Quercus Pubescens Research Articles

Mapping Coverage and Typology Based on Function and Spatial Configuration of Forests in Latium Region, Central Italy

Among the land use–land cover products, tree cover maps are essential tools for assessing forest functionality and ecosystem services, and implementing sustainable forest management. By combining open-source and ancillary high-resolution cartographic datasets, this study aims to map trees and forests in the Latium region in central Italy, highlighting their spatial configuration, function, and forest typology. The main findings show that trees cover 44.2% of the regional land area. Forests cover 508,056 ha, forming the core matrix of the Latium mountain landscape, providing significant ecological and socio-economic value for forest management and the regional wood supply chain. Although trees outside the forest represent only 3.1% of regional tree cover, they play a crucial role in enhancing ecological connectivity and landscape resilience. Approximately 2% of the tree and forest cover occurs in urban areas, contributing significantly to climate regulation and air quality in densely populated environments. The dominant forest types in Lazio include Turkey oak, temperate broadleaf, beech, downy oak, and Holm oak, which together account for 58.6% of the total tree cover. The accuracy tests confirm the feasibility of using open-source data for reliable, cost-effective forest mapping. Regular updates of these maps can support continuous monitoring and promote sustainable forest management practices.

Phytochemical and Bioactivity Evaluation of Bee Pollen and Androecia of Castanea, Salix, and Quercus Species.

Qualitative and quantitative differences in the chemical composition between bee pollen originated from Castanea sativa (Türkiye and Slovenia), Salix spp. (Türkiye and Slovenia), and Quercus spp. (Türkiye) and androecia of Castanea sativa, Salix alba, and Quercus pubescens (apetalous trees) were evaluated for the first time by new high-performance thin-layer chromatography (HPTLC) and ultra-performance liquid chromatography (UPLC) methods using marker compounds. N1,N5,N10-tricaffeoylspermidine was isolated, and its structure was elucidated by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). It was the main and the marker compound common to bee pollen (≈3-41 mg/g) and androecia (≈3-6 mg/g) samples. To the best of our knowledge, this is the first report of the identification of N1,N5,N10-tricaffeoylspermidine in bee pollen originated from Salix spp. and androecia of C. sativa, S. alba, and Q. pubescens. The botanical origins of bee pollen were determined via phytochemical profiling using HPTLC-image analyses showing that bee pollen from the same botanical source had almost identical profiles regardless of collection location, geographical differences, and the bee race. In vitro tests and HPTLC-effect-directed analyses (EDAs) were performed to assess antioxidant and xanthine oxidase (XO) inhibitory activities of bee pollen, androecia, and N1,N5,N10-tricaffeoylspermidine. HPTLC-EDA combined with image analyses was used for comparing the activities of bee pollen, androecia, N1,N5,N10-tricaffeoylspermidine, and also other marker compounds (quercetin, myricitrin, hyperoside, quercitrin, isoquercitrin, and rutin). The remarkable bioactivity of N1,N5,N10-tricaffeoylspermidine was for the first time evaluated by HPTLC-EDA and in vitro tests. This is the first study performing HPTLC-XO inhibitory activity analyses on the HPTLC NH2 F254S plates. Further bioactivity studies on botanically and chemically well-characterized bee pollen samples are needed to aid in the use of bee pollen-containing supplements in the prevention and treatment of diseases.

The influence of post-glacial migration and hybridization on the gene pool of marginal Quercus pubescens populations in Central Europe.

