Genetic relationships among species and cultivars of Pistacia using RAPDs and AFLPs

The genus Pistacia (Anacardiaceae) includes 11 species divided into four sections, according to leaf characters and nut morphology. Recently two monophyletic groups have been proposed by using cpDNA,Lentiscus and Terebinthus,containing evergreen and deciduous species, respectively. In the present work molecular markers, derived from two different methods, RAPD and AFLP, are used to study the relationships of native and introducedPistacia species present in Greece. According to the cophenetic correlation coefficients best results for both methods were obtained by using the Jaccard algorithm and the UPGMA clustering method. However, phenograms were constructed using the NJ method (r_cs= 0.987 for RAPDs and r_cs= 0.982 for AFLP), since it is less sensitive against varying mutation rates. Correlation among the genetic similarity (GS) matrices for the two methods was high(r = 0.941). The AFLP and RAPD phenograms were comparable with minor clustering differences. According to our results, two main branches are obtained, one containing the evergreen species P. lentiscusand the resin producing P. lentiscuscv. Chia (cultivated only in the island of Chios), and the other branch containing the deciduous species P. terebinthus,P. palaestina and P. vera, P. chinensis was clustered either with the evergreen species (RAPD data) or with the deciduous species (AFLP data). P. palaestina is clustered to P. terebinthus and can be considered as a subspecies of P. terebinthus, since its GS values are close or smaller than GS values of entries within species (P. vera). The four female cultivars were found to have a narrow genetic basis, probably related to cultivar ‘Nazareth‘ with Syrian origin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and phylogenetic relationships among accessions of hop, Humulus lupulus, as determined by amplified fragment length polymorphism fingerprinting compared with pedigree data

DNA fingerprinting using amplified fragment length polymorphisms (AFLPs) was successfully employed to detect genetic relationships and variability among 90 hop cultivars and breeding lines comprising a collection of the world's hop germplasm. Seven AFLP primer combinations produced a total of 347 fragments of which 151 (43.5%)) were polymorphic. One‐hundred and thirty informative, highly reproducible DNA polymorphisms were used to estimate the genetic similarity (GS) which varied between 1.0 (e.g. ‘Saazer’ vs. ‘Tettnanger’) and 1.17 (‘Columbus’ vs. ‘Tettnanger’, ‘Spalter’ and ‘Saazer’). UPGMA (unweighted pair‐group method with arithmetic averages) clustering revealed two main clusters, reflecting the two main sources of origin and the two main breeding objectives: one cluster of mainly European origin representing the aroma pool and a second cluster associating accessions with European germplasm infiltrated by wild American genes with less aroma quality, but a higher bittering potential. Each main branch was composed of four or three subclusters with subgroups, respectively. Assignment of almost all genotypes in the dendrogram was consistent with the pedigree data as far as they are known. Consequently, AFLPs are shown to be suitable for assessing the genetic variability in hop germplasm and are useful for describing the genetic relationships among cultivars and accessions, which allows phylogenetic questions to be addressed.     

Genetic diversity of late blight resistant and susceptible Indian potato cultivars revealed by RAPD markers

Twenty-four tetraploid Indian potato cultivars were characterized by using RAPD markers to assess diversity within and between late blight resistant and susceptible cultivars. Sixty-four random decamer primers generated802 fragments, ranging in size from 60–3200 bp, with 96.4% fragment polymorphism. Shannon's index of diversity was used to quantify the degree of variability present within and between the variety types. Most of the diversity was detected within variety types, with 88% of variation being within and 12% being between the resistant and susceptible cultivars. No clear groupings based on late blight resistance and susceptibility or kinship was reflected on the dendogram. The late blight resistant cultivars exhibited higher variability compared to susceptible cultivars and they were more dispersed on the PCO plot. This revised version was published online in August 2006 with corrections to the Cover Date.     

