Air quality in post-mining towns: tracking potentially toxic elements using tree leaves.

In this study, leaves of the evergreen holm oak Quercus ilex were used to assess airborne contamination of potentially toxic elements (PTEs) at five towns located on the slopes of the Mt. Amiata (central Italy), an area with a long history of mining and, more recently, an important district for the industrial exploitation of geothermal energy. PTE composition and covariance of washed and unwashed Q. ilex leaves of three different ages (6, 12 and 24 month-old) were used to identify atmospheric inputs of PTEs at residential areas, evaluate long-term adsorption and retention of PTEs by the leaves, thus providing an indication of potential human exposure. Moreover, the determination of foliar concentrations of major elements (C, N, S and P) allowed an assessment of the nutritional status of the investigated urban tree stands which excluded the existence of stress condition caused by air pollution or other disturbances. Results indicated that overall Pb, Cu, and Cd concentration were low in the investigated urban sites, if compared with similar studies conducted in larger Italian cities, denoting a low contribution of vehicular traffic to the atmospheric pathway. The five urban settlements were characterized by a specific profile of elements (Al, Ba, Hg and Sb) enriched in unwashed leaves, resulting from the distinct geochemical characteristics of the area and from diffuse (i.e., urban activity) and point sources of PTEs emission (i.e., brownfields, geothermal power plants). The latter sources primarily govern the distribution of Hg, whose contamination was found to be very localized close to a major abandoned mining area. Our data provided quantitative evidence of the spectrum of PTEs potentially impacting resident population and may prove useful in support of follow-up instrumental monitoring campaigns of air quality, as well as for human health and ecological risk assessments.

Increasing temperature and vapour pressure deficit lead to hydraulic damages in the absence of soil drought.

Temperature (T) and vapour pressure deficit (VPD) are important drivers of plant hydraulic conductivity, growth, mortality, and ecosystem productivity, independently of soil water availability. Our goal was to disentangle the effects of T and VPD on plant hydraulic responses. Young trees of Fagus sylvatica L., Quercus pubescens Willd. and Quercus ilex L. were exposed to a cross-combination of a T and VPD manipulation under unlimited soil water availability. Stem hydraulic conductivity and leaf-level hydraulic traits (e.g., gas exchange and osmotic adjustment) were tracked over a full growing season. Significant loss of xylem conductive area (PLA) was found in F. sylvatica and Q. pubescens due to rising VPD and T, but not in Q. ilex. Increasing T aggravated the effects of high VPD in F. sylvatica only. PLA was driven by maximum hydraulic conductivity and minimum leaf conductance, suggesting that high transpiration and water loss after stomatal closure contributed to plant hydraulic stress. This study shows for the first time that rising VPD and T lead to losses of stem conductivity even when soil water is not limiting, highlighting their rising importance in plant mortality mechanisms in the future.

The D165H Polymorphism of QiMYB-like-1 Is Linked to Interactions between Tannin Accumulation, Herbivory and Biogeographical Determinants of Quercus ilex.

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.

Multiomics Molecular Research into the Recalcitrant and Orphan Quercus ilex Tree Species: Why, What for, and How.

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.

Proteomics Data Analysis for the Identification of Proteins and Derived Proteotypic Peptides of Potential Use as Putative Drought Tolerance Markers for Quercus ilex.

Drought is one of the main causes of mortality in holm oak (Quercus ilex) seedlings used in reforestation programs. Although this species shows high adaptability to the extreme climate conditions prevailing in Southern Spain, its intrinsic genetic variability may play a role in the differential response of some populations and individuals. The aim of this work was to identify proteins and derived proteotypic peptides potentially useful as putative markers for drought tolerance in holm oak by using a targeted post-acquisition proteomics approach. For this purpose, we used a set of proteins identified by shotgun (LC-MSMS) analysis in a drought experiment on Q. ilex seedlings from four different provenances (viz. the Andalusian provinces Granada, Huelva, Cadiz and Seville). A double strategy involving the quantification of proteins and target peptides by shotgun analysis and post-acquisition data analysis based on proteotypic peptides was used. To this end, an initial list of proteotypic peptides from proteins highly represented under drought conditions was compiled that was used in combination with the raw files from the shotgun experiment to quantify the relative abundance of the fragment's ion peaks with the software Skyline. The most abundant peptides under drought conditions in at least two populations were selected as putative markers of drought tolerance. A total of 30 proteins and 46 derived peptides belonging to the redox, stress-related, synthesis,-folding and degradation, and primary and secondary metabolism functional groups were thus identified. Two proteins (viz., subtilisin and chaperone GrpE protein) were found at increased levels in three populations, which make them especially interesting for validation drought tolerance markers in subsequent experiments.

