Amplified Fragment Length Polymorphism Data Research Articles

English

Six Egyptian pomegranate (Punica granatum L.) cultivars were characterized by fruit characteristics (physical and chemical) and two molecular markers; Inter simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP). Genetic diversity of the pomegranate genotypes was evaluated. Physical fruit traits were determined (weight, volume and diameter), calyx [diameter, length (mm) and Carpels number], fruit firmness (Newton), peel as (weight and thickness), arils weight (g), volume of juice (ml), seeds [fresh and dry weight (g)], and color parameter of (fruit skin, internal peel, arils, juice and seeds). The chemical traits such as soluble solids contents (SSC), vitamin C content, anthocyanin content, pH, and titratable acidity (TA) were assessed and wide variations were observed in each of these traits among the studied cultivars. The genetic variability and relationships among six Egyptian pomegranate cultivars were tested using ISSR and AFLP analyses. The level of polymorphism across cultivars was 53 and 90.7% as revealed by ISSR and AFLP, respectively. ISSR and AFLP revealed different genetic similarities among the six pomegranate cultivars. Each analysis differs not only in its underlying principle, but also in their in-formativeness with regard to the type and amount of polymorphism detected. Genetic similarity matrices estimated from ISSR and AFLP data, showed similarity coefficients to range from 0.77 to 0.94 and 0.33 to 0.73, respectively. ISSR and AFLP characterized the six pomegranate cultivars by a large number of unique markers being 23 and 46 unique markers, respectively. The fruit weight ranged between 479.4 to 185 g of Ê»Nab El Gamalʼ and Ê»Assuityʼ, the firmness was 79.98 of Ê»Nab El Gamalʼ and 71.84 Newton of Ê»Manfaloutyʼ cv. The fruit peel thickness varied from 0.6 mm Ê»Arabyʼ, Ê»Hegazyʼ and Ê»Wardiʼ to 0.3 mm Ê»Assuityʼ. The arils weight ranged from 87.5 to 275 g of Ê»Assuityʼ and Ê»Nab El Gamalʼ cvs. The percentage net of arils weight/ fruit weight was the highest (59.34% of Ê»Manfaloutyʼ cv). The juice volume ranged from 62.41 to 71.81 ml/100 g arils for Ê» Wardiʼ and Ê»Nab El Gamalʼ cvs. The SSC content ranged between 16.01 Ê»Hegazyʼ and 12.55% Ê»Assuityʼ. V.C. content ranged from 3.21 to 14 mg. vitamin/100 ml juice of Ê»Nab El Gamalʼ and Ê»Assuityʼ. The anthocyanin content ranged from 1.47 to 10.03 for Ê»Arabyʼ and Ê»Hegazyʼ. The pH values varied from 3.3 (Wardi) to 2.9 (Araby). The Egyptain cultivars of pomegranate have a wide variation in the morphological and chemical characteristics for many uses of fresh fruit and of industry purpose.   Key words: Morphological and chemical fruits characterization, pomegranate, inter simple sequence repeat (ISSR), DNA markers, amplified fragment length polymorphism (AFLP).

A Permutation Test of Genetic Association between Salmonella Isolated On-farm and At-abattoir using Amplified Fragment Length Polymorphism

Pork and pork products have been identified as a significant source of Salmonella infection, which is a major public health concern. The contamination of Salmonella on pork can happen both on farms (before slaughter) and at abattoirs (after slaughter). Salmonella isolates were collected from both feces on farms and lymph nodes in the abattoir to determine if contamination at abattoirs can be linked back to the farms of origin. Molecular subtyping of the isolated Salmonella was performed using amplified fragment length polymorphism (AFLP), a Polymerase chain reaction-based, high-throughput, relatively inexpensive method. In this paper, we develop a permutation test for the genetic association of Salmonella isolated on-farm and at-abattoir using the AFLP data. Simulation studies show that the proposed permutation test controls the type I error rate appropriately as well as possesses high power. An application of the proposed permutation test to the real Salmonella ALFP data results in a p-value of 0.038 which shows strong evidence of association between Salmonella isolated on-farm and at-abattoir.

