Evaluation of the relevance of using RAPD-analysis for revealing the phylogenic connections between cultivars of drum wheat (T. durum Desf.)

A comparison of similarity indices between 64 durum wheat cultivars calculated using pedigree analysis and RAPD method showed a correspondence between these two approaches to estimation of genetic diversity. The associations between the results of RAPD clustering and coefficients of parentage (chi 2 test) and the coefficient of correlation between similarity matrices were statistically significant. However, the correlation was rather weak while pedigree analysis and RAPD method did not yield completely identical estimates of genetic diversity in the set of cultivars studied.     

A Comparison of AFLP and RAPD Markers for Genetic Diversity and Cultivar Identification in Cotton

AFLP and RAPDmarkers were employed in sixteen diploid cotton (Gossypium sp) cultivars for genetic diversity estimation and cultivar identification. Polymorphism information content (PIC) and percent polymorphism were found to be more for AFLP markers as compared to RAPD markers. Average Jaccard’s genetic similarity index was found to be almost similar using either AFLP or RAPD markers. All the cultivars could be distinguished from one another using AFLP markers and also by the combined RAPD profiles. Cultivar identification indicators like resolving power, marker index and probability of chance identity of two cultivars suggested the usefulness of AFLP markers over the RAPD markers. AFLP and RAPD analyses revealed limited genetic diversity in the studied cultivars. Cluster analysis of both RAPD and AFLP data produced two clusters, one containing cultivars of G. herbaceum and another containing cultivars of G. arboreum species. Highly positive correlation between cophenetic matrices using RAPD and AFLP markers was observed. AFLP markers were found to be more efficient for genetic diversity estimation, polymorphism detection and cultivar identification.     

利用分子标记和形态学性状检测的陆地棉栽培品种遗传多样性

利用RAPD,ISSR和SSR3种分子标记方法和2年田间实验对国内外36个陆地棉栽培品种的遗传多样性进行了研究,以3种分子标记在36个品种之间扩增的282条多态性位点所赋值的0,1数据,采用Nei和Li的方法,计算的品种成对相似系数从0．5745到0．9291,其品种平均数从0．6547到0．7524,又以2年品种表现的性状平均数经正态标准化后,采用欧氏距离计算了成对品种的遗传距离。分别以相似系数和传距离矩阵,采用类平均法进行聚类分析,其聚类结果把供试品种大致分为国外品种,新疆品种,早熟类型品种和我国的中熟棉品种等几个类群,类内进一步分组表明,分子标记确定的传关系基本上与品种系谱的种质系统一致,但并不能按系谱或种植生态区域简单地归属,尽管分子标记数据计算的相似系数矩阵和表现型计算的遗传距离矩阵存在极显著的相关关系（r=-0.335),但以遗传距离进行聚类分析的类内分组的组间特征不明显,分子标记是检测类内品种间遗传差异的有效方法,研究结果为棉花育种亲本选配提供了理论依据。     

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。     

Comparative analysis of seeded and vegetative biotype buffalograsses based on phylogenetic relationship using ISSRs,SSRs, RAPDs,and SRAPs

Buffalograss [Buchloe dactyloides (Nutt.) Englem.] is the only native grass that is being used extensively as a turfgrass in the Great Plains region. Its low-growth habit, drought resistance, and low-maintenance requirement make it attractive as a turfgrass species. Our objective was to obtain an overview on the genetic relatedness among and within seeded and vegetative biotype buffalograsses using inter-simple sequence repeats (ISSRs), random amplified polymorphic DNA (RAPDs), sequence-related amplified polymorphisms (SRAPs), and simple sequence repeats (SSRs) markers that were derived from related species (maize, pearl millet, sorghum, and sugarcane). Twenty individuals per cultivar were genotyped using 30 markers from each marker system. All buffalograss cultivars were uniquely fingerprinted by all four marker systems. Mean genetic similarities were estimated at 0.52, 0.51, 0.62, and 0.57 using SSRs, ISSRs, SRAPs, and RAPDs, respectively. Two main clusters separating the seeded-biotype from the vegetative-biotype cultivars were produced using UPGMA analysis. Further subgroupings were unequivocal. The Mantel test resulted in a very good fit (SRAP=0.92, ISSR=0.90) to good fit (RAPD=0.86, SSR=0.88) of cophenetic values. Comparing the four marker systems to each other, RAPD and SRAP similarity indices were highly correlated (r=0.73), while Spearmans rank correlation coefficient between RAPDs and SSRs was r=0.24 and between ISSRs and SSRs was r=0.66. A genotype-assignment analytical approach might be useful for cultivar identification and property rights protection. Polymorphic SRAPs were abundant and demonstrated genetic diversity among closely related cultivars.Communicated by B. FriebeA contribution of the University of Nebraska Agricultural Research Division, Lincoln, Nebraska 68583. Journal Series No. 14398.     