Quercus ilex pollen allergen, Que i 1, responsible for pollen food allergy syndrome caused by fruits in Spanish allergic patients.

BACKGROUND: Pollen food allergy syndrome (PFAS) related to PR10 from vegetables is common in northern Europe, whereas in Mediterranean countries PFAS has been preferentially associated with profilins. However, there are pollen-allergic patients reactive to Bet v 1 in birch-free regions. Since it cannot be the primary sensitizer, there has to be another culprit. Quercus ilex is a good candidate as it belongs to the order Fagales. This order includes trees with highly sensitizing pollen such as alder, hazel, hornbeam, oak and chestnut because of the presence of PR10 allergens. PR10 allergens have indeed been described in other Quercus species. OBJECTIVE: Our goals were to determine the rate of sensitization to Q. ilex in central Spain and the associated frequency of PFAS; secondly to identify and clone the Q. ilex allergen PR10. METHODS: We included 224 allergic patients with respiratory symptoms to estimate the rate of sensitization. A skin prick test (SPT) and ImmunoCAP were performed. A total of 38 Q. ilex-sensitized patients were tested using Western blotting to determine the rate of Que i 1. Peptides from Que i 1 were analysed by MALDI-TOF/TOF and Orbitrap LC-MSMS. The Que i 1 sequence was first obtained from the Holm oak transcriptome then cloned and expressed in bacteria. RESULTS: 59.8% of pollen-allergic children were sensitized to Q. ilex. We described and cloned the Q. ilex PR10, Que i 1, which has a sensitization rate of 60.5% and was recognized by 65.4% patients reporting PFAS. CONCLUSION AND CLINICAL RELEVANCE: Sensitization to Q. ilex pollen has increased significantly since 1995. This sensitization could be important, as the presence of PFAS in this population is higher than in patients not sensitized to Q. ilex. The first Q. ilex allergen has been described and is related to PFAS in Spanish patients sensitized to PR10 but not exposed to birch pollen.

Resource manipulation through experimental defoliation has legacy effects on allocation to reproductive and vegetative organs in Quercus ilex.

BACKGROUND AND AIMS: In plants, high costs of reproduction during some years can induce trade-offs in resource allocation with other functions such as growth, survival and resistance against herbivores or extreme abiotic conditions, but also with subsequent reproduction. Such trade-offs might also occur following resource shortage at particular moments of the reproductive cycle. Because plants are modular organisms, strategies for resource allocation to reproduction can also vary among hierarchical levels. Using a defoliation experiment, our aim was to test how allocation to reproduction was impacted by resource limitation. METHODS: We applied three levels of defoliation (control, moderate and intense) to branches of eight Quercus ilex trees shortly after fruit initiation and measured the effects of resource limitation induced by leaf removal on fruit development (survival, growth and germination potential) and on the production of vegetative and reproductive organs the year following defoliation. KEY RESULTS: We found that defoliation had little impact on fruit development. Fruit survival was not affected by the intense defoliation treatment, but was reduced by moderate defoliation, and this result could not be explained by an upregulation of photosynthesis. Mature fruit mass was not affected by defoliation, nor was seed germination success. However, in the following spring defoliated branches produced fewer shoots and compensated for leaf loss by overproducing leaves at the expense of flowers. Therefore, resource shortage decreased resource allocation to reproduction the following season but did not affect sex ratio. CONCLUSIONS: Our results support the idea of a regulation of resource allocation to reproduction beyond the shoot scale. Defoliation had larger legacy effects than immediate effects.

Variation in seed traits among Mediterranean oaks in Tunisia and their ecological significance.