In Central Europe, the drought-tolerant downy oak (Quercus pubescens) is at the northern edge of its natural distribution range, often growing in small and spatially isolated populations. Here, we elucidate how the population genetic structure of Central European Q. pubescens was shaped by geographic barriers, genetic drift and introgression with the closely related sessile oak (Q. petraea). 27 Q. pubescens populations from the northern margin of Q. pubescens' natural distribution range were sampled. Based on 16 nuclear microsatellite markers (nSSRs), Bayesian clustering and distance-based analyses were performed to determine the intraspecific genetic structure and to identify genetic barriers. To identify drivers of introgression with Q. petraea, generalised linear models were applied to link levels of introgression with environmental conditions. To track post-glacial migration routes, the spatial distribution of haplotypes based on 8 chloroplast microsatellite markers (cpSSRs) was investigated. Based on nSSRs, the study populations of Q. pubescens were divided into a western and an eastern genetic cluster. Within these clusters, more pronounced genetic substructure was observed in the west, probably due to a rugged topography and limited gene flow. Introgression from Q. petraea was more prevalent at wetter and north-exposed sites and in the west. The identified cpSSR haplotypes followed known migration pathways. Our results suggest two late-glacial refugia in or near the southwestern Alps and the southeastern Alps as potential sources for post-glacial migration. Although some genetic exchange is evident in Northern Italy, south of the Alps, the two clusters remain distinct at a large scale. Landscape features and introgression with Q. petraea shaped the genetic substructure at a smaller scale. Our study provides a comprehensive overview of the genetic structure of Q. pubescens in Central Europe, relevant for conservation.

Differentiation and Taxonomic Identification of Roburoid Oaks in the Caucasian and Crimean Regions Using Nuclear Microsatellite Markers

The inter- and intra-specific structure of genetic variability was studied using 18 microsatellite loci (nSSRs) in closely related roburoid oaks in the Crimean-Caucasian region.The seven most widespread Quercus taxa in the region were studied in 29 morphologically pure populations from different parts of the North Caucasus, Transcaucasia, Crimea and northeastern Europe. Most taxa were studied using nSSR markers for the first time. Among the 492 trees studied, Bayesian clustering method implemented in STRUCTURE identified clusters corresponding to the pedunculate oak Quercus robur, the Hartwiss oak Q. hartwissiana, the Caucasian oak Q. macranthera, the downy oak Q. pubescens and three subspecies of sessile oak: Q. petraea ssp. petraea, Q. petraea ssp. iberica, Q. petraea ssp. medwediewii. Geographic structure was identified within Q. robur, Q. pubescens and Q. p. ssp. petraea. The 18 nSSR loci used are efficient in the taxonomic assignment of individuals, and identifying hybrids. The close relationship between the “long-pedunculate” roburoid oaks (Q. robur and Q. hartwissiana) is shown, with a greater difference from other species. For one of the subspecies of sessile oak, widespread in the North Caucasus and Crimea Q. petraea ssp. medwediewii (syn. Q. calcarea), or limestone oak, significant differences from other taxa were found, reaching the species level. The assumption of a possible hybrid origin of this taxon as a result of hybridization of Q. petraea and Q. pubescens is not confirmed by genetic analysis. The other two subspecies of Q. petraea (Q. p. ssp. petraea and Georgian oak Q. p. ssp. iberica) are differentiated to a lesser extent and are related to each other, which confirms the legitimacy of distinguishing two geographically isolated taxa at the rank of subspecies. The highest variability was observed in Q. pubescens (He = 0.777). In Q. p. ssp. medwediewii variability was lower than in other widespread taxa (He = 0.652), and was approximately at the level of variability of Q. hartwissiana (He = 0.633) and Q. macranthera (He = 0.659). Clear differentiation of taxa by nuclear markers shows the limited introgression in closely related oak species in the Caucasus and Crimea. The identified genetic clusters can be used as reference groups for further population genetic studies of oaks in the Crimean-Caucasian region.

Reduced soil moisture drives leaf anatomical shifts more than chronically elevated temperatures in European temperate trees.

Chronic reductions in soil moisture combined with high air temperatures can modify tree carbon and water relations. However, little is known about how trees acclimate their foliar structure to the individual and combined effects of these two climate drivers. We used open-top chambers to determine the multi-year effects of chronic air warming (+5 °C) and soil moisture reduction (-50%) alone and in combination on the foliar anatomy of two European tree species. We further investigated how these climate drivers affected the relationship between foliar anatomy and physiology/chemistry in young downy oak and European beech trees. After 4 years, reduced soil moisture led to development of thinner leaves with a narrower epidermis and lower gas exchange for oak and beech, but to a lesser extent in the latter. In contrast, prolonged warming did not affect the anatomical and physiological/chemical traits in either species. Warming also did not exacerbate the impacts of dry soils, highlighting soil moisture as the key driver in leaf anatomical shifts. While soil moisture altered oak foliar anatomy, and the physiology and chemistry of both species, our work revealed a limited acclimation potential towards more drought- and heat-tolerant leaves as conditions become drier and warmer, suggesting potentially high vulnerability of both species to future climate predictions.

