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Small abandoned mining areas of Fe and Mn oxides located in the Portuguese sector of the Iberian Pyrite Belt (SW of Europe) have been converted into agrosilvopastoral systems with very few environmental management measures after their closure. Although at the landscape scale, no visible differences were observed between the former mining intervention areas and adjacent areas, it is essential to assess the state and environmental risk of the soil–plant system, especially in the herbaceous pastures grazed by domestic animals. This was carried out in the Ferragudo mining area, where an agrosilvopastoral system, composed of holm oak and dryland pasture, had been established after the closure of the mine at ≈45 years. The soils presented neutral pH and variable fertility degree. The pseudo-total soil concentrations of Cu, Mo, and Zn exceeded the Portuguese limit values established for agriculture use (>180 mg Cu/kg; >8.2 mg Mo/kg; 349 mg Zn/kg), but their soil available fractions were small (<8.4% of the pseudo-total concentrations). Trees and herbaceous plants showed good development, and the concentrations of the elements (except Mn) were considered normal or sufficient. For Mn, most of the plant samples exceeded phytotoxic Mn values, but no visual signs of phytotoxicity were observed. Only the concentrations of Fe and K in the shoots of some herbaceous samples exceeded the maximum tolerable levels for cattle and sheep, so the risk to animals can be considered small since other sources are present in animal feed. In general, this agrosilvopastoral system did not pose a significant environmental risk. [ABSTRACT FROM AUTHOR]

Hyperspectral imagery and advanced classification techniques can significantly enhance remote sensing's role in forest monitoring. Thanks to recent missions, such as the Italian Space Agency's PRISMA (PRecursore IperSpettrale della Missione Applicativa—Hyperspectral PRecursor of the Application Mission), hyperspectral data in narrow bands spanning visible/near infrared to shortwave infrared are now available. In this study, hyperspectral data from PRISMA were used with the aim of testing the applicability of PRISMA with different band sizes to classify tree species in highly biodiverse forest environments. The Serre Regional Park in southern Italy was used as a case study. The classification focused on forest category classes based on the predominant tree species in sample plots. Ground truth data were collected using a global positioning system together with a smartphone application to test its contribution to facilitating field data collection. The final result, measured on a test dataset, showed an F1 greater than 0.75 for four classes: fir (0.81), pine (0.77), beech (0.90), and holm oak (0.82). Beech forests showed the highest accuracy (0.92), while chestnut forests (0.68) and a mixed class of hygrophilous species (0.69) showed lower accuracy. These results demonstrate the potential of hyperspectral spaceborne data for identifying trends in spectral signatures for forest tree classification. [ABSTRACT FROM AUTHOR]

We examined the distribution of termites along an elevational gradient in the western Rif Mountains of northern Morocco. Sampling followed standardized protocols at nine forest sites (125, 325, 488, 715, 850, 1150, 1316, 1544, and 1700 masl), all except the lowest site within the Intercontinental Mediterranean Biosphere Reserve. We found a total of three termite species, with Reticulitermes grassei (Rhinotermitidae) being the most abundant, followed by Kalotermes flavicollis (Kalotermitinae) and Amitermes sp. (Terminidae). Comparing microhabitat preferences, we found R. grassei and Amitermes sp. exclusively in buried wood and under rocks, and K. flavicollis exclusively in dead tree trunks and surface wood. The findings reveal the distribution of microhabitats along an elevation gradient and the corresponding preferences of termite species across these microhabitats. We found that total termite density decreased significantly with elevation (rs = −0.40; p < 0.001), but with distinct differences among species. Kalotermes flavicollis and Amitermes sp. showed a strong negative correlation between density and elevation. In contrast, for R. grassei, the most common termite, we found no significant correlation. Termite density also varied among forest types, with a non-significant trend (p = 0.075) of higher density in the cork oak forest (hits/m2 = 0.06) compared to holm oak (0.04) and fir–cedar (0.02) forests. This research contributes to the limited knowledge of termite diversity and ecology in northern Africa and western Mediterranean forests. [ABSTRACT FROM AUTHOR]

Acute Oak Decline (AOD), a bacterial disease previously known in Northern and Central Europe, has recently been reported in Salento (a Mediterranean coastal region of Southern Italy), where holm oak trees exhibiting AOD-like symptoms have tested positive for infection with AOD-related bacteria such as Brenneria goodwinii and Gibbsiella quercinecans. Sampling symptomatic trees, strains BLEC23 (B. goodwinii) and GSAC47 (G. quercinecans) were isolated and identified by partial 16S rRNA and gyrB gene sequencing. Pathogenicity tests demonstrate that these bacteria induce wood necrosis when inoculated in excised branches, providing details for the etiology of AOD in Italy. Phylogenetic analysis indicated a substantial genetic similarity between the Italian strains and those found in various European and non-European countries. These findings leave a space open to the possibility that the bacteria involved in AOD are much more widespread in Europe than the findings indicate, but that their presence is frequently hidden. [ABSTRACT FROM AUTHOR]

Fires affect forest dynamics in seasonally dry regions such as the Mediterranean Basin. There, fire impacts on tree growth have been widely characterized in conifers, particularly pine species, but we lack information on broadleaf tree species that sprout after fires. We investigated post-fire radial growth responses in two coexisting Mediterranean hardwood species (the evergreen Quercus ilex, the deciduous Celtis australis) using tree-ring width data. We compared growth data from burnt and unburnt stands of each species subjected to similar climatic, soil and management conditions. We also calculated climate–growth relationships to assess if burnt stands were also negatively impacted by water shortage, which could hinder growth recovery. Tree-ring data of both species allowed us to quantify post-fire growth enhancements of +39.5% and +48.9% in Q. ilex and C. australis, respectively, one year after the fire. Dry spring climate conditions reduced growth, regardless of the fire impact, but high precipitation in the previous winter enhanced growth. High June radiation was negatively related to the growth of unburnt Q. ilex and burnt C. australis stands, respectively. Post-fire growth enhancement lasted for five years after the fire and it was a transitory effect because the growth rates of burnt and unburnt stands were similar afterwards. [ABSTRACT FROM AUTHOR]

Forests play a key role in the global carbon (C) cycle through multiple interactions between above-ground and soil microbial communities. Deeper insights into the soil microbial composition and diversity at different spatial scales and soil depths are of paramount importance. We hypothesized that in a homogeneous above-ground tree cover, the heterogeneous distribution of soil microbial functional diversity and processes at the small scale is correlated with the soil's chemical properties. From this perspective, in a typical Mediterranean holm oak (Quercus ilex L.) peri-urban forest, soil carbon dioxide (CO2) emissions were measured with soil chambers in three different plots. In each plot, to test the linkage between above-ground and below-ground communities, soil was randomly sampled along six vertical transects (0–100 cm) to investigate soil physico-chemical parameters; microbial processes, measured using Barometric Process Separation (BaPS); and structural and functional diversity, assessed using T-RFLP and qPCR Real Time analyses. The results highlighted that the high spatial variability of CO2 emissions—confirmed by the BaPS analysis—was associated with the microbial communities' abundance (dominated by bacteria) and structural diversity (decreasing with soil depth), measured by H′ index. Bacteria showed higher variability than fungi and archaea at all depths examined. Such an insight showed the clear ecological and environmental implications of soil in the overall sustainability of the peri-urban forest system. [ABSTRACT FROM AUTHOR]

Biodiversity monitoring is key for understanding the delivery of ecosystem functions and services. Mediterranean forests and woodlands harbor many characteristic species of the Mediterranean vascular flora, and hence, they are a good surrogate for detecting changes in biodiversity linked to global change. In this work, we present a database resulting from the study of vascular plant diversity in multi-scale plots of 0.1 ha, measured around the first decade of this century and located in Mediterranean forest environments. Diversity profiles are calculated from Hill numbers (0, 1 and 2) for local (α) and regional (ϒ) diversity, as well as a multiplicative calculation of differential diversity (β). The main Mediterranean forests sampled had a medium coverage of 51% and stand dominant height of 10.6 m, and they were monospecific in two-thirds of cases. Local diversity reaches its highest values (around 78 species per 0.1 ha) in Holm oak dehesas, with values below 50 species for the most productive forest stands dominated by species of the genus Pinus. As regards the contribution to regional diversity, broadleaf formations contribute the most, with stone pine forests and dehesas in an intermediate position, and pine forests contributing the lowest in species richness terms. [ABSTRACT FROM AUTHOR]

