Impact of coppicing on microclimate and understorey vegetation diversity in an ancient Mediterranean oak forest.

Coppicing is one of the oldest silvicultural practices and is still widely applied to produce renewable energy from broadleaf forests. However, the consequences on microclimate and understorey vegetation are still poorly understood, especially in Mediterranean oak forests. With the ongoing changes in the climate system and global biodiversity loss, a better understanding of how the forest temperature buffering capacity and below-canopy plant community are impacted by coppicing is crucial. Here we quantify microclimate and understorey vegetation changes in adjacent ancient coppice-with-standards and high forest stands dominated by oaks in Italy, where these systems have been applied for a long time. Air and soil temperatures were recorded for 2.5 years, and nested vegetation plots were used to analyse coppicing effects on species composition, taxonomic, phylogenetic, and functional diversity. Coppicing significantly reduced the forest temperature buffering capacity. The mean of the daily maximum temperatures over the entire period was 1.45 °C higher in the coppiced sites, whereas the mean of the daily minimum temperatures was 0.62 °C lower than in the high forest. Coppicing increased understorey species richness by favouring generalist taxa, but significantly decreased the proportion of forest specialists. The understorey community in coppiced forests consisted of more warm-adapted species. Moreover, coppicing also led to a loss of phylogenetic evenness and to shifts in diversity and community weighted mean Leaf Dry Matter content, pointing to habitat filtering and acclimation processes. In sum, we show that coppicing affects microclimate and understory vegetation in a direction that can exacerbate the effects of climate change, negatively affecting the oak forest specialist flora and its phylogenetic evenness.

Topography modulates climate sensitivity of multidecadal trends of holm oak decline.

Forest decline events have increased worldwide over the last decades being holm oak (Quercus ilex L.) one of the tree species with the most worrying trends across Europe. Since this is one of the tree species with the southernmost distribution within the European continent, its vulnerability to climate change is a phenomenon of enormous ecological importance. Previous research identified drought and soil pathogens as the main causes behind holm oak decline. However, despite tree health loss is a multifactorial phenomenon where abiotic and biotic factors interact in time and space, there are some abiotic factors whose influence has been commonly overlooked. Here, we evaluate how land use (forests versus savannas), topography, and climate extremes jointly determine the spatiotemporal patterns of holm oak defoliation trends over almost three decades (1987-2014) in Spain, where holm oak represents the 25% of the national forested area. We found an increasing defoliation trend in 119 out of the total 134 holm oak plots evaluated, being this defoliation trend significantly higher in forests compared with savannas. Moreover, we have detected that the interaction between topography (which covariates with the land use) and summer precipitation anomalies explains trends of holm oak decline across the Mediterranean region. While a higher occurrence of dry summers increases defoliation trends in steeper terrains where forests dominate, an inverse relationship was found in flatter terrains where savannas are mainly located. These opposite relationships suggest different causal mechanisms behind decline. Whereas hydric stress is likely to occur in steeper terrains where soil water holding capacity is limited, soil waterlogging usually occurs in flatter terrains what increases tree vulnerability to soil pathogens. Our results contribute to the growing evidence of the influence of local topography on forest resilience and could assist in the identification of potential tree decline hotspots and its main causes over the Mediterranean region.

Quercus ilex L. dieback is genetically determined: Evidence provided by dendrochronology, δ13C and SSR genotyping.

Quercus ilex L. dieback has been reported in several Mediterranean forests, revealing different degree of crown damages even in close sites, as observed in two Q. ilex forest stands in southern Tuscany (IT). In this work, we applied a novel approach combining dendrochronological, tree-ring δ13C and genetic analysis to test the hypothesis that different damage levels observed in a declining (D) and non-declining (ND) Q. ilex stands are connected to population features linked to distinct response to drought. Furthermore, we investigated the impact of two major drought events (2012 and 2017), that occurred in the last fifteen years in central Italy, on Q. ilex growth and intrinsic water use efficiency (WUEi). Overall, Q. ilex showed slightly different ring-width patterns between the two stands, suggesting a lower responsiveness to seasonal climatic variations for trees at D stand, while Q. ilex at ND stand showed changes in the relationship between climatic parameters and growth across time. The strong divergence in δ13C signals between the two stands suggested a more conservative use of water for Q. ilex at ND compared to D stand that may be genetically driven. Q. ilex at ND resulted more resilient to drought compared to trees at D, probably thanks to its safer water strategy. Genotyping analysis based on simple-sequence repeat (SSR) markers revealed the presence of different Q. ilex populations at D and ND stands. Our study shows intraspecific variations in drought response among trees grown in close. In addition, it highlights the potential of combining tree-ring δ13C data with SSR genotyping for the selection of seed-bearing genotypes aimed to preserve Mediterranean holm oak ecosystem and improve its forest management.