Analysis of genetic diversity and relationships in East African banana germplasm

The genetic diversity and phylogenetic relationships of 29 East African highland banana (Musa spp.) cultivars and two outgroup taxa, M. acuminata Calcutta 4 and Agbagba were surveyed by RAPD analysis. A genetic similarity matrix was established based on the presence or absence of polymorphic amplified fragments. Phylogenetic relationships were determined by UPGMA cluster analysis. RAPDs showed that the highland bananas are closely related with a narrow genetic base. Nevertheless, there were sufficient RAPD polymorphisms that were collectively useful in distinguishing the cultivars. The dendrogram was divisible into a major cluster composed of all the AAA highland banana cultivars and Agbagba (AAB) and a minor cluster consisting of Kisubi (AB), Kamaramasenge (AB) and Calcutta 4 (AA). Several subgroups are recognized within the major cluster. RAPD data did not separate beer and cooking banana cultivars. Our study showed that RAPD markers can readily dissect genetic differences between the closely related highland bananas and provide a basis for the selection of parents for improvement of this germplasm. Received: 28 June 2000 / Accepted: 1 August 2000     

Molecular genetic diversity of the French-American grapevine hybrids cultivated in North America.

French-American hybrid grapevines are most popular in eastern and mid-western North America: they are hardy cultivars derived from crosses between the European Vitis vinifera and American wild vines. The aim of this study was to characterize their genetic background using 6 microsatellite (SSR) markers and a set of 33 diagnostic RAPD markers. The latter were reproducible with different PCR thermal cyclers. Two SSR loci were found to be synonymous, VrZAG47 and VVMD27. The DNA profile frequencies estimated for each cultivar were much lower with multi-locus SSR data than that obtained from multi-fragment RAPD data. There was no significant correlation between the multi-locus DNA profile frequencies derived from SSRs and those from RAPDs. Estimates of genetic diversity derived from SSRs were generally higher and the average similarity between cultivars was generally lower than values reported for subgroups of V. vinifera, in accordance with expectations for hybrid cultivars. The phenetic relationships depicted by UPGMA (unweighted pair-group method with arithmetic averaging) and neighbor-joining analyses of microsatellite data were congruent and, to a large extent, in agreement with the known pedigree or history of each cultivar. A major dichotomy was observed between one group where the known genetic background was dominated by the North American Vitis riparia and Vitis labrusca, and another one where the genetic background was dominated by the European V. vinifera. Two Kulhmann varieties thought to be synonymous were found to be different, though closely related.     

Molecular Characterization of Brinjal (Solanum melongena L) Cultivars using RAPD and ISSR Markers

Molecular characterization of 19 advanced cultivars and landraces of brinjal was carried out using RAPD and ISSR markers. Twenty-nine RAPD primers generated a total of 240 amplified fragments, while 23 anchored and non-anchored ISSR primers produced 299 fragments. Of these, 66 (27.5%) RAPD and 56 (18.73%) ISSR fragments were polymorphic. All the cultivars could be distinguished based on RAPD and/or ISSR profiles. A set of two RAPD primers, OPW 11 and OPX 07, was adequate to distinguish all the 19 cultivars. On the other hand, a minimum of ten ISSR primers were required to achieve the same result. Eleven cultivars could be identified by the unique presence or absence of one to four markers. The correlation between primer Rp and the number of cultivars distinguished by RAPD was r = 0.873, while that for ISSR it was r = 0.327. The correlation between PIC of primer and the number of cultivars distinguished was r = 0.324 for RAPD, while for ISSR primers it was r = ? 0.066. The probability of chance identity between two cultivars for RAPD and ISSR markers was calculated as 8.94×10^?4 and 2.25×10^?2, respectively. The average Jaccard’s similarity coefficient between cultivars based on combined RAPD and ISSR data was estimated to be 0.919. The UPGMA analysis grouped the cultivars into three main clusters with significant bootstrap support. While the cultivars bred at Indian Agricultural Research Institute, New Delhi formed one sub-cluster; others did not show a prominent region-based clustering.     

