Chlorophyll fluorescence responses to temperature and water availability in two co-dominant Mediterranean shrub and tree species in a long-term field experiment simulating climate change

A rain exclusion experiment simulating drought conditions expected in Mediterranean areas for the following decades (15% decrease in soil moisture) is being conducted since 1999 in a Mediterranean holm oak forest to study its response to the forecasted climatic changes for the coming decades. The maximum PSII quantum yield of primary photochemistry (Fv/Fm) was measured in Quercus ilex, and Phillyrea latifolia, the co-dominant species of the studied forest, from 1999 to 2009 in four plots: two of them were control plots and the other two plots received the rain exclusion treatment. In both species, the Fv/Fm values were highly dependent on air temperatures, and in a second term, in water availability. P. latifolia was the species with the larger decrease in Fv/Fm values induced by low air temperatures, while in hot seasons, the Fv/Fm values in P. latifolia were even higher than in Q. ilex. Rainfall exclusion decrease Fv/Fm values significantly only in few monitoring dates. The most drought resistant species P. latifolia was more affected by the experimental rainfall exclusion than Q. ilex that instead lost number of leaves per tree. There was a synergic effect of drought stress and winter cold in P. latifolia not observed in Q. ilex, but a more conservative strategy in P. latifolia maintaining leaves with a down-regulation of the linear photosynthetic electron transport. These results indicate that, although other physiological and reproductive strategies at whole plant level must be also taken into account, the warmer and drier environment expected for the following decades could favour the species more sensitive to cold and more resistant to drought, the shrub P. latifolia, in detriment of the tree Q. ilex as already observed in the field after severe heat-drought episodes.     

Tissue-water relations of two co-occurring evergreen Mediterranean species in response to seasonal and experimental drought conditions

Tissue-water relations were used to characterize the responses of two Mediterranean co-occurring woody species (Quercus ilex L. and Phillyrea latifolia L.) to seasonal and experimental drought conditions. Soil water availability was reduced 15% by partially excluding rain throughfall and lateral flow (water runoff). Seasonal and experimental drought elicited physiological and morphological adaptations other than osmotic adjustment: both species showed large increases in cell-wall elasticity and decreased saturated-to-dry-mass ratio. Increased elasticity (lower elastic modulus) resulted in concurrent decreases in relative water content at turgor loss. In addition, P. latifolia showed significant increases in apoplastic water fraction. Decreased saturated-to-dry-mass ratio and increased apoplastic water fraction were accompanied by an increased range of turgor maintenance, which indicates that leaf sclerophyllous traits might be advantageous in drier scenarios. In contrast, the degree of sclerophylly (as assessed by the leaf mass-to-area ratio) was not related to tissue elasticity. An 15% reduction in soil water availability resulted in significant reductions in diameter growth when compared to control plants in both species. Moreover, although P. latifolia underwent larger changes in tissue water-related traits than Q. ilex in response to decreasing water availability, growth was more sensitive to water stress in P. latifolia than in Q. ilex. Differences in diameter growth between species might be partially linked to the effects of cell-wall elasticity and turgor pressure on growth, since Q. ilex showed higher tissue elasticity and higher intrinsic tolerance to water deficit (as indicated by lower relative water content at turgor loss) than P. latifolia.     

Tree growth,mortality, and above-ground biomass accumulation in a holm oak forest under a five-year experimental field drought