Tuber aestivum, commonly known as the summer truffle, is typically found in various parts of Europe where it grows naturally. However, its presence in Portugal was not confirmed until now. The first fruit bodies were collected in April 2024 at stone pine stands (Alenquer and Arruda dos Vinhos, Lisbon) and in June at holm oak stands (Salir, Faro). These specimens are characterized by hypogeous, subglobose, black ascomata with a peridium surface covered with pyramidal warts. Ascopores are subglobose-to-broadly ellipsoid, distinctively ornamented, usually 1–6 per asci. According to the results of the internal transcribed spacer (ITS) rDNA sequence analysis, these specimens form a well-supported group within the Aestivum clade, with T. aestivum being the closest phylogenetic taxon. This remarkable discovery opens up new opportunities for truffle exploitation in Portugal thanks to the summer truffle's gastronomical value and high market prices. [ABSTRACT FROM AUTHOR]

Quercus ilex L., an evergreen oak species typical of the western and central Mediterranean basin, is facing decline and dieback episodes due to the increase in the severity and frequency of heat waves and drought events. Studying xylogenesis (the wood formation process) is crucial for understanding how trees respond with their secondary growth to environmental conditions and stress events. This study aimed to characterize the wood formation dynamics of Quercus ilex and their relationship with the meteorological conditions in an area experiencing prolonged drought periods. Cambial activity and xylem cell production were monitored during the 2019 and 2020 growing seasons in a Q. ilex forest located at the Vesuvius National Park (southern Italy). The results highlighted the significant roles of temperature and solar radiation in stimulating xylogenesis. Indeed, the correlation tests revealed that temperature and solar radiation positively influenced growth and cell development, while precipitation had an inhibitory effect on secondary wall formation. The earlier cell maturation in 2020 compared to 2019 underscored the impact of global warming trends. Overall, the trees studied demonstrated good health, growth and adaptability to local environmental fluctuations. This research provides novel insights into the intra-annual growth dynamics of this key Mediterranean species and its adaptation strategies to climatic variability, which will be crucial for forest management in the context of climate change. [ABSTRACT FROM AUTHOR]

The development of new natural antibiotics is considered as the heart of several investigations in the nutraceutical field. In this work, leaves of Quercus ilex L. treated by tropospheric ozone (O3) and nitrogen (N) deposition, exhibited a clear antimicrobial efficacy against five multi-drug resistant (MDR) bacterial strains (two gram-positive and three gram-negative). Under controlled conditions, it was studied how simulated N deposition influences the response to O3 and the antibacterial and antioxidant activity, and antioxidant performance. The extraction was performed by ultra-pure acetone using two different steps. A higher antioxidant activity was measured in the presence of interaction between O3 and N treatments on Quercus leaves. At the same time, all organic extracts tested have shown bacteriostatic activity against all the tested strains with a MIC comprised between 9 and 4 micrograms/mL, and a higher antioxidant efficacy shown by spectrophotometric assay. Stronger antimicrobial activity was found in the samples treated with O3, whereas N-treated plants exhibited an intermediate antibacterial performance. This performance is related to the stimulation of the non-enzymatic antioxidant system induced by the oxidative stress, which results in an increase in the production of antimicrobial bioactive compounds. [ABSTRACT FROM AUTHOR]

Xylophagous insects play a crucial role in forest ecosystems, contributing to population dynamics. The "Cerambyx complex" (CC) constitutes an emerging pest in Mediterranean oak woodlands. We studied the fragmentation and connectivity of holm and cork oak stands in Andalusia (Spain), and the relationships with the current dispersion of CC, as well as the effect on the connectivity and dispersion patterns with the implementation of nests of a predator bird (Garrulus glandarius) to reduce insect populations in highly connected areas. The Kernel Density Estimation (KDE) was used to assess the spatial distribution of CC. Connectivity was assessed using graphs theory (Graphab 2.6) to characterize the importance of patches and linkages for contributing to dispersal. We selected the Eurasian jay (G. glandarius) as a reference bird species to generate "barriers" to the dispersion of the CC. We used the probability of connectivity (PC) and the flux (F) to compare the effect of the introduction of Eurasian jay nets. Results showed an increasing trend in the distribution and incidence of CC during the period 2001–2016, resulting in 7.3% and 13.1% mortality rates for Q. ilex and Q. suber, respectively. The connectivity model using only Q. ilex and Q. suber forests as reference habitats was not enough to explain the distribution of CC. The value of PC and F metrics decreased by 38.09% and 20.59% by introducing 300 nests of Eurasian jay. Our methodology provides a pest management tool using connectivity metrics, which can be integrated with other variables to control pest outbreaks and pest dispersion. [ABSTRACT FROM AUTHOR]

Numerous studies have shown the multiple benefits of plant biodiversity and the crucial role of residents' awareness of conservation and land management. Plant biodiversity was investigated in an analytical monitoring report conducted jointly with the local population engaged in livestock activities and young residents who were shifting toward hospitality and tourism. The study area, covering over 800 km2 in Sardinia, is half forested and attracts interest in its landscape. During our research, genotypes of Quercus ilex L., with sweet acorns rich in polyphenols, as the oldest ecosystem services in these rural communities, were discovered. Collaborative monitoring focused attention on the benefits of plants in different habitats as follows: 53% were known to livestock farmers as food for local breeds of goats and pigs, 15% were official, and 13% were endemic. They had also been used for human nutrition within the Mediterranean diet and attracted interest for their use in landscapes and gardens. This study analyzes numerical data from critical and educational perspectives. These data serve as indicators of ecosystem health for the purpose of sustainable management policies and attest to collaborative monitoring as a tool for analyzing human activities and the necessary balance between profit and biodiversity conservation, given the current challenging climate change conditions. [ABSTRACT FROM AUTHOR]

Holm oak (Quercus ilex) is considered to be one of the major structural elements of Mediterranean forests and the agrosilvopastoral Spanish "dehesa", making it an outstanding example of ecological and socioeconomic sustainability in forest ecosystems. The exotic Phytophthora cinnamomi is one of the most aggressive pathogens of woody species and, together with drought, is considered to be one of the main drivers of holm oak decline. The effect of and response to P. cinnamomi inoculation were studied in the offspring of mother trees from two Andalusian populations, Cordoba and Huelva. At the two locations, acorns collected from both symptomatic (damaged) and asymptomatic (apparently healthy) trees were sampled. Damage symptoms, mortality, and chlorophyll fluorescence were evaluated in seedlings inoculated under humid and drought conditions. The effect and response depended on the population and were more apparent in Huelva than in Cordoba. An integrated proteomic and metabolomic analysis revealed the involvement of different metabolic pathways in response to the pathogen in both populations, including amino acid metabolism pathways in Huelva, and terpenoid and flavonoid biosynthesis in Cordoba. However, no differential response was observed between seedlings inoculated under humid and drought conditions. A protective mechanism of the photosynthetic apparatus was activated in response to defective photosynthetic activity in inoculated plants, which seemed to be more efficient in the Cordoba population. In addition, enzymes and metabolites of the phenylpropanoid and flavonoid biosynthesis pathways may have conferred higher resistance in the Cordoba population. Some enzymes are proposed as markers of resilience, among which glyoxalase I, glutathione reductase, thioredoxin reductase, and cinnamyl alcohol dehydrogenase are candidates. [ABSTRACT FROM AUTHOR]