Comparing RAPD and AFLPTM analysis in discrimination and estimation of genetic similarities among apple (Malus domestica Borkh.) cultivars

Forty-one apple (Malus × domestica Borkh.) cultivars were screened for RAPD (Random Amplified Polymorphic DNA) and AFLP(Amplified Fragment Length Polymorphism) markers. RAPD analysis was performed with 35 arbitrary 10-mer primers, selected from 60 primers tested (kits A, C and E, Operon Technologies, Inc.). Of a total of 362bands observed, 208 (57.5%) were polymorphic. Three-hundred-and-eighty-one AFLP fragments were obtained with 8primer combinations, of which 218 (57.2%) were polymorphic. Cultivars differentiated through mutation were included in this study and showed identical patterns when analysed with both RAPD and AFLP analysis. The estimated genetic relationships were correlated (r = 73.7%) between the analysis with the two different markers. UPGMA analysis was performed and dendrograms were constructed using either the data apart from each(RAPD and AFLP) method or combined in a single joint matrix. The relationships among the forty-one studied cultivars were basically consistent with the known lineage and geographic origins of the cultivars. The four Portuguese cultivars included in this study clustered together and diverged from the other cultivars. Apparently they constitute an independent genetic pool, which could be of interest for apple plant breeders. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic relationships and diversity among Tibetan wheat, common wheat and European spelt wheat revealed by RAPD markers

An endemic hexaploid wheat found in Tibet, China was taxonomically classified as a subspecies in common wheat, i.e. Triticum aestivum ssp. tibetanum. Seven accessions of the Tibetan wheat, 22 cultivars of common wheat and 17 lines of spelt wheat were used for RAPD analysis to study the genetic relationships of the Tibetan wheat with common wheat and spelt wheat, and to assess the genetic diversity (GD) among and within the taxa. RAPD polymorphism was found to be much higher within spelt wheat and the Tibetan wheat than within common wheat. The GD value between the Tibetan wheat and common wheat is lower than that between the Tibetan wheat and spelt wheat. The result of cluster analysis showed that the 46 genotypes were distinctly classified into two groups. Group 1 included all European spelt wheat lines, while group 2 includes all Chinese common wheat and the Tibetan wheat accessions. However, the Tibetan wheat was substantially differentiated from Chinese common wheat at a lower hierarchy. Our results support an earlier classification of the Tibetan wheat as a subspecies in common wheat. European spelt wheat and the Tibetan wheat showed much higher genetic diversity than Chinese common wheat, which could be used to diversify the genetic basis for common wheat breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular diversity in Indian sugarcane cultivars as revealed by randomly amplified DNA polymorphisms

Genetic diversity in 28 prominent Indian sugarcane varieties cultivated under a wide range of agroclimatic conditions, was studied using 25 RAPD markers. The mean genetic distance among the 28 varieties was only 29.31%, implying that a large part of the genome is similar among the varieties. This probably arises from the lack of parental diversity, with few clones which are themselves related, contributing to the parentage of these varieties. The parentage of the varieties did not contribute significantly to the clustering pattern. Varieties belonging to the same parentage were grouped under different clusters while varieties from different parentages were grouped under the same cluster. The tropical and subtropical identities of the varieties also did not contribute to the clustering pattern as individual clusters included varieties from both tropics and subtropics. This shows that genetically similar varieties are present in both the regions. Among the varieties, Co 7717 was found to be totally distinct and divergent from rest of the varieties. The study reveals the limited genetic base of the current Indian commercial varieties and the need to diversify the genetic base by using new sources from the germplasm. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pedigree, RAPD and simplified AFLP-based assessment of genetic relationships among Avena sativa L. cultivars

