Deep phylogeographic splits but no taxonomic structure in the disjointly distributed Draba pacheri (Brassicaceae), a subendemic of the Eastern Alps

Draba (Brassicaeae), a model group for diversification and evolution in Arctic and mountain habitats, is taxonomically challenging and many of its species are insufficiently investigated. One such species is D. pacheri, an endemic of the eastern European Alps and the western Carpathians (here presumably extinct). Several hypotheses exist with respect to the phylogenetic position and the taxonomy of this species, but none of these has ever been tested using molecular data. In this article we examine (i) DNA sequence data to assess the phylogenetic position of D. pacheri within the genus and (ii) AFLP fingerprint data as well as morphometric data to address whether this species can be divided taxonomically into species or subspecies. DNA sequence data firmly place D. pacheri within the Core Draba Group III, whose internal relationships are, however, insufficiently resolved to precisely identify the closest relative of D. pacheri. AFLP data identify several genetically divergent lineages corresponding to geographically distinct regions. Although these lineages are congruent with hypotheses distinguishing either two species (D. pacheri s. str., D. norica) or one species with several subspecies, the lack of clear morphological separation, both with respect to the entire set of traits and single presumably diagnostic characters such as trichome morphology, renders recognition of a single species D. pacheri, as suggested previously, the best taxonomic solution. The deep and geographically strongly structured splits of D. pacheri likely are the result of isolation in several Pleistocene refugia and warrant that conservation efforts should involve populations from each of the main geographic subgroups.

AFLP MARKER IN GENETIC DIVERSITYASSESSMENT OF FIG (Ficus carica L.) POPULATIONS IN KURDISTAN REGION – IRAQ.

In this study, the genetic relatedness of 12 cultivars of fig from different populations in Kurdistan region- Iraq were analyzed using eleven AFLP primers pairs combinations by using the technology of molecular analysis the DNA. Genetic similarity matrices were produced for the AFLP data to calculate genetic distances among their cultivars. Genetic similarity coefficient ranged from 0.1261 to 0.3905. The lowest genetic similarity was observed between Kola and Gala Zard (0.1261). The Hejeera Rash and Shela cultivars were most similar ones with a coefficient of 0.3905. Clustering based on AFLP data for the 12 fig cultivars was identified at the 0.32 similarity level. In the developed dendogram two main groups were found, the first one combined Ketek and Shela together, while the second group contained two sub group Shingaly and Benatty combined together, while in the other sub group cluster three other sub-group were identified. The results of this study may help in the formulation of appropriate strategies for conservation and cultivar improvement in figs, for which limited knowledge of the genetic diversity is available.

Hybridization in the Temperate Bamboos (Poaceae: Bambusoideae: Arundinarieae): A Phylogenetic Study Using AFLPs and cpDNA Sequence Data

Abstract—The temperate bamboos are a taxonomically difficult group with nearly 600 species in approximately 30 genera and at least 12 constituent lineages. In this study, phylogenetic relationships were explored using amplified fragment length polymorphism (AFLP) data in comparison with a phylogeny based on plastid DNA sequences, with an emphasis onArundinariaof North America and its allies in East Asia (theArundinariaclade). Molecular analyses involved 248 individuals in 10 genera and 60 species. Hybridization was detected both within and among genera. Comparative analyses indicated hybrid origins for species in several widespread and well-known genera, includingHibanobambusa,Sasaella, andSemiarundinaria. Evidence also indicated thatPseudosasa japonica(the type species ofPseudosasa) is an intergeneric hybrid involvingPleioblastusandSasamorpha. In addition, cryptic hybrids were detected within and amongPleioblastus,Sasa, andSasamorpha. After accounting for hybrids, phylogenetic analyses of AFLP data provided resolution for core lineages in theArundinariaclade, includingPleioblastussensu stricto,Sasas. s., andSasamorpha.AFLP data also provided evidence for the monophyly of the North American cane bamboos (Arundinaria, three species) but failed to identify their closest relative among the East Asian taxa. The broader evolutionary implications of hybridization in the temperate bamboos are discussed along with recommendations for future studies.