A holm oak forest was exposed to an experimental drought during 5 years to elucidate the growth responses of the dominant species Quercus ilex, Arbutus unedo and Phillyrea latifolia. Soil water availability was partially reduced, about 15% as predicted for this area for the next decades by GCM and ecophysiological models, by plastic strips intercepting rainfall and by ditch exclusion of water runoff. The stem diameter increment was highly correlated with annual rainfall in all species, and drought treatment strongly reduced the diameter increment of Q. ilex (41%) and specially of A. unedo (63%), the species showing higher growth rates. Stem mortality rates were highly correlated with previous stem density, but drought treatment increased mortality rates in all species. Q. ilex showed the highest mortality rates (9% and 18% in control and drought plots, respectively), and P. latifolia experienced the lowest mortality rates (1% and 3% in control and drought plots, respectively). Drought strongly reduced the increment of live aboveground biomass during these 5 years (83%). A. unedo and Q. ilex experienced a high reduction in biomass increment by drought, whereas P. latifolia biomass increment was insensitive to drought. The different sensitivity to drought of the dominant species of the holm oak forest may be very important determining their future development and distribution in a drier environment as expected in Mediterranean areas for the next decades. These drier conditions could thus have strong effects on structure (species composition) and functioning (carbon uptake and biomass accumulation) of these Mediterranean forests.     

Establishment of co‐existing Mediterranean tree species under a varying soil moisture regime

We investigated the responses of two co‐existing Mediterranean trees with different regeneration strategies (Phillyrea latifolia seedlings and Quercus ilex sprouts) to experimental drought below the forest canopy. We considered different recruitment stages and used leaf isotopic discrimination to estimate water use efficiency (WUE) and nitrogen availability and use. Drought decreased the emergence and survival of seedlings and sprouts. Survival and growth of older saplings were not influenced by drought. Seedling emergence of P. latifolia was higher than Q. ilex sprout production, but Q. ilex sprouts had higher survival and growth rates. These differences disappeared in the sapling stage. Carbon isotopic discrimination suggested that Q. ilex sprouts had higher WUE than P. latifolia seedlings. Drought increased WUE of recruits, particularly in Q. ilex. Water use regulation increased with ageing, particularly in P. latifolia. Q. ilex had higher δ¹⁵N values than P. latifolia; these were also higher under drier soil conditions. Current year seedlings had higher δ¹⁵N than saplings, particularly in P. latifolia, suggesting they exploit superficial soil layers. These results suggest that sprouts obtain benefit from resources stored in parent plants. At earlier stages, they perform better than seedlings. This response is not coupled to adult vulnerability to drought for these species, revealing the difficulty of predicting species dynamics during climate change.     

Contribution of physiological and morphological adjustments to drought resistance in two Mediterranean tree species

Plant water potential (ψ), its components, and gas exchange data of two Mediterranean co-occurring woody species (Quercus ilex L. and Phillyrea latifolia L.) were measured in response to seasonal changes in water availability over two consecutive years. The relative contribution of physiological and morphological adjustments to drought resistance was assessed through Principal Component Analyses. There were large adjustments in stomatal conductance (∼36 % of accounted variance). Net photosynthetic rate and water use efficiency were closely tuned to water availability and accounted for ∼17 % of variance. The slope of the water potential vs. relative water content (dψ/dRWC₀) below zero pressure potential increased as a result of seasonal and ontogenic increases in apoplastic water fraction and accounted for ∼20 % variance. This tolerance mechanism was accompanied by an increased range of positive pressure potential, suggesting a functional role of sclerophylly in these Mediterranean evergreens. Similarly, changes in the slope of dψ/dRWC in the range of positive pressure potential (∼13 % of accounted variance) were associated to variations in cell wall elasticity and resulted in lower RWC at zero pressure potential. When considering the species studied separately, the results indicated the primary role of stomatal regulation in the drought resistance of Qilex, while increased apoplastic water fraction had a major contribution in the drought resistance of P. latifolia. This research was supported by Spanish CICYT grants CLI99-0479 and REN-2002-00633. L.S. acknowledges the financial support from Ministerio de Ciencia y Tecnologia (“Ramon y Cajal” program, Spain). An erratum to this article is available at .     