BACKGROUND AND AIMS: Oaks are the foundation and dominant tree species of most Mediterranean forests. As climate models predict dramatic changes in the Mediterranean basin, a better understanding of the ecophysiology of seed persistence and germination in oaks could help define their regeneration niches. Tunisian oaks occupy distinct geographical areas, which differ in their rainfall and temperature regimes, and are thus a valuable model to investigate relationships between seed traits and species ecological requirements. METHODS: Seed morphological traits, desiccation sensitivity level, lethal freezing temperature, embryonic axis and cotyledon sugar and lipid composition, and seed and acorn germination rates at various constant temperatures were measured in Quercus canariensis, Q. coccifera, Q. ilex and Q. suber, using seeds sampled in 22 Tunisian woodlands. KEY RESULTS: Only faint differences were observed for desiccation sensitivity in the oak species studied. By contrast, the species differed significantly in sensitivity to freezing, germination rates at low temperature and base temperature. Quercus ilex and Q. canariensis, which occur at high elevations where frost events are frequent, showed the lowest freezing sensitivity. A significant correlation was found between hexose contents in the embryonic axis and freezing tolerance. Significant interspecific differences in the time for seeds to germinate and the time for the radicle to pierce the pericarp were observed. The ratio of pericarp mass to acorn mass differed significantly among the species and was negatively correlated with the acorn germination rate. Quercus coccifera, which is frequent in warm and arid environments, showed the highest acorn germination rate and synchrony. CONCLUSIONS: Seed lethal temperature, seed germination time at low temperatures, the ratio of pericarp mass to acorn mass and the embryonic axis hexose content appeared to be key functional traits that may influence the geographical ranges and ecological requirements of Mediterranean oaks in Tunisia.

Glyphosate treatments for weed control affect early stages of root colonization by Tuber melanosporum but not secondary colonization.

The cultivation of the ectomycorrhizal fungus Tuber melanosporum has considerably spread in recent years throughout the world. During the first years of truffle cultivation, weed control is a key practice to improve the establishment of host trees and the proliferation of the fungus in the soil. Glyphosate is nowadays the most commonly used herbicide in Spanish truffle orchards. We explored the effect of glyphosate on the proliferation of T. melanosporum mycorrhizae, on extraradical mycelium and on the inoculum potential of T. melanosporum spores in greenhouse experiments using Quercus ilex seedlings as host plants. No detrimental effect on the secondary infection of T. melanosporum was found after three sequential glyphosate applications in young seedlings during one vegetative period. Instead, a change in the distribution of fine roots and T. melanosporum mycorrhizae along soil depth was observed. On the other hand, results indicate that high application rates of glyphosate hinder the infectivity of T. melanosporum spore inoculum, without apparent impact on the host performance. Our results suggest that glyphosate has the potential to jeopardise the role of the soil spore bank as inoculum source for the colonisation of new roots, also raising the question of whether glyphosate could hinder the presumed role of spores in sexual mating.

Dissecting the Seed Maturation and Germination Processes in the Non-Orthodox Quercus ilex Species Based on Protein Signatures as Revealed by 2-DE Coupled to MALDI-TOF/TOF Proteomics Strategy.

Unlike orthodox species, seed recalcitrance is poorly understood, especially at the molecular level. In this regard, seed maturation and germination were studied in the non-orthodox Quercus ilex by using a proteomics strategy based on two-dimensional gel electrophoresis coupled to matrix-assisted laser desorption ionization/time of flight (2-DE-MALDI-TOF).Cotyledons and embryo/radicle were sampled at different developmental stages, including early (M1-M3), middle (M4-M7), and late (M8-M9) seed maturation, and early (G1-G3) and late (G4-G5) germination. Samples corresponding to non-germinating, inviable, seeds were also included. Protein extracts were subjected to 2-dimensional gel electrophoresis (2-DE) and changes in the protein profiles were analyzed. Identified variable proteins were grouped according to their function, being the energy, carbohydrate, lipid, and amino acid metabolisms, together with protein fate, redox homeostasis, and response to stress are the most represented groups. Beyond the visual aspect, morphometry, weight, and water content, each stage had a specific protein signature. Clear tendencies for the different protein groups throughout the maturation and germination stages were observed for, respectively, cotyledon and the embryo axis. Proteins related to metabolism, translation, legumins, proteases, proteasome, and those stress related were less abundant in non-germinating seeds, it related to the loss of viability. Cotyledons were enriched with reserve proteins and protein-degrading enzymes, while the embryo axis was enriched with proteins of cell defense and rescue, including heat-shock proteins (HSPs) and antioxidants. The peaks of enzyme proteins occurred at the middle stages (M6-M7) in cotyledons and at late ones (M8-M9) in the embryo axis. Unlike orthodox seeds, proteins associated with glycolysis, tricarboxylic acid cycle, carbohydrate, amino acid and lipid metabolism are present at high levels in the mature seed and were maintained throughout the germination stages. The lack of desiccation tolerance in Q. ilex seeds may be associated with the repression of some genes, late embryogenesis abundant proteins being one of the candidates.