Application of RAPD markers for characterization of γ-ray-induced rose mutants and assessment of genetic diversity

Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard’s coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.     

Analysis of genetic similarity detected by AFLP and coefficient of parentage among genotypes of sugar cane ( Saccharum spp.)

Despite the economical importance of sugar cane, until the present-date no studies have been carried out to determine the correlation of the molecular-based genetic similarity (GS) and the coefficient of parentage (f)-estimates generated for cultivars. A comprehensive knowledge of the amount of genetic diversity in parental cultivars, could improve the effectiveness of breeding programmes. In this study, amplified fragment length polymorphism (AFLP) and pedigree data were used to investigate the genetic relationship in a group of 79 cultivars (interspecific hybrids), used as parents in one of the Brazilian breeding programmes, and four species of Saccharum (Saccharum sinense, Saccharum barberi and two of Saccharum officinarum) . The objectives of this study were to assess the level of genetic similarity among the sugar-cane cultivars and to investigate the correlation between the AFLP-based GS and f, based on pedigree information. Twenty one primer combinations were used to obtain the AFLP molecular markers, generating a total of 2,331 bands, of which 1,121 were polymorphic, with a polymorphism rate, on average, of 50% per primer combination. GSs were determined using Jaccard’s similarity coefficient, and a final dendrogram was constructed using an unweighted pair-group method using arithmetic average (UPGMA). AFLP-based GS ranged from 0.28 to 0.89, with a mean of 0.47, whereas f ranged from 0 to 0.503, with a mean of 0.057. Cluster analysis using GS divided the genotypes into related subgroups suggesting that there is important genetic relationship among the cultivars. AFLP-based GS and f were significantly correlated (r = 0.42, P < 0.001), thus the significance of this r value suggests that the AFLP data may help to more-accurately quantify the degree of relationship among sugar-cane cultivars. Received: 27 November 2000 / Accepted: 27 April 2001     

长江、黄河流域两棉区陆地棉品种的遗传多样性比较研究

从300个随机引物中筛选到带型清晰且重复性强的41个多态性引物,用于长江、黄河流域两棉区的91陆地棉品种的RAPD标记分析。分析采用Jaccard‘s相似系数,使用NTSYS－pcl.80数据分析软件,非加权组平均法（UPGMA）聚类。来自长江流域棉区的23个陆地棉品种产生了84个多态性位点,品种之间的平均相似系数为0．631。而来自黄河流域棉区的68个陆地棉品种产生了96个多态性位点,品种之间的平均相似系数为0．632.两棉区品种的相似系数矩阵比较及成对相似系数分布的比较均表明,长江流域和黄河流域棉区陆地棉品种的遗传多样性水平相当。共同的基础种质资源、相同的育种目标及相近的育种方法和策略可能是造成两大棉区品种遗传多样性水平相当的重要因素。     

Genetic diversity among Turkish local grape accessions (Vitis vinifera L.) using RAPD markers

To estimate genetic relationships among 46 local grape cultivars, RAPD analysis was performed with 25 decamer primers selected from a total of 60 primers. Genetic relationships among these cultivars were determined by calculating similarity indexes, from which a dendogram was derived. There was high genetic variation among the cultivars, with values of genetic diversity ranging from 0.553 to 0.952 using the Jaccard coefficient. UPGMA analysis of a distance matrix produced a dendogram with six clusters. The relatively high genetic similarity ratios observed for the cultivars was also reflected in the dendogram. In general, no relationship was encountered between the genetic similarity ratios of the cultivars and the results of previous ampelographic analyses.     