Upside down and the game of C allocation.

Non-structural carbohydrates (NSCs) represent the primary carbon (C) reserves and play a crucial role for plant functioning and resilience. Indeed, these compounds are involved in the regulation between C supply and demand, and in the maintenance of hydraulic efficiency. NSCs are stored in parenchyma of woody organs, which is recognized as a proxy for reserve storage capacity of tree. Notwithstanding the importance of NSCs for tree physiology, their long-term regulation and trade-offs against growth were not deeply investigated. This work evaluated the long-term dynamics of mature tree reserves in stem and root, proxied by parenchyma features, and focusing on the trade off and interplay between the resources allocation in radial growth and reserves in stem and coarse root. In a Mediterranean beech forest, NSCs content, stem and root wood anatomy analysis, and eddy covariance data, were combined. The parenchyma fraction (RAP) of beech root and stem was different, due to differences in axial parenchyma (AP) and narrow ray parenchyma (nRP) fractions. However, these parenchyma components and radial growth showed synchronous inter-annual dynamics between the two organs. In beech stem, positive correlations were found among soluble sugars content and nRP, and among starch content and the AP. Positive correlations were found among Net Ecosystem Exchange (NEE) and AP of both organs. In contrast, NEE was negatively correlated to radial growth of root and stem. Our results suggest a different contribution of stem and roots to reserves storage, and a putative partitioning in the functional roles of parenchyma components. Moreover, a long-term trade-off of C allocation between growth and reserve pool was evidenced. Indeed, in case of C source reduction, trees preferentially allocate C towards reserves pool. Conversely, in high productive years, growth represents the major C sink.

Above- and belowground interplay: Canopy CO2 uptake, carbon and nitrogen allocation and isotope fractionation along the plant-ectomycorrhiza continuum.

In forests, mycorrhizal fungi regulate carbon (C) and nitrogen (N) dynamics. We evaluated the interplay among ectomycorrhizas (ECM), ecosystem C fluxes, tree productivity, C and N exchange and isotopic fractionation along the soil-ECM-plant continuum in a Mediterranean beech forest. From bud break to leaf shedding, we monitored: net ecosystem exchange (NEE, a measure of the net exchange of C between an ecosystem and the atmosphere), leaf area index, stem growth, N concentration, δ13 C and δ15 N in rhizosphere soil, ectomycorrhizal fine root tips (ERT), ECM-free fine root portions (NCR) and leaves. Seasonal changes in ERT relative biomass were strictly related to NEE and mimicked those detected in the radial growth. The analysis of δ13 C in ERT, leaves and NCR highlighted the impact of canopy photosynthesis on ERT development and an asynchronous seasonal C allocation strategy between ERT and NCR at the root tips level. Concerning N, δ15 N of leaves was negatively related to that of ERT and dependent on seasonal 15 N differences between ERT and NCR. Our results unravel a synchronous C allocation towards ERT and tree stem driven by the increasing NEE in spring-early summer. Moreover, they highlighted a phenology-dependent 15 N fractionation during N transfer from ECM to their hosts. This evidence, obtained in mature beech trees under natural conditions, may improve the knowledge of Mediterranean forests functionality.

Amplified Drought Alters Leaf Litter Metabolome, Slows Down Litter Decomposition, and Modifies Home Field (Dis)Advantage in Three Mediterranean Forests.

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.

Three Millennia of Vegetation, Land-Use, and Climate Change in SE Sicily.

