The Balkan mountain ranges are major hotspots of genetic diversity and endemism, yet many species remain poorly studied. One such species is Alyssum bosniacum, a narrow endemic of the Central Dinaric Alps. To fill this gap, we examined 143 individuals from 15 populations across the species’ range using flow-cytometric ploidy determination, amplified fragment length polymorphisms (AFLPs), nuclear microsatellites, and chloroplast DNA sequences. Microsatellite data revealed two genetic clusters, showing moderate differentiation and relatively high diversity. AFLP profiles indicated shallow but geographically structured variation, while chloroplast haplotypes showed limited divergence and regional clustering. Our data suggest possible persistence in multiple microrefugia within the Central Dinaric Alps, although further evidence is needed to confirm this scenario. Despite range fragmentation, genetic variation within the population remains high, indicating evolutionary resilience and supporting the species’ long-term future population stability under current conditions.
Hybridization and polyploidization are the main drivers of diversification in Sorbus, and have generated numerous evolutionary lineages across Europe. Newly derived polyploid lineages usually reproduce via apomixis and represent novel genetic and morphological entities, usually circumscribed as distinct species. Whereas Sorbus has been thoroughly studied in Central and Western Europe, its diversity in the Balkans remains less explored. This is especially the case for Sorbus subgen. Tormaria that includes derivates of crosses between S. subgen. Aria and S. torminalis. Here, we report the discovery of a triploid population of S. subgen. Tormaria in south-western Bosnia and Herzegovina. We performed amplified fragment length polymorphism fingerprinting, evaluation of nuclear microsatellites, plastid DNA sequencing, flow cytometric ploidy level estimation, reproduction mode screening, and morphological characterization to disentangle its relationship to the previously reported diploid populations from Bosnia and Herzegovina and congeners from Central Europe, as well as to both parents. The data revealed that the aforementioned population includes mostly triploid facultative apomicts that are genetically divergent from other taxa. We therefore describe it as a new species, Sorbus hercegovinae, and provide its description, distribution data, and conservation status. Our study highlights the Balkan Peninsula as one of the hotspots of whitebeam diversity.
axiomFP.py is an open-source software developed to diagnose ploidy level and call quality for samples genotyped on Affymetrix Axiom SNP arrays by making frequency plots of normalized SNP call positions among SNPs meeting specific clustering parameters. This research outlines the methods employed in the development of the software, and presents the results obtained through its application on a dataset of mixed ploidy apple (Malus spp.) cultivars and germplasm accessions. The tools required to prepare the input files and operate the software are also described. The frequency plots generated by the software require a visual inspection to assess ploidy and call quality. The results have been validated using the available ploidy data, as well as flow cytometry, and have shown complete accuracy. The software is available on GitHub at https://github.com/allmiraria/axiomFP.
While 38% of tree species are at risk of extinction worldwide, their inventory and occurrence at ecologically and biogeographically meaningful scales is lacking in many parts of the world, including the biodiversity-rich Mediterranean region. Here, we provide presence/absence, extinction risk, biogeography and genetic diversity data of trees in 39 climatically and ecologically Mediterranean territories (so-called “botanical territories”) in North Africa, Western Asia and Southern Europe. The inventory includes 496 species and 147 subspecies from 50 families and 111 genera, including 48 species and 8 subspecies previously not considered as trees. We show that native tree species distribution is highly skewed across the tree of life with a few species-rich families such as the Rosaceae and the majority with less than 1% of all species. Endemism was not evenly distributed among botanical territories and neither was extinction risk, an assessment of which was lacking in almost half of the species. While no geographic trends were detectable, species richness was found to be positively correlated with botanical territory area and, when standardized by area, with habitat heterogeneity. Information on genetic diversity was lacking in two thirds of the species inventoried and mostly focused on species with economic importance. Our data are open access and can be used by researchers and stakeholders for a wide range of purposes, including conservation and restoration. Our findings identified major native tree richness hotspots as well as key knowledge gaps and biases related to extinction risk and genetic diversity. Our findings also emphasize the importance of increased collaboration to support the conservation of Mediterranean forest trees.
