The formation of coccoliths, intricate calcium carbonate scales that cover the cells of unicellular marine microalgae, is a highly regulated biological process. For decades, scientists have tried to elucidate the cellular, chemical, and structural mechanisms that control the precise mineralogy and shape of the inorganic crystals. Transmission electron microscopy was pivotal in characterizing some of the organelles that orchestrate this process. However, due to the difficulties in preserving soluble inorganic phases during sample preparation, only recently, new intracellular ion-pools were detected using state-of-the-art cryo X-ray and electron microscopy techniques. Here, we combine a completely non-aqueous sample preparation procedure and room temperature electron microscopy, to investigate the presence, cellular location, and composition, of mineral phases inside mineral forming microalga species. This methodology, which fully preserves the forming coccoliths and the recently identified Ca-P-rich bodies, allowed us to identify a new class of ion-rich compartments that have complex internal structure. In addition, we show that when carefully choosing heavy metal stains, elemental analysis of the mineral phases can give accurate chemical signatures of the inorganic phases. Applying this approach to mineral forming microalgae will bridge the gap between the low-preservation power for inorganic phases of conventional chemical-fixation based electron microscopy, and the low-yield of advanced cryo techniques.

The potential impacts of species colonization on the structure and functioning of ecosystems are poorly understood. We propose a novel approach for understanding the consequences of habitat colonization, highlighting the influence of colonists on the availability of limiting resources to resident species. We studied how colonization of dry oak woodlands by pines (Pinus halepensis) is affecting water stress of resident oaks (Quercus calliprinos). We monitored predawn leaf water potential (PLWP) of oaks monthly for 2 years. Using maximum likelihood and multi-model inference, we evaluated how the PLWP of oaks was affected by pine colonists. The influence of colonizing pines on PLWP of resident oaks varied in time and space from negative to positive depending on season, oak size, pine size, and proximity to pines presence. The water stress of oaks increased along the dry season (- 1.5 to - 4.5 MPa), with small oaks becoming more severely stressed than large ones (up to 60% difference). During the dry season, neighboring pine colonists increased the water stress of oaks (up to - 0.4 MPa difference), but during the wet season, they reduced the water stress mainly for large oaks. Our findings indicate that pine colonization differentially affects water limitation for resident oaks with implications for future development and regeneration. The influence of pine colonists shifted from positive to negative along an increasing water stress gradient, contrary to predictions by the stress gradient hypothesis. Our work demonstrates how colonization by non-resident species can influence key ecosystem processes through the redistribution of limiting resources. Identifying these processes is fundamental for understanding the consequences of colonization, mitigating these influences, and predicting future change in the structure and function of ecosystems.

Supplementation of broiler breeder hens with beneficial additives bears great potential for affecting nutrient deposition into the fertile egg. Guanidinoacetate (GAA) is the endogenous precursor of creatine that is used as a feed additive for improving cellular energy metabolism in animal nutrition. In the present study, we have investigated whether GAA supplementation in broiler breeder feed affects creatine deposition into the hatching egg and molecular mechanisms of creatine transport and synthesis within hens and their progeny. For this, broiler breeder hens of 47 wk of age were supplemented with 0.15% GAA for 15 wk, and samples from their tissues, hatching eggs and progeny were compared with those of control, nonsupplemented hens. A significant increase in creatine content was found within the yolk and albumen of hatching eggs obtained from the GAA group, compared with the control group. The GAA group exhibited a significant increased creatine transporter gene expression compared with the control group in their small intestines and oviduct. In GAA group progeny, a significant decrease in creatine transporter expression at embryonic day 19 and day of hatch was found, compared with control group progeny. At the day of hatch, creatine synthesis genes (arginine glycine amidinotransferase and guanidinoacetate N-methyltransferase) exhibited significant decrease in expression in the GAA group progeny compared with control group progeny. These results indicate that GAA supplementation in broiler breeder feed increases its absorbance and deposition into hatching eggs, subsequently affecting GAA and creatine absorbance and synthesis within broiler progeny.