Rainfall exclusion and thinning can alter the relationships between forest functioning and drought.

Increasing drought caused by the ongoing climate change, and forest management by thinning that aims at mitigating its impact, may modify the current relationships between forest functions and drought intensity and preclude our ability to forecast future ecosystem responses. We used 15 yr of data from an experimental rainfall exclusion (-27% of rainfall) combined with thinning (-30% stand basal area) to investigate differences in the drought-function relationships for each component of above-ground net primary productivity (ANPP) and stand transpiration in a Mediterranean Quercus ilex stand. Rainfall exclusion reduced stand ANPP by 10%, mainly because of lowered leaf and acorn production, whereas wood production remained unaffected. These responses were consistent with the temporal sensitivity to drought among tree organs but revealed an increased allocation to wood. Thinning increased wood and acorn production and reduced the sensitivity of standing wood biomass change to drought. Rainfall exclusion and thinning lowered the intercept of the transpiration-drought relationship as a result of the structural constraints exerted by lower leaf and sapwood area. The results suggest that historical drought-function relationships can be used to infer future drought impacts on stand ANPP but not on water fluxes. Thinning can mitigate drought effects and reduce forest sensitivity to drought.

Proteomics, Holm Oak (Quercus ilex L.) and Other Recalcitrant and Orphan Forest Tree Species: How do They See Each Other?

Proteomics has had a big impact on plant biology, considered as a valuable tool for several forest species, such as Quercus, Pines, Poplars, and Eucalyptus. This review assesses the potential and limitations of the proteomics approaches and is focused on Quercus ilex as a model species and other forest tree species. Proteomics has been used with Q. ilex since 2003 with the main aim of examining natural variability, developmental processes, and responses to biotic and abiotic stresses as in other species of the genus Quercus or Pinus. As with the progress in techniques in proteomics in other plant species, the research in Q. ilex moved from 2-DE based strategy to the latest gel-free shotgun workflows. Experimental design, protein extraction, mass spectrometric analysis, confidence levels of qualitative and quantitative proteomics data, and their interpretation are a true challenge with relation to forest tree species due to their extreme orphan and recalcitrant (non-orthodox) nature. Implementing a systems biology approach, it is time to validate proteomics data using complementary techniques and integrate it with the -omics and classical approaches. The full potential of the protein field in plant research is quite far from being entirely exploited. However, despite the methodological limitations present in proteomics, there is no doubt that this discipline has contributed to deeper knowledge of plant biology and, currently, is increasingly employed for translational purposes.

Tree species identity and diversity drive fungal richness and community composition along an elevational gradient in a Mediterranean ecosystem.

Ecological and taxonomic knowledge is important for conservation and utilization of biodiversity. Biodiversity and ecology of fungi in Mediterranean ecosystems is poorly understood. Here, we examined the diversity and spatial distribution of fungi along an elevational gradient in a Mediterranean ecosystem, using DNA metabarcoding. This study provides novel information about diversity of all ecological and taxonomic groups of fungi along an elevational gradient in a Mediterranean ecosystem. Our analyses revealed that among all biotic and abiotic variables tested, host species identity is the main driver of the fungal richness and fungal community composition. Fungal richness was strongly associated with tree richness and peaked in Quercus-dominated habitats and Cistus-dominated habitats. The highest taxonomic richness of ectomycorrhizal fungi was observed under Quercus ilex, whereas the highest taxonomic richness of saprotrophs was found under Pinus. Our results suggest that the effect of plant diversity on fungal richness and community composition may override that of abiotic variables across environmental gradients.

Growth reduction after defoliation is independent of CO2 supply in deciduous and evergreen young oaks.