Determination of relatedness and geographical movements ofPistacia vera (Pistachio; Anacardiaceae) germplasm by RAPD analysis

The pistachio tree (Pistacia vera) has long been cultivated in south-central Asia and throughout the Mediterranean region of southern Europe, north Africa and the Middle East. We examined genetic diversity and patterns of relatedness amongfifteen P. vera cultivars, representing germplasm originating from throughout this range, by using Random Amplified Polymorphic DNA (RAPD) markers. The resulting data were used to construct a similarity matrix and to perform a UPGMA cluster analysis. These analyses revealed two major clusters of P. vera germplasm: a Mediterranean cluster, which includes cultivars originatingfrom the Mediterranean region of Europe, north Africa and the Middle East; and an Iranian-Caspian cluster, comprising germplasm originating from locations east of the Zagros mountains plus ’Peters’, a cultivar selected as a seedling of unknown origin in the U.S.A. The data presented here, in combination with historical and geographical records, support the hypothesis that pistachio cultivation originated in or near the present natural range of the species and was spread by cultivation to the Mediterranean region of the Middle East. This more limited pool was the germplasm source for subsequent spread of pistachio by cultivation throughout the Mediterranean region.     

松花型花椰菜主要品种鉴定的分子标记分析

利用RAPD、ISSR和SRAP 3种分子标记对我国南方地区松花型花椰菜主栽品种进行鉴定,分析了品种间的遗传多样性。3种标记共产生370条扩增带,238条为多态性条带,其多态率为64.32%。其中只有SRAP标记的引物m e1/em1可将20个品种全部鉴别。遗传相似系数分析表明,松花型花椰菜品种之间的亲缘关系较近,遗传背景比较狭窄。聚类分析表明品种间的亲缘关系与熟性、地理分布相关。研究表明,分子标记能有效地应用于花椰菜品种鉴定,且综合多种分子标记分析品种间的遗传多样性将更加准确可靠。     

Detection of genetic diversity in closely related bread wheat using microsatellite markers

Wheat microsatellites (WMS) were used to estimate the extent of genetic diversity among 40 wheat cultivars and lines, including mainly European elite material. The 23 WMS used were located on 15 different chromosomes, and revealed a total of 142 alleles. The number of alleles ranged from 3 to 16, with an average of 6.2 alleles per WMS. The average dinucleotide repeat number ranged from 13 to 41. The correlation coefficient between the number of alleles and the average number of repeats was only slight (r _s = 0.55). Based on percentage difference a dendrogram is presented, calculated by the WMS-derived data. All but two of the wheat cultivars and lines could be distinguished. Some of the resulting groups are strongly related to the pedigrees of the appropriate cultivars. Values for co-ancestry (f) of 179 pairs of cultivars related by their pedigrees (f0.1) averaged 0.29. Genetic similarity (GS) based on WMS of the same pairs averaged 0.44. The rank correlation for these pairs was slight, with r _s = 0.55, but highly significant (P<0.001). The results suggest that a relatively small number of microsatellites can be used for the estimation of genetic diversity and cultivar identification in elite material of hexaploid bread wheat.     

Molecular Analysis of Rhynchosporium secalis Population Collected from Barley Cultivars having Different Resistance Specificities

Molecular analysis was performed to detect genetic diversity in 106 Rhynchosporium secalis isolates collected from different regions of Canada using random amplified polymorphic DNA (RAPD) markers. The isolates collected from barley cultivars having different resistance specificity to R. secalis and grown in geographically distinct regions, exhibited reproducible variation for 2–3 polymorphic PCR products per decamer primer. Analysis of 1960 RAPD markers data obtained with five primers formed 5 groups with different genetic similarity. High genetic variation was observed in R. secalis isolates obtained from resistant and susceptible cultivars of barley. Isolates collected from susceptible cultivars showed a tendency to group together, whereas isolates from resistant cultivars were divergent. R. secalis isolates infecting different barley cultivars released as resistant to the barley scald formed a specific group with UPGMA, even though all these isolates were collected from the same epidemiological region. Analysis of 15 isolates collected from one resistant cultivar Duke formed three clusters with low bootstrap values indicating high genetic diversity among the isolates present on a single host cultivar.     