Performance comparison of two reduced-representation based genome-wide marker-discovery strategies in a multi-taxon phylogeographic framework

Multi-locus genetic data are pivotal in phylogenetics. Today, high-throughput sequencing (HTS) allows scientists to generate an unprecedented amount of such data from any organism. However, HTS is resource intense and may not be accessible to wide parts of the scientific community. In phylogeography, the use of HTS has concentrated on a few taxonomic groups, and the amount of data used to resolve a phylogeographic pattern often seems arbitrary. We explore the performance of two genetic marker sampling strategies and the effect of marker quantity in a comparative phylogeographic framework focusing on six species (arthropods and plants). The same analyses were applied to data inferred from amplified fragment length polymorphism fingerprinting (AFLP), a cheap, non-HTS based technique that is able to straightforwardly produce several hundred markers, and from restriction site associated DNA sequencing (RADseq), a more expensive, HTS-based technique that produces thousands of single nucleotide polymorphisms. We show that in four of six study species, AFLP leads to results comparable with those of RADseq. While we do not aim to contest the advantages of HTS techniques, we also show that AFLP is a robust technique to delimit evolutionary entities in both plants and animals. The demonstrated similarity of results from the two techniques also strengthens biological conclusions that were based on AFLP data in the past, an important finding given the wide utilization of AFLP over the last decades. We emphasize that whenever the delimitation of evolutionary entities is the central goal, as it is in many fields of biodiversity research, AFLP is still an adequate technique.

Investigation of Genetic Diversity in Afghan Bread Wheat Genotypes Using SSR and AFLP Markers

Genetic diversity assessment is the principle component for conservation and characterization of germplasm. Genetic diversity study of Afghan bread wheat genotypes is a first step to identify and to select high performance genotypes and distribute to wheat breeding programs. The main objective of this study is to investigate of genetic diversity in 35 Afghan bread wheat genotypes by using Simple Sequence Repeat (SSR) and Amplified Fragment Length Polymorphism (AFLP) markers. DNA extraction according to Cetyl Trimethyl Ammonium Bromide (CTAB) method was conducted and the total genomic DNA was isolated from each variety. Sixty-four SSR primer markers were used and eighteen EcoRI+(N)/MseI+(N) primer combinations with their primer sequences were used for selective polymerase chain reaction (PCR) amplification. Every SSR and AFLP fragment was scored as present (1) or absent (0) within all genotypes under study. Marker/ Value ratio of pairwise genetic distance between genotypes according to the SSRs data was from 0.508 to 0.691 with an average distance of 0.599. Relatively different grouping pattern in comparison to AFLP data observed through cluster analysis. Both types of molecular markers (AFLP and SSR) used in this research proved to be suitable for investigating genetic diversity in the genotypes of Afghan bread wheat, however, AFLP markers gave better view of genetically relationships among genotypes than the SSR markers. The grouping generated by AFLP data showed a special agreement with the origin regions of genotypes (Ariana-07 and Mazar-99 originating from the north of Afghanistan, Lalmi-03 and Kabul-02. Large number of DNA bands identified with AFLP markers might provide a better estimation of genetic similarity than those of SSR markers.

Genetic diversity in natural populations of noble crayfish (Astacus astacus L.) in north-western Poland on the basis of combined SSR and AFLP data

Background Conservation of noble crayfish (Astacus astacus) populations is becoming particularly important since the number of individuals is rapidly declining across the distribution range of the species in Europe. Five crayfish populations in northwestern Poland have been constantly monitored for two decades. However, the genetic structure of these populations has not been analysed, although this information is important to devise effective conservation strategies. Methods Noble crayfish were collected in the autumn of 2014 by scuba diving in Lakes Graniczne, Babinki, Biwakowe, Sęki and Kwisno, all of which are situated in the Bytów Lakeland of northwestern Poland. Genetic diversity of the five populations was assessed based on allele variability in nine SSR regions and six AFLP primer combinations. Results Microsatellite results analysed with AMOVA showed that the diversity between populations corresponds to 18% of total variability, which was confirmed by similar results obtained using AFLP. Additionally, significant genetic diversity was revealed by high average FST values. All of the studied crayfish populations significantly deviated from the expected Hardy–Weinberg genetic equilibrium and were characterised by negative values of inbreeding coefficient (FIS). Discussion The invariably negative inbreeding coefficients (FIS) suggest a low number of mating individuals, a possible consequence of the phenomenon known as genetic bottleneck. However, additional comprehensive analyses are needed to assess the genetic structure, origin and vulnerability of the remaining populations of noble crayfish in the Bytów Lakeland of northwestern Poland, which have high conservation value and are particularly important as a live genetic bank for breeding and restitution purposes.