First records of moss-feeding aphids (Hemiptera: Sternorrhyncha: Aphididae) from Hungary

Four species of moss-feeding aphids were captured by Berlese samples collected in Heves county, NE Hungary. These data are establishing the first Hungarian records of moss-dwelling aphids, all the four species and three genera are reported from the country for the first time. Muscaphis (Aspidaphium) eschirichi (Börner, 1939) was yielded from a colony of Palustriella commutata (Hedw.) Ochyra collected from rock in a karst spring outlet, Muscaphis (Muscaphis) musci Börner, 1933 was yielded from colonies of Hypnum cupressiforme Hedw. collected from bark of Quercus pubescens Wild. in an open karst forest, Pseudacaudella rubida (Börner, 1939) was yielded from both of the above mosses, while a Myzodium sp. was yielded from a colony of Hypnum cupressiforme mixed with a single stem of Brachythecium salebrosum (Hoffm. ex F.Weber & D.Mohr) Schimp., collected from bark of dead trunk of Quercus cerris L. in a mixed deciduous forest. All moss species are new host records for the corresponding species. Myzodium sp. explain some distinctive characters different from the only European species of its genus and probably belong to an undescribed species, but we are avoiding of describing it on the basis of the single adult captured. Distinctive morphological and biological characteristics of the Hungarian specimens of the four aphid species are discussed and illustrated. A list of the accompanying taxa yielded from the Berlese samples are listed in a table.

Decline of soil volatile organic compounds from a Mediterranean deciduous forest under a future drier climate

Biogenic volatile organic compounds (BVOCs) are crucial for ecosystem functioning, atmospheric chemistry and climate. While modulation of BVOC emissions from living vegetation with biotic and abiotic factors is well documented, how these factors drive soil BVOC emissions remain less understood, particularly in Mediterranean forests. To fill this gap, this pioneer study investigates whether BVOC fluxes from natural soil covered by litter (referred to as forest soil) vary under natural and amplified long-term water stress (35% annual rain exclusion over the past 10 years) in a deciduous oak Mediterranean forest (Quercus pubescens Willd.) compared to natural climate conditions. This aim has only been tackled in a single evergreen Mediterranean forest so far. Using proton transfer reaction time of flight mass spectrometer (PTR-ToF-MS) we also provide, for the first time, a detailed diurnal cycle of soil BVOCs in relation to air temperature, air humidity, and biotic factors including soil respiration and litter content in lignin, cellulose and hemicellulose. The main results revealed that forest soil represents a source of most BVOCs (e.g., acetaldehyde, acetone, acrolein, hexanol, monoterpenes) with maximum values at mid-day (42 μgC.m−2. h−1) in response to higher temperatures while it acts as a clear sink of isoprene. Total soil BVOC emission rates, together with soil respiration, decreased by 43% after a decade of partial rain restriction. These results will contribute to enhance further modeling of soil BVOC emissions under various climate scenarios both at regional and global scales.

A decade of rain exclusion in a Mediterranean forest reveals trade-offs of leaf chemical defenses and drought legacy effects