Leaf plasticity in response to light of three evergreen species of the Mediterranean maquis

Morphological, anatomical, biochemical and physiological traits of sun and shade leaves of adult Quercus ilex, Phillyrea latifolia and Pistacia lentiscus shrub species co-occurring in the Mediterranean maquis at Castelporziano (Latium) were studied. Fully expanded sun leaves had 47% (mean of the three species) greater leaf mass area (LMA) and 31% lower specific leaf area (SLA) than shade leaves. Palisade parenchyma thickness contributed on an average 42% to the total leaf thickness, spongy layer 43%, upper epidermal cells 5%, and upper cuticle thickness 3%. Stomatal size was greater in sun (25.5 μm) than in shade leaves (23.6 μm). Total chlorophyll content per fresh mass was 71% greater in shade than in sun leaves, and nitrogen content was the highest in sun (13.7 mg g⁻¹) than in shade leaves (11.8 mg g⁻¹). Difference of net photosynthetic rates (P _N) between sun and shade leaves was 97% (mean of the three species). The plasticity index (sensu Valladares et al., New Phytol 148:79–91, 2000a) was the highest for physiological leaf traits (0.86) than for morphological, anatomical and biochemical ones. Q. ilex had the highest plasticity index of morphological, anatomical and physiological leaf traits (0.37, 0.28 and 0.71, respectively) that might explain its wider ecological distribution. The higher leaf plasticity of Q. ilex might be advantageous in response to varying environmental conditions, including global change.     

Seasonal ultrasonic acoustic emissions of <Emphasis Type="Italic">Quercus ilex</Emphasis> L. trees in a Mediterranean forest

Ultrasonic acoustic emissions were measured in Quercus ilex trees of a Mediterranean forest in Catalonia (NE Spain) each season from summer of 2004 to autumn of 2005. Acoustic emissions were maximum during hot and dry summer periods. Acoustic emissions started below 17% soil moisture, 0.85 RWC, and 2.5 MPa leaf water potential. They were negatively correlated with soil moisture and leaf water potential. The relationship between acoustic emissions and leaf water potential was the strongest, indicating that xylem tension is the most important factor inducing both cavitation (acoustic emissions) and a decrease in leaf water potential. Future increase of xylem cavitation derived from climate change may result in growth and survival limitations for this species in the drier southern limits of its current distribution.     

Increasing drought decreases phosphorus availability in an evergreen Mediterranean forest

Mediterranean ecosystems are water-limited and frequently also nutrient-limited. We aimed to investigate the effects of increasing drought, as predicted by GCM and eco-physiological models for the next decades, on the P cycle and P plant availability in a Mediterranean forest. We conducted a field experiment in a mature evergreen oak forest, establishing four drought-treatment plots and four control plots (150 m² each). After three years, the runoff and rainfall exclusion reduced an overall 22% the soil moisture, and the runoff exclusion alone reduced it 10%. The reduction of 22% in soil moisture produced a decrease of 40% of the accumulated aboveground plant P content, above all because there was a smaller increase in aerial biomass. The plant leaf P content increased by 100 ± 40 mg m⁻² in the control plots, whereas it decreased by 40 ± 40 mg m⁻² in the drought plots. The soil Po-NaHCO₃ (organic labile-P fraction) increased by 25% in consonance with the increase in litterfall, while the inorganic labile-P fraction decreased in relation to the organic labile-P fraction up to 48%, indicating a decrease in microbial activity. Thus, after just three years of slight drought, a clear trend towards an accumulation of P in the soil and towards a decrease of P in the stand biomass was observed. The P accumulation in the soil in the drought plots was mainly in forms that were not directly available to plants. These indirect effects of drought including the decrease in plant P availability, may become a serious constraint for plant growth and therefore may have a serious effect on ecosystem performance.     