This study presents the first Late Holocene marine pollen record (core ND2) from SE Sicily. It encompasses the last 3000 years and is one of the most detailed records of the south-central Mediterranean region in terms of time resolution. The combined approach of marine palynology and historical ecology, supported by independent palaeoclimate proxies, provides an integrated regional reconstruction of past vegetational dynamics in relation to rapid climatic fluctuations, historical socio-economic processes, and past land-use practices, offering new insights into the vegetation history of SE Sicily. Short-term variations of sparse tree cover in persistently open landscapes reflect rapid hydroclimatic changes and historical land-use practices. Four main phases of forest reduction are found in relation to the 2.8 ka BP event, including the Late Antique Little Ice Age, the Medieval Climate Anomaly, and the Little Ice Age, respectively. Forest recovery is recorded during the Hellenistic and Roman Republican Periods, the Early Middle Ages, and the last century. Agricultural and silvicultural practices, as well as stock-breeding activities, had a primary role in shaping the current vegetational landscape of SE Sicily.

Seed and seedling interactions in three tree species from Mediterranean forests as a knowledge base for ecological restoration.

The ecosystems of the Mediterranean regions are severely threatened by human activity, and although we have made progress in physical restoration measures, little is known about the interactions between the plants of these biomes. The objective of this study is to contribute to document interactions between seeds and seedlings of three woody species native to Chile (P. chilensis, Q. saponaria and A. caven), which could be used for restoration actions (e.g., after forest fires). In a first experiment, we evaluated the germination response, the initial elongation and the interactions between the seedlings that germinate exposed to the chemical compounds of the other species. In a second experiment, we compared the survival and growth of seedlings in monospecific versus bispecific mixtures, using a substrate similar to that which is present after a wildfire. Seed extracts of teguments promoted germination of P. chilensis, but cotyledons and whole seed extracts inhibited germination of only one species, Q. saponaria, with very high intensity. The effects of the extracts on initial seedling elongation were more variable, including five inhibitions and one facilitation. Negative effects on germination and elongation included two cases of autotoxicity. The survival of seedlings grown in mixtures showed only two differences between monospecific and bispecific mixtures, both positive, constituting a reciprocal effect between two species. Only in one case was there a significant difference in seedling growth, which was root growth inhibition. Indeed, these results reveal an interaction between species, the type and intensity of which varies according to the condition of the seed or seedling. The negative effect found in seedling root growth reflects an allelopathic interaction that conditions a vital aspect for the establishment of these species, so this information is an opportunity to improve the establishment conditions in future reforestation projects, by avoiding particular species or promoting their proportion in plantation mixtures, either by planting or direct seeding.

Amplified Drought and Seasonal Cycle Modulate Quercus pubescens Leaf Metabolome.

The intensification of summer drought expected with climate change can induce metabolism modifications in plants to face such constraints. In this experiment, we used both a targeted approach focused on flavonoids, as well as an untargeted approach, to study a broader fraction of the leaf metabolome of Quercus pubescens exposed to amplified drought. A forest site equipped with a rainfall exclusion device allowed reduction of natural rainfall by ~30% over the tree canopy. Leaves of natural drought (ND) and amplified drought (AD) plots were collected over three seasonal cycles (spring, summer, and autumn) in 2013 (the second year of rain exclusion), 2014, and 2015. As expected, Q. pubescens metabolome followed a seasonal course. In the summer of 2015, the leaf metabolome presented a shifted and early autumnal pattern because of harsher conditions during this year. Despite low metabolic modification at the global scale, our results demonstrated that 75% of Quercus metabolites were upregulated in springs when trees were exposed to AD, whereas 60 to 73% of metabolites (93% in summer 2015), such as kaempferols and quercetins, were downregulated in summers/autumns. Juglanin, a kaempferol pentoside, as well as rhododendrin derivatives, were upregulated throughout the year, suggesting an antioxidant ability of these metabolites. Those changes in terms of phenology and leaf chemistry could, in the end, affect the ecosystem functioning.

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

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

Impact of successive spring frosts on leaf phenology and radial growth in three deciduous tree species with contrasting climate requirements in central Spain.