Use of fluorescent-based amplified fragment length polymorphism to resolve phylogenetic relationships of Colchicum species from Turkey

The study of phylogenetic relationships between 14 Colchicum taxa spread throughout Turkey was performed using a fluorescent-based amplified fragment length polymorphism (AFLP) technique. Five primer pair combinations were used in AFLP reactions. The data set was analyzed statistically using the NTSYS 2.1 software, and the neighbor-joining and maximum parsimony methods were implemented to generate phylogenetic trees. These analyses clustered the samples into 3 main clades. Both the neighbor-joining and maximum parsimony analyses resulted in similar topologies. Furthermore, supporting the phylogenetic trees, a similar grouping of 14 taxa was generated by principal component analysis. The AFLP analysis with 5 primer combinations was carried out to assess 14 taxa. Fragment sizes ranged from 54 to 462 bp in length for each primer combination. The average was 166 fragments per primer pair, primer B2 generated the highest number of bands (200), and primer B3 produced the lowest number of bands (112). A total of 834 polymorphic bands were scored. The cophenetic correlation coefficient between the data matrix and the cophenetic matrix for AFLP data was 0.72. Based on this molecular data, we concluded that the genetic diversity among these Turkish accessions is relatively high.

Genetic structure of various Magnaporthe oryzae populations in Iran and Uruguay

Rice blast, caused by Magnaporthe oryzae is one of the most destructive rice diseases worldwide. The genetic variability of 55 M. oryzae isolates from Iran and 32 isolates from Uruguay was analyzed using amplified fragment length polymorphism (AFLP). Cluster analysis using different grouping methods and similarity coefficients, based on the AFLP data from 679 monomorphic and polymorphic bands generated with eight primer combinations, was performed. The resulted grouping of the isolates revealed 4 separate AFLP lineages among a total of 87 isolate, designated as A1, A2, B, and C. Within each AFLP lineage, three or more clones were detected with a genetic similarity of 100 %. Overall genetic similarity was greater than 78 % between Iranian and Uruguayan populations. Evidence of gene flow between the Iranian and Uruguayan populations obtained from indica rice cultivars was observed as one Iranian isolate grouped lineage 1A together with Urugyayan isolates. Analysis of molecular variance (AMOVA) revealed that rice varietal type, sampling year and geographic region were the dominant factors determining genetic structure of M. oryzae populations; but rice cultivar had not significant effect.

Multilocus analyses indicate a mosaic distribution of hybrid populations in ground squirrels (genusIctidomys)

DNA sequence data from mitochondrial cytochrome-b (Cytb) and Y-linked structural maintenance of chromosomes (SmcY) genes were combined with 478 nuclear loci obtained from amplified fragment length polymorphisms (AFLP) to assess the extent of hybridization and genetic spatial structure of populations in two hybridizing species of ground squirrel (Ictidomys parvidens and Ictidomys tridecemlineatus). Based on AFLP analyses of 134 individuals from 28 populations, 10 populations were identified that possessed hybrid individuals. Overall estimates of FST values revealed strong support for population structure in the Cytb data set; however, analyses of the SmcY gene and the AFLP data indicated ongoing gene flow between species. Pairwise FST comparisons of populations were not significant for the SmcY gene; although they were significant for the Cytb gene, indicating that these populations were structured and that gene flow was minimal. Therefore, gene flow between I. parvidens and I. tridecemlineatus appeared to be restricted to populations that exhibited hybridization. In addition, the fragmented nature of the geographic landscape suggested limited gene flow between populations. As a result, the distributional pattern of interspersed parental and hybrid populations were compatible with a mosaic hybrid zone model. Because ground squirrels display female philopatry and male-biased dispersal, the ecology of these species is compatible with this hypothesis.