Tuber melanosporum is an ascomycete that forms ectomycorrhizal (ECM) symbioses with a wide range of host plants, producing edible fruiting bodies with high economic value. The quality of seedlings in the early symbiotic stage is important for successful truffle cultivation. Numerous bacterial species have been reported to take part in the truffle biological cycle and influence the establishment of roots symbiosis in plant hosts and the development of the carpophore. In this work, three different bacteria formulations were co-inoculated in Quercus ilex L. seedlings two months after T. melanosporum inoculation. At four months of bacterial application, the T. melanosporum ECM root tip rate of colonization and bacterial presence were assessed using both morphological and molecular techniques. A 2.5-fold increase in ECM colonization rate was found in the presence of Pseudomonas sp. compared to the seedlings inoculated only with T. melanosporum. The same treatment caused reduced plant growth either for the aerial and root part. Meanwhile, the ECM colonization combined with Bradyrhizobium sp. and Pseudomonas sp. + Bradyrhizobium sp. reduced the relative density of fibrous roots (nutrient absorption). Our work suggests that the role of bacteria in the early symbiotic stages of ECM colonization involves both the mycorrhizal symbiosis rate and plant root development processes, both essential for improve the quality of truffle-inoculated seedlings produced in commercial nurseries. [ABSTRACT FROM AUTHOR]

The emergence of new plant diseases is an increasingly important concern. Climate change is likely to be among the factors causing most of the emerging diseases endangering forest and tree heritage around the world. Such diseases may be caused by latent pathogens or microorganisms cryptically associated with plants. The shift from a non-pathogenic to a pathogenic stage may depend on physiological alterations of the host, environmental changes, and/or stress factors. In some woods of the Salento Peninsula (Apulia Region, Italy), sudden declines of holm oak plants (Quercus ilex L.) have been observed since 2016. The morphological and molecular characterization of representative fungal isolates associated with cankers and necrosis in declining plants indicated that these isolates belong to the Botryosphaeriaceae family, and the most frequent species were Diplodia corticola and Diplodia quercivora, followed by Neofusicoccum vitifusiforme. In artificially inoculated young holm oak plants, both D. corticola and D. quercivora species produced intense and severe subcortical and leaf margin necrosis. N. vitifusiforme, although less aggressive, induced the same symptoms. Our research, in addition to confirming the involvement of D. corticola in olm oak decline, represents the first report of D. quercivora as a new pathogen of Q. ilex in Italy. Furthermore, to the best of our knowledge, we also found N. vitifusiforme as a new pathogen of Q. ilex. [ABSTRACT FROM AUTHOR]

Urbanization and urban regeneration can significantly impact cultural heritage, but a greater knowledge of the past natural and historical features is needed to value the past and understand the present. The lost Villa Peretti Montalto in Rome, once located in the area that corresponds to the current front side of Termini station, deserves great attention due to its cultural value. This work aimed to provide a floristic and functional reconstruction of the gardens of the villa during the XVI and XVII Centuries. From several bibliographic and iconographic sources, a critical analysis and interpretation of plant names was conducted. A list of 87 species and their location in the different garden sectors, during different periods with their specific uses, is provided. The arboreal design was made by classical species in the Roman context, like Cupressus sempervirens, Pinus pinea, Quercus ilex, and Ulmus glabra. In addition, ancient lost varieties of fruit trees (mainly Pyrus communis and Ficus carica) and several species of conservation interest were found. The knowledge of the ancient flora in historical gardens could be a key tool in urban greenery planning and touristic and cultural valorization. [ABSTRACT FROM AUTHOR]

The aim of our work is to develop a methodology to identify the areas most prone to natural rockfall retention through the integrated use of remote sensing data. The area chosen as a case study is located in Campania (Italy) nearby Mount San Liberatore. In this area, which is itself geomorphologically predisposed to landslide risk, there are several rockfall risk hotspots, so defined because of the high exposed value constituted by an articulated infrastructure network located along the northwest slope of the mountain. The area is largely covered by dense vegetation, of which holm oak is the most representative type, characterized by a taproot apparatus that, giving it strength and stability, makes it an ideal tree for slope protection. Based on high-resolution multispectral satellite images, vegetation indices (VIs) were calculated to estimate health status, approximate age, average height, robustness, and vigor. Morphometric parameters suitable for describing slope dynamics were also calculated, derived from LiDAR data. The classification of areas with similar characteristics was carried out using Self-Organizing Maps. The results made it possible to identify all those areas where there is a greater contribution of protective forests in the mitigation of rockfall risk and, consequently, to identify areas to carry out a combined strengthening of protective actions. [ABSTRACT FROM AUTHOR]

Diplodia corticola is one of the most aggressive fungal pathogens of Quercus species involved in the decline of Mediterranean oak forests. In this study, three strains of D. corticola associated with holm (Quercus ilex) and cork (Quercus suber) oak trees exhibiting dieback symptoms and cankers in Algeria were selected to investigate the production of secondary metabolites. Metabolomic analyses revealed the production of several known compounds, such as sphaeropsidins, diplopyrones and diplofuranones. Moreover, the comparative investigation of secondary metabolites produced by the analyzed strains with different degrees of virulence revealed possible implications of these compounds in the fungal virulence. In particular, sphaeropsidins seem to be the main phytotoxic compounds of D. corticola involved in the infections of Quercus species, with a possible synergistic influence of the less representative compounds in the fungal virulence. [ABSTRACT FROM AUTHOR]

Plant water status can be assessed through leaf spectral reflectance in the near-infrared (NIR), the "water bands", considering indices that include the reflectance at a band absorbed by water over and another one as reference. We have assessed i/ the accuracy of reflectance at 1450, 1599 and 1940 nm without reference bands and ii/ the potential use of leaf water content index (LWCI) for the estimation of plant water status in holm oak, the main host plant for black truffle cultivation. We demonstrated that contact measurements of leaf reflectance in the "water bands" constitute an accurate and non-invasive estimator of relative water content (RWC) in holm oak, despite the absence of a reference wavelength, probably due to the low variation in leaf thickness under dehydration. The use of a reference wavelength, which is needed for remote sensing, diminished the accuracy of RWC estimation. Contrastingly, LWCI increased the accuracy of RWC estimation as well as a reference wavelength were used. However, LWCI required the reflectance value at full turgor, diminishing its potential for implementation at field level. In conclusion, this technique would allow the continuous monitoring of the physiological state of holm oak and intelligent water control in truffle cultivation. [ABSTRACT FROM AUTHOR]

Holm oaks (Quercus ilex L.) can suffer severe infection by the oomycete Phytophthora cinnamomi Rands; the production of more tolerant plants is, therefore, required. Embryo formation is a key period in the establishment of epigenetic memory. Somatic embryos from three holm oak genotypes were elicited, either over 3 days or 60 days, with methyl-jasmonate, salicylic acid (SA), β-aminobutyric acid (BABA), or benzothiadiazole (all at 50 μM and 100 μM), or 10% and 30% of a filtered oomycete extract (FILT10 and FILT30) to activate plant immune responses. The number of embryos produced and conversion rate under all conditions were recorded. Some elicited embryos were then exposed to P. cinnamomi in dual culture, and differential mycelial growth and the progression of necrosis were measured. The same was performed with the roots of germinated embryos. Within genotypes, significant differences were seen among the elicitation treatments in terms of both variables. Embryos and roots of 60-day BABA, SA, or FILT10 treatments inhibited mycelium growth. The 3-day BABA (either concentration) and 60-day FILT10 induced the greatest inhibition of necrosis. Mycelium and necrosis inhibition were compared with those of tolerant trees. Both inhibitions might be a defense response maintained after primed embryo germination, thus increasing the likelihood of tolerance to infection. [ABSTRACT FROM AUTHOR]

