Genetic diversity in mung bean germplasm revealed by RAPD markers

Knowledge of the genetic relationships among landraces is useful to gene bank managers because it permits a better organization of the crop's gene pool management, more efficient sampling of the available germplasm resources and better access to useful genetic variation for breeders. Genetic diversity of 19 landraces of the cultivated mung bean, Vigna radiate, and three weedy and wild relatives including Vigna mungo, Vigna luteola and Vigna radiate var. sublobata, was investigated at the DNA level with the random amplified polymorphic DNA (RAPD) procedure. Sixty random decamer primers were employed in amplification reactions; 28 of these were informative and yielded 246 bands, of which 229 were polymorphic with a mean of 8.2 bands per primer. A genetic distance matrix based on Nei and Li coefficient was converted to a dendrogram and a two-dimensional plot using multidimensional scaling (MDS). The accessions studied were separated into three main clusters, which included V. radiate landraces, V. mungo and V. luteola, respectively. The variation of this cluster supports the view that the genetic distance of V. mungo and V. luteola varies considerably from the accession VO2955 (V. radiata). The multidimensional scaling plot confirmed that V. mungo, V. luteola and most of the accessions of V. radiata formed distinct clusters with no overlap, and two mung bean accessions (PI177493 and VO4134–1 from Turkey and India, respectively) were genetically distant from other V. radiata landraces. V. radiata and V. mungo are positioned in separate botanical species and V. radiata var. sublobata is classified within other V. radiata landraces. Based on the limited range of accessions tested, the approach holds promise for the classification of mung bean germplasm, identification of mung bean landraces and applications of molecular markers to mung bean breeding.     

Genetic diversity in European perennial ryegrass cultivars investigated with RAPD markers

Perennial ryegrass (Lolium perenne L.) is the most important grass species for temperate grassland agriculture. The genetic relationship and distance among cultivars is largely unknown but of great interest for breeding programmes. The objectives of this study were to (i) investigate the molecular variation and structure of population cultivars, (ii) describe the relationship among cultivars in terms of the modified Rogers’ distance, and (iii) determine the minimum sample size required for characterization of cultivars of L. perenne using random amplified polymorphic DNA (RAPD) markers. A total of 22 ryegrass cultivars, mainly of European origin, were investigated with RAPD markers. The minimum sample size required for the characterization of cultivars was about 20 individuals per population. An analysis of molecular variance (AMOVA) revealed a much larger genetic variation within cultivars (66%) than between them (34%).     

Genetic diversity and relationships among Lablab purpureus genotypes evaluated using RAPD as markers

Summary Genetic variation in 40 accessions of Lablab purpureus was evaluated using random amplified polymorphic DNA as markers. A high level of genetic variation in this species was detected but this was mainly restricted to the difference between cultivated and wild forms. Of the cultivated genotypes, genetic variation among Asian collections was significantly higher than that among African collections. The three most divergent cultivated genotypes were all from Asia. Four of the five wild accessions, two from Zimbabwe and the other two from Zambia, were closely related. The other one, CPI 31113 collected from Uganda, was highly divergent. The two commercial forage varieties used in Australia, Rongai from Kenya and Highworth from India, were not very different.     

Genetic diversity for RAPD markers between cultivated and wild accessions of Coffea arabica

Summary Random amplified polymorphic DNA (RAPD) markers have been successfully employed to analyse the genetic diversity among cultivated and subspontaneous accessions of Coffea arabica. The narrow genetic base of commercial cultivars was confirmed. On the other hand, a relatively large genetic diversity was observed within the germplasm collection demonstrating the importance of collecting missions. Results suggested an East-West differentiation in Ethiopia, the primary centre of diversification of C. arabica. The large heterosis effect reported in intergroup hybrids could be related to such genetic differentiation. RAPD method appeared to be effective in resolving genetic variations and in grouping germplasm in C. arabica.     