Rear-edge tree populations forming the equatorward limit of distribution of temperate species are assumed to be more adapted to climate variability than central (core) populations. However, climate is expected to become more variable and the frequency of climate extremes is forecasted to increase. Climatic extreme events such as heat waves, dry spells and spring frosts could become more frequent, and negatively impact and jeopardize rear-edge stands. To evaluate these ideas, we analyzed the growth response of trees to successive spring frosts in a mixed forest, where two temperate deciduous species, Fagus sylvatica L. (European beech) and Quercus petraea (Matt.) Liebl. (sessile oak), both at their southernmost edge, coexist with the Mediterranean Quercus pyrenaica Willd. (Pyrenean oak). Growth reductions in spring-frost years ranked across species as F. sylvatica > Q. petraea > Q. pyrenaica. Leaf flushing occurred earlier in F. sylvatica and later in Q. pyrenaica, suggesting that leaf phenology was a strong determinant of spring frost damage and stem growth reduction. The frost impact depended on prior climate conditions, since warmer days prior to frost occurrence predisposed to frost damage. Autumn Normalized Difference Vegetation Index data showed delayed leaf senescence in spring-frost years and subsequent years as compared with pre-frost years. In the studied forest, the negative impact of spring frosts on Q. petraea and especially on F. sylvatica growth, was considerably higher than the impacts due to drought. The succession of four spring frosts in the last two decades determined a trend of decreasing resistance of radial growth to frosts in F. sylvatica. The increased frequency of spring frosts might prevent the expansion and persistence of F. sylvatica in this rear-edge Mediterranean population.

Sampling forest soils to describe fungal diversity and composition. Which is the optimal sampling size in mediterranean pure and mixed pine oak forests?

Soil sampling is a critical step affecting perceived fungal diversity, however sampling optimization for high-throughput-DNA sequencing studies have never been tested in Mediterranean forest ecosystems. We identified the minimum number of pooled samples needed to obtain a reliable description of fungal communities in terms of diversity and composition in three different Mediterranean forests (pine, oak, and mixed-pine-oak). Twenty soil samples were randomly selected in each of the three plots per type. Samples obtained in 100 m2 plots were pooled to obtain mixtures of 3, 6, 10, 15, 20 samples, and sequenced using Illumina MiSeq of fungal ITS2 amplicons. Pooling three soil samples in Pinus and Quercus stands provided consistent richness estimations, while at least six samples were needed in mixed-stands. ß-diversity decreased with increasing sample pools in monospecific-stands, while there was no effect of sample pool size on mixed-stands. Soil sample pooling had no effect over species composition. We estimate that three samples would be already optimal to describe fungal richness and composition in Mediterranean pure stands, while at least six samples would be needed in mixed stands.

Crown defoliation decreases reproduction and wood growth in a marginal European beech population.

BACKGROUND AND AIMS: Abiotic and biotic stresses related to climate change have been associated with increased crown defoliation, decreased growth and a higher risk of mortality in many forest tree species, but the impact of stresses on tree reproduction and forest regeneration remains understudied. At the dry, warm margin of species distributions, flowering, pollination and seed maturation are expected to be affected by drought, late frost and other stresses, eventually resulting in reproduction failure. Moreover, inter-individual variation in reproductive performance versus other performance traits (growth, survival) could have important consequences for population dynamics. This study investigated the relationships among individual crown defoliation, growth and reproduction in a drought-prone population of European beech, Fagus sylvatica. METHODS: We used a spatially explicit mating model and marker-based parentage analyses to estimate effective female and male fecundities of 432 reproductive trees, which were also monitored for basal area increment and crown defoliation over 9 years. KEY RESULTS: Female and male fecundities varied markedly between individuals, more than did growth. Both female fecundity and growth decreased with increasing crown defoliation and competition, and increased with size. Moreover, the negative effect of defoliation on female fecundity was size-dependent, with a slower decline in female fecundity with increasing defoliation for the large individuals. Finally, a trade-off between growth and female fecundity was observed in response to defoliation: some large trees maintained significant female fecundity at the expense of reduced growth in response to defoliation, while some other defoliated trees maintained high growth at the expense of reduced female fecundity. CONCLUSIONS: Our results suggest that, while decreasing their growth, some large defoliated trees still contribute to reproduction through seed production and pollination. This non-coordinated decline of growth and fecundity at individual level in response to stress may compromise the evolution of stress-resistance traits at population level, and increase forest tree vulnerability.

Traditional and TLS-based forest inventories of beech and pine forests located in Sila National Park: A dataset.