Understanding how fire severity affects resprouting plants during post-fire regeneration is key to anticipating Mediterranean vegetation vulnerability in a context of increasingly large fires with high intensity and severity due to climate change. Here, we monitored the water status, leaf gas exchange, and plant growth in holm oaks (Quercus ilex) of central Spain burned with different fire severity throughout the first post-fire year. The Q. ilex burned with high severity (HB+) showed higher water potential and shoot growth than those burned with low severity (LB+) or unburned (B−), especially during spring and summer. In summer, resprouting HB+ and LB+ plants exhibited higher carbon assimilation than unburned ones. Moreover, we also found that plants with higher water availability and growth, i.e., HB+ individuals, had higher specific leaf area and lower water use efficiency. Overall, our study shows that holm oak forests exhibit high plasticity to fire and that Q. ilex burned with high severity have a faster short-term regeneration than those burned with low severity. However, this rapid regeneration is based on a less conservative water-use strategy, which could jeopardize their populations in case of extreme drought events increasingly common in the current context of climate change. [ABSTRACT FROM AUTHOR]

The fungal biodiversity associated with a stable plant community appears to vary from year to year. To analyse the annual behaviour in the formation of fruiting bodies, a statistical study of the data obtained for 4 years (2009–2012) in an apparently undisturbed Mediterranean ecosystem dominated by the holm oak (Quercus ilex L. subsp. ballota (Desf.) Samp.), located in the Midwest of the Iberian Peninsula, was carried out. These data were related to the main meteorological variables. The 150 species collected showed a significant annual, monthly, and weekly difference in their fruiting during the collection period. All this implies a variation in the annual fungal fruiting which can modify the moment of when maximum peaks of fruiting appear, their duration, and the number of species that compose them. In addition, the results make it possible to establish an annual behaviour pattern, with sporocarp formation throughout the year and four fruiting groups (two of them in the dry season). They also allow for inferring a possible response to climate change, with a delay in the fruiting of the autumn-winter group and earlier fruit bearing in the winter-spring group. [ABSTRACT FROM AUTHOR]

Long-term monitoring of microbial communities in the rhizosphere of post-fire forests is currently one of the key knowledge gaps. Knowing the time scale of the effects is indispensable to aiding post-fire recovery in vulnerable woodlands, including holm oak forests, that are subjected to a Mediterranean climate, as is the case with forests that are found in protected areas such as the Sierra Nevada National and Natural Park in southeastern Spain. We took rhizosphere soil samples from burned and unburned holm oak trees approximately 3, 6, and 9 years after the 2005 fire that devastated almost 3500 ha in southeastern Spain. We observed that the prokaryotic communities are recovering but have not yet reached the conditions observed in the unburned forest. A common denominator between this fire and other fires is the long-term persistence of three ecosystem recovery biomarkers—specifically, higher proportions of the genera Arthrobacter, Blastococcus, and Massilia in soil microbial communities after a forest fire. These pyrophilous microbes possess remarkable resilience against adverse conditions, exhibiting traits such as xerotolerance, nitrogen mineralization, degradation of aromatic compounds, and copiotrophy in favorable conditions. Furthermore, these biomarkers thrive in alkaline environments, which persist over the long term following forest fires. The relative abundance of these biomarkers showed a decreasing trend over time, but they were still far from the values of the control condition. In conclusion, a decade does not seem to be enough for the complete recovery of the prokaryotic communities in this Mediterranean ecosystem. [ABSTRACT FROM AUTHOR]

Integrative taxonomic studies of macrofungal diversity in the Brijuni National Park (Istria County, Croatia) led to the discovery of a second species of Inocybe (Agaricales, Inocybaceae) new to science. Inocybe istriaca sp. nov. is described on the basis of morphological, ecological, and multigene phylogenetic analyses, and its placement within the family Inocybaceae is discussed. The combination of most important morphological characters that distinguish I. istriaca from the other similar Inocybe species are smooth, (sub)amygdaliform, (sub)phaseoliform, or ellipsoid basidiospores (ca. 8.5–12 × 5–7 μm), large basidia (36–45 × 9–15 μm), mostly (sub)fusiform and weakly thick-walled (up to 1.5 μm) metuloid pleurocystidia, and lamellar edge and stipe apex partially covered by a dark resinous substance. The species was collected on the edge of grassland and Mediterranean evergreen holm oak (Quercus ilex) forest. In this study, a total of 14 DNA sequences from four Inocybe species were generated. Two-gene (ITS, LSU) and four-gene (ITS, LSU, rpb2, tef1) phylogenetic analyses confirmed the status of I. istriaca as an independent species. [ABSTRACT FROM AUTHOR]

Ectomycorrhizas (ECM) are a common symbiotic association between fungi and various plant species in forest ecosystems, affecting community assemblages at the landscape level. ECMs benefit host plants by increasing the surface area for nutrient uptake, defending against pathogens, and decomposing organic matter in the soil. ECM-symbiotic seedlings are also known to perform better in conspecific soils than other species unable to carry the symbiosis, in a process referred to as plant-soil feedback (PSF). In this study, we tested the effects of different leaf litter amendments on ECM and non-ECM seedlings of Quercus ilex inoculated with Pisolithus arrhizus and how they altered the litter-induced PSF. Our experiment showed that the ECM symbiont induced a shift from negative PSF to positive PSF in Q. ilex seedlings by analysing plant and root growth parameters. However, non-ECM seedlings performed better than ECM seedlings in a no-litter condition, indicating an autotoxic effect when litter is present without ECM symbionts. Conversely, ECM seedlings with litter performed better at different decomposition stages, suggesting a possible role of the symbiosis of P. arrhizus and Q. ilex in recycling autotoxic compounds released from conspecific litter, transforming them into nutrients that are transferred to the plant host. [ABSTRACT FROM AUTHOR]

Acorns, frequently left uncollected in the fields, have been a part of the traditional medicine of different cultures. Among the different properties associated with them, their antimicrobial potential is of particular importance. However, this characterization has long been superficial and has not ventured into other topics such as biofilm inhibition. Thus, the current work aimed to characterize the antimicrobial and antibiofilm potential of an array of phenolic rich extracts attained from acorns, two different acorn varieties Q. ilex and Q. suber, considering the fruit and shell separately, fresh and after heat-treating the acorns to aid in the shelling process. To accomplish this, the extracts' capacity to inhibit an array of different microorganisms was evaluated, the minimum bactericidal concentration (MBC) was determined, time-death curves were drawn whenever an MBC was found and the antibiofilm potential of the most effective extracts was drawn. The overall results showed that Gram-positive microorganisms were the most susceptible out of all the microorganisms tested, with the shell extracts being the most effective overall, exhibiting bactericidal effect against S. aureus, B. cereus and L. monocytogenes as well as being capable of inhibiting biofilm formation via the two S. aureus strains. The attained results demonstrated that acorn extracts, particularly shell extracts, pose an interesting antimicrobial activity which could be exploited in an array of food, cosmetic and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Stem diameter growth in living trees refers to the invariably increase in dimension over a given period. However, reversible stem diameter decrease could occur, related to water movement in the vessels within the wood (on a daily basis) and to tree water deficit and depletion of stored water (on a seasonal basis). Recently, the perennial decrement in stem diameter size has been related to a tree decline pathway, and eventually resulting in tree death. In this study, we analysed stem diameter intra-annual growth dynamics of holm oak (Quercus rotundifolia Lam.) trees for two full growing seasons in distinct woodlands, Laborela and Aldeia dos Fernandes, in southern Portugal (Ourique district). Our focus was to assess stem diameter growth patterns and trends in holm oak trees in endangered woodlands with high tree mortality and to question if perennial decrement. Holm oaks in Laborela were much more sensitive in their stem diameter variations than in Aldeia dos Fernandes and, on average, their stem diameter decreased continuously by 1.0 mm along the 2-year study period, with a slightly higher annual decrease in the first study year (0.6 mm.yr−1). In addition, on average, trees had a higher decrease in stem sectional area of 5.8 cm2 in Laborela against a decrease of 3.7 cm2 in Aldeia dos Fernandes, where the stem diameter of holm oaks only decreased in the first study year (0.1 mm.yr−1). In each study area, the repeated- measures ANOVA showed that tree size effect influenced the stem diameter variations in contrast to tree crown defoliation. Trees were, on average, relatively smaller in Laborela, with DBH = 35.5 cm against DBH = 40.6 cm in Aldeia dos Fernandes, and were highly sensitive in their stem diameter decrement along two consecutives full growing seasons. This is the first exploratory study on assessing the stem diameter fluctuations on holm oaks to address a decline pathway. Our results revealed that holm oaks can survive during two consecutive growing seasons, with a continuous decrease in their stem diameters, which might indicate one tree's decline pathway. [ABSTRACT FROM AUTHOR]