Increasing aridity in the Mediterranean region will result in longer and recurrent drought. These changes could strongly modify plant defenses, endangering tree survival. We investigate the response of chemical defenses from central and specialized metabolism in Quercus pubescens Willd. to future Mediterranean drought using a long-term drought experiment in natura where trees have been submitted to amplified drought (~ –30% annual precipitation) since April 2012. We focused on leaf metabolites including chlorophylls and carotenoids (central metabolism) and flavonols (specialized metabolism). Measurements were performed in summer from 2016 to 2022. Amplified drought led to higher concentrations of total photosynthetic pigments over the 2016–2022 period. However, it also led to lower AZ/VAZ and flavonol concentrations. Additionally, chemical defenses of Q. pubescens responded to previous precipitation where low precipitation 1 year and/or 2 years preceding sampling was associated to low concentrations of VAZ, flavonol and high neoxanthin concentrations. Our study indicates that the decline of flavonol concentration under long-term drought is counterbalanced by a higher production of several central metabolites. Such results are potentially due to an adjustment in tree metabolism, highlighting the importance of performing long-term experimental studies in natura for assessing drought legacy effects and thus forest adaptation to climate change.

Interpreting the shifts in forest structure, plant community composition, diversity, and functional identity by using remote sensing-derived wildfire severity

BackgroundWildfires are increasingly impacting ecosystems worldwide especially in temperate dry habitats, often interplaying with other global changes (e.g., alien plant invasions). Understanding the ecological consequences of wildfires is crucial for effective conservation and management strategies. The aim of this study was to investigate the impacts of wildfire severity on plant community (both the canopy trees and herbaceous layer) and alien plant invasion, combining field observations and remotely sensed data.We conducted an observational study in the Karst forests (North-East Italy) 1 year after the large wildfire which affected the area in 2022. We assessed the impact through 35 field plots (200 m2 each) distributed among different fire severity (i.e., the loss of organic matter) classes assessed using the differenced normalized burn ratio (dNBR) calculated from satellite images. In each plot, tree species, diameter, vitality, resprouting capacity, and seedling density were measured. In addition, herb species richness (taxonomical diversity) was quantified, and plant cover was visually estimated. Functional diversity was also assessed considering six functional traits retrieved from databases.ResultsSome woody species (e.g., Quercus pubescens) showed a higher resistance to the fire (i.e., lower mortality rate), while others showed a higher resilience (i.e., recovery after fire through resprouting or seedlings, e.g., Cotinus coggygria). The transition to a shrub-dominated community (i.e., Cotinus coggygria) where fire severity was the highest underlines the dynamic nature of the post-fire succession. We detected a significant variation in the herbaceous plant community composition, diversity, and functional identity (i.e., community-weighted mean of trait) along the fire severity gradient. In particular, high-fire severity areas exhibited higher species richness compared to low-severity or unburned areas. Total alien plant cover increased with fire severity, while native cover remained constant. We also found shifts in species that enhance traits related to germination potential and growth strategy.ConclusionsOur results highlight the vulnerability of the forest stands to an increase in wildfire severity, resulting in significant mortality and changes in tree community structure. This study contributes to the understanding of ecological processes after wildfires using a novel remote sensing approach in a temperate forest, emphasizing the need for conservation strategies aimed at mitigating high severity wildfires.

Application timing affects the efficacy of Bacillus thuringiensis subsp. kurstaki against Tortrix viridana in deciduous oak forests

The green oak leaf roller moth, Tortrix viridana, is one of the main defoliators of deciduous oaks in the Mediterranean region. Although aerial applications of Btk-based insecticides represent the most effective method to control the larval populations of this pest at a large spatial scale, the optimal time window for sprayings has not yet been defined. Accordingly, experimental trials were conducted in a Quercus pubescens forest district in Sardinia (Italy) involving both large-scale aerial treatments and small-scale applications from the ground. The aim of our work was to study how different larval development stages and/or sprouting phenological phases of the host trees affect Btk efficacy. Aerial sprayings were conducted in 2003 and 2004 in two different areas of about 300 ha, selected according to altitude and exposure to simulate three different application timings against different larval instars and sprouting phenological phases. From-the-ground applications were carried out in 2021 in a Q. pubescens forest stand by applying Btk-based insecticides at different timings, and the overall Q. pubescens phenological stage and the distribution of T. viridana larval development stages were assessed accordingly. The effectiveness of Btk applications was evaluated by estimating larval mortality 7, and 14 days after aerial applications, and 7, 14 and 21 days after ground applications. Defoliation due to T. viridana was also evaluated at the end of larval development in all years. In both 2003 and 2004, the average larval mortality due to Btk exceeded 80 % two weeks after aerial applications across all application timings. Damage caused by T. viridana varied with Btk application timing, with the later applications showing a higher defoliation than other timings. In ground application experiments, larval mortality significantly increased when Btk was sprayed against a larval population primarily composed of IV instars, which were feeding on sprouts with leaves that were spread out and had not fully developed internodes, thus resulting in a significative reduction of insect damage. Our findings indicate that, in accordance with the strict synchrony between the larval development and the oak flushing, the optimal timing for Btk application against T. viridana can be defined based on the bud burst phenological phase. In particular, the prevalence of sprout with expanding leaf surface allowing insecticidal droplet interception and the simultaneous presence of susceptible larval instars (within the IV), resulted in an increased treatment efficacy.

