Are Foliar Nutrition Status and Indicators of Oxidative Stress Associated with Tree Defoliation of Four Mediterranean Forest Species?

Mediterranean forest ecosystems in Croatia are of very high significance because of the ecological functions they provide. This region is highly sensitive to abiotic stresses such as air pollution, high sunlight, and high temperatures alongside dry periods; therefore, it is important to monitor the state of these forest ecosystems and how they respond to these stresses. This study was conducted on trees in situ and focused on the four most important forest species in the Mediterranean region in Croatia: pubescent oak (Quercus pubescens Willd.), holm oak (Quercus ilex L.), Aleppo pine (Pinus halepensis Mill.) and black pine (Pinus nigra J. F. Arnold.). Trees were selected and divided into two groups: trees with defoliation of >25% (defoliated) and trees with defoliation of ≤25% (undefoliated). Leaves and needles were collected from selected trees. Differences in chlorophyll content, hydrogen peroxide content, lipid peroxidation and enzyme activity (superoxide dismutase, catalase, ascorbate peroxidase, non-specific peroxidase), and nutrient content between the defoliated and undefoliated trees of the examined species were determined. The results showed that there were significant differences for all species between the defoliated and undefoliated trees for at least one of the examined parameters. A principal component analysis showed that the enzyme ascorbate peroxidase can be an indicator of oxidative stress caused by ozone. By using oxidative stress indicators, it is possible to determine whether the trees are under stress even before visual damage occurs.

Impact of ground-level ozone on Mediterranean forest ecosystems health.

Given the high ozone concentrations observed in the Mediterranean region during summer, it is crucial to extend our knowledge on the potential ozone impacts on forest health with in situ studies, especially to protect typical endemic forests of the Mediterranean basin. This study is focused on ozone measurements and exposures over the Eastern Adriatic coast and on the calculation of different O3 metrics, i.e., accumulated exposure AOT40 (AOT40dir, AOT40ICP, AOT40pheno) and stomatal O3 fluxes with an hourly threshold of uptake (Y) to represent the detoxification capacity of trees (PODY, with Y = 0, 1, 2 nmol O3 m-2 s-1) used for forest protection. Finally, we provide an assessment of the relationships between the forest response indicators and environmental variables. Passive ozone measurements and monitoring of forest health indicators, namely growth and crown defoliation, were performed for Quercus ilex, Quercus pubescens, Pinus halepensis, and Pinus nigra forests. Results showed that, for all the analysed species, ozone levels were close to reached the upper plausibility limits for passive monitoring of air quality at forest sites (100 ppb), with the highest values found on P. halepensis in the summer period. O3 metrics based on exposure were found to be higher in pine plots than in oak plots, while the highest values of uptake-based metrics were found on P. nigra. Regarding relationships between environmental variables and forest-health response indicators, the crown defoliation was significantly correlated with the soil water content at various depth while the tree growth was correlated with the different O3 metrics. The most important predictors affecting tree growth of Q. pubescens and Q. ilex were AOT40pheno and AOT40dir and POD0 for P. nigra.

Genetic Structure and Relationships among Wild and Cultivated Grapevines from Central Europe and Part of the Western Balkan Peninsula.

The genetic diversity and relationship between wild (Vitis vinifera L. subsp. sylvestris (Gmel.) Hegi and cultivated (V. vinifera L. subsp. vinifera) grapevine in the western Balkan region and Central Europe have not been studied together previously, although this area has a rich viticultural past. Here, we studied wild grapevine populations sampled from their natural habitats in several countries of the western Balkan region and Central Europe. Their genetic diversity and structure were compared to cultivars that are traditionally in use in this region. A sample set of 243 accessions was genotyped at 20 nuclear microsatellite loci, including 167 sylvestris and 76 diverse vinifera cultivars. The genetic diversity of the wild grapevines was lower than that of cultivars by all genetic parameters. Both hierarchical and nonhierarchical clustering methods differentiated two main groups, indicating clear separation between wild and cultivated vines but also revealed clear gene flow between the cultivated and wild gene pools through overlaps and admixed ancestry values in the graphs. There was greater affinity to the wild grapes in Central European cultivars than in Balkan cultivars. Fine arrangement of the structure among cultivated grapevines showed differentiation among Central European and Balkan cultivars. These results confirm the divergence of wild grapes from vinifera and highlight the "crossroad" role of the western Balkan peninsula in the broader context of European viticulture.

Assessment of wild grapevine (Vitis vinifera ssp. sylvestris) chlorotypes and accompanying woody species in the Eastern Adriatic region.

The Eastern Adriatic region, encompassing Croatia and Bosnia and Herzegovina, is considered an important area of natural populations of wild grapevines (Vitis vinifera ssp. sylvestris). The wild grapevine arises in the Eastern Adriatic region in a contact zone of the EU-Mediterranean and the sub-Mediterranean characterized by typical karst relief. This study focuses on the chloroplast DNA (cpDNA) analysis of wild grapevines and the biodiversity of accompanying woody species to better understand the genetic variation of the sylvestris populations of the Eastern Adriatic region and to investigate how this variation fits within today's wild grapevine distribution in the European continent. The allelic variation at nine cpDNA microsatellite loci of wild individuals was used to characterize haplotype diversity in 53 individuals from four population sites. All individuals were grouped into two chlorotypes: A and D, D being the rare haplotype among wild populations on the European continent. In total, 52 woody plant species were identified. However, the studied vegetation structures have been affected by permanent human pressure on natural resources and the preservation status of the collection sites. Based on our results, we conclude that the investigated areas were probably shelter zones for wild grapevine preservation during the unfavorable glaciation era.