Two molecular marker techniques: RAPD and simplified ^PstIAFLP have been compared in order to decide on, which technique is better suited to genetic characterization of oat (Avena sativa L.) cultivars. It was investigated, if the same pattern of variability is revealed by two approaches and whether the observed molecular variability reflects pedigree-based relationships. Polymorphic RAPD and ^PstIAFLP markers were sufficient to distinguish all analysed cultivars, demonstrating the usefulness of both methods for cultivar identification. Genetic similarity estimates derived from RAPD, simplified ^PstIAFLP and combined RAPD and ^PstIAFLP data were compared with coefficients of parentage (COP). Molecular markers-based mean genetic similarities were considerably greater than mean COP value. Correlation coefficients between COP and genetic similarities calculated from RAPD, ^PstIAFLP and combined molecular data were very low and not significant. A better correlation (0.50) was found between similarity estimates derived from RAPD and ^PstIAFLP markers. Four separate dendrograms were constructed based on pedigree and molecular analyses using a neighbor-joining algorithm (NJ). The dendrograms were compared and found to be topologically different. The results of this study showed, that both molecular techniques can be conveniently used for genetic characterization of oat cultivars, however ^PstIAFLP would be the method of choice due to the higher efficiency and reproducibility. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic relationships of macadamia cultivars and species revealed by AFLP markers

World production of macadamia nuts is based on two species, the smooth shell Macadamia integrifolia Maiden and Betche, and the rough shell Macadamiatetraphylla L.A.S. Johnson, and their hybrids. One hundred and five AFLP markers were used to analyze 26 macadamia accessions representing four species: M. integrifolia, M. tetraphylla,M. ternifolia, and M. hildebrandii as well as a wild relative,Hicksbeachia pinnatifolia (rose nut).Each macadamia accession showed distinct AFLP fingerprints indicating a significant level of genetic variation in this macadamia germplasm collection. The fourMacadamia species included in this study were clearly separated using cluster analysis with AFLP marker data. Based on a single accession, the separation of M. ternifolia from M. integrifoliasuggested the relatively distant genetic relationship between these two species and casts doubts on the notion that the M. ternifolia may be a variant of M. integrifolia. Within the major cluster ofM. integrifolia, nine established smooth shell cultivars were separated into two sub-clusters, suggesting the heterozygous nature of the original gene pool that had contributed to macadamia variety improvement programs. M. hildebrandii and H. pinnatifoliaformed a distinct cluster and share dramatically less genetic similarity with the other Macadamia species. Additional data would be needed to clarify the phylogenetic nature and status of M. hildebrandii in the genus Macadamia. This revised version was published online in July 2006 with corrections to the Cover Date.     

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 variation among Southern African cultivated peanut (Arachis hypogaea L.) genotypes as revealed by AFLP analysis

The amplified fragment length polymorphism (AFLP) technique, employing two different rare cutters, EcoRI and MluI in combination with the frequent cutter MseI, was used to assess genetic diversity and relationships among 21 closely related cultivated Southern African peanut genotypes. A dendrogram was constructed using Jaccard's coefficient and the UPGMA clustering method. Low levels of polymorphism (on average 2.78%) were detected. Results indicated that both EcoRI/MseI and MluI/MseIAFLP enzyme combinations efficiently detected polymorphism within closely related cultivated peanut, although the EcoRI/MseI enzyme combination detected more fragments per primer combination (on average 67.8) as opposed to29.7 by the MluI/MseI enzyme combination. All 21 genotypes could be uniquely distinguished from each other with a minimum of three MluI/MseI primer combinations. Genetic data correlated well with known species and pedigree data, dividing the 21 genotypes into two main groups corresponding to the two subspecies of Arachis hypogaea namely fastigiata and hypogaea. Divisions within the two main groups correlated with botanical types and pedigree data. This is the first report where MluI/MseI primer combinations were used on cultivated peanut and also the first successful detection of polymorphisms between closely related cultivated peanut genotypes worldwide. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity in cultivated Osteospermum as revealed by random amplified polymorphic DNA analysis

A collection of 28 Osteospermum clones and cultivated varieties of different origin were evaluated by random amplified polymorphic DNA (RAPD) analysis. All the clones were identified by 12 decamers selected from a set of 30. This is the first characterization by molecular markers of the genetic material of Osteospermum. The level of genetic diversity among genotypes was assayed and all the accessions tested were then classified into six groups by UPGMA cluster analysis; the clustering of genotypes using the RAPD data proved to be in accordance with their breeding group origin. RAPD analysis can therefore be a useful tool for evaluating genetic variability in other Osteospermum germplasm collections for breeding purposes and for protecting intellectual property rights of improved varieties.     