Hydraulic plasticity and water use regulation act to maintain the hydraulic safety margins of Mediterranean trees in rainfall exclusion experiments.

Hydraulic failure due to xylem embolism has been identified as one of the main mechanisms involved in drought-induced forest decline. Trees vulnerability to hydraulic failure depends on their hydraulic safety margin (HSM). While it has been shown that HSM globally converges between tree species and biomes, there is still limited knowledge regarding how HSM can adjust locally to varying drought conditions within species. In this study, we relied on three long-term partial rainfall exclusion experiments to investigate the plasticity of hydraulic traits and HSM for three Mediterranean tree species (Quercus ilex L., Quercus pubescens Willd., and Pinus halepensis Mill.). For all species, a homeostasis of HSM in response to rainfall reduction was found, achieved through different mechanisms. For Q. ilex, the convergence in HSM is attributed to the adjustment of both the turgor loss point (Ψtlp) and the water potential at which 50% of xylem conductivity is lost due to embolism (P50). In contrast, the maintenance of HSM for P. halepensis and Q. pubescens is related to its isohydric behavior for the first and leaf area adjustment for the latter. It remains to be seen whether this HSM homeostasis can be generalized and if it will be sufficient to withstand extreme droughts expected in the Mediterranean region.

Integration of ecological indicators to assess a multitemporal impact of cement industries

The present study evaluated an integrated biomonitoring approach based on three different bioindicators: tree rings, lichens, and beetles in a complex environment (urban-industrial-forest). In Central Italy, four sampling sites were selected to assess the anthropogenic impact of cement plants taking into account (1) long-term exposure (1988–2020) through the analysis of trace elements in tree rings of Quercus pubescens; (2) medium-term exposure (2020–2021) through the analysis of trace elements in thalli (outermost portions) of the lichen Xanthoria parietina; (3) short-term exposure in spring 2021 through the bioaccumulation and evaluation of sample vitality in transplants of the lichen Evernia prunastri and a periodic survey of entomological biodiversity carried out during spring and summer 2021. Trace elements of industrial origin were found in tree rings, with different levels of accumulation between 1988 and 2020 and a maximum in 2012. Native thalli of the lichen X. parietina showed an overall low bioaccumulation of trace elements except for Cr, probably reflecting the influence of national lockdown measures. The transplants of E. prunastri showed a weak stress response in the industrial and urban sites, but not in the forest, and identified Tl and V as the main elements contributing to atmospheric contamination, with peaks at the industrial sites. Concerning the beetles, a significantly lower number of species was found at the Semonte industrial site.

Developing Oak Buds Produce Volatile Emissions in Response to Herbivory by Freshly Hatched Caterpillars.