Accelerated microbial degradation following previous repeated applications of the same pesticide, or another pesticide of a similar chemical structure, is a known phenomenon. Currently there is limited information regarding accelerated degradation of sulfonylurea (SU) herbicides. This study is aimed to evaluate the effect of repeated SU applications on the degradation rate of the SU herbicide sulfosulfuron in soil. The effect of repeated applications of sulfosulfuron on its degradation was assessed in two soils, using a sorghum root elongation bioassay. The effect of consecutive applications of sulfonylurea herbicides over the course of three to four seasons was further examined in controlled environment and a field study. Degradation of sulfosulfuron was determined following its application to soil samples from the field or a controlled environment, by measuring sulfosulfuron residues using liquid chromatography-tandem mass spectrometry. Following the repeated application of sulfosulfuron in the bioassay, the time to reduce sorghum root growth by 50% was shortened by up to 31.6%. However, consecutive application of SUs in the controlled environment had no effect on sulfosulfuron degradation rate constant. Yet, sulfosulfuron degradation rate was enhanced by a factor of 1.35 following consecutive application of SUs in the field, compared to untreated control soil. The data confirm that sulfosulfuron degradation could be enhanced due to repeated sulfosulfuron applications, thus potentially reducing its herbicidal efficacy.

Diplotaxis erucoides and Erucaria hispanica are common weeds of the Mediterranean region; they infest various habitats including cultivated fields and roadsides. In several fields across Israel, farmers have reported on poor control of D. erucoides and E. hispanica plants using acetolactate synthase (ALS) inhibitors. Greenhouse experiments were conducted to determine the effect of various ALS inhibitors on plants from two potentially resistant D. erucoides and E. hispanica populations. Additionally, alternative management strategies using auxinic herbicides were studied. Plants from both populations exhibited resistance to all tested ALS inhibitors, up to 20-fold the label field rate, as compared with ALS sensitive populations of D. erucoides and E. hispanica. Sequencing of the ALS gene revealed Trp574 to Leu substitution in ALS-resistant D. erucoides plants, whereas a Pro197 to Ser substitution was detected in ALS-resistant E. hispanica plants. Although high levels of resistance were observed in individuals from both putative resistant populations, sensitive individuals were also detected, suggesting the evolution of resistance in these two populations is still in progress. Auxinic herbicides, 2,4-D, and mecoprop-P, provided excellent control of plants from both ALS-resistant populations. This study documents and confirms the first case of evolution of resistance to ALS inhibitors in D. erucoides and E. hispanica populations.

The discovery of giant viruses infecting eukaryotes from diverse ecosystems has revolutionized our understanding of the evolution of viruses and their impact on protist biology, yet knowledge on their replication strategies and transcriptome regulation remains limited. Here, we profile single-cell transcriptomes of the globally distributed microalga Emiliania huxleyi and its specific giant virus during infection. We detected profound heterogeneity in viral transcript levels among individual cells. Clustering single cells based on viral expression profiles enabled reconstruction of the viral transcriptional trajectory. Reordering cells along this path unfolded highly resolved viral genetic programs composed of genes with distinct promoter elements that orchestrate sequential expression. Exploring host transcriptome dynamics across the viral infection states revealed rapid and selective shutdown of protein-encoding nuclear transcripts, while the plastid and mitochondrial transcriptomes persisted into later stages. Single-cell RNA-seq opens a new avenue to unravel the life cycle of giant viruses and their unique hijacking strategies.

Background and Aims The effect of the timing of the initiation of irrigation during the growing season on Vitis vinifera L. Cabernet Sauvignon vines was investigated during 5 successive years. Methods and Results Five irrigation initiation thresholds based on measurement of midday stem water potential (psi( s)) were examined: at Budburst, -0.6 MPa, -0.8 MPa, -1.0 MPa and - 1.2 MPa. Midday psi( s), gas exchange parameters [stomatal conductance (g( s)) and net assimilation rate (A( n))] and leaf area index (LAI) were measured weekly. At harvest, 12 vines per replicate (48 per treatment) were separately harvested, the yield was determined and the number of bunches per vine recorded. During the winter period, the number of shoots per vine and the pruning mass of 12 vines per replicate were recorded separately. Vines in the early irrigation treatments (Budburst, -0.6 MPa) displayed improved psi( s), g( s) and A( n), accompanied by vigorous vegetative growth and high yield. The increased yield was derived from enlarged berry size and an increased number of bunches. In contrast, vines in the late irrigation treatments (-1.0 MPa, -1.2 MPa) exhibited low psi( s), g( s) and A( n) values combined with depressed vegetative growth and reduced yield. The depression effect on late irrigation vines was cumulative and became more pronounced as trial years advanced. Our results emphasise the crucial role of water availability during springtime (vegetative growth period) on vine development, physiological performance and yield parameters. Conclusions Early irrigation initiation during spring, in which most vegetative growth processes occur, resulted in the enhancement of all vegetative and physiological parameters. Postponing irrigation initiation, however, until advanced stages of the growing season was followed by diminished vegetative growth, reduced physiological performance and decreased yield. Significance of the Study Our results provide insights into the effect of the timing of irrigation initiation on vegetative growth, physiological parameters and yield components in field-grown Cabernet Sauvignon grapevines.