The distribution and abundance of the various truffle species are influenced by the climate, soil, and vegetation conditions. The setting of these characteristics is necessary for the potential species' cultivation. Here, we describe the ecological characteristics of Moroccan truffles and desert truffles and their associations with host plants. We also determine the climate and soil parameters relating to the geographic distribution and fructification of truffles and desert truffles. In contrast to truffles, which are found in sub-humid environments, desert truffles are found in semi-arid and arid regions of Morocco. The dissemination of desert truffles in the Mamora forest and oriental regions of Morocco is typically linked to the presence of Helianthemum sp., whereas the existence of truffles (Tuber spp.) in the Middle Atlas depends on the subsistence of Quercus ilex and Q. faginea. The truffles' and desert truffles' fructification depends mainly on the precipitation frequency. Terfezia arenaria and Tuber oligospermum, the two major desert truffles of Mamora forest, require an annual rainfall of 435 mm on average in slightly acidic soil. While the oriental and Highland desert truffles, namely Terfezia boudieri, T. claveryi, and Tirmania spp., require an annual precipitation average of 123 to 267 mm and a high CaCO3 content. Otherwise, there is Tuber aestivum, localized in humid regions with a rainfall rate of more than 650 mm, and found under calcareous soil rich in organic matter with the presence of potential host plants, such as oaks, cedars, and pines. Our findings open up the possibility of successful cultivation of truffles and desert truffles having an economic interest through understanding their ecological requirements in Morocco. [ABSTRACT FROM AUTHOR]

Root rot affecting holm oak is a cause of high ecological and economic losses in the Iberian Peninsula, highlighting the relevance of developing disease control methods. The aim of this work was to assess the effect of a biological treatment composed of beneficial organisms (Trichoderma complex, T-complex) on holm oak seedlings infected by Phytophthora cinnamomi in two contrasted holm oak ecotypes, one considered highly susceptible (HU) and another considered tolerant to the pathogen (GR). For this purpose, a complete multifactorial test was carried out in a greenhouse, and seedlings were monitored for survival analysis and morphological and physiological attribute evaluation. Mortality began earlier in the susceptible (HU) than in the tolerant (GR) ecotype, and survival showed different trends due to the inoculation with beneficial microorganisms depending on the ecotype of the plants. The tolerant ecotype showed a high survival rate and better response to the treatment with beneficial microorganisms. GLM showed that the main reason for differences between treatments was ecotype, followed by T-complex and irrigation, and a weak interaction between ecotype and P. cinnamomi was found. The linear relationship between photosynthesis (A) and transpiration (Tr) showed an increase in the A/Tr rates for infected and inoculated plants under drought conditions for the GR ecotype. The tolerant ecotype was benefited more by the beneficial microorganism treatment. The understanding of the genetic diversity of Q. ilex and water stress influence on the efficacy of biological treatments against root rot provides useful information to develop environmentally friendly disease control methods to address the holm oak decline. [ABSTRACT FROM AUTHOR]

This study identified the compounds obtained from four native Dehesa plants, which were holm oak, elm, blackberry and white rockrose, and evaluated their ability to inhibit the growth and production of aflatoxins B1 and B2 of two strains of mycotoxigenic Aspergillus flavus. For this purpose, phenolic compounds present in the leaves and flowers of the plants were extracted and identified, and subsequently, the effect on the growth of A. flavus, aflatoxin production and the expression of a gene related to its synthesis were studied. Cistus albidus was the plant with the highest concentration of phenolic compounds, followed by Quercus ilex. Phenolic acids and flavonoids were mainly identified, and there was great variability among plant extracts in terms of the type and quantity of compounds. Concentrated and diluted extracts were used for each individual plant. The influence on mold growth was not very significant for any of the extracts. However, those obtained from plants of the genus Quercus ilex, followed by Ulmus sp., were very useful for inhibiting the production of aflatoxin B1 and B2 produced by the two strains of A. flavus. Expression studies of the gene involved in the aflatoxin synthesis pathway did not prove to be effective. The results indicated that using these new natural antifungal compounds from the Dehesa for aflatoxin production inhibition would be desirable, promoting respect for the environment by avoiding the use of chemical fungicides. However, further studies are needed to determine whether the specific phenolic compounds responsible for the antifungal activity of Quercus ilex and Ulmus sp. produce the antifungal activity in pure form, as well as to verify the action mechanism of these compounds. [ABSTRACT FROM AUTHOR]

Climate extremes such as cold spells are becoming more frequent as climate variability increases. However, few studies have evaluated the impacts of winter cold spells on forest cover, tree growth and leaf and sapwood non-structural carbohydrate (NSC) concentrations. We analyzed changes in tree cover using remote sensing data and compared the radial growth of coexisting and defoliated Pinus halepensis trees and non-defoliated P. halepensis and Pinus pinaster trees. We also compared NSC concentrations in leaves and sapwood of defoliated and non-defoliated P. halepensis and Quercus ilex trees. In January 2021, a rapid drop in temperatures led to minimum values (−21.3 °C) in eastern Spain and triggered canopy defoliation in several planted (P. halepensis) and native (Q. ilex) tree species. The cold spell led to a decrease in forest cover in the most defoliated stands and reduced radial growth of defoliated P. halepensis and sapwood NSC concentrations in P. halepensis and Q. ilex, particularly starch. Prior to the cold spell, defoliated P. halepensis trees significantly (p < 0.05) grew more (2.73 ± 1.70 mm) in response to wetter winter conditions than non-defoliated P. halepensis (2.29 ± 1.08 mm) and P. pinaster (1.39 mm) trees. Those P. halepensis individuals which grew faster at a young age were less resilient to the winter cold spell in later years. The study stands showed a high recovery capacity after the cold spell, but the Mediterranean drought-avoiding P. halepensis was the most affected species. [ABSTRACT FROM AUTHOR]

Firewood extraction by mule forwarding is still common in oak coppices in Central and Southern Italy. This is due to the scarce presence of aerial extraction systems such as cable yarders. Considering the importance of forest soil for all ecosystem services, the evaluation of the disturbance that a given extraction system has on the forest soil is a fundamental aspect in the framework of sustainable forest management. Therefore, this study was developed to assess the disturbance caused to the physicochemical and biological features of soil and to coppice after mule logging according to the standards of silvicultural treatment, as well as the recovery time needed after the logging intervention. Four cutting blocks located in Central Italy represented the study area, one cutting block represented the unharvested control, while the others were logged 3 years (CB-2019), 8 years (CB-2014) and 10 years (CB-2012) prior to the field surveys. In each harvested cutting block the soil was subdivided into disturbed soil (DIST—mule trails) and low disturbance soil (LD—area within the harvested cutting block not affected by mule passage). This experimental design assessed the disturbance caused by logging operations by mules (DIST soil) and the silvicultural treatment (LD soil) to soil physicochemical (bulk density, penetration resistance, shear resistance, and soil organic matter) and biological properties (soil microarthropod community evaluated with the QBS-ar index). The results revealed a significant disturbance in the mule trails for all the investigated variables. The disturbance was particularly strong for the QBS-ar index, with values which were lower than half of those of the control area. Furthermore, no recovery process was evident even after 10 years from the logging interventions. Instead, values of the various parameters became worse with time after harvesting. On the other hand, no marked disturbance was revealed in LD soil, except for a significant decrease in soil organic matter. Although this is a preliminary evaluation that needs to be confirmed with further study, this trial suggested that mule logging cannot be considered a fully low-impact approach to forest operations and that studies with a longer time span after harvesting are needed to assess the recovery process in the mule trails. [ABSTRACT FROM AUTHOR]