Reduced productivity of trees after defoliation might be caused by limited carbon (C) availability. We investigated the combined effect of different atmospheric CO2 concentrations (160, 280 and 560 ppm) and early season defoliation on the growth and C reserves (nonstructural carbohydrates (NSC)) of saplings of two oak species with different leaf habits (deciduous Quercus petraea and evergreen Quercus ilex). In both species, higher CO2 supply significantly enhanced growth. Defoliation had a strong negative impact on growth (stronger for Q. ilex), but the relative reduction of growth caused by defoliation within each CO2 treatment was very similar across all three CO2 concentrations. Low CO2 and defoliation led to decreased NSC tissue concentrations mainly in the middle of the growing season in Q. ilex, but not in Q. petraea. However, also in Q. ilex, NSC increased in woody tissues in defoliated and low-CO2 saplings towards the end of the growing season. Although the saplings were C limited under these specific experimental conditions, growth reduction after defoliation was not directly caused by C limitation. Rather, growth of trees followed a strong allometric relationship between total leaf area and conductive woody tissue, which did not change across species, CO2 concentrations and defoliation treatments.

Recent climate hiatus revealed dual control by temperature and drought on the stem growth of Mediterranean Quercus ilex.

A better understanding of stem growth phenology and its climate drivers would improve projections of the impact of climate change on forest productivity. Under a Mediterranean climate, tree growth is primarily limited by soil water availability during summer, but cold temperatures in winter also prevent tree growth in evergreen forests. In the widespread Mediterranean evergreen tree species Quercus ilex, the duration of stem growth has been shown to predict annual stem increment, and to be limited by winter temperatures on the one hand, and by the summer drought onset on the other hand. We tested how these climatic controls of Q. ilex growth varied with recent climate change by correlating a 40-year tree ring record and a 30-year annual diameter inventory against winter temperature, spring precipitation, and simulated growth duration. Our results showed that growth duration was the best predictor of annual tree growth. We predicted that recent climate changes have resulted in earlier growth onset (-10 days) due to winter warming and earlier growth cessation (-26 days) due to earlier drought onset. These climatic trends partly offset one another, as we observed no significant trend of change in tree growth between 1968 and 2008. A moving-window correlation analysis revealed that in the past, Q. ilex growth was only correlated with water availability, but that since the 2000s, growth suddenly became correlated with winter temperature in addition to spring drought. This change in the climate-growth correlations matches the start of the recent atmospheric warming pause also known as the 'climate hiatus'. The duration of growth of Q. ilex is thus shortened because winter warming has stopped compensating for increasing drought in the last decade. Decoupled trends in precipitation and temperature, a neglected aspect of climate change, might reduce forest productivity through phenological constraints and have more consequences than climate warming alone.

An ecosystem services approach to the ecological effects of salvage logging: valuation of seed dispersal.

Forest disturbances diminish ecosystem services and boost disservices. Because post-disturbance management intends to recover the greatest possible value, selling timber often prevails over other considerations. Ecological research has shown diverse effects of salvage logging, yet such research has focused on the biophysical component of post-disturbance ecosystems and lacks the link with human well-being. Here we bridge that gap under the ecosystem services framework by assessing the impact of post-fire management on a non-timber value. By employing the replacement cost method, we calculated the value of the post-fire natural regeneration of Holm oaks in southern Spain under three post-fire management options by considering the cost of planting instead. The value of this ecosystem service in non-intervention areas doubled that of salvage-logged stands due to the preference for standing dead trees by the main seed disperser. Still, most of the value resulted from the resprouting capacity of oaks. The value of this and other ecosystem services should be added to traditional cost/benefit analyses of post-disturbance management. We thus call for a more holistic approach to salvage logging research, one that explicitly links ecological processes with human well-being through ecosystem services, to better inform decision-makers on the outcomes of post-disturbance management.

Quercus ilex L.: How season, Plant Organ and Extraction Procedure Can Influence Chemistry and Bioactivities.