Silica deposition in plants is a common phenomenon that correlates with plant tolerance to various stresses. Deposition occurs mostly in cell walls, but its mechanism is unclear. Here we show that metabolic processes control the formation of silica aggregates in roots of sorghum (Sorghum bicolor L.), a model plant for silicification. Silica formation was followed in intact roots and root segments of seedlings. Root segments were treated to enhance or suppress cell wall biosynthesis. The composition of endodermal cell walls was analysed by Raman microspectroscopy, scanning electron microscopy and energy-dispersive X-ray analysis. Our results were compared with in vitro reactions simulating lignin and silica polymerization. Silica aggregates formed only in live endodermal cells that were metabolically active. Silicic acid was deposited in vitro as silica onto freshly polymerized coniferyl alcohol, simulating G-lignin, but not onto coniferyl alcohol or ferulic acid monomers. Our results show that root silica aggregates form under tight regulation by endodermal cells, independently of the transpiration stream. We raise the hypothesis that the location and extent of silicification are primed by the chemistry and structure of polymerizing lignin as it cross-links to the wall.

A novel hyperspectral-physiological system that monitors plants dynamic response to abiotic alterations was developed. The system is a sensor-to-plant platform which can determine the optimal time of day during which physiological traits can be successfully identified via spectral means. The directly measured traits include momentary and daily transpiration rates throughout the daytime and daily and periodical plant weight loss and gain. The system monitored and evaluated pepper plants response to varying levels of potassium fertilization. Significant momentary transpiration rates differences were found between the treatments during 07:00-10:00 and 14:00-17:00. The simultaneous frequently measured high-resolution spectral data provided the means to correlate the two measured data sets. Significant correlation coefficients between the spectra and momentary transpiration rates resulted with a selection of three bands (rho 523, rho 697 and rho 818nm) that were used to capture transpiration rate differences using a normalized difference formula during the morning, noon and the afternoon. These differences also indicated that the best results are not always obtained when spectral (remote or proximal) measurements are typically preformed around noon (when solar illumination is the highest). Valuable information can be obtained when the spectral measurements are timed according to the plants' dynamic physiological status throughout the day, which may vary among plant species and should be considered when planning remote sensing data acquisition.

Plants optimize water use and carbon assimilation via transient regulation of stomata resistance and by limiting hydraulic conductivity in a long-term response of xylem anatomy. We postulated that without effective hydraulic regulation plants would permanently restrain water loss and photosynthetic productivity under salt stress conditions. We compared wild-type tomatoes to a transgenic type (TT) with impaired stomatal control. Gas exchange activity, biomass, starch content, leaf area and root traits, mineral composition and main stems xylem anatomy and hydraulic conductivity were analyzed in plants exposed to salinities of 1 and 4 dS m(-1) over 60 days. As the xylem cannot easily readjust to different environmental conditions, shifts in its anatomy and the permanent effect on plant hydraulic conductivity kept transpiration at lower levels under unstressed conditions and maintained it under salt-stress, while sustaining higher but inefficient assimilation rates, leading to starch accumulation and decreased plant biomass, leaf and root area and root length. Narrow conduits in unstressed TT plants were related to permanent restrain of hydraulic conductivity and plant transpiration. Under salinity, TT plants followed the atmospheric water demand, sustained similar transpiration rate from unstressed to salt-stressed conditions and possibly maintained hydraulic integrity, due to likely impaired hydraulic regulation, wider conduits and higher hydraulic conductivity. The accumulation of salts and starch in the TT plants was a strong evidence of salinity tolerance via osmotic regulation, also thought to help to maintain the assimilation rates and transpiration flux under salinity, although it was not translated into higher growth.