Holm oak is the dominant tree species in the Mediterranean climate. Currently, worrisome degradation of its ecosystems has been observed, produced, among other factors, by changes in land use, extreme weather events, forest fires, climate change, and especially the increasingly frequent episodes of high tree mortality caused by "oak decline", which has brought with it a social concern that transcends the productive interest. Breeding and conservation programs for this species are necessary to ensure the prevalence of these ecosystems for future generations. Biotechnological tools such as somatic embryogenesis (SE) have great potential value for tree improvement and have been shown to be highly efficient in the propagation and conservation of woody species. One challenge to this approach is that SE induction in holm oak has not yet been optimized. Here, we present a new reproducible procedure to induce SE in holm oak; we evaluated the responsiveness of different initial explants exposed to different types, concentrations, and durations of auxin. SE rates were significantly improved (37%) by culturing nodal segments for two weeks in induction medium. In addition, a significant auxin–genotype interaction was observed. [ABSTRACT FROM AUTHOR]

The accumulation in the leaves and young stems of phenolic compounds, such as hydrolyzable and condensed tannins, constitutes a defense mechanism of plants against herbivores. Among other stressing factors, chronic herbivory endangers Quercus ilex, a tree playing a central role in Mediterranean forests. This work addressed the connections between the chemical defenses of Q. ilex leaves and their susceptibility to herbivory, quantitative traits whose relationships are modulated by environmental and genetic factors that could be useful as molecular markers for the selection of plants with improved fitness. A search for natural variants detected the polymorphism D165H in the effector domain of QiMYB-like-1, a TT2-like transcription factor whose family includes members that control the late steps of condensed tannins biosynthesis in different plant species. QiMYB-like-1 D165H polymorphism was screened by PCR-RFLP in trees from six national parks in Spain where Q. ilex has a relevant presence, revealing that, unlike most regions that match the Hardy-Weinberg equilibrium, homozygous plants are over-represented in "Monfragüe" and "Cabañeros", among the best examples to represent the continental Mediterranean (cM) ecosystem. Accordingly, the averages of two stress-related quantitative traits measured in leaves, herbivory index and accumulation of condensed tannins, showed asymmetric distributions depending on the clustering of trees based on ecological and genetic factors. Thus, the impact of herbivory was greater in managed forests with a low density of trees from the cM region, among which QiMYB-like-1 D165 homozygotes stand out, whereas condensed tannins accumulation was higher in leaves of QiMYB-like-1 H165 homozygotes from low-density forests, mainly in the Pyrenean (Py) region. Besides, the correlation between the contents of condensed tannins and total tannins vanished after clustering by the same factors: the cM region singularity, forest tree density, and QiMYB-like-1 genotype, among which homozygous shared the lowest link. The biogeographical and genetic constraints that modulate the contribution of condensed tannins to chemical defenses also mediated their interactions with the herbivory index, which was found positively correlated with total phenolics or tannins, suggesting an induction signal by this biotic stress. In contrast, a negative correlation was observed with condensed tannins after tree clustering by genetics factors where associations between tannins were lost. Therefore, condensed tannins might protect Q. ilex from defoliation in parks belonging to the cM ecosystem and carrying genetic factor(s) linked to the QiMYB-like-1 D165H polymorphism. [ABSTRACT FROM AUTHOR]

The intra- and interdomain phyllosphere microbiome features of Quercus ilex L. in a Mediterranean context is reported. We hypothesized that the main driver of the phyllosphere microbiome might be the season and that atmospheric pollutants might have a co-effect. Hence, we investigated the composition of epiphytic bacteria and fungi of leaves sampled in urban and natural areas (in Southern Italy) in summer and winter, using microscopy and metagenomic analysis. To assess possible co-effects on the composition of the phyllosphere microbiome, concentrations of particulate matter and polycyclic aromatic hydrocarbons (PAHs) were determined from sampled leaves. We found that environmental factors had a significative influence on the phyllosphere biodiversity, altering the taxa relative abundances. Ascomycota and Firmicutes were higher in summer and in urban areas, whereas a significant increase in Proteobacteria was observed in the winter season, with higher abundance in natural areas. Network analysis suggested that OTUs belonging to Acidobacteria, Cytophagia, unkn. Firmicutes(p), Actinobacteria are keystone of the Q. ilex phyllosphere microbiome. In addition, 83 genes coding for 5 enzymes involved in PAH degradation pathways were identified. Given that the phyllosphere microbiome can be considered an extension of the ecosystem services offered by trees, our results can be exploited in the framework of Next-Generation Biomonitoring. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The integrated analysis of different omic layers can provide new knowledge not provided by their individual analysis. This approach is also necessary to validate data and reveal post-transcriptional and post-translational mechanisms of gene expression regulation. In this work, we validated the possibility of applying this approach to non-model species such as Quercus ilex. Transcriptomics, proteomics, and metabolomics from Q. ilex seedlings subjected to drought-like conditions under the typical summer conditions in southern Spain were integrated using a non-targeted approach. Two integrative approaches, PCA and DIABLO, were used and compared. Both approaches seek to reduce dimensionality, preserving the maximum information. DIABLO also allows one to infer interconnections between the different omic layers. For easy visualization and analysis, these interconnections were analyzed using functional and statistical networks. We were able to validate results obtained by analyzing the omic layers separately. We identified the importance of protein homeostasis with numerous protease and chaperones in the networks. We also discovered new key processes, such as transcriptional control, and identified the key function of transcription factors, such as DREB2A, WRKY65, and CONSTANS, in the early response to drought. [ABSTRACT FROM AUTHOR]

The flowering period of plants is a critical time since it determines their reproductive success. Flowering is controlled by different factors including genetic regulation and environmental conditions. In the Mediterranean area, favourable conditions usually occur in spring, when most plant species flower including those of the Mediterranean Quercus genus. This paper reveals and analyses an unusual and lesser-known phenomenon occurring in the two main Mediterranean agroforestry ecosystems of South Europe, the Mediterranean forest and "dehesa", that is, a second flowering occurring in autumn for the species Quercus ilex subsp. ballota (holm oak). The continuous pollen monitoring of the atmosphere in the city of Cordoba (southern Spain) for 25 years, together with field phenological observations in the area, has indicated that, apart from the main pollination period in spring, secondary flowerings also occasionally occur in this area, specifically in autumn. The present work examines these uncommon pollination events detected in the autumns of certain years with the aim of determining the main environmental factors that influence and control them. During the 25-year study period, there were 7 years in which a secondary Quercus flowering was detected in the area from the second half of October until the end of November. The univariate statistical analysis of the influence of environmental variables determined that the meteorological conditions in September were the most influential. Low mean temperatures, together with record rainfall in that month, led to autumn flowering events. The phenological characteristics of the spring pollen season were also influential. In the years with a shorter spring, the Quercus pollen season tended to present autumn flowerings. A multivariate adaptive regression splines (MARS) model was built to explain the effects of the different variables on the occurrence of autumn pollination. The results indicated that the combined effect of three predicting variables, September rainfall, the length of the spring pollen season, and the end of the spring pollen season, explained 92% of the variance. The validation showed a strong relationship between the expected and the observed autumn pollen concentrations. Therefore, the present analysis of a long-term pollen database revealed that the main causes of this unusual second flowering in autumn were strongly related to climate change, i.e., strong dry summers and warm autumns. In addition, the results showed that the phenomenon was more frequent in the years with low pollination during spring due to different meteorological events potentiated by climate change, such as dryness or heavy rain episodes, as a way of ensuring acorn crops. The results explain how this unusual and lesser-known phenomenon in agroforestry dynamics is related to the adaptation to climate change and the main factors that are driving it, as well as the potential consequences for these important and endangered Mediterranean ecosystems. [ABSTRACT FROM AUTHOR]