AFLP Polymorphisms Allow High Resolution Genetic Analysis of American Tegumentary Leishmaniasis Agents Circulating in Panama and Other Members of the Leishmania Genus

American Tegumentary Leishmaniasis is caused by parasites of the genus Leishmania, and causes significant health problems throughout the Americas. In Panama, Leishmania parasites are endemic, causing thousands of new cases every year, mostly of the cutaneous form. In the last years, the burden of the disease has increased, coincident with increasing disturbances in its natural sylvatic environments. The study of genetic variation in parasites is important for a better understanding of the biology, population genetics, and ultimately the evolution and epidemiology of these organisms. Very few attempts have been made to characterize genetic polymorphisms of parasites isolated from Panamanian patients of cutaneous leishmaniasis. Here we present data on the genetic variability of local isolates of Leishmania, as well as specimens from several other species, by means of Amplified Fragment Length Polymorphisms (AFLP), a technique seldom used to study genetic makeup of parasites. We demonstrate that this technique allows detection of very high levels of genetic variability in local isolates of Leishmania panamensis in a highly reproducible manner. The analysis of AFLP fingerprints generated by unique selective primer combinations in L. panamensis suggests a predominant clonal mode of reproduction. Using fluorescently labeled primers, many taxon-specific fragments were identified which may show potential as species diagnostic fragments. The AFLP permitted a high resolution genetic analysis of the Leishmania genus, clearly separating certain groups among L. panamensis specimens and highly related species such as L. panamensis and L. guyanensis. The phylogenetic networks reconstructed from our AFLP data are congruent with established taxonomy for the genus Leishmania, even when using single selective primer combinations. Results of this study demonstrate that AFLP polymorphisms can be informative for genetic characterization in Leishmania parasites, at both intra and inter-specific levels.

Genetic relationships of North American bears (Ursus) inferred from amplified fragment length polymorphisms and mitochondrial DNA sequences

The three species of bears in North America, polar bears (Ursus maritimus Phipps, 1774), brown bears (Ursus arctos L., 1758), and black bears (Ursus americanus Pallas, 1780), have differentiated morphologies and nuclear and mitochondrial genomes. An exception is a paraphyletic mitochondrial DNA relationship and some nuclear gene lineages common to polar bears and a population of brown bears from islands in southeast Alaska. In this study, we quantified the genetic relationships of extant brown bears and black bears from Alaska and Montana, and polar bears from Alaska, with amplified fragment length polymorphisms (AFLP) and mtDNA cytochrome-b sequences. Bayesian cluster analyses of the AFLP data show each species is distinct. All brown bears, including those from the islands in southeast Alaska, cluster separately from polar bears, and black bears cluster separately from brown bears and polar bears. The mtDNA of polar bears and southeast Alaska island brown bears is paraphyletic as reported previously, but the species have different haplotypes. These data indicate that extant populations of brown bears and polar bears have separate nuclear and mitochondrial gene pools and are supported as species under the genetic species concept.

Análisis morfológico y genético de <i>Triplaris guayaquilensis</i> Wedd (Polygonaceae): un árbol nativo de Ecuador