Vegetation structure is a key determinant of species distribution and diversity. Compared to traditional methods, the use of Terrestrial Laser Scanning (TLS) has allowed massive amounts of point cloud data collected for quantifying three-dimensional habitat properties at increasing spatial and temporal scales. We used TLS to characterize the forest plots across a broad range of forest structural diversity, located in the Sila National Park, South Italy. The dataset reports data collected in 24 15-m-radius circular plots, 12 of which were dominated by beech (Fagus sylvatica L.) and 12, by black pine (Pinus nigra subsp. laricio). In detail, this work provides dataset of i) plot-level attributes calculated from raw data, such as the number of trees, ii) tree-level data, comprising a total of 1709 trees, with information related to field-based forest inventory such as the diameter at breast height (DBH), and iii) plot-level information related to the time for conducting both traditional field- and TLS-based forest inventories. Compared to traditional methods, the use of TLS allows a very high-resolution quantification of the 3D forest structural properties, also reducing the time for conducting forest inventories.

Forest fire induces short-term shifts in soil food webs with consequences for carbon cycling.

We tested for fire-induced (5-6 years post-fire) changes in the structure and functioning of the soil food web along a 3000-km north-south transect across European Russia, spanning all major forest types in the northern hemisphere outside the tropics. The total biomass of the detrital food web, including microbes and invertebrates, was not affected by fire. However, fire reduced the biomass of microfauna and mites, but had no impact on mesofauna or macrofauna. Fire also reduced rates of carbon (C) mobilisation by soil biota. Our results demonstrate that fire-induced shifts in soil food webs have significant short-term effects on forest soil C cycling, but that these effects vary across forest types and geographic locations.

Resilience of Spanish forests to recent droughts and climate change.

A widespread increase in forest cover is underway in northern Mediterranean forests because of land abandonment and decreased wood demand, but the resilience of these successional forests to climate change remains unresolved. Here we use 18-year time series of canopy greenness derived from satellite imagery (NDVI) to evaluate the impacts of climate change on Spain's forests. Specifically, we analyzed how NDVI was influenced by the climatic water balance (i.e. Standardized Precipitation-Evapotranspiration Index, SPEI), using monthly time-series extracted from 3,100 pixels of forest, categorized into ten forest types. The forests increased in leaf area index by 0.01 per year on average (from 1.7 in 2000 to 1.9 in 2017) but there was enormous variation among years related to climatic water balance. Forest types varied in response to drought events: those dominated by drought-avoiding species showed strong covariance between greenness and SPEI, while those dominated by drought-tolerant species showed weak covariance. Native forests usually recovered more than 80% of greenness within the 18 months and the remainder within 5 years, but plantations of Eucalyptus were less resilient. Management to increase the resilience of forests-a key goal of forestry in the Mediterranean region-appears to have had a positive effect: canopy greenness within protected forests was more resilient to drought than within non-protected forests. In conclusion, many of Spain's successional forests have been resilient to drought over the past 18 years, from the perspective of space. Future studies will need to combine remote sensing with field-based analyses of physiological tolerances and mortality processes to understand how Mediterranean forests will respond to the rapid climate change predicted for this region in the coming decades.

Environmental Drivers of Microbial Functioning in Mediterranean Forest Soils.

Mediterranean forests own distinct characteristics resulting from climate, soil, and vegetation that affect soil microbial communities' assembly and their associated functions. We initiated a multi-scalar analysis of environmental drivers of soil functioning to (1) identify pertinent factorial scales and (2) determine the relative importance of soil, vegetation, and geoclimate influences in shaping soil microbial functions across the French Mediterranean forests. Soil samples (0-15 cm) were collected from 60 forest sites and soil physicochemical and microbiological properties were assessed across different factorial scales i.e., bioclimates, slope exposures, and forest stands. Patterns in microbial catabolic potential (i.e., extracellular enzymes and microbial respiration) and carbon (C) source utilization (i.e., catabolic-level physiological profiling) were partitioned between vegetation cover, soil characteristics, and geoclimate components. Our results reveal that the catabolic potential of soil microbes was strongly influenced by the forest stands and mainly relied on ammonium and nitrate contents. In contrast, variation in C source utilization was mainly explained by vegetation cover. Soil metabolic capacities of microorganisms and resulting C dynamics were largely constrained by climate parameters, which suggests potentially important consequences for soil C storage. Our study revealed diverse structuration patterns between the catabolic potential and the carbon source utilization of soil microbial communities, and gives insights into the underlying mechanisms of soil microbial functioning in Mediterranean forests.