The effect of four lignocellulosic waste fillers on the thermal and mechanical properties of biocomposites was investigated. Powdered licorice root, palm leaf, holm oak and willow fillers were melt compounded with polypropylene at two different weight contents, i.e., 10 and 30, and then injection molded. A commercially available maleated coupling agent was used to improve the filler/matrix interfacial adhesion at 5 wt.%. Composites were subjected to chemical (FTIR-ATR), thermal (TGA, DSC, DMA) and mechanical (tensile, bending and Charpy impact) analyses coupled with a morphological investigation by scanning electron microscopy. Although similarities among the different formulations were noted, holm oak fillers provided the best combination of thermal and mechanical performance. In particular, at 30 wt.% content with coupling agent, this composite formulation displayed remarkable increases in tensile strength and modulus, flexural strength and modulus, of 28% and 110%, 58% and 111%, compared to neat PP, respectively. The results imply that all these lignocellulosic waste fillers can be used successfully as raw materials for biocomposites, with properties comparable to those featured by other natural fillers. [ABSTRACT FROM AUTHOR]

Holm oak (Quercus ilex subsp. ballota (Desf.) Samp.) bark is a commonly used remedy to treat gastrointestinal disorders, throat and skin infections, hemorrhages, and dysentery. It has also been previously reported that its methanol extracts possess antibacterial activity, which can be related to the richness of Quercus spp. extracts in phenolic compounds, such as flavonoids and tannins. However, there is no information on the antifungal (including oomycete) properties of the bark from Q. ilex or its subspecies (ilex and ballota). In this work, we report the characterization of the aqueous ammonia extract of its bark by FTIR and GC-MS and the results of in vitro and ex situ inhibition tests against three phytopathogens. The main phytochemical components identified were inositols (19.5%), trans-squalene (13%), 4-butoxy-1-butanol (11.4%), gulopyranose (9.6%), lyxose (6.5%), 2,4-dimethyl-benzo[H]quinoline (5.1%), catechol (4.5%), and methoxyphenols (4.2%). The efficacy of the extract in controlling forest phytopathogens was tested in vitro against Fusarium circinatum (responsible for pitch canker of Pinus spp.), Cryphonectria parasitica (which causes chestnut blight), and Phytophthora cinnamomi (which causes 'root and crown rot' in a variety of hosts, including Castanea, conifers, Eucalyptus, Fagus, Juglans, Quercus, etc.), obtaining EC90 values of 322, 295, and 75 μg·mL−1, respectively, much lower than those attained for a commercial strobilurin fungicide (azoxystrobin). The extract was further tested ex situ against P. cinnamomi on artificially inoculated, excised stems of 'Garnem' almond rootstock, attaining complete protection at a dose of 782 μg·mL−1. The results suggest that holm oak bark extract may be a promising source of bioactive compounds against invasive forest pathogens, including the oomycete that is causing its decline, the so-called 'seca' in Spain. [ABSTRACT FROM AUTHOR]

The Mediterranean vegetation belt on the eastern Adriatic covers an area of nearly 15,000 km2. It is comprised of forest stands that can be divided into three vegetation zones based on the presence of certain plant species within each: sub-Mediterranean, eu-Mediterranean, and steno-Mediterranean. The dominant ecological factors result in the domination of specific tree species within the floral composition between these vegetation zones. The aim of this study was to collect climate data from 38 weather stations over a 30-year period to compare climate data and bioclimate properties in the area of these three vegetation zones. The results confirmed statistically significant differences between the main climatic elements and most bioclimatic indices between the vegetation zones. Cooler and more humid conditions were found in the sub-Mediterranean zone, warmer and somewhat drier conditions in the eu-Mediterranean zone, and particularly pronounced warm and dry conditions in the steno-Mediterranean zone. However, the analysis of the main components for researching climate parameters showed that the mean annual air temperature, average minimum air temperature of the coldest month of the year and continentality index, length of the dry season, and de Martonne aridity index contribute the most to the grouping of vegetation in forest stands in the Mediterranean vegetation belt of the eastern Adriatic. [ABSTRACT FROM AUTHOR]

In Mediterranean ecosystems, the projected rainfall reduction of up to 30% may alter plant–soil interactions, particularly litter decomposition and Home Field Advantage (HFA). We set up a litter transplant experiment in the three main forests encountered in the northern part of the Medi-terranean Basin (dominated by either Quercus ilex, Quercus pubescens, or Pinus halepensis) equipped with a rain exclusion device, allowing an increase in drought either throughout the year or concentrated in spring and summer. Senescent leaves and needles were collected under two precipitation treatments (natural and amplified drought plots) at their "home" forest and were left to decompose in the forest of origin and in other forests under both drought conditions. MS-based metabolomic analysis of litter extracts combined with multivariate data analysis enabled us to detect modifications in the composition of litter specialized metabolites, following amplified drought treatment. Amplified drought altered litter quality and metabolomes, directly slowed down litter decomposition, and induced a loss of home field (dis)advantage. No indirect effect mediated by a change in litter quality on decomposition was observed. These results may suggest major alterations of plant–soil interactions in Mediterranean forests under amplified drought conditions. [ABSTRACT FROM AUTHOR]

Hybridization and introgression between cork oak (Quercus suber) and holm oak (Q. ilex) have traditionally been reckoned as undesirable processes, since hybrid individuals lack the profitable bark characteristics of cork oak. Nevertheless, a systematic and quantitative description of the bark of these hybrids at the microscopic level, based on a significant number of individuals, is not available to date. In this work we provide such a qualitative and quantitative description, identifying the most relevant variables for their classification. Hybrids show certain features intermediate between those of the parent species (such as phellem percentage in the outer bark, which was approximately 40% as a mean value for hybrids, 20% in holm oak and almost 99% in cork oak), as well as other unique features, such as the general suberization of inactive phloem (up to 25% in certain individuals), reported here for the first time. These results suggest a relevant hybridization-induced modification of the genetic expression patterns. Therefore, hybrid individuals provide a valuable material to disentangle the molecular mechanisms underpinning bark development in angiosperms. [ABSTRACT FROM AUTHOR]

The holm oak (Quercus ilex L.) is the dominant tree species of the Mediterranean forest and the Spanish agrosilvopastoral ecosystem, "dehesa." It has been, since the prehistoric period, an important part of the Iberian population from a social, cultural, and religious point of view, providing an ample variety of goods and services, and forming the basis of the economy in rural areas. Currently, there is renewed interest in its use for dietary diversification and sustainable food production. It is part of cultural richness, both economically (tangible) and environmentally (intangible), and must be preserved for future generations. However, a worrisome degradation of the species and associated ecosystems is occurring, observed in an increase in tree decline and mortality, which requires urgent action. Breeding programs based on the selection of elite genotypes by molecular markers is the only plausible biotechnological approach. To this end, the authors' group started, in 2004, a research line aimed at characterizing the molecular biology of Q. ilex. It has been a challenging task due to its biological characteristics (long life cycle, allogamous, high phenotypic variability) and recalcitrant nature. The biology of this species has been characterized following the central dogma of molecular biology using the omics cascade. Molecular responses to biotic and abiotic stresses, as well as seed maturation and germination, are the two main objectives of our research. The contributions of the group to the knowledge of the species at the level of DNA-based markers, genomics, epigenomics, transcriptomics, proteomics, and metabolomics are discussed here. Moreover, data are compared with those reported for Quercus spp. All omics data generated, and the genome of Q. ilex available, will be integrated with morphological and physiological data in the systems biology direction. Thus, we will propose possible molecular markers related to resilient and productive genotypes to be used in reforestation programs. In addition, possible markers related to the nutritional value of acorn and derivate products, as well as bioactive compounds (peptides and phenolics) and allergens, will be suggested. Subsequently, the selected molecular markers will be validated by both genome-wide association and functional genomic analyses. [ABSTRACT FROM AUTHOR]