Genetic diversity of Tainan-white maize inbred lines and prediction of single cross hybrid performance using RAPD markers

To evaluate the genetic diversity of 13 maize inbred lines, and to determine the correlation between genetic distance and single cross hybrid performance, we employed the randomly amplified polymorphic DNA(RAPD)- a PCR-based technique. Six of these lines came from the Taichung population, and others derived from seven different sites. Forty different primers were used to give a total of 646 reproducible amplification products, 547 (84.7%) of them being polymorphic. Genetic divergence was determined using Jaccard's similarity coefficient, and a final dendrogram was constructed by UPGMA (unweighted pair-group method with arithmetical averages) cluster analysis in the CLUSTER procedure of the SAS system. The RAPD analysis was a useful tool in determining the extent of genetic diversity among Tainan-white maize inbred lines in the present case. Cluster analysis showed that the 13 inbred lines could be classified into distinct heterotic groups. There was no significant linear regression of grain dry weight heterosis value and mean performance of hybrids on genetic distance. And their coefficients of determination(R²) are small, so that predictive value is limited. The present results showed that the Jaccard's similarity coefficients based on RAPD data cannot be used to precisely predict the F₁ hybrids yield performance and heterosis value. This revised version was published online in August 2006 with corrections to the Cover Date.     

利用RAPD标记对彩色棉遗传多样性的分析

以新彩棉1号、2号和71份彩色棉材料为基础,利用RAPD标记对彩棉品种之间分子标记遗传差异进行研究。从240个随机引物中筛选出10个引物能在73份彩棉材料间扩增出稳定性较好的多态性片段。73份彩棉材料的欧氏遗传距离在1.00～4.79之间。按类平均距离法进行聚类分析表明,这些彩色棉材料可分为7组,品种间的遗传关系与品种自身的系谱有关,大多数品种的遗传基础比较狭窄。     

Genetic diversity of Cannabis sativa germplasm based on RAPD markers

Random amplified polymorphic DN A (RAPD) markers were generated from 13 cultivars and accessions of Cannabis sativa L. Approximately 200 fragments generated by 10 primers of arbitrary sequence were used to assess the level of DNA variation. Statistical analysis was performed using the Dice coefficient of similarity and principal coordinate analysis. The grouping of the accessions according to the cluster analysis was in good agreement with their origin and lines with common ancestors were grouped together. Principal coordinates 1 and 2 revealed a clear separation of Italian and Hungarian germplasm and a third group, including a mixture of genotypes coming from different places; the third coordinate separated the Korean group which is probably the most divergent germplasm. Variability within the two cultivars ‘Carmagnola’ and ‘Fibranova1’ was also shown, suggesting good possibilities for long–term selection work. RAPD markers provide a powerful tool for the investigation of genetic variation in cultivars/accessions of hemp.     

Genetic relationships in cultivars of hop, Humulus lupulus L., determined by RAPD analysis

Random amplified polymorphic DNA (RAPD) was used to evaluate the genetic variability and relationship of 65 hop cultivars from all the major hop-growing regions in the world. Twenty-eight selected random primers used in the RAPD reaction generated an average of 38.6%) polymorphic fragments, which was sufficient to produce 47 different RAPD profiles among the cultivars examined. The level of genetic variability was much higher than previously reported. Genetic similarity was estimated and UPGMA cluster analysis was performed using the RAPD data. Cluster analysis separated the cultivars into genetically related RAPD groups which were compared with pedigree data and grouping of the hop cultivars by essential oil type. The RAPD groups, strongly supported by pedigree data, gave more precise information on the level and distribution of genetic variability within hop cultivars than characterization by essential oils. Cultivars were divided into American and European groups, supporting the distinction between two geo-graphically distinct hop germplasms. Five genetically distinct groups revealed differences within the European germplasm, reflecting past hop breeding practices which have been adopted in different regions. The use of RAPD markers for hop germplasm characterization and genetic diversity study is discussed.     