http://dx.doi.org/10.5902/198050989286En éste trabajo, seis poblaciones nativas (55 árboles) de Triplaris guayaquilensis Wedd (Fernán Sánchez), una de las principales species forestales de Ecuador, se sometieron al análisis morfológico y genético con AFLPs (polimorfismos en la longitud de los fragmentos amplificados). Las poblaciones se colectaron a través de dos macro-sitios (Litoral Central: Quevedo, Ventanas, la Guayas; Estribaciones de los Andes: la Maná, Patricia Pilar, Pichincha). Las poblaciones exhibieron las siguientes características: fuste recto (66 %); forma de la copa irregular y redonda (50 %); ángulo de inserción de las ramas de 0° a 30° (86 %). Cuatro características cualitativas (forma del fuste, tipo de terminación de hojas, ancho de hojas y pubescencia de las hojas) y cuatro cuantitativas (altura comercial del árbol, área basal, volumen comercial y volumen total) fueron las más explicativas de acuerdo con el análisis de componentes principales (ACP). El ACP separó las poblaciones en dos grupos: uno incluyó poblaciones del Litoral Central, con características morfológicas alta y positivamente correlacionadas con la producción de madera y, el otro, con poblaciones con crecimiento de árboles menor provenientes de las estribaciones de los Andes. Las poblaciones del Litoral Central mostraron los mayores valores de diversidad genetic y los marcadores AFLP separaron dichas poblaciones con base en el macrositio de origen. Para un valor K = 2 el análisis Bayesiano separó las poblaciones de FS en dos grupos: dos poblaciones de la región Litoral Central y las otras cuatro de la región de las estribaciones de los Andes (3) y una del Litoral Central (La Guayas). Para valores K mayores fue evidente la fragmentación genética de las poblaciones de acuerdo con el origen pues para K = 5 se formaron cuatro grupos: uno incluyó poblaciones de Quevedo y Ventanas y otro de La Guayas (Litoral), un tercer grupo incluyó árboles de La Mana y Pichincha y un cuarto grupo, de Patricia Pilar (estribaciones de los Andes). Los resultados sugieren la constante y efectiva recombinación genética o flujo genetic entre y dentro de poblaciones de Fernán Sánchez con una tendencia clara hacia la diferenciación genética.

Genetic, cytological and morphological differentiation within the Balkan‐CarpathianSesleria rigidasensu Fl. Eur. (Poaceae): A taxonomically intricate tetraploid‐octoploid complex

AbstractReconstruction of relationships among populations of the morphologically polymorphic and taxonomically intricateSesleria rigidasensu Fl. Eur. based on Amplified Fragment Length Polymorphisms (AFLPs) revealed four clearly differentiated genetic groups that did only partly follow recent taxonomic concepts, but were strictly allopatric. While some of the previously described taxa constitute distinct genetic entities, others have no taxonomic value. Synthesizing our AFLP data with ploidy‐level information obtained from all genetically investigated individuals as well as with chromosome counts revealed that tetraploid individuals prevail, while octoploids occur only withinS. filifolia. Lack of AFLP divergence between tetra‐ and octoploids suggests an autopolyploid origin of the latter. The genetic differentiation pattern was reflected by morphological differentiation, allowing for a taxonomic revision of the constituents ofS. rigidasensu Fl. Eur. resulting in recognition of the four speciesS. achtarovii,S. filifolia,S. rigida, andS. serbica.

First Report of Sequence Type 1, Pathotype A Xanthomonas citri pv. citri from Lime and Lemon Fruit Originating from Bangladesh.