Unravelling genetic diversity and cultivar parentage in the Danish apple gene bank collection

Characterization of apple germplasm is important for conservation management and breeding strategies. A set of 448 Malus domestica accessions, primarily of local Danish origin, were genotyped using 15 microsatellite markers. Ploidy levels were determined by flow cytometry. Special emphasis was given to pedigree reconstruction, cultivar fingerprinting and genetic clustering. A reference set of cultivars, mostly from other European countries, together with a private nursery collection and a small set of Malus sieversii, Malus sylvestris and small-fruited, ornamental Malus cultivars, was also included. The microsatellite markers amplified 17–30 alleles per loci with an average degree of heterozygosity at 0.78. We identified 104 (23%) duplicate genotypes including colour sports. We could infer first-degree relationships for many cultivars with previously unknown parentages. STRUCTURE analysis provided no evidence for a genetic structure but allowed us to present a putative genetic assembly that was consistent with both PCA analysis and parental affiliation. The Danish cultivar collection contains 10% duplicate genotypes including colour sports and 22% triploids. Many unique accessions and considerable genetic diversity make the collection a valuable resource within the European apple germplasm. The findings presented shed new light on the origin of Danish apple cultivars. The fingerprints can be used for cultivar identification and future management of apple genetic resources. In addition, future genome-wide association studies and breeding programmes may benefit from the findings concerning genetic clustering and diversity of cultivars.     

Genetic studies on tissue culture response and some agronomical traits in Egyptian bread wheat

The main effort of wheat breeder is the detection of genes and to merge them in a particular genotype using most suitable combination. Five Egyptian cultivars of bread wheat (Triticum aestivum L.) were crossed in a half diallel mating design to produce 10 crosses. The genetic potential of embryogenic callus (EC%), plant regeneration (RGP%) response and its association with heading date (HD) and grain yield per plant (GY/P) were investigated. The results showed that GY/P was significantly and positively correlated with EC% and RGP%. The combining ability analysis showed that the magnitudes of general combining ability (GCA) were higher than those of specific combining ability (SCA) for both tissue culture response and agronomic traits. The promising crosses which exhibited desirable SCA effects, showed also high useful heterosis for all studied traits. The magnitudes of additive genetic variance (σ²A) were larger than those of non-additive ones (σ²D) for all studied traits except for number of days to heading. The estimates of narrow sense heritability were 84.56%, 82.13%, 43.46% and 70.28% for the percentage of EC%, RGP%, HD and GY/P, respectively. The genetic similarity percents based on RAPD markers ranged from 76% to 93% between the cultivars. The UPGMA cluster analysis revealed that the cultivars could be divided into two main clusters. The range of Euclidean distances based on morphological characters among the cultivars was relatively wide (4.37–27.87), indicating relatively high amount of phenotypic variation. A significant positive correlation between Euclidean distance and RAPD distance (0.72⁷) was found.     

Identification of genetically diverse genotypes for photoperiod insensitivity in soybean using RAPD markers

Most of the Indian soybean varieties were found to be highly sensitive to photoperiod, which limits their cultivation in only localized area. Identification of genetically diverse source of photoperiod insensitive would help to broaden the genetic base for this trait. Present study was undertaken with RAPD markers for genetic diversity estimation in 44 accessions of soybean differing in response to photoperiod sensitivity. The selected twenty-five RAPD primers produced a total of 199 amplicons, which generated 89.9 % polymorphism. The number of amplification products ranged from 2 to 13 for different primers. The polymorphism information content ranged from 0.0 for monomorphic loci to 0.5 with an average of 0.289. Genetic diversity between pairs of genotypes was 37.7% with a range of 3.9 to 71.6%. UPGMA cluster analysis placed all the accessions of soybean into four major clusters. No discernable geographical patterns were observed in clustering however; the smaller groups corresponded well with pedigree. Mantel’s test (r = 0.915) indicates very good fit for clustering pattern. Two genotypes, MACS 330 and 111/2/1939 made a very divergent group from other accessions of soybean and highly photoperiod insensitive that may be potential source for broadening the genetic base of soybean for this trait.Key words: Genetic diversity, Photoperiod, RAPD, Soybean, UPGMA