Genetic diversity and phylogenetic relationships among the annual Cicer species as revealed by isozyme polymorphism

Summary There are few estimates of genetic variability within and among populations of the nine annual Cicer species and for the wild species this information is based on few accessions. The present study was undertaken to examine genetic variation within and between annual Cicer species. One hundred and thirty-nine accessions of nine annual Cicer species were used for electrophoretic analysis at ICARDA. High levels of polymorphism in all eight wild annual Cicer species was found. This is in contrast to earlier research which had shown high polymorphism only in C. reticulatum. Cicer reticulatum had the highest proportion of polymorphic loci. However, for the cultigen, among 14 loci assayed, only two were polymorphic, ADH and PGD2. The nine species formed four phylogenetic groups based on the neighbor-joining method. The first group comprised C. arietinum, C. Reticulatum and C. echinospermum, the second C. bijugum, C. judaicum and C. pinnatifidum, the third C. chorassanicum and C. yamashitae; and the fourth group consisted of one species, C. cuneatum. The phylogenetic tree developed from the neighbor-joining technique illustrated that C. reticulatum is the probable progenitor of C. arietinum and that C. echinospermum split off from a common ancestor at an earlier stage in the evolutionary history of Cicer. Genetic diversity data showed that the greatest diversity was within C. reticulatum and the lowest with the cultigen, C. arientinum. With the exception of C. reticulatum, genetic diversity increased with genetic distance from the cultigen. Little geographic variation in genetic diversity was found.     

The phylogenetic relationship between different Cichorium intybus cultivars and cultivar groups, as revealed by RAPDs

The phylogenetic relationship between different Cichorium intybus cultivars and cultivar groups was studied. A total of 127 random amplified polymorphic DNA markers have been used to distinguish between 51 Cichorium intybus cultivars belonging to three different cultivar groups. A principal component analysis was performed on the complete data set and a clear grouping of chicory cultivar groups was observed. The three chicory groups tested (root chicory, witloof and pain de sucre) were separated from one another in the principal component plots. The first two axes accounted for 43.9% of the total variation. Although the chicory used in traditional forcing in soil and the red chicory cultivars were related more to each other and to the chicory cultivars from the witloof group, they still formed separate groups in the principal component plot.     

Variation in Capsicum annuum revealed by RAPD and AFLP markers

Genetic relationships were examined among thirty-four pepper (Capsicum annuum) cultivars of different types. Two types of PCR-based markers were used, RAPD and AFLP, and their relative effectiveness was compared. A dendrogram based on RAPD markers separated the large-fruited sweet cultivars from the small-fruited pungent peppers, and the former group showed less divergence than the latter. The percentage of polymorphic markers was lower for AFLP than for RAPD markers (13 and 22% respectively). However, AFLP primers amplified on average six times more products than RAPD markers. The average numbers of polymorphic products per primer were 1.6 and 6.5 for RAPD and AFLP primers, respectively, i.e., AFLP primers were four times more efficient than RAPD primers in their ability to detect polymorphism in pepper. While four blocky type cultivars were indistinguishable by RAPD, two AFLP primer pairs were sufficient to distinguish the four cultivars from each other. This revised version was published online in July 2006 with corrections to the Cover Date.     

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 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 diversity in cornsalad (Valerianella locusta) and related species as determined by AFLP markers

Fifteen amplified fragment length polymorphism (AFLP) EcoRI/MseI‐based primer combinations with five selective bases (Eco RI‐ANN, MseI‐CN) were used to estimate genetic diversity among 45 line varieties of cultivated cornsalad and 19 genebank accessions classified into nine different species related to cornsalad. Polymorphic fragments were scored for calculation of Jaccard's coefficient of genetic similarity (GS). The average GS estimate in elite germplasm (GS = 0.90) was substantially higher than in exotic germplasm (GS = 0.47). UPGMA‐cluster analysis revealed genetic relationships among recently bred varieties, old varieties and genebank accessions. Analysis of molecular variance indicated almost threefold variability within sets compared with between sets due to a high level of polymorphism among wild species. Sources for increasing genetic diversity in elite germplasm of cornsalad were suggested and a duplicate among the genebank accessions was detected. AFLPs could be considered a powerful tool for genetic diversity estimation in cornsalad germplasm and are recommended for systematic fingerprinting of remaining cornsalad species.     