Asiatic canker caused by Xanthomonas citri pv. citri, a quarantine pest in several countries (including the European Union), strongly impacts both national citrus markets in tropical and subtropical areas and international trade. This bacterium induces erumpent, callus-like lesions often with a water-soaked margin in a wide range of citrus species causing premature fruit drop and twig dieback. Long distance dispersal is mainly through infected propagative material and the role of fruit is still debated. During inspection of imported limes (C. aurantifolia) and lemons (C. limon) from Bangladesh from 2006 to 2009, canker-like infected fruits were intercepted by the UK plant health service. Typical corky lesions were surface sterilized and comminuted in 0.1% peptone solution. Suspensions were plated onto nutrient dextrose (ND) and yeast dextrose chalk (YDC) plates for bacterial isolation. After incubation for 3 to 7 days at 25°C, typical Xanthomonas-like yellow colonies were purified for identification. Identification of 18 isolates as Xanthomonas was carried out initially by fatty acid methyl ester (FAME) analysis. Identification at the species level (X. citri) was completed by sequencing of the gyrase B gene (4). PCR (3) was used to confirm the identity of these isolates using X. citri pv. citri CFBP 2525 as the positive control and distilled water as the negative control. The expected DNA fragment was only obtained from all of the bacterial isolates using primer pair 4/7 (3). Multilocus sequence analysis (MLSA) of four housekeeping genes (atpD, dnaK, efp, and gyrB) identified isolates from Bangladesh as two sequence types of X. citri pv. citri, ST1 (n = 5; GenBank Accession Nos. FJ376118, FJ376168, FJ376216, and FJ376251) and ST2 (n = 13; EU333904, EU333907, EU333910, and FJ376357), but not as any other xanthomonad pathogenic to citrus (2). Amplified fragment length polymorphism (AFLP) analysis of all X. citri pv. citri isolates from Bangladesh and additional reference isolates from pathotype A, A*, Aw and X. citri pv. aurantifolii (2) using Sac I/Msp I and four primer pairs (unlabelled MspI + 1 (A, C, T, or G) primers and 5'-labeled - SacI + C primer for the selective amplification step) confirmed identification as X. citri pv. citri. All five ST1 isolates grouped as a single cluster by AFLP, although not strongly supported by bootstrap analysis. Evolutionary genome divergences (EGD) computed from AFLP data ranged from 0.0000 to 0.0097 (median EGD 0.0055) suggested a relatively wide diversity within isolates originating from Bangladesh (median EGD from a worldwide pathotype A collection [n = 73] 0.0028) (2). When inoculated to Mexican lime SRA 140 and grapefruit cv. Duncan using a detached leaf assay (2), all the Bangladesh isolates produced typical extensive canker lesions on both species whereas the negative control (10 mM Tris buffer pH 7.2) did not, and Koch's postulates were fulfilled. To our knowledge, this is the first report of pathotype A assigned to ST1 by MLSA. All strains previously assigned to ST1 displayed a narrow host range (pathotype A*) (2). Our results further identify the Indian subcontinent as an area of relatively wide genetic diversity of X. citri pv. citri (1).

Can gene flow among populations counteract the habitat loss of extremely fragile biotopes? An example from the population genetic structure in Salix daphnoides

Wild river gravel banks (RGB) represent an extremely fragile biotope that is significantly endangered by human activities due to its fragmentation over the past century. The consequences of such processes were studied using the endangered violet willow (Salix daphnoides) at the westernmost foothills of the Carpathian Mountains. We quantified population genetic characteristics for 14 ecologically and demographically characterised populations using simple sequence repeats (SSR) and amplified fragment length polymorphism (AFLP) markers. We found a significant correlation between the biotope, sex, age and genetic structure of the populations. The natural RGB populations revealed high genotypic variability when using SSR markers, in contrast to low genotypic variability of the populations of other biotopes of Cirsium wet meadows (CWM) and Ash-alder forests (AAF) that consisted of one to two clones at each site. High heterozygosity (Hobs = 0.428–0.532) was similar across all natural (RGB) populations; however, these populations were deficient in heterozygotes (FIS/ρIS > 0). All RGB populations exhibited moderate to very significant genetic differentiation for microsatellites, despite the fact that the AFLP data showed strong differentiation only between CWM and AAF populations. Division into clusters by Structure confirmed consistent geographic groups for the RGB populations. According to our results, the strong decrease in previously continuous and large natural habitats for the violet willow is slightly counterbalanced by among-population gene flow. However, the survival of natural populations is tightly linked to the presence of river gravel banks and bars. Strict protection of this habitat is therefore essential for the conservation of the species.

Hierarchical Population Genetic Structure in a Direct Developing Antarctic Marine Invertebrate

