Tree species with wide distributions often exhibit different levels of genetic structuring correlated to their environment. However, understanding how environmental heterogeneity influences genetic variation is difficult because the effects of gene flow, drift and selection are confounded. We investigated the genetic variation and its ecological correlates in a wind-pollinated Mediterranean tree species, Fraxinus angustifolia Vahl, within a recognised glacial refugium in Croatia. We sampled 11 populations from environmentally divergent habitats within the Continental and Mediterranean biogeographical regions. We combined genetic data analyses based on nuclear microsatellite loci, multivariate statistics on environmental data and ecological niche modelling (ENM). We identified a geographic structure with a high genetic diversity and low differentiation in the Continental region, which contrasted with the significantly lower genetic diversity and higher population divergence in the Mediterranean region. The positive and significant correlation between environmental and genetic distances after controlling for geographic distance suggests an important influence of ecological divergence of the sites in shaping genetic variation. The ENM provided support for niche differentiation between the populations from the Continental and Mediterranean regions, suggesting that contemporary populations may represent two divergent ecotypes. Ecotype differentiation was also supported by multivariate environmental and genetic distance analyses. Our results suggest that despite extensive gene flow in continental areas, long-term stability of heterogeneous environments have likely promoted genetic divergence of ashes in this region and can explain the present-day genetic variation patterns of these ancient populations.

Species with wide ranges often exhibit morphological variability that is mostly shaped by ecological and evolutionary processes, and the resulting diversity is frequently reflected in intraspecific taxonomy. Dianthus sylvestris in the Balkan Peninsula encompasses six subspecies (D. sylvestris subspp. alboroseus, bertisceus, kozjakensis, nodosus, sylvestris and tergestinus), the identification of which is problematic due to a great morphological variability and complex patterns of environmental variation. To explore morphological variation of described subspecies and to investigate how their morphology varies in relation to environmental variations across their geographical distribution, we evaluated 97 populations of D. sylvestris for variability in 25 morphological characters and 20 environmental variables. Number and shape of epicalyx scales, calyx length and petal denticulation captured the greatest extent of morphological variability between populations. Morphometric analyses revealed a continuous variability in quantitative morphological characters and an absence of clear morphological differentiation between the investigated subspecies. However, two main morphologically distinct entities, the north-western Balkan populations and the south-eastern Balkan populations, were identified and they were split approximately along the Neretva River valley. In addition, our results indicate that environment, rather than geography, plays a potentially more important role in shaping the morphological variability of this taxon. Overall, our results do not support the recognition of the described subspecies or the currently accepted taxonomic concept, instead pointing to a continuous morphological variability of D. sylvestris in the Balkan Peninsula.

Croatia, like many other transition countries has undergone radical changes in its nature protection models. This paper discusses a historical overview, present situation and future possibilities for nature conservation in Croatia. A conservative top-down approach to nature protection was applied in the past in Croatia and is now being replaced by a prevalent bottom-up approach. Social context is crucial to introducing participatory conservation, therefore special concern is given to the perception of the local population towards protected area management in Istria as a case study in Croatia. Survey data were used to assess the conservation knowledge of local populations and their perception towards Protected Areas (PAs), leadership activities and management authorities in Istria County. This paper examines the perceptions of 313 residents living in and around six natural PAs located in Istria. The results revealed a moderate general knowledge about PAs in Istria and environmental issues, and a low awareness of institutions managing PAs, eagerness to participate in the activities of PAs and general support for the conservation cause. Understanding the perception of local residents enables the creation of feasible, long-term strategies for the implementation of participatory conservation. The research identifies the need for greater human, technical and financial efforts to strengthen the management capabilities of local agencies responsible for PAs. The process of participatory conservation optimization in Croatia is underway and world experiences must be observed in order to create a congruent, site-specific model with the best possible results.

The noble crayfish, Astacus astacus, is an indigenous European freshwater species. Its populations show significant declines caused by anthropogenic pressure on its habitats, climate change and the spread of invasive species. Diminishing populations’ trends and loss of genetic diversity highlight the need for effective conservation that will ensure their long-term survival. We combined population genetics and species distribution modelling (SDM) to reveal the impact of climate change and invasive species on the noble crayfish, and to guide future conservation programs of current populations. Our study showed that Croatian populations of A. astacus harbour an important part of species genetic diversity and represent significant genetic reservoir at the European level. The SDM results predicted substantial reductions of suitable habitats for A. astacus by the 2070; only 13% of its current potential distribution is projected to remain stable under pessimistic Representative Concentration Pathway (RCP 8.5) emission scenario. Moreover, most of the populations with high genetic diversity are located in the areas predicted to become unsuitable, and consequently have a high probability of being lost in the future. Further, SDM results also indicated considerable decrease of future habitat suitability for invasive crayfish species in Croatia, suggesting that climate change poses a major threat to already endangered A. astacus. The obtained results help in the identification of populations and areas with the highest conservation value which should be given the highest priority for protection. In order to preserve present diversity in areas that are predicted as suitable, we propose assisted migration and repopulation approaches, for enhancing populations’ size and saving maximum genetic variability. The result of our research emphasizes once again the benefits of multidisciplinary approach in the modern biodiversity conservation.

The Balkan Peninsula played an important role in the evolution of many Mediterranean plants and served as a major source for post-Pleistocene colonisation of central and northern Europe. Its complex geo-climatic history and environmental heterogeneity significantly influenced spatiotemporal diversification and resulted in intricate phylogeographic patterns. To explore the evolutionary dynamics and phylogeographic patterns within the widespread eastern Mediterranean and central European species Aurinia saxatilis, we used a combination of phylogenomic (restriction-site associated DNA sequencing, RADseq) and phylogenetic (sequences of the plastid marker ndhF) data as well as species distribution models generated for the present and the Last Glacial Maximum (LGM). The inferred phylogenies retrieved three main geographically distinct lineages. The southern lineage is restricted to the eastern Mediterranean, where it is distributed throughout the Aegean area, the southern Balkan Peninsula, and the southern Apennine Peninsula, and corresponds to the species main distribution area during the LGM. The eastern lineage extends from the eastern Balkan Peninsula over the Carpathians to central Europe, while the central lineage occupies the central Balkan Peninsula. Molecular dating places the divergence among all the three lineages to the early to middle Pleistocene, indicating their long-term independent evolutionary trajectories. Our data revealed an early divergence and stable in situ persistence of the southernmost, eastern Mediterranean lineage, whereas the mainland, south-east European lineages experienced more complex and turbulent evolutionary dynamics triggered by Pleistocene climatic oscillations. Our data also support the existence of multiple glacial refugia in southeast Europe and highlight the central Balkan Peninsula not only as a cradle of lineage diversifications but also as a source of lineage dispersal. Finally, the extant genetic variation within A. saxatilis is congruent with the taxonomic separation of peripatric A. saxatilis subsp. saxatilis and A. saxatilis subsp. orientalis, whereas the taxonomic status of A. saxatilis subsp. megalocarpa remains doubtful.