The genetic, morphological and taxonomic diversity of the genus Sorbus is due to homoploid and polyploid hybridisation, autopolyploidy and apomixis, which also influence the production and diversity of secondary metabolites, especially flavonoids. The aim of this study was to investigate the relationships and variations of flavonoids in terms of hybrid origin and ploidy level between the parental species and their hybrid derivatives. The sampling design included leaf material of the following Sorbus accessions from ten natural localities: parental taxa (di-, tri- and tetraploids of S. aria; diploid S. torminalis and S. aucuparia) and their di-, tri- and tetraploid hybrid derivatives from crosses of S. aria × S. torminalis (subg. Tormaria) as well as the tetraploid S. austriaca and S. bosniaca, which originate from crosses of S. aria × S. aucuparia (subg. Soraria). We analysed the flavonoid profiles from the leaf fractions by LC-MS. A total of 23 flavonoids were identified, including apigenin and luteolin derivatives, which distinguish the hybrid groups from each other. This profiling highlights the distinctiveness of the Tormaria and Soraria accessions and emphasises the potential of the subg. Tormaria for further research on bioactive compounds in biological studies.
Polyploidy is a powerful mechanism driving genetic, physiological, and phenotypic changes among cytotypes of the same species across both large and small geographic scales. These changes can significantly shape population structure and increase the evolutionary and adaptation potential of cytotypes. Alyssum moellendorfianum, an edaphic steno-endemic species with a narrow distribution in the Balkan Peninsula, serves as an intriguing case study. We conducted a comprehensive analysis of genetic diversity and population structure across the species’ range, employing an array of genetic techniques (nuclear microsatellites, amplified fragment length polymorphisms, and plastid DNA sequences), flow cytometry (FCM), morphometry, and pollen analysis. The study reveals two genetic lineages: spatially distributed diploid and tetraploid cytotypes. Clear divergence between diploids and tetraploids was shown by AFLP, while plastid DNA sequences confirmed private haplotypes in each of the studied populations. Higher genetic diversity and allelic richness following the north-south pattern were documented in tetraploids compared to diploids, as indicated by nuclear microsatellites. Morphometric analysis via principal component analysis (PCA) and canonical discriminant analysis (CDA) did not reveal any divergence between diploid and tetraploid cytotypes. Nonetheless, a distinction in pollen size was clearly observed. The results suggest an autopolyploid origin of tetraploids from diploid ancestors. Despite the population fragmentation in a very small geographic range, these populations harbour high genetic diversity, which would allow them to remain stable if natural processes remain undisturbed.
The Balkan Peninsula is one of Europe?s major hotspots for plant biodiversity. Although the rich flora of the Balkans has been exten?sively studied from many points of view, genome size studies have received comparatively little attention. This paper contributes to the online available Genome Size Database of Balkan Flora (GeSDaBaF; http://www.pmf.unsa.ba/gesdabaf) and updates this database with new bibliographic data on the C-values for plants in the Balkan region. Additionally, 2C DNA values determined by flow cytometry are reported for 87 accessions, encompassing 82 taxa (70 species, 11 subspecies, and one hybrid taxon, 14 of which are endemic to the Balkans and two with amphi-adriatic distribution) across 63 genera and 31 families. The newly obtained genome size values include the first data for one genus, 25 species, eight subspecies, and one hybrid taxon. These new data represent 9.99% of the 821 taxa for which nuclear DNA content has been assessed in the Balkans to date, highlighting existing knowledge gaps for future research.
Objective: Iris species are widely used in pharmaceutical and cosmetic applications owing to their high content of bioactive compounds with anti-inflammatory and antimicrobial properties. This study aimed to investigate the potential antibacterial effect of crude extracts (aqueous, 50% and 80% ethanol) of three Iris species ( I. pumila , while I. reichenbachii and I. illyrica are endemic) from Bosnia and Herzegovina against the multiresistant bacterial strain methicillin-resistant Staphylococcus aureus subsp. aureus ATCC 33591 (MRSA strain). Materials and Methods: The antimicrobial compounds in the crude extracts were identified using High-performance liquid chromatography (HPLC), and their effects on the MRSA strain were tested using agar well diffusion and broth microdilution method. The binding affinities were analysed using molecular docking simulations. Results: We identified bioactive targeted compounds in these extracts, mainly flavonoids named isorhamnetin, hesperidin, quercetin, fisetin, genistein, and kaempferol. Antibacterial assays showed that extracts of all three Iris species inhibited MRSA. The binding affinity analysis showed that isorhamnetin and hesperidin had the highest affinity scores, stronger (isorhamnetin) or the same (hesperidin) as the positive control ceftobiprole. Conclusion: This in vitro and in silico study showed that Iris species represent a valuable source of bioactive compounds that can be used against multidrug-resistant strains such as MRSA. The potential use of these agents in multiple drugs is warranted, and further evaluation for human application is needed.
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