Early to mid-Pleistocene ostracod δO and δC in the central Tibetan Plateau: Implication for Indian monsoon change

As one of the pioneering projects of the Chinese Environmental Scientific Drilling (CESD) Program, a 206.5 m long sediment core (CE) was retrieved from Co Ngoin (Co = lake) Basin in the central Tibetan Plateau. Limnic records of the abundance of ostracod shells and their oxygen and carbon isotope measurements, together with the geochemical and the sedimentological investigations, were carried out on the interval between 143 and 52 m core depths, corresponding to ca. 2.01–0.84 Myr. Distinct oscillations of the abundance and the chemical compositions of ostracod shells have yielded the original information about variations in lakewater chemistry in association with the change in the Indian monsoon over the period of 2.01 to 0.84 Myr. The climate conditions during the Early Pleistocene were characterized by frequent oscillations and the ostracod records had expressed its synchronous response to these oscillations. A major climate change occurred in the Co Ngoin area at about 0.93 Myr. A pronounced fluctuation in ostracod δ¹⁸O and δ¹³C during the Mid-Pleistocene transition (MPT) is possibly associated with an increase in continental ice sheets. The compositional and the sedimentological characters of the immediately overlying sediments imply a shallow aquatic environment and also a harsh (salinity?) condition beyond the tolerance limits for ostracod survival, resulting in the disappearance of the Qinghaicypris crassa Huang 1979 around 0.84 Myr.     

Organic matter rain rates, oxygen availability, and vital effects from benthic foraminiferal δC in the historic Skagerrak, North Sea

The sediment cores 225514 and 225510 were recovered from 420 and 285 m water depth, respectively. They were investigated for their benthic foraminiferal δ¹³C during the last 500 years. Both cores were recovered from the southern flank of the Skagerrak. The δ¹³C values of Uvigerina mediterranea and other shallow infaunal species in both cores indicate that organic matter rain rates to the seafloor varied around a mean value until approximately AD 1950 after which they increased. This increase might result from changes in the North Atlantic Current System and a co-occurring persistently high North Atlantic Oscillation index state in the 1980s to 1990s, rather than from anthropogenic eutrophication. Using δ¹³C mean values of multiple species, we reconstruct δ¹³C gradients of dissolved inorganic carbon (DIC) within pore waters for the time periods AD 1500 to 1950 and AD 1950 to 2000. The calculated δ¹³C_DIC ranges, interpreted as indicating total organic matter remineralization due to respiration, are generally bigger in Core 225514 than in Core 225510. Since mean δ¹³C values of U. mediterranea suggest that organic matter rain rates were similar at both locations, differences in total organic matter remineralization are attributed to differing oxygen availability. However, oxygen concentrations in the overlying bottom water masses are not likely to have differed significantly. Thus, we suggest that organic matter remineralization was controlled by oxygen availability within the sediments, reflecting strong differences in sedimentation rates at the two investigated core sites. Based on the assumptions that tests of benthic foraminiferal species inhabiting the same microhabitat depth should show equal δ¹³C values unless they are affected by vital effects and that Globobulimina turgida records pore water δ¹³C_DIC, we estimate microhabitat-corrected vital effects for several species with respect to G. turgida: > 0.7‰ for Cassidulina laevigata, > 1.3‰ for Hyalinea balthica, and > 0.7‰ for Melonis barleeanus. Melonis zaandami seems to closely record pore water δ¹³C_DIC.     

Tissue-specific variation of δC in mature canopy trees in a temperate forest in central Europe

Stable carbon isotope analysis has become a key tool in functional ecology, yet considerable natural variability often limits the interpretations. In this study we document the spatial, taxonomic, temporal and tissue-specific δ¹³C variability in 10 tree species of a temperate European forest. The Swiss Canopy Crane provided access to the three dimensional space within 55 trees 30–35 m high representing the genera Acer, Carpinus, Fagus, Prunus, Quercus, Tilia, Abies, Larix, Picea and Pinus. The results from six broad-leaved and four conifer species (seven deciduous, three evergreen) documented that the species effect was not significant in contrast to tissue-specific and spatial differences in the canopy. Year-to-year differences were not large but still significant.