Understanding the relationship between life-history variation and population structure in marine invertebrates is not straightforward. This is particularly true of polar species due to the difficulty of obtaining samples and a paucity of genomic resources from which to develop nuclear genetic markers. Such knowledge, however, is essential for understanding how different taxa may respond to climate change in the most rapidly warming regions of the planet. We therefore used over two hundred polymorphic Amplified Fragment Length Polymorphisms (AFLPs) to explore population connectivity at three hierachical spatial scales in the direct developing Antarctic topshell Margarella antarctica. To previously published data from five populations spanning a 1500 km transect along the length of the Western Antarctic Peninsula, we added new AFLP data for four populations separated by up to 6 km within Ryder Bay, Adelaide Island. Overall, we found a nonlinear isolation-by-distance pattern, suggestive of weaker population structure within Ryder Bay than is present over larger spatial scales. Nevertheless, significantly positive F st values were obtained in all but two of ten pairwise population comparisons within the bay following Bonferroni correction for multiple tests. This is in contrast to a previous study of the broadcast spawner Nacella concinna that found no significant genetic differences among several of the same sites. By implication, the topshell's direct-developing lifestyle may constrain its ability to disperse even over relatively small geographic scales.

Combining population genetics, species distribution modelling and field assessments to understand a species vulnerability to climate change

AbstractClimate change is recognized as a major threat to biodiversity. Multidisciplinary approaches that combine population genetics and species distribution modelling to assess these threats and recommend conservation actions are critical but rare. Combined, these methods provide independent verification and a more compelling case for developing conservation actions. This study integrates these data streams together with field assessments and spatial analyses to develop future genetic resource management recommendations. The study species was Callistemon teretifolius (Needle Bottlebrush), a shrub species endemic to the Mount Lofty and Flinders Ranges, South Australia, and potentially vulnerable to climate change. Chloroplast microsatellite and Amplified Fragment Length Polymorphism data were combined with species distribution modelling (MaxEnt), spatial analysis and field assessment to evaluate climate change vulnerability. Two major genetic groups were identified (Mount Lofty and Flinders Ranges). Populations in the Flinders Ranges, especially the Southern Flinders Ranges exhibited the highest genetic diversity, indicating a possible genetic refugium. Lower genetic diversity to the south in the Mount Lofty Ranges and north in the Gammon Ranges may be due to post‐glacial expansion into these areas from the Flinders Ranges or loss of alleles. Low levels of contemporary gene flow were identified, which suggests Callistemon teretifolius may have a limited capacity to respond to climate change through migration. Range restrictions were predicted for all future climates, especially in the north. It is likely that C. teretifolius will be adversely affected by climate change, due to limited gene flow, predicted range restriction and loss of suitable habitat. The Southern Flinders Ranges should be a priority for conservation because it contains the highest number of individuals and genetic diversity. We recommend monitoring and adaptive management involving restoration in the Southern Flinders Ranges, potentially incorporating genetic translocations from other areas to capture diversity, to assist C. teretifolius to adapt to climate change.

Molecular evidence for a natural diploid hybrid between Miscanthus sinensis (Poaceae) and M. sacchariflorus

The hybrid origin of Miscanthus purpurascens has previously been proposed, primarily because of its intermediate morphology. In this study, phylogenies based on the DNA sequences from the internal transcribed spacer region of nuclear ribosomal DNA (nrDNA ITS), on the DNA sequences of the trnL intron and trnL-F intergenic spacer of chloroplast DNA, and on amplified fragment length polymorphism (AFLP) fingerprinting confirm that M. purpurascens originated through homoploid hybridization between M. sinensis and M. sacchariflorus. Two different types of ITS sequences were identified from almost all plants of M. purpurascens. One type was found to be closely related to M. sinensis and the other to M. sacchariflorus. Miscanthus purpurascens was found to possess many M. sinensis- and M. sacchariflorus-specific AFLP bands but no band specific to itself. Clustering with the Unweighted Pair Group Method with Arithmetic Mean and principal coordinate analysis based on the AFLP data also demonstrated that M. purpurascens is an approximate intermediate of the two species. In addition, M. purpurascens has the plastid genome of M. sinensis or M. sacchariflorus, suggesting that either species could be its maternal parent. All specimens of M. purpurascens and its coexisting parental species are identified as diploids (2n = 2x = 38). Possible mechanisms of natural hybridization, hybrid status, chloroplast DNA recombination, and evolutionary implications of this hybridization are also discussed.