Reciprocal hybridization between diploid Ficaria calthifolia and tetraploid Ficaria verna subsp. verna: evidence from experimental crossing, genome size and molecular markers

Ficaria is a taxonomically intriguing polyploid complex with high morphological variability. Both hybridization and polyploidization have been suggested as the main evolutionary forces behind the high morphological variability in this genus; however, detailed studies are lacking. In Central Europe, two Ficaria taxa (diploid F. calthifolia and tetraploid F. verna subsp. verna) occasionally co-occur in local sympatry, which might result in hybridization. We investigated sympatric populations of the two Ficaria taxa using flow cytometry, chromosome counts, AFLP analysis and plastid DNA sequencing; we also performed experimental homoploid and heteroploid crosses to determine the frequency and direction of hybrid triploid formation, an alternative route of triploid origin (autopolyploidy) and the possibility of a one-step neoallotetraploid origin. Sympatric populations were composed of three genetic clusters corresponding to diploid F. calthifolia (2n = 16), tetraploid F. verna subsp. verna (2n = 32) and triploid plants (2n = 24). The holoploid genome size and AFLP data suggest a hybrid origin of the triploids, thereby making their formation via autopolyploidization in F. calthifolia unlikely. The triploid populations are monoclonal and of independent origin. In contrast, the parental populations exhibit high genotypic diversity and frequent sexual reproduction, including those of predominantly asexual F. verna subsp. verna. Experimental crossing confirmed that both parental taxa produce fertile seeds via a sexual pathway, but not by apomixis, and that both serve as pollen acceptors in heteroploid crosses, which is consistent with the plastid sequencing. However, hybridization is asymmetric, with maternal-excess crosses being significantly more successful. No signs of neoautotetraploidization or neoallotetraploidization were detected. In summary, recent gene flow between the studied Ficaria taxa is either limited or absent.

Additional information on Meloidogyne polycephannulata and its proposal as a junior synonym of M. incognita

Summary The type population of Meloidogyne polycephannulata is synonymised with M. incognita based on morphological and morphometric characters, as well as biochemical, molecular and phylogenetic studies. Morphological variability and a wide host range were reported for M. incognita during its first description and later re-description. Meloidogyne polycephannulata was described in Brazil from specimens collected in a carrot field (type population). The esterase phenotype (Est) characterised for this species was identical to the phenotype Est I2 of M. incognita, the most ubiquitous phenotype used for diagnostics. Morphological and morphometric characters of the descriptions of the two nominal species showed major similarities, as well as variability within the range of variation detected in M. incognita. In PCR assays, three SCAR markers species-specific for M. incognita (incK14 F/R, Mi/FR and incB06 F/R) amplified the same fragments of 399 bp, 955 bp and 1200 bp, respectively, for populations in both species. In phylogenetic studies based either on concatenated sequences of ITS1-5.8S-ITS2, D2-D3 rRNA, mitochondrial COII regions or on RAPD and AFLP data, the populations of both species grouped in the same clade with high bootstrap support. Altogether, these results provide congruent evidence that the M. polycephannulata type isolate deposited at the Embrapa Cryopreserved National Collection of Root-knot Nematodes is not a valid species but rather a junior synonym of M. incognita.

Evaluation of genetic variations and phylogeny of the most popular pear (Pyrus communis L.) cultivars in Duhok city using AFLP markers

Introduction: Genotyping and evaluation of genetic variation and polymorphic information content of the locally cultivated pear (Pyrus communis L.) might play an important role in building the genetic bank. These are also immensely important for present and future pear breeding program in the region. Methods: In the current study, AFLP markers have been employed to estimate the level of genetic diversity and to assess the phylogeny among the seven most popular pear cultivars in Duhok city. Results: Eight selective primer combinations generated a total of 653 AFLP fragments from which 445 (68.2%) fragments were polymorphic. The number of visible amplified products per primer combination were varied and ranged from 66 to 96 bands. The highest percentage of polymorphism (78.4%) was observed by the primer pair P174/M182, while the lowest percentage of polymorphism (58.6%) was observed by the primer pair P174/M100. The highest PIC (0.85) was obtained with the primer combination P174/M182, while, the lowest PIC (0.49) was obtained by the primer combination P174/M307. The genetic distance was ranged from 0.1348 (between Danimarki and Amreki cultivars) to 0.3131 (between Italy and Zaafaran2 cultivars). Based on the AFLP data, all the seven pear genotypes were successfully clustered into two separate clusters (C1 and C2) with an out-group of Itali cultivar. Conclusions: Overall, it can be concluded that there was high polymorphism among the studied genotypes. Also, it can be stated that the AFLP was a reliable and a powerful technique in genotyping and discriminating of respective pear cultivars.