Genetic variability among elite red clover ( Trifolium pratense L.) parents used in Chile as revealed by RAPD markers

Red clover (Trifolium pratense L.) is one of the main forage species of temperate regions. Cultivars of red clover are heterogeneous which makes their genetic analysis difficult. We applied RAPDs (Random Amplifed Polymorphic DNA) in order to assess the genetic relationship and levels of genetic variability existing among a group of 16 elite red clover parents organised in four subsets of 4 parents each. Out of 55 primers 21 provided reproducible results. A total of 135 reliable and polymorphic RAPD bands were detected which were used to estimate genetic distances among pair-wise combinations of elite parents. Nei and Li's similarity values ranged from 0.60 to 0.77, with a mean of 0.66, which reflects a rather high genetic variability among the genotypes evaluated. Lower levels of genetic variability, as detected by polymorphic loci and mean heterogeneity values, were detected in a subset of parents selected for resistance to the stem nematode. Cluster analyses resolved the different sets of parents in a manner consistent with what is known from their breeding origins. An Analysis of Molecular Variance detected substantial levels of variation within subsets of parents. RAPDs represent a valuable source of genetic information for red clover breeding programmes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic base of Indian potato selections as revealed by pedigree analysis

Pedigree analysis of the 77 advanced Indian potato selections showed that their origin could be traced to only 49 ancestors out of which 29 were exotic, which accounted for 69.52% of the total genomic constitution and maximum contribution (40.65%) was by 10 ancestors from U.K. Breeders tendency was to involve adapted advanced clones as immediate parents. Coefficient of relationship between pair of selections ranged from 0 to 0.68. The success of the parents used was not related to region-specific choice of parents. The findings are discussed in the context of genetic uniformity and the relevance of having separate breeding programmes for the three major areas of potato cultivation in India. Attempt has also been made to give some directions for choosing selections for use as parents and release as cultivars based on their coefficient of relationship so as to broaden the genetic base of the future potato cultivars.     

Identification of RAPD markers linked to the Ns locus in potato

Using the RAPD method and bulked segregant analysis we identified four RAPD markers linked to a dominant gene Ns, responsible for a hypersensitive reaction of potato (Solanum tuberosum L.) to potato virus S (PVS) infection. The markers OPE15⁵⁵⁰, OPJ13⁵⁰⁰, OPG17⁴⁵⁰ and OPH19⁹⁰⁰ were found to be closely linked to the Ns gene in diploid potato clones. They are situated at 2.6, 3.3, 4.6 and 6.6 cM from Ns, respectively. As a source of the gene, clone G-LKS 678147/60, which is known to carry Ns transferred from S. tuberosum ssp. andigena was used. These RAPD markers were not amplified in resistant tetraploid clones containing Ns derived from the clone MPl65 118/3, also having an andigenum origin. This suggests that there may be two separate loci of Ns in the sources identified, or different alleles with the same specificity at a single locus, or that the genetic background of tetraploids tested results in different RAPD amphlification patterns.     

河北省和中棉所育成陆地棉品种的遗传多样性分析

分别选取河北省历年育成的 1 9个陆地棉品种 ,中国农业科学院棉花研究所自 2 0世纪 70年代末到 90年代中期选育的品种中的 1 6个陆地棉品种 ,利用 RAPD分子标记研究各自育成品种的遗传多样性。 41个带型清晰、重复性好的多态性引物用于 RAPD分析 ,河北省育成的 1 9个品种得到 6 6个多态性位点 ,而中棉所育成的 1 6个品种扩增到 6 4个多态性位点。分析采用 Jaccard' s相似系数 ,使用 NTSYS- pc1 .80数据分析软件 ,非加权组平均法 (UPGMA)聚类。成对相似系数比较表明 ,中棉所在 2 0世纪 70年代末到 90年代中期育成的品种的遗传多样性水平高于河北省育成的品种。河北省育成的 1 9个品种的遗传基础很狭窄 ,斯字棉和岱字棉是河北省棉花两个最重要的基础种质 ,其中的 1 5个品种在聚类图上可以被分为两类 ,分别属于岱字棉和斯字棉系统。中棉所育成的 1 6个品种在聚类图上可被划分为三个类群。     

Genetic relationships between cultivated and wild olives of Corsica and Sardinia using RAPD markers