Genetic diversity and taxonomic relationships within the genus Lens as revealed by allozyme polymorphism

Summary A survey of allozyme polymorphism at 11 loci was carried out on 439 accessions from the genus Lens. This comprised 153 Lens culinaris subsp. orientalis, 35 L. odemensis, 117 L. ervoides, 32 L. nigricans, 2 of a differentiated cytotype of L. nigricans and 100 landrace accessions of the cultivated lentil (L. culinaris subsp. culinaris), from 10 different countries. The aim of the survey was to determine intra-specific genetic diversity and species relationships, based on phylogenetic and phenetic analyses, particularly regarding the position of L. odemensis and the differentiated cytotype of L. nigricans. Diversity was described by three statistics. The level of diversity in the cultivated taxon was lower than in any of the wild species according to two of these statistics, the percentage of polymorphic loci and mean number of alleles per locus. For the third measure (Nei's mean genetic diversity) it was only greater than L. ervoides. Genetic diversity statistics of the wild species indicated differences in the nature of between-population genetic diversity within the different taxa. Phylogenetic analysis suggests that L. odemensis and L. ervoides evolved from a common ancestor, and L. culinaris subsp. orientalis subsequently evolved from L. odemensis. Phenetic analysis, however, places L. odemensis closer to L. culinaris subsp. orientalis than to L. ervoides. Nei's mean genetic distance of L. odemensis from both L. culinaris subsp. culinaris (0.204) and L. culinaris subsp. orientalis (0.110) was greater than the distance between them (0.062). This evidence is not conclusive in determining whether L. odemensis should retain its specific status. Further crossability studies should be carried out on a range of genotypes to assess the potential for gene flow. The evidence presented shows the differentiated cytotype of L. nigricans to be quite distinct from other L. nigricans accessions, both phenetically and phylogenetically. This indicates that the differentiated cytotype of L. nigricans may constitute a new taxon. Discriminant function analysis reveals that isozymes may be useful in validating species classification.     

Identification of commercial chicory cultivars for hydroponic forcing and their phenetic relationships revealed by random amplified polymorphic DNAs and amplified fragment length polymorphisms

One‐hundred and twenty‐four amplified fragment length polymorphism (AFLP) and 49 random amplified polymorphic DNA (RAPD) markers have been used to distinguish between 20 and 23 commercial chicory cultivars, respectively. These were all Cichorium intybus var. foliosum F₁ hybrids, currently used in hydroponic forcing. Five‐hundred and twenty RAPD primers (OPERON) were tested, of which 156 resulted in reproducible patterns and 26 yielded polymorphisms. Two‐hundred and fifty‐six AFLP primer‐combinations were tested and six combinations were selected for identification purposes. Similarity indices were measured and clustering has been done using pairwise comparison. Both types of marker provide similar conclusions. Two major clusters are formed, representing late and early cultivars. All cultivars were identified using 10 informative RAPD primers or three AFLP primer combinations. A low degree of polymorphism was detected between some early cultivars, suggesting a narrow genetic base in their breeding strategy.     

利用RAPD和SSR分析36个贵州主要玉米种质的比较研究

利用RAPD和SSR两种标记方法研究了36个玉米自交系的遗传多样性,并对这两种分子标记系统进行了比较.利用筛选出的22条RAPD引物,检测到了148条有多态性的带;利用筛选出的34对SSR引物,检测到158个等位基因.RAPD和SSR分子标记均有很高的多态性,RAPD多态性带比例为95.95%,SSR位点检测出的平均等位基因数位4.65.RAPD分子标记结果将36个玉米自交系划分为6大类,SSR分子标记将其划分为5大类.与系谱分析基本一致,两种分子标记划分的结果也相似.研究认为,RAPD、SSR两种分子标记系统均适合于玉米种质的遗传多样性研究,但SSR更可取.