Our analysis confirmed a significant difference between δ¹³C of foliage collected in the upper and lower canopy, but revealed no systematic differences with respect to azimuthal directions in tree crowns of the broad-leaved trees, as opposed to the conifers, which show clear differences between the sun-exposed and the shaded side. Tissue-specific differences were significant, despite surprisingly similar mean values for most tissue types. Such tissue effect was largely due to young branch xylem, which exhibited a systematic less negative deviation from the other tissue types. These findings were consistent across the species tested and provided some guidelines towards a representative sampling strategy for ¹³C analysis.     

Antinoceptive effect of triterpenoid α,β-amyrin in rats on orofacial pain induced by formalin and capsaicin

The effects of α,β-amyrin, a pentacyclic triterpene isolated from Protium heptaphylum was investigated on rat model of orofacial pain induced by formalin or capsaicin. Rats were pretreated with α,β-amyrin (10, 30, and 100 mg/kg, i.p.), morphine (5 mg/kg, s.c.) or vehicle (3% Tween 80), before formalin (20 μl, 1.5%) or capsaicin (20 μl, 1.5 μg) injection into the right vibrissa. In vehicle-treated controls, formalin induced a biphasic nociceptive face-rubbing behavioral response with an early first phase (0–5 min) and a late second phase (10–20 min) appearance, whereas capsaicin produced an immediate face-rubbing (grooming) behavior that was maximal at 10–20 min. Treatment with α,β-amyrin or morphine significantly inhibited the face-rubbing response in both test models. While morphine produced significant antinociception in both phases of formalin test, α,β-amyrin inhibited only the second phase response, more prominently at 30 mg/kg, in a naloxone-sensitive manner. In contrast, α,β-amyrin produced much greater antinociceptive effect at 100 mg/kg in the capsaicin test, which was also naloxone-sensitive. These results provide first time evidence to show that α,β-amyrin attenuates orofacial pain atleast, in part, through a peripheral opioid mechanism but warrants further detailed study for its utility in painful orofacial pathologies.     

陕西省3种主要树种叶片、凋落物和土壤N、P化学计量特征

以陕西省29个县(市)39个样点的刺槐、辽东栎和油松林为研究对象,分析比较不同树种乔木叶片、凋落物与土壤N、P化学计量特征及其与经纬度、海拔、年均温度和年降水等环境因子间关系的异同以及三者之间可能存在的关系,以期为认识陕西省主要森林树种养分限制状况、制定合理的植被管理和恢复措施提供理论依据。结果表明:3树种叶片N、P含量及比值均为刺槐辽东栎油松,与叶片相比,凋落物中N、P含量变化幅度较小,为刺槐辽东栎油松,N∶P比值为油松辽东栎刺槐。10—20 cm与0—10 cm土层相比,3树种中除辽东栎中P含量差异不显著外,其它指标N、P含量及N∶P比值均显著下降(P0.05)。刺槐、辽东栎和油松叶片N、P含量与土壤N、P含量均没有显著相关性,以刺槐、辽东栎和油松3种植物叶片为总体来说,P含量与土壤P含量显著正相关(P0.05)。叶片N、P含量均大致表现出随着年均温度和年降水的增加而增加,随着经纬度的增加而降低的趋势,这一点在刺槐叶中最为明显。凋落物N含量随着年均温度和年降水的增加而增加,随着纬度和经度的增加而降低;P含量随着年降水和经度的增加而降低;N∶P比值随着年均温度和年降水的增加而增加,随着纬度的增加而降低。研究区内,土壤N、P含量随着纬度、海拔的增加和年均温度、年降水、经度的降低而增加,N∶P比值则呈相反的趋势。3树种土壤N、P含量及N∶P比值中,P含量比N含量受环境影响更大,且0—10 cm和10—20 cm土层N、P含量及N∶P比值与各环境因子的关系基本一致。     

