivars and their parental lines was subjected to RAPD screening with 50 primers. Two of four primers which detected polymorphism between the parents tested, generated paternal-specific RAPDs, enabling a clear distinction to be made between hybrids and their maternal parents. In addition, combination of the polymorphic DNA products generated by these primers exhibited hybrid-specific patterns, enabling each cultivar to be identified. This result indicates the practical usefulness of RAPD markers in hybrid-tomato-seed purity-control tests and cultivar identification. The approach is advantageous in its rapidity and simplicity, particularly as an alternative for those cultivars for which lengthy and costly phenotypic tests are currently used.     

野生小豆种质资源遗传多样性的RAPD分析

利用RAPD分子标记技术对16份不同来源地的野生小豆种质进行基因组DNA多态性分析,25个随机引物共扩增出240个DNA片段,其中197条谱带表现出多态性。对基于数字化处理后的多态性谱带进行聚类,16个野生小豆种质可划分为4个类群：日本类群、中国类群、不丹类群和尼泊尔类群,材料的归组与其来源地有显著的相关性。     

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。     

陆两优996种子纯度的SRAP指纹图谱鉴定

利用270对SRAP引物对陆两优996和2个亲本的DNA指纹图谱，获得能在父、母本间表现出多态性的特异SRAP标记引物有54对，其中差异性条带136个。利用Me3Em8引物，条带在杂交种完全互补，对200株样本进行了纯度鉴定，结果鉴定纯度为97.50%，与田间种植鉴结果97.50%(海南鉴定)一致，表明SRAP标记技术适用于种子纯度鉴定。     

杂交油葵A15种子纯度的RAPD鉴定

从杂交油葵A15及其亲本中提取基因组DNA,用180个RAPD随机引物进行扩增,从中筛选出3个可将亲本和子代区分开的引物OPD09、OPD12和OPK12。OPD09产生亲本互补的特征带OPD09-1470bp、OPD09-870bp;OPD12产生母本特征带OPD12-1230bp,OPK12产生父本特征带OPK12-1540bp、OPK12-940bp,上述谱带均在子代中出现。以单引物(OPD09)和双     

Random amplified polymorphic DNA (RAPDs) markers for genetic analysis in micropropagated plants of Robinia pseudoacacia L.

Four years' old micropropagated plants regenerated by enhanced axillary branching from shoot buds of a single genotype of Robinia pseudoacacia were characterized by RAPDs. Random amplified polymorphic DNA analysis was carried outusing 19 random 10-mer DNA primers and 286 RAPD bands were examined which showed 30% polymorphism. Similarity indices ranged from 0.86 to 0.96 among different plants based on RAPD data. The UPGMA dendrogram was constructed based on similarity indices which showed clustering of different plants into subgroups based on similarity values. Our results suggest that somaclonal DNA sequence variations are present even when organized cultures such as shoot buds were used as explant for micro-propagation. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic variation within and between two cultivars of red clover (Trifolium pratense L.): Comparisons of morphological,isozyme, and RAPD markers

Summary Morphological, isozyme and random amplified polymorphic DNA (RAPD) markers were used to estimate genetic variation within and between cultivars of red clover (Trifolium pratense L.), an important temperate forage legume. Two cultivars of red clover, Essi from Europe and Ottawa from Canada, were evaluated. Six monogenic morphological characters were observed for 80 plants from each of these two cultivars. All six morphological loci were polymorphic in the cultivar Essi whereas only four loci were polymorphic in the cultivar Ottawa. Forty plants from each cultivar were assayed for isozyme markers. A total of 21 enzyme-coding loci with 43 alleles was detected using twelve enzyme systems. Thirteen and nine of these loci were polymorphic in Essi and Ottawa, respectively. The mean number of alleles per locus was 1.81 in Essi and 1.67 in Ottawa. Seventeen random 10-mer primers were screened for RAPD markers. Nine primers which gave clear and consistent amplified products were used to assay 20 individuals from each cultivar. Each primer gave from 7 to 20 amplified bands with an average of 14.8 bands per primer. One hundred and eight of 116 putative loci were polymorphic in Essi and 90 of 98 loci were polymorphic in Ottawa. High within-cultivar variation was observed in both cultivars using both isozyme and RAPD markers. This high polymorphism makes these markers useful for germplasm characterization and genetic studies in red clover.     