Tree xylem water isotope analysis by Isotope Ratio Mass Spectrometry and laser spectrometry: A dataset to explore tree response to drought.

Water isotopes from plant xylem and surrounding environment are increasingly used in eco-hydrological studies. Carrière et al. [1] analyzed a dataset of water isotopes in (i) the xylem of three different tree species, (ii) the surrounding soil and drainage water and (iii) the underlying karst groundwater, to understand tree water uptake during drought in two different Mediterranean forests on karst setting. The xylem and soil water were extracted by cryogenic distillation. The full dataset was obtained with Isotope Ratio Mass Spectrometry (IRMS) and Isotope Ratio Infrared Spectrometer (IRIS), and included 219 measurements of δ2H and δ18O. Prompted by unexpected isotopic data characterized by a very negative deuterium excess, a subsample of 46 xylem samples and 9 soil water samples were double checked with both analytical techniques. IRMS and IRIS analyses yielded similar data. Therefore, the results reveal that laser spectrometry allows an accurate estimation of xylem and soil water isotopes. The dataset highlights a strong 2H depletion in xylem water for all species. Deuterium does not seem adequate to interpret ecological processes in this dataset given the strong fractionation.

Litter traits and rainfall reduction alter microbial litter decomposers: the evidence from three Mediterranean forests.

The objective of the study was to evaluate changes in microbial communities with the predicted arrival of new species to Mediterranean forests under projected intensification of water stress conditions. For that, litter from three Mediterranean forests dominated respectively by Quercus pubescens Willd., Quercus ilex L. and Pinus halepensis Mill. were collected, and placed to their 'home' forest but also to the two other forests under natural and amplified drought conditions (i.e. rainfall reduction of 30%). Quantitative PCR showed that overall, actinobacteria and total bacteria were more abundant in Q. pubescens and Q. ilex than in P. halepensis litter. However, the abundance of both groups was dependent on the forest sites: placement of allochthonous litter to Q. pubescens and P. halepensis forests (i.e. P. halepensis and Q. pubescens, respectively) increased bacterial and fungal abundances, while no effect was observed in Q. ilex forest. P. halepensis litter in Q. pubescens and Q. ilex forests significantly reduced actinobacteria (A/F) and total bacteria (B/F) to fungi ratios. The reduction of rainfall did not influence actinobacteria and bacteria but caused an increase of fungi. As a result, a reduction of A/F ratio is expected with the plant community change towards the dominance of spreading P. halepensis under amplified drought conditions.

Allelopathic effects of volatile organic compounds released from Pinus halepensis needles and roots.

The Mediterranean region is recognized as a global biodiversity hotspot. However, over the last decades, the cessation of traditional farming in the north part of the Mediterranean basin has given way to strong afforestation leading to occurrence of abandoned agricultural lands colonized by pioneer expansionist species like Pinus halepensis. This pine species is known to synthesize a wide range of secondary metabolites, and previous studies have demonstrated strong allelopathic potentialities of its needle and root leachates. Pinus halepensis is also recognized to release significant amounts of volatile organic compounds (VOC) with potential allelopathic effects that have never been investigated. In this context, the objectives of the present study were to improve our knowledge about the VOC released from P. halepensis needles and roots, determine if these VOC affect the seed germination and root growth of two herbaceous target species (Lactuca sativa and Linum strictum), and evaluate if soil microorganisms modulate the potential allelopathic effects of these VOC. Thirty terpenes were detected from both, needle and root emissions with ß-caryophyllene as the major volatile. Numerous terpenes, such as ß-caryophyllene, δ-terpinene, or α-pinene, showed higher headspace concentrations according to the gradient green needles < senescent needles < needle litter. Seed germination and root growth of the two target species were mainly reduced in presence of P. halepensis VOC. In strong contrast with the trend reported with needle leachates in literature, we observed an increasing inhibitory effect of P. halepensis VOC with the progress of needle physiological stages (i.e., green needle < senescent needle < needle litter). Surprisingly, several inhibitory effects observed on filter paper were also found or even amplified when natural soil was used as a substrate, highlighting that soil microorganisms do not necessarily limit the negative effects of VOC released by P. halepensis on herbaceous target species.