Patterns of genetic diversity in natural populations of Paspalum agamic complexes

Paspalum has many multiploid species displaying a wide range of ploidy levels and reproductive systems including apomixis. However, not much is known about the genetic structure of natural populations of the apomictic species of Paspalum. The aim of this work was to evaluate the genetic diversity of several natural populations belonging to five species of Paspalum. A total of 13 populations were analyzed using amplified fragment length polymorphism (AFLP). The AFLP data revealed maximal genotypic diversity and significant levels of genetic diversity in diploid and mixed diploid–tetraploid populations of P. denticulatum and P. rufum, where all individuals represent different genotypes. This may be mainly due to the reproductive system of diploid members and the gene flow from diploids to polyploids. The pure populations of tetraploids consist of either multiple genotypes (P. nicorae) or of one dominant genotype with a few deviated genotypes (P. denticulatum and P. lividum). Here, the main source of variability may be the residual sexuality, which continues generating new genotypic combinations. The hexaploid populations of P. buckleyanum consist of a single AFLP genotype and each population represents a particular genotype suggesting that populations arose from independent polyploidization events. This study represents one of the first reports of genetic diversity in natural populations of several Paspalum agamic complexes. Apomixis in these five species may be acting as a successful method for the dispersion of better adapted genotypes.

Genetic divergence among Brazilian garlic cultivars based on morphological characters and AFLP markers

Outside its centers of origin, garlic propagates only asexually. Since asexual reproduction leads to the absence of meiotic recombination, the main garlic cultivars available for cultivation have arisen from the accumulation of somatic mutations in early cultivars. Thus, it is common for a single clone to have different names in different regions. This study aimed to evaluate the genetic diversity of 20 garlic cultivars by using morphological characters and amplified fragment length polymorphism (AFLP) markers to identify possible duplicate cultivars. We assessed 28 morphological characters related to the leaves, bulbs, and bulbils of the garlic plant and divided them into two categories: quantitative and qualitative (14 characters each). For molecular marker-based analysis, we used three AFLP primer combinations. Genetic divergence was calculated using the Jaccard coefficient; the cultivars were grouped using unweighted pair-group mean analysis. The average genetic divergence detected using the morphological characters was 2.30 (range, 0.45-4.70). Plant height and coat adhesion exhibited the highest divergence among the cultivars. The average genetic diversity based on AFLP data was 43% (range, 0-79%). Dendrograms derived from both techniques divided the cultivars into two groups: noble and semi-noble. Together with the divergence within groups, the correlation between morphological and molecular data suggested that the cultivars in the noble group had greater phenotypic stability than those in the semi-noble group. Analysis of Jonas and Quitéria cultivars using these two techniques revealed only slight differences, suggesting that these cultivars may be clones or have a high degree of kinship.

Molecular Genetic Diversity in the Turkish National Melon Collection and Selection of a Preliminary Core Set

Turkey is a secondary center of diversity for melon (Cucumis melo) and is home to a variety of regional morphotypes. This diversity is housed in a national germplasm repository with more than 500 accessions. Molecular genetic variability of 209 melon genotypes from 115 accessions of this collection was characterized using amplified fragment length polymorphisms (AFLPs). Ten AFLP primer combinations yielded 279 reproducible fragments, which were used for dendrogram and principal coordinate analyses. These analyses showed two major clusters of Turkish melons: one group contained highly similar genotypes (maximum Dice dissimilarity coefficient of 0.18), whereas the other group was genetically more diverse (maximum dissimilarity 0.41). Although average dissimilarity was low (0.13), a broad range of genetic diversity was observed in the collection. A marker allele richness strategy was used to select a core set of 20 genotypes representing the allelic diversity of the AFLP data. The core set had double the average diversity (0.26) of the entire set and represented the major morphotypes present in the collection. Molecular genetic diversity of the core set was further validated using simple sequence repeat marker data (116 polymorphic fragments), which confirmed that the selected core set retained high levels of molecular genetic diversity.