In situ measurement of fine root water absorption in three temperate tree species—Temporal variability and control by soil and atmospheric factors

Miniature heat balance-sap flow gauges were used to measure water flows in small-diameter roots (3–4 mm) in the undisturbed soil of a mature beech–oak–spruce mixed stand. By relating sap flow to the surface area of all branch fine roots distal to the gauge, we were able to calculate real time water uptake rates per root surface area (J_s) for individual fine root systems of 0.5–1.0 m in length. Study aims were (i) to quantify root water uptake of mature trees under field conditions with respect to average rates, and diurnal and seasonal changes of J_s, and (ii) to investigate the relationship between uptake and soil moisture θ, atmospheric saturation deficit D, and radiation I. On most days, water uptake followed the diurnal course of D with a mid-day peak and low night flow. Neighbouring roots of the same species differed up to 10-fold in their daily totals of J_s (<100–2000 g m⁻² d⁻¹) indicating a large spatial heterogeneity in uptake. Beech, oak and spruce roots revealed different seasonal patterns of water uptake although they were extracting water from the same soil volume. Multiple regression analyses on the influence of D, I and θ on root water uptake showed that D was the single most influential environmental factor in beech and oak (variable selection in 77% and 79% of the investigated roots), whereas D was less important in spruce roots (50% variable selection). A comparison of root water uptake with synchronous leaf transpiration (porometer data) indicated that average water fluxes per surface area in the beech and oak trees were about 2.5 and 5.5 times smaller on the uptake side (roots) than on the loss side (leaves) given that all branch roots <2 mm were equally participating in uptake. Beech fine roots showed maximal uptake rates on mid-summer days in the range of 48–205 g m⁻² h⁻¹ (i.e. 0.7–3.2 mmol m⁻² s⁻¹), oak of 12–160 g m⁻² h⁻¹ (0.2–2.5 mmol m⁻² s⁻¹). Maximal transpiration rates ranged from 3 to 5 and from 5 to 6 mmol m⁻² s⁻¹ for sun canopy leaves of beech and oak, respectively. We conclude that instantaneous rates of root water uptake in beech, oak and spruce trees are above all controlled by atmospheric factors. The effects of different root conductivities, soil moisture, and soil hydraulic properties become increasingly important if time spans longer than a week are considered.     

Seasonal and annual changes in leaf δ13C in two co-occurring Mediterranean oaks: relations to leaf growth and drought progression

1. Changes of δ¹³C and its relation to leaf development, biochemical content and water stress were monitored over a 2 year period in two co-occurring Mediterranean oak species: the deciduous Quercus pubescens and the evergreen Quercus ilex .
2. The time course of leaf δ¹³C showed different patterns in the two species. Young Q. pubescens leaves had a high δ¹³C and a marked decrease occurred during leaf growth. In contrast, leaves at budburst and maturity did not differ significantly in the case of Q. ilex . We suggest that the difference between δ¹³C of young leaves was linked to differential use of reserves of carbon compounds in the two species.
3. δ¹³C values of mature leaves were negatively correlated with minimum seasonal values of predawn water potential, suggesting that a functional adjustment to water resources occurred.
4. There was a significant correlation between individual δ¹³C values for two successive years. This interannual dependence showed that δ¹³C rankings between trees were constant through time.     

The absorption, fluorescence and phosphorescence of phalloidin and α-amanitin

The peptides, phalloidin and α-amanitin, contain two unusual indole-derived chromophores, whose absorption, fluorescence and phosphorescence in aqueous solutions are described. Fluorescence is very weak, but phosphorescence at 77 K is intense. Phalloidin seems to undergo adiabatic photodissociation in alkaline solutions and to fluorescence from the indole anion form. In contrast, neither the phenolic nor the indolic protons of α-amanitin undergo photodissociation. The pK_a value of the phenolic hydroxy group of α-amanitin is 9.71 at 23°C.     