Plant responses to damage by insectivorous herbivores are well-documented in mature leaves. The resulting herbivore-induced plant volatiles (HIPVs) protect the plant by attracting carnivorous arthropods and even some insectivorous vertebrates, to parasitize or consume the plant invaders. However, very little is known about plant production of HIPVs in developing buds, particularly when herbivorous insects are too small to be considered a prey item. It is additionally unclear whether plants respond differently to generalist and specialist chewing insects that overlap in distribution. Therefore, we compared HIPV production of Downy oak (Quercus pubescens Willd.) buds infested with freshly hatched caterpillars of Tortrix viridana (specialist) and Operophtera brumata (generalist), against uninfested buds. Of the compounds identified in both years of the experiment, we found that (Z)-hex-3-enyl acetate, (E)-β-ocimene, acetophenone, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl salicylate, α-copaene, α-humulene, (E)-caryophyllene, and (E,E)-α-farnesene appeared to be higher in infested buds compared to controls. We found no difference in HIPV production between the specialist and the generalist herbivores. Production of HIPVs was also associated with leaf damage, with higher HIPV production in more severely attacked buds. Thus, our study shows that oak trees already start responding to insect herbivory before leaves are developed, by producing compounds similar to those found in damaged mature leaves. Future work should focus on how Downy oak may benefit from initiating alarm cues at a time when carnivorous arthropods and insectivorous vertebrates are unable to use herbivorous insects as host or food.

Coordination between degree of isohydricity and depth of root water uptake in temperate tree species

In an increasingly dry environment, it is crucial to understand how tree species use soil water and cope with drought. However, there is still a knowledge gap regarding the relationships between species-specific stomatal behaviour, spatial root distribution, and root water uptake (RWU) dynamics.Our study aimed to investigate above- and below-ground aspects of water use during soil drying periods in four temperate tree species that differ in stomatal behaviour: two isohydric tracheid-bearing conifers, Scots pine and Norway spruce, and two more anisohydric deciduous species, the diffuse-porous European beech, and the ring-porous Downy oak. From 2015 to 2020, soil-tree-atmosphere-continuum parameters were measured for each species in monospecific forests where trees had no access to groundwater. The hourly time series included data on air temperature, vapor pressure deficit, soil water potential, soil hydraulic conductivity, and RWU to a depth of 2 m. Analysis of drought responses included data on stem radius, leaf water potential, estimated osmotically active compounds, and drought damage.Our study reveals an inherent coordination between stomatal regulation, fine root distribution and water uptake. Compared to conifers, the more anisohydric water use of oak and beech was associated with less strict stomatal closure, greater investment in deep roots, four times higher maximum RWU, a shift of RWU to deeper soil layers as the topsoil dried, and a more pronounced soil drying below 1 m depth. Soil hydraulic conductivity started to limit RWU when values fell below 10−3 to 10−5 cm/d, depending on the soil. As drought progressed, oak and beech may also have benefited from their leaf osmoregulatory capacity, but at the cost of xylem embolism with around 50 % loss of hydraulic conductivity when soil water potential dropped below −1.25 MPa.Consideration of species-specific water use is crucial for forest management and vegetation modelling to improve forest resilience to drought.

Whole-genome screening for near-diagnostic genetic markers for four western European white oak species identification

Key messageMining genome-wide DNA sequences enabled the discovery of near-diagnostic markers for species assignment in four European white oaks (Quercus petraea (Matt.) Liebl., Quercus pubescens Willd., Quercus pyrenaica Willd., and Quercus robur L.) despite their low interspecific differentiation. Near-diagnostic markers are almost fully fixed in one species and absent in the three others. As a result, only a handful of markers are needed for species identification, making this genetic assay a very promising operational taxonomic assignment procedure in research and forestry.ContextIdentifying species in the European white oak complex has been a long-standing concern in taxonomy, evolution, forest research, and management. Quercus petraea (Matt.) Liebl., Q. robur L., Q. pubescens Willd., and Q. pyrenaica Willd. are part of this species complex in western temperate Europe and hybridize in mixed stands, challenging species identification.AimsOur aim was to identify near-diagnostic single-nucleotide polymorphisms (SNPs) for each of the four species that are suitable for routine use and rapid diagnosis in research and applied forestry.MethodsWe first scanned existing whole-genome and target-capture data sets in a reduced number of samples (training set) to identify candidate diagnostic SNPs, i.e., genomic positions being characterized by a reference allele in one species and by the alternative allele in all other species. Allele frequencies of the candidates SNPs were then explored in a larger, range-wide sample of populations in each species (validation step).ResultsWe found a subset of 38 SNPs (10 for Q. petraea, 7 for Q. pubescens, 9 for Q. pyrenaica, and 12 for Q. robur) that showed near-diagnostic features across their species distribution ranges with Q. pyrenaica and Q. pubescens exhibiting the highest (0.876) and lowest (0.747) diagnosticity, respectively.ConclusionsWe provide a new, efficient, and reliable molecular tool for the identification of the species Q. petraea, Q. robur, Q. pubescens, and Q. pyrenaica, which can be used as a routine tool in forest research and management. This study highlights the resolution offered by whole-genome sequencing data to design near-diagnostic marker sets for taxonomic assignment, even for species complexes with relatively low differentiation.