Evaluation of the genetic relationship between Fritillaria species from Turkey’s flora using fluorescent-based AFLP

In this study, the genetic relationships among 12 Fritillaria taxa collected from different regions of Turkey were investigated using the amplified fragment length polymorphism (AFLP) technique. Seven primer pair combinations were used for the analysis. The neighbor-joining and principal coordinate analysis methods were implemented to identify the taxonomy of the species. The analyses generated congruent results with each other and with the previous studies. The neighbor-joining analysis clustered the species into 3 groups, while the principal coordinates analysis did not support separation of the third group. The subgenera Fritillaria and Petilium diverged from each other with strong support in all analyses. The neighbor-joining tree further divided the members of subgenus Fritillaria into 2 subgroups. Statistical analysis of the data set using NTSYS 2.1 showed a very good fit between the data matrix and the cophenetic matrix for the AFLP data with an r-value of 0.91. Generating a high polymorphic band profile, this analysis presented the feasibility of using AFLP for a genetic relationship analysis of Fritillaria. This primary report on AFLP analysis of Fritillaria species could facilitate achieving a deeper understanding of genetic relationships between the species.

Genetic diversity and species delimitation in the cultivated and wild Guinea yams (Dioscorea spp.) from Southwest Ethiopia as determined by AFLP (amplified fragment length polymorphism) markers

Yams (Dioscorea spp.) rank as the fourth most important root and tuber crop after potatoes, cassava and sweet potatoes. They are an economic crop in most of the tropics especially in West Africa, which produces over 95 % of the world output. Despite their cultural and economic importance there is taxonomic confusion regarding Guinea yams. The currently used classification scheme, which relies on vegetative and inflorescence characters, does not consistently delimit species boundaries between members of Guinea yams (D. cayenensis Lam.–D. rotundata Poir. complex), their wild relatives (D. abyssinica Kunth and D. praehensilis Benth.,) and D. sagittifolia Pax. Establishing the taxonomic identity of the germplasm and understanding the systematic relationships among crops is vital to the management of genetic resources and the utilization of accessions. In this study, amplified fragment length polymorphism (AFLP) genetic fingerprinting was used to evaluate and characterize 43 individual plants, belonging to different populations of wild and cultivated Guinea yams. The three primer combinations used in the AFLP analyses generated 158 scorable bands, with an overall polymorphism of 78 %. Ordination and cluster analyses of AFLP data failed to produce any clear species boundary between either the Guinea yam accessions or between them and their wild relatives. The average genetic similarity between the study individuals of Guinea yams and their wild relatives ranged from 60 to 100 %. The first, second and third principal coordinates axes cumulatively account for 77.45 % of the total variation. AFLP analyses also revealed a higher genetic divergence among cultivated Guinea yam accessions of the Sheko cultivars. Ordination and cluster analysis using UPGMA revealed no clear species boundaries between members of the complex. Thus, the taxonomy of these “species” needs to be revisited using other markers.

Automated Masking of AFLP Markers Improves Reliability of Phylogenetic Analyses

The amplified fragment length polymorphisms (AFLP) method has become an attractive tool in phylogenetics due to the ease with which large numbers of characters can be generated. In contrast to sequence-based phylogenetic approaches, AFLP data consist of anonymous multilocus markers. However, potential artificial amplifications or amplification failures of fragments contained in the AFLP data set will reduce AFLP reliability especially in phylogenetic inferences. In the present study, we introduce a new automated scoring approach, called “AMARE” (AFLP MAtrix REduction). The approach is based on replicates and makes marker selection dependent on marker reproducibility to control for scoring errors. To demonstrate the effectiveness of our approach we record error rate estimations, resolution scores, PCoA and stemminess calculations. As in general the true tree (i.e. the species phylogeny) is not known, we tested AMARE with empirical, already published AFLP data sets, and compared tree topologies of different AMARE generated character matrices to existing phylogenetic trees and/or other independent sources such as morphological and geographical data. It turns out that the selection of masked character matrices with highest resolution scores gave similar or even better phylogenetic results than the original AFLP data sets.