In order to ensure the genetic diversity of the Olea europaea complex,it is necessary to characterize the cultivated varieties and the wildpopulations. In the present study, we focused on the olives growing on twoMediterranean islands, Corsica and Sardinia. On these two islands, there areolives with many denominations, as well as forests of oleasters. Here, it wasproposed to determine the relationships among cultivated and wild olives.Some Italian denominations were studied in addition to assess the influenceof the mainland on the two islands in this respect.The 59 RAPD markers obtained showed the existence of manysynonymous, and homonymous. A dendrogram was constructed using theUPGMA method and a FCA was carried out. The results of these twoanalyses showed the existence of a genetic divergence between the oleastersand the cultivated varieties. They suggest that some of the Corsicanvarieties were probably selected from local wild forms, contrary to theSardinian varieties. They also show that there are feral forms growing onboth islands, which result from hybridization between oleasters andvarieties.     

RAPD and SCAR markers for resistance to acochyta blight in lentil

Resistance to ascochyta blight of lentil (Lens culinaris Medikus),caused by the fungus Ascochyta lentis, is determined by a single recessive gene, ral ₂, in the lentil cultivar Indian head. Sixty F₂ individuals from a cross between Eston (susceptible) and Indian head (resistant) lentil were analyzed for the presence of random amplified polymorphic DNA (RAPD) markers linked to the ral ₂gene, using bulked segregant analysis (BSA). Out of 800 decanucleotide primers screened, two produced polymorphic markers that co-segregated with the resistance locus. These two RAPD markers, UBC227₁₂₉₀and OPD-10₈₇₀, flanked and were linked in repulsion phase to the gene ral ₂ at 12 cm and 16 cm, respectively. The RAPD fragments were converted to SCAR markers. The SCAR marker developed from UBC227₁₂₉₀ could not detect any polymorphism between the two parents or in the F₂. The SCAR marker developed from OPD-10₈₇₀ retained its polymorphism. The polymorphic RAPD marker UBC227₁₂₉₀ and the SCAR marker developed from OPD-10₈₇₀ can be used together in a marker assisted selection program for ascochyta blight resistance in lentil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of the genetic diversity among elite tea (Camellia sinensis var. sinensis) accessions using RAPD markers

The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan and Taiwan was examined with RAPD-PCR (Random Amplified Polymorphic DNA Polymerase Chain Reaction) markers. Out of the 50 primers screened, 17 primers generated 58 polymorphic and reproducible bands. A minimum of 3 primers was sufficient to distinguish all the 27 accessions studied. The Shannon's index used to partition diversity into inter- and intra-group, revealed that 71 percent of variability resided within groups and 29 percent between groups. Diversity was greatest within the Korean group followed by Taiwan and Japan. The relatively high diversity observed in Korea might reflect the larger genetic base of its plantations while the low diversity in Japan could be explained by the long and intensive tea selection programme in this country. A dendrogram based on the UPGMA-link method using Jaccard's distances and multivariate Factorial correspondence analysis clustered the tea accessions into two main groups, regrouping the Taiwan cultivars on the one side and the Korean and Japanese accessions on the other side. This suggests that the Taiwan tea studied here may have a different origin from that of Korea and Japan. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variation among cultivars of red clover (Trifolium pratense L.) detected by RAPD markers amplified from bulk genomic DNA

Summary The use of random amplified polymorphic DNA (RAPD) markers obtained from bulked samples was investigated for cultivar identification in red clover. Pooled samples were examined in order to minimize variation within cultivars. To determine the appropriate number of individuals to include in the bulked samples representing each cultivar, DNA samples from two, three, four, five, ten and twenty individuals were pooled. Twenty was found to be an appropriate number of red clover individuals per bulk in order to amplify only the DNA sequences shared among most individuals in each cultivar. Fourteen 10-mer primers were used to amplify genomic DNA from combined leaf samples of 15 red clover cultivars from European, Japanese and North American origins. A total of 79 amplified products, of which 55 were polymorphic, was obtained. Cultivar-specific bands were observed with 13 primers. The amplification patterns obtained from two primers could distinguish all 15 red clover cultivars. Rogers' genetic distances for all 105 pairwise comparisons were calculated to evaluate relationships among these cultivars. Cluster analysis based on these genetic distances separated these 15 cultivars into three groups, with two of the groups consisting of a single Japanese cultivar each, while the third group included cultivars from European, North American, and Japanese origins.     