Leaf morpho-anatomical adjustments in a Quercus pubescens forest after 10 years of partial rain exclusion in the field.

In the Mediterranean region, a reduction of annual precipitation and a longer and drier summer season are expected with climate change by the end of the century, eventually endangering forest survival. To cope with such rapid changes, trees may modulate their morpho-anatomical and physiological traits. In the present study, we focused on the variation in leaf gas exchange and different leaf morpho-anatomical functional traits of Quercus pubescens Willd. in summer using a long-term drought experiment in natura consisting of a dynamic rainfall exclusion system where trees have been submitted to amplified drought (AD) (~-30% of annual precipitation) since April 2012 and compared them with trees under natural drought (ND) in a Mediterranean forest. During the study, we analyzed net CO2 assimilation (An), stomatal conductance (gs), transpiration (E), water-use efficiency (WUE), stomatal size and density, density of glandular trichomes and non-glandular trichomes, thickness of the different leaf tissues, specific leaf area and leaf surface. Under AD, tree functioning was slightly impacted, since only An exhibited a 49% drop, while gs, E and WUE remained stable. The decrease in An under AD was regulated by concomitant lower stomatal density and reduced leaf thickness. Trees under AD also featured leaves with a higher non-glandular trichome density and a lower glandular trichome density compared with ND, which simultaneously limits transpiration and production costs. This study points out that Q. pubescens exhibits adjustments of leaf morpho-anatomical traits which can help trees to acclimate to AD scenarios as those expected in the future in the Mediterranean region.

Interactive effects of soil moisture, air temperature and litter nutrient diversity on soil microbial communities and Folsomia candida population

Soil organisms play a key role in carbon and nutrient cycling in forest ecosystems. While soil organisms are strongly influenced by litter chemistry and are highly sensitive to abiotic conditions, little is known about the interactive effects of these two factors. To address this gap in knowledge, we conducted a 10 week microcosm experiment in which we simulated the effects of climate change on soil ecology. More specifically, we studied relationships among litter nutrient concentration, microbial biomass, Collembola demographic parameters, and litter decomposition, exploring the potential impacts of increasing air temperature and decreasing soil moisture. To develop a gradient of nutrient concentrations, we created six tree litter mixtures with materials gathered from Quercus pubescens and its companion species. In contrast to microbes, we observed that Collembola abundance and litter decomposition were interactively affected by soil moisture and air temperature: the negative effect of increasing air temperature on Collembola abundance was amplified by reduced soil moisture, whereas the positive effect of increasing air temperature on litter decomposition disappeared under reduced soil moisture conditions. In contrast to fungi, the response of bacterial biomass and Collembola abundance to litter nutrient concentration was dependent on abiotic conditions. More specifically, the relationships between nutrients, especially calcium and magnesium, and bacterial biomass and Collembola abundance were less robust or disappeared under drier or warmer conditions. In conclusion, our findings underscore that ongoing climate change could affect soil organisms directly as well as indirectly, by altering their responses to litter nutrient concentrations. In addition, we found that nutrient‐rich habitats might be more affected than nutrient‐poor habitats by altered climatic conditions.