Forest ecosystems are some of the largest carbon (C) reservoirs on earth. Pinus halepensis Mill., Quercus ilex L. and Quercus pubescens Willd. represent the dominant tree cover in the Mediterranean forests of southern France. However, their contributions to the French and global forest C and nitrogen (N) stocks are frequently overlooked and inaccurately quantified and little is known about to what extent the ongoing climate change can alter these stocks. We quantified the soil organic C (SOC) and N (SN) stocks in Mediterranean forests dominated by these tree species and evaluated to what extent an experimental precipitation reduction (about −30% yearly) affects these stocks and the litter decomposition efficiency. Litter mass losses were 55.7, 49.8 and 45.7% after 24 months of decomposition in Q. ilex, Q. pubescens and P. halepensis forests, respectively, and were 19% lower under drier climatic conditions. The SOC stocks were 14.0, 16.7 and 18.5 Mg ha−1 and the SN stocks were 0.70, 0.93 and 0.88 Mg ha−1 in Q. ilex, Q. pubescens and P. halepensis forests, respectively. The shallowness and stoniness of these Mediterranean forests could explain these limited stocks. By distinguishing the organic from the organo–mineral layer, we showed 74% less SOC in the organic layer of the P. halepensis forest under drier conditions, while no difference was detected in the organo–mineral layer or in the two oak forests. This last finding deserves further investigation and points out the necessity to distinguish the organic from the organo–mineral layer to detect the first impacts of climate change on SOC stocks. [ABSTRACT FROM AUTHOR]

Land-use change is the main driver of biodiversity loss in the Mediterranean basin. New socio-economic conditions produced a rewilding process so that cultural landscapes are being invaded by more natural habitats. We analyze the effects of landscape change on the demography and the spatial distribution of Crocidura russula in six protected areas of the western Mediterranean basin. The study was conducted in the period 2008–2020 on 19 live trapping plots representing the three main natural habitats of the area (scrubland, pinewood, and holm oak woodland). We used a multiscale approach to ensure that the scale of response matched landscape structure (from plot to landscape) using either vegetation profiles (LiDAR) and land use data obtained from years 2007 and 2017. Statistical models (multiple-season single-species occupancy models) showed that C. russula populations were strongly associated to habitat features at the plot level. These models were used to predict occupancy at sampling units for the whole study area (850 km2), showing contrasting trends that shifted at relatively small spatial scales (expansions and retractions of species ranges). Parks showing extreme scrubland encroachment (−8% of area) and afforestation (+6%) significantly reduced habitat suitability for shrews and reductions in occupancy (−5%). Results would indicate faster changes in the spatial distribution of the target species than previously expected on the basis of climate change, driven by fast landscape changes. [ABSTRACT FROM AUTHOR]

Climatic conditions are extremely important for the start and spread of forest fires. Flammability and the spread of fire are sensitive to the interactions of variables within ecosystems, such as weather, fuel, and topography. Certain variables are highly significant for forest fires and are applied in various models, particularly the moisture content of potential fuel, and its flammability or combustibility. However, such these models cannot determine the true values of the required variables, unlike actual empirical research. Areas with the highest number of fires show significant variability and differences in air temperature, humidity, and precipitation. These factors have a strong influence on flammability, combustibility, and moisture content of Mediterranean species. This study analyses the flammability and combustibility of two Mediterranean species important for the Adriatic area in Croatia: climazonal holm oak (Quercus ilex L.) and Aleppo pine (Pinus halepensis Mill.) as an important conifer for afforestation and reforestation. The results of linear correlation coefficients of flammability of these species at the studied localities show a statistically significant and very strong correlation between flammability and the moisture content of the tested sample, with the exception of Aleppo pine on the island of Rab. The analysis of variance of combustibility showed that there were no statistically significant differences between nearly all variables tested. The results indicate the significant effect of live fuel moisture content on the flammability and combustibility of natural fuels. [ABSTRACT FROM AUTHOR]

To verify the responses of visible foliar injury (VFI), we exposed seedlings of three oak species for 4.5 months in an open air facility, using differing ozone (O3) and drought treatments: O3 (three levels from ambient to ×1.4 ambient), and drought (three levels of irrigation from 40% to 100% field capacity). We related the accumulated phytotoxic O3 dose (POD1) and cumulative drought index (CDI) to the O3 and drought VFI and assessed growth increment (height, diameter, leaf number), biomass (of all organs), and physiological parameters: net photosynthesis per plant (Pn), photosynthetic nitrogen (PNUE) and phosphorus use efficiency (PPUE)). The results indicated that an increase in POD1 promoted O3 VFI in Quercus robur and Quercus pubescens, while Quercus ilex was asymptomatic. The POD1-based critical level at the onset of O3 VFI was lower for Q. robur than for Q. pubescens (12.2 vs. 15.6 mmol m−2 POD1). Interestingly, drought reduced O3 VFI in Q. robur but increased it in Q. pubescens. Both O3 and drought were detrimental to the plant biomass. However, Q. robur and Q. pubescens invested more in shoots than in roots, while Q. ilex invested more in roots, which might be related to a hormetic mechanism. Pn, PNUE and PPUE decreased in all species under drought, and only in the sensitive Q. robur (PPUE) and Q. pubescens (PNUE) under O3. This study confirms that POD1 is a good indicator to explain the development of O3 VFI and helps a differential diagnosis of co-occurring drought and O3 VFI in oak forests. [ABSTRACT FROM AUTHOR]

Afforestation can improve hydrological processes, such as infiltration, in basins and, therefore, reduce the impact on human populations of floods, soil erosion, landslides, droughts, and climate variation. The aim of this work was to analyze how afforestation and other changes in land use influence infiltrability and the evolution of soils. Infiltration rates, soil water repellency, and physical and chemical properties of sandy loam soils were measured in four types of land: native holm oak forest, afforested 20-year-old pine forest, shrubs, and grasslands. Non-forest covers are the result of the degradation of native oak forests for centuries, while the pine afforestation in this study took place on a perennial wet mountain pasture (cervunalito). Our results show that soil infiltration rates are much higher in pine afforestation areas (857.67 mm·h−1) than in holm oak forest (660.67 mm·h−1), grasslands (280.00 mm·h−1), or shrubs (271.67 mm·h−1). No statistically significant differences in fertility, organic matter content, bulk density, or effective porosity were found between afforestation areas and other types of cover; however, pine afforestation improved the drainage of the soil, as its infiltration rate was higher than that of the native holm oak forest. [ABSTRACT FROM AUTHOR]

Holm oak populations are severely affected by oak decline syndrome, and reliable methods of conserving the plant material are required. A vitrification-based cryopreservation method was used for the first time for the long-term conservation of holm oak embryogenic cultures. Successful cryopreservation was achieved after determining the best developmental stage of the somatic embryos used and the optimal incubation period in plant vitrification solution 2 (PVS2). Embryos were recovered from individual nodular embryogenic structures (NES) derived from four embryogenic lines after preculture on a medium containing 0.3 M sucrose, incubation in PVS2 vitrification solution for 15 min at 25 °C and direct immersion in liquid nitrogen (LN). Embryo recovery rates of 16.7–63.3% were obtained after cryostorage for four years in LN. In addition to the embryo developmental stage and the PVS2 treatment time, the genotype can also significantly affect embryo recovery after LN storage. There were no significant differences in plant regeneration or polyploid stability between somatic embryos and plants derived from control embryos (not cryopreserved) and cryopreserved embryos. The findings indicate that embryo proliferation, plant conversion and polyploid stability are maintained in material recovered from the vitrification solution and subsequently cryopreserved. [ABSTRACT FROM AUTHOR]