Evaluation of random amplified polymorphic DNA (RAPD) markers in Hevea brasiliensis

The applicability of random amplified polymorphic DNA (RAPD) markers in the cultivated rubber tree, Hevea, was evaluated using 43 decamer oligonucleotide primers in a set of 24 clones selected in different South-East Asian countries. A total of 220 0.35–3.5 kb DNA fragments were amplified, of which 111 were polymorphic. Of these, 80 fragments (RAPD markers) which were repeatable and clearly scorable across all genotypes were used to estimate genetic distances among the clones tested. The estimated genetic distances ranged from 0.05 (RRII 308 and PB 5/51) to 0.75 (RRIC 100 and SCATC 88–13). A mean genetic distance of 0.5 indicates a rather high genetic variability among the tested clones. As expected, because of the breeding history of Hevea, UPGMA cluster analysis and Principal Coordinate Analysis (PCoA) indicated the absence of a distinct geographical grouping. The possible application of RAPD markers for clone identification and also for analysis of genetic relationships among Hevea clones is discussed.     

澳洲棉种遗传多样性的RAPD分析

利用ＲＡＰＤ（ＲａｎｄｏｍｌｙＡｍｐｌｉｆｉｅｄＰｏｌｙｍｏｒｐｈｉｃＤＮＡ）对斯特提棉（Ｇ．ｓｔｕｒｔｉａｎｕｍ）、南岱华棉（Ｇ．ｎａｎｄｅｗａｒｅｎｓｅ）、鲁滨逊氏棉（Ｇ．ｒｏｂｉｎｓｏｎｉ）、澳洲棉（Ｇ．ａｕｓｔｒａｌｅ）、比克氏棉（Ｇ．ｂｉｃｋｉ）和奈尔逊氏棉（Ｇ．ｎｅｌｓｏｎｉ）进行了研究,结果表明：６个澳洲棉种具有丰富的遗传多样性,在这６个澳洲棉种中,澳洲棉与鲁滨逊氏棉、南岱华棉与斯特提棉具有较近的亲缘关系。聚类分析发现,鲁宾逊氏棉和比克氏棉是两个较为特殊的棉种。此外,本文对比克氏棉和木槿组（Ｈｉｂｉｓ－ｃｏｉｄｅａ）的其它棉种的染色体组的归属问题进行了讨论     

Genetic diversity of soybean germplasm resistant to Heterodera glycines

Information on the genetic background of soybean [Glycine max (L.) Merr.] germplasm resistant to Soybean Cyst Nematode (SCN) (Heterodera glycines Ichinohe) is essential for developing plant breeding strategies, however, pedigree information for SCN resistant germplasm is not always available. Using restriction fragment length polymorphisms (RFLPs) to fingerprint different soybean lines is a useful means of determining genetic relationships and identifying unique genotypes. In this study, 102 soybean genomic probes with five different restriction enzymes were used to detect DNA variations and investigate genetic relationships among 56 soybean plant introductions (PIs) and cultivars. Among 510 probe/enzyme combinations, a total of 1707 hybridization bands were generated, of which 501 were polymorphic. A cluster dendrogram and a principal component analysis (PCA) plot diagram were constructed using DNA fingerprinting of the 56 soybean lines. Based on the clustering and PCA analyses, two major clusters comprising 15 groups were identified for the PIs and cultivars in the dendrogram. Reactions to different race isolates of SCN were distinguishable among different groupings of the clusters and PCA results, and various origins of the PIs and the pedigree information of the cultivars could be associated with the different clusters. Additional information on the appropriate number of probes used to detect genetic diversity more efficiently was elucidated through this research. We believe that the genetic relationships determined among these 56 soybean lines could provide useful information in identifying unique sources of genes for SCN resistance. This revised version was published online in July 2006 with corrections to the Cover Date.