Drought legacies in mixed Mediterranean forests: Analysing the effects of structural overshoot, functional traits and site factors

Previous favorable climate conditions stimulate tree growth making some forests more vulnerable to hotter droughts. This so-called structural overshoot may contribute to forest dieback, but there is little evidence on its relative importance depending on site conditions and tree species because of limited field data. Here, we analyzed remote sensing (NDVI) and tree-ring width data to evaluate the impacts of the 2017 drought on canopy cover and growth in mixed Mediterranean forests (Fraxinus ornus, Quercus pubescens, Acer monspessulanum, Pinus pinaster) located in southern Italy. Legacy effects were assessed by calculating differences between observed and predicted basal area increment (BAI). Overall, the growth response of the study stands to the 2017 drought was contingent on site conditions and species characteristics. Most sites presented BAI and canopy cover reductions during the drought. Growth decline was followed by a quick recovery and positive legacy effects, particularly in the case of F. ornus. However, we found negative drought legacies in some species (e.g., Q. pubescens, A. monspessulanum) and sites. In those sites showing negative legacies, high growth rates prior to drought in response to previous wet winter-spring conditions may have predisposed trees to drought damage. Vice versa, the positive drought legacy found in some F. ornus site was linked to post-drought growth release due to Q. pubescens dieback and mortality. Therefore, we found evidences of structural drought overshoot, but it was restricted to specific sites and species. Our findings highlight the importance of considering site settings such as stand composition, pre-drought conditions and different tree species when studying structural overshoot. Droughts contribute to modify the composition and dynamics in mixed forests.

Macro- and Microelements and the Impact of Sub-Mediterranean Downy Oak Forest Communities on Their Composition in Rainwater

In this study, we analyzed the content of chemical elements in rainwater and investigated the influence of forest cover on the composition of precipitation. The results obtained showed that the concentration of some elements in the rainwater collected under the forest canopy was higher than that in the open area. As part of the work, we calculated the enrichment factor and examined the sources of chemical elements in rainwater. We found that all macro-elements had increased values of the enrichment factors compared to the supporting elements of the Earth’s crust. Ca had the highest value. The values of the remaining elements (Sr, Pb, Mn, Cr, Ba, V, Fe) indicated their lithogenic and anthropogenic origins. We noted that the enrichment factor under the forest canopy was significantly lower than in the open area, indicating the dilution of these elements during water passage through the canopy. Elements such as Zn, Co, Cu, and Ni also had high enrichment factors, which indicate their anthropogenic origin. In the open area, most elements had an inverse relationship with pH, except for the alkali metals Na, Mg, and Ca, which had a positive relationship with the pH value. The concentration of K was not dependent on pH. In rainwater that had passed through the forest canopy, the concentrations of Na, Mg, and Ca were also not dependent on pH, while the concentration of K had an inverse relationship with pH. As the concentration of heavy metals in rainwater increases, the role of Na, Mg, and Ca in the process of water neutralization decreases.

An efficient protocol for extracting thylakoid membranes and total leaf proteins from Posidonia oceanica and other polyphenol-rich plants

BackgroundThe extraction of thylakoids is an essential step in studying the structure of photosynthetic complexes and several other aspects of the photosynthetic process in plants. Conventional protocols have been developed for selected land plants grown in controlled conditions. Plants accumulate defensive chemical compounds such as polyphenols to cope with environmental stresses. When the polyphenol levels are high, their oxidation and cross-linking properties prevent thylakoid extraction.ResultsIn this study, we developed a method to counteract the hindering effects of polyphenols by modifying the grinding buffer with the addition of both vitamin C (VitC) and polyethylene glycol (PEG4000). This protocol was first applied to the marine plant Posidonia oceanica and then extended to other plants synthesizing substantial amounts of polyphenols, such as Quercus pubescens (oak) and Vitis vinifera (grapevine). Native gel analysis showed that photosynthetic complexes (PSII, PSI, and LHCII) can be extracted from purified membranes and fractionated comparably to those extracted from the model plant Arabidopsis thaliana. Moreover, total protein extraction from frozen P. oceanica leaves was also efficiently carried out using a denaturing buffer containing PEG and VitC.ConclusionsOur work shows that the use of PEG and VitC significantly improves the isolation of native thylakoids, native photosynthetic complexes, and total proteins from plants containing high amounts of polyphenols and thus enables studies on photosynthesis in various plant species grown in natural conditions.