Quercus species have a plethora of applications, either in wine and wood industries, in human and animal nutrition or in human health. In order to improve the knowledge on this genus, the aim of the present study was to correlate, for the first time, the phenolic composition of different Quercus ilex L. plant tissues (leaves in two maturation stages, acorns, teguments and cotyledons) and different extraction procedures with scavenging and anticholinesterase activities. The hydromethanolic and aqueous extracts obtained showed strong radical scavenging activity against DPPH, superoxide anion radical and nitric oxide radical, leaves exhibiting higher total phenolic content and revealing the best antioxidant properties, followed by tegument and acorns. Concerning the phenolic profile, fifteen compounds were identified and quantified by HPLC-DAD, ranging from 1568.43 to 45,803.16 mg/kg dried extract. The results indicate that Q. ilex can be a source of strong antioxidant phenolic compounds with possible interest for food and pharmaceutical industries.

Shifting demographic conflicts across recruitment cohorts in a dynamic post-disturbance landscape.

Seed dispersal effectiveness, which measures the number of adult plant individuals produced by seed dispersal, is the product of the number of seeds dispersed and the probability a seed produces an adult. Directed dispersal to certain habitat types may enhance some stages of recruitment but disfavor others, generating demographic conflicts in plant ontogeny. We asked whether temporal changes in habitat features may affect the distribution of seedlings recruited from dispersed acorns, and whether this could induce shifts in the life-stage conflicts experienced by successive cohorts of naturally recruited plants. As early successional habitats are characterized by rapid change, we used a burnt pine stand in southern Spain to monitor the recruitment and performance of a major tree species (Quercus ilex) across 7 yr in four types of post-fire habitats. These differed in structure and included patches of unburnt forest and three management alternatives of burnt trees: logging, partial cutting, and nonintervention. Young oaks that resprouted after the fire were mainly located near acorn sources, while new seedlings initially emerged mostly in habitats with standing snags due to habitat selection by European jays, Garrulus glandarius, for dispersal. The dead pines gradually collapsed and attracted less dispersal, so subsequent seedling cohorts mainly recruited within patches of unburnt pines. These live pines enhanced the survival of the oaks located beneath their canopy but greatly reduced their growth as compared to the other post-fire habitats, thus representing a demographic conflict that was absent elsewhere. As a consequence of the directional shift in the habitat where seedlings recruited, successive seedling cohorts experienced a gradual improvement in their likelihood of survival but a reduction in growth. The progressive intensification of this life-stage conflict hinged on the reduction of vertical structures in the habitat with standing burnt pines. Recruitment success thus involved temporal variation in the habitat where recruitment occurred, likely resulting from changes in the direction of seed dispersal, and spatial variation in habitat suitability for seedling establishment and growth. Temporal changes in habitat structure can indirectly change the environment in which recruitment occurs, and consequently seed dispersal effectiveness, by shifting the direction of seed dispersal.

Explanatory ecological factors for the persistence of desiccation-sensitive seeds in transient soil seed banks: Quercus ilex as a case study.

BACKGROUND AND AIMS: Dominant tree species in northern temperate forests, for example oak and beech, produce desiccation-sensitive seeds. Despite the potentially major influence of this functional trait on the regeneration and distribution of species under climate change, little is currently known about the ecological determinants of the persistence of desiccation-sensitive seeds in transient soil seed banks. Knowing which key climatic and microsite factors favour seed survival will help define the regeneration niche for species whose seeds display extreme sensitivity to environmental stress METHODS: Using the Mediterranean Holm oak (Quercus ilex) forest as a model system, an in situ time-course monitoring of seed water status and viability was performed during the unfavourable winter season in two years with contrasting rainfall, at an instrumented site with detailed climate records. In parallel, the characteristics of the microhabitat and their influence on the post-winter water status and viability of seeds were investigated in a regional survey of 33 woodlands representative of the French distribution of the species. KEY RESULTS: Time-course monitoring of seed water status in natural conditions confirmed that in situ desiccation is the main abiotic cause of mortality in winter. Critical water contents could be reached in a few days during drought spells. Seed dehydration rates were satisfactorily estimated using integrative climate proxies including vapour pressure deficit and potential evapotranspiration. Seed water status was therefore determined by the balance between water uptake after a rainfall event and water loss during dry periods. Structural equation modelling of microhabitat factors highlighted the major influence of canopy openness and resulting incident radiation on the ground. CONCLUSIONS: This study provides part of the knowledge required to implement species distribution models which incorporate their regeneration niche. It is an important step forward in evaluating the ecological consequences of increasing winter drought and environmental filtering due to climate change on the regeneration of the most dominant Mediterranean tree species.