Antibacterial activity of α-mangostin against vancomycin resistant Enterococci (VRE) and synergism with antibiotics

alpha-Mangostin, isolated from the stem bark of Garcinia mangostana L., was found to be active against vancomycin resistant Enterococci (VRE) and methicillin resistant Staphylococcus aureus (MRSA), with MIC values of 6.25 and 6.25 to 12.5 microg/ml, respectively. Our studies showed synergism between alpha-mangostin and gentamicin (GM) against VRE, and alpha-mangostin and vancomycin hydrochloride (VCM) against MRSA. Further studies showed partial synergism between alpha-mangostin and commercially available antibiotics such as ampicillin and minocycline. These findings suggested that alpha-mangostin alone or in combination with GM against VRE and in combination with VCM against MRSA might be useful in controlling VRE and MRSA infections.     

γ -Ray-Induced DNA Damage and Repair in Methanosarcina barkeri

The capacity of the mesophilic archaeon, Methanosarcina barkeri (DSM 804) for DNA double strand break repair following⁶⁰Co- γ irradiation was investigated. The genome (1.9 Mb) of Methanosarcina barkeri was largely fragmented and was found to be repaired on incubation in medium under anaerobic conditions at 37°C for 4 h. To get an insight into its repair process a set of inhibitors were used. The methanogenesis inhibitor, bromoethanesulfonate showed partial inhibition of repair in Methanosarcina barkeri but not in Escherichia coli or human peripheral blood mononuclear cells. The Methanosarcina barkeri cells could also partially repair the DNA damage in a non-nutrient medium. Arabinosine-CTP, a nucleoside analogue and a polymerase inhibitor, completely inhibited repair in this archaeon. Arabinosine-CTP did not affect DSB (double-strand break) repair in human peripheral blood mononuclear cells but completely inhibited repair in Escherichia coli (a bacterium). The involvement of polymerase indicates recombination to be the underlying mechanism in DSB repair of Methanosarcina barkeri. 3-Aminobenzamide, a poly (ADP-ribose) polymerase inhibitor, completely inhibited repair in this archaeon as well as in eukarya but not in Escherichia coli showing the involvement of poly (ADP-ribose) polymerase in the DSB repair of Methanosarcina barkeri.     

The combined effects of a long‐term experimental drought and an extreme drought on the use of plant‐water sources in a Mediterranean forest

Vegetation in water‐limited ecosystems relies strongly on access to deep water reserves to withstand dry periods. Most of these ecosystems have shallow soils over deep groundwater reserves. Understanding the functioning and functional plasticity of species‐specific root systems and the patterns of or differences in the use of water sources under more frequent or intense droughts is therefore necessary to properly predict the responses of seasonally dry ecosystems to future climate. We used stable isotopes to investigate the seasonal patterns of water uptake by a sclerophyll forest on sloped terrain with shallow soils. We assessed the effect of a long‐term experimental drought (12 years) and the added impact of an extreme natural drought that produced widespread tree mortality and crown defoliation. The dominant species, Quercus ilex, Arbutus unedo and Phillyrea latifolia, all have dimorphic root systems enabling them to access different water sources in space and time. The plants extracted water mainly from the soil in the cold and wet seasons but increased their use of groundwater during the summer drought. Interestingly, the plants subjected to the long‐term experimental drought shifted water uptake toward deeper (10–35 cm) soil layers during the wet season and reduced groundwater uptake in summer, indicating plasticity in the functional distribution of fine roots that dampened the effect of our experimental drought over the long term. An extreme drought in 2011, however, further reduced the contribution of deep soil layers and groundwater to transpiration, which resulted in greater crown defoliation in the drought‐affected plants. This study suggests that extreme droughts aggravate moderate but persistent drier conditions (simulated by our manipulation) and may lead to the depletion of water from groundwater reservoirs and weathered bedrock, threatening the preservation of these Mediterranean ecosystems in their current structures and compositions.