枣树不同品种遗传多样性的RAPD分析

为了验证现有枣树材料的亲缘关系,本研究利用RAPD标记对21种枣树品种的遗传多样性进行了分析。结果表明：从36个RAPD引物中,筛选出20个可扩增出清晰且多态性高的条带的引物用于RAPD-PCR扩增,共获得185条DNA片段,其中173条具有多态性,多态率达93.51%,平均每个引物可以扩增出9.25条。利用DPS软件建立了21种枣树品种的聚类分析树状图,聚类分析结果显示：在相似系数82.48处所有枣树品种被聚为一类,在65.98处21种枣树品种被分为两大类群。     

Genetic Diversity of Cultivated Chinese Chive Germplasm by ISSR and RAPD Markers

利用RAPD(Random Amplified Polymorphic DNA)和ISSR(Inter-simple Sequence Repeat)两种分子标记技术对20份韭菜栽培品种进行了遗传多样性研究。结果表明，筛选后选用的12个ISSR引物和15个RAPD引物分析分别产生了258和101条扩增产物带，其中多态性条带(即20个韭菜品种中一个或多个但不是全部具有的带)分别为132和40条，分别占总数的51.2%和39.6%。，也就是说12个ISSR引物和15个SSR引物对韭菜不同品种的扩增可分别产生51     

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     

石榴基因组DNA的提取及RAPD引物筛选

以山东省枣庄市峄城区中国石榴种质资源圃内石榴品种为材料,采取改良CTAB法提取石榴基因组DNA,经RAPD-PCR扩增筛选引物。结果表明：CTAB法提取的石榴基因组DNA谱带清晰,纯度高,浓度大,可以用于RAPD分析。从50条随机引物中筛选出多态性高、重复性好的多态性引物20条。20条引物对10个品种扩增结果共产生180个位点,多态性位点为134,多态性比率为74.4%。     

Random amplified polymorphic DNA variation among German and Dutch asparagus cultivars

DNA polymorphism among nine cultivars of Asparagus officinalis L. was measured using random amplified polymorphic DNA (RAPD). Of 69 reproducible amplification products from 12 arbitrary decamer primers, 49 RAPD markers were polymorphic and could be used to distinguish six German and three Dutch asparagus cultivars. Even with very small sample sizes, genetic similarity measurements based on the RAPD data allowed accurate grouping of the nine cultivars into distinct clusters, with the exception of two individuals which clustered to closely related varieties. Two German cultivars showed high genetic similarity and were distinct from the remaining German varieties. The German and Dutch cultivars were clearly separated by a relatively large genetic distance.     

Analysis of molecular variance and population structure in southern Indian finger millet genotypes using three different molecular markers

The genetic relationship among 42 genotypes of finger millet collected from different geographical regions of southern India was investigated using random amplified polymorphic DNA (RAPD), inter simple sequence repeats (ISSR), and simple sequence repeats (SSR) markers. Ten RAPD primers produced 111 polymorphic bands. Five ISSR primers produced a total of 61 bands. Of these, 23 bands were polymorphic. The RAPD and ISSR fingerprints revealed 71.3 and 37.4% polymorphic banding patterns, respectively. Thirty-six SSR primers yielded 83 scorable alleles in which 62 were found to be polymorphic. Out of 36 SSR primers used, 14 primers (46.6%) produced polymorphic bands. The SSR primer UGEP7 produced a maximum number of six alleles. Mean polymorphic information content (PIC) of RAPD, ISSR and SSR were 0.44, 0.28, and 0.14, respectively. Molecular variances among the population were 2, 11, and 1% for RAPD, ISSR, and SSR markers, respectively. SSR produced 99% molecular variance within individuals. RAPD and ISSR markers produced a low level of molecular variance within individuals. The STRUCTURE (model-based program) analysis revealed that the 42 finger millet genotypes could be divided into a maximum of four subpopulations. Based on the Bayesian statistics, each RAPD and SSR marker produced three subpopulations (K=3), while ISSR marker showed four subpopulations (K=4). This study revealed that RAPD and SSR markers could narrow down the analysis of population structure and it may form the basis for finger millet breeding and improvement programs in the future.     

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。