Eragrostis tef (Zucc.) Trotter (tef) is a small annual grain, panicle-bearing, C-4 cereal crop native to Ethiopia, where it is a major staple food. The objectives of the present study were to characterize the responses of two tef genotypes to escalating nitrogen (N) levels in terms of shoot, root, and grain biomass production, N concentration and uptake, and to determine an optimum N range at which tef performance is maximized. The N was applied in the irrigation water (Fertigation) in order to provide a consistent concentration of N in the root zone. A second goal was to test the feasibility of growing tef in the hot, arid conditions of the Northern Negev Desert. Two experiments were carried out in the Gilat Research Station (Negev region, Israel), each testing two different genotypes of tef (405B and 406W), and each including five replicates for each treatment. In the winter of 2015-2016, tef plants were grown in perlite filled pots in a walk-in plastic-covered tunnel. Five different N treatments were applied through fertigation, meaning the fertilizer was applied with the irrigation water (10, 20, 40, 80, 120 mg L-1). All other nutrients were applied at the same sufficient rate. In the summer of 2016, tef plots were sown in open-field and applied with four different rates of N fertilization (0, 30, 60, 120 mg L-1). Biomass of the different plant parts, SPAD values, N, P and K concentration, and the lodging index were recorded in each experiment. The harvest index was also calculated. Optimum N fertigation concentration in both experiments was between 40 and 80 mg L-1, under which the time to flowering was decreased, and yield and grain protein concentration were optimized. Underfertilization caused a decrease in overall plant growth, whereas overfertilization caused an increase in vegetative growth at the expense of grain yield. Potassium uptake increased along with increased N availability, whereas P uptake did not. The fertilization rate will always need to take into account local soil and climate conditions. The field experiment also pointed to low harvest index as a major limitation on tef cultivation in the Northern Negev.

Artemisinin-based drugs are the most effective medicine for the malaria treatment. To date, the main method of artemisinin production is its extraction from wormwood plants Artemisia annua L. Due to the limitation of this source, considerable efforts are now directed to the development of methods for artemisinin production using heterologous expression systems. Artemisinin is a sesquiterpene lactone, synthesized through the cyclization of farnesyl diphosphate involved in other sesquiterpene biosynthetic systems. Chrysanthemum species as well as A. annua, belong to Asteraceae family, and had been characterized by containing highly content of sesquiterpenes and their precursors. This makes chrysanthemum a promising target for the production of artemisinin in heterologous host plants. Chrysanthemum (C. morifolium Ramat.) was transformed by Agrobacterium tumefaciens carrying with the binary vectors p1240 and p1250, bearing artemisinin biosynthesis genes coding: amorpha-4,11-diene synthase, artemisinic aldehyde D11(13) reductase, amorpha-4,11-diene monooxygenase (p1240 was targeted to the mitochondria and p1250 was targeted to the cytosol), cytochrome P450 reductase from A. annua, as well as yeast truncated 3-hydroxy-3-methylglutarylcoenzyme A reductase. This study obtained 8 kanamycin-resistant lines after transformation with the p1240 and 2 lines from p1250. All target genes were detected in 2 and 1 transgenic lines of the 2 vectors. The transformation frequency of all target genes were 0.33% and 0.17% for p1240 and p1250, relative to the total transformed explant numbers. RT-PCR analysis revealed the transcription of all transferred genes in two lines obtained after transformation with the p1240 vector, confirming the possibility of transferring genetic modules encoding entire biochemical pathways into the chrysanthemum genome. This holds promise for the development of a chrysanthemum-based expression system to produce non-protein substances, such as artemisinin.

Self-supporting plants and climbers exhibit differences in their structural and biomechanical properties. We hypothesized that such fundamental differences originate at the level of the material properties. In this study, we compared three non-woody members of the Solanales exhibiting different growth habits: (1) a self-supporting plant (potato,Solanum tuberosum), (2) a trailing plant (sweet potato,Ipomoea batatas), and (3) a twining climber (morning glory,Ipomoea tricolor). The mechanical properties investigated by materials analyses were combined with structural, biochemical, and immunohistochemical analyses. Generally, the plants exhibited large morphological differences, but possessed relatively similar anatomy and cell wall composition. The cell walls were primarily composed of hemicelluloses (similar to 60%), with alpha-cellulose and pectins constituting similar to 25% and 5-8%, respectively. Immunohistochemistry of specific cell wall components suggested only minor variation in the occurrence and localization between the species, although some differences in hemicellulose distribution were observed. According to tensile and flexural tests, potato stems were the stiffest by a significant amount and the morning glory stems were the most compliant and showed differences in two- and three-orders of magnitude; the differences between their effective Young's (Elastic) modulus values (geometry-independent parameter), on the other hand, were substantially lower (at the same order of magnitude) and sometimes not even significantly different. Therefore, although variability exists in the internal anatomy and cell wall composition between the different species, the largest differences were seen in the morphology, which appears to be the primary determinant of biomechanical function. Although this does not exclude the possibility of different mechanisms in other plant groups, there is apparently less constraint to modifying stem morphology than anatomy and cell wall composition within the Solanales.

Main conclusion The phloem-mobile protein SlCyp1 traffics to distant parts of the shoot to regulate its gravitropic response. In addition, SlCyp1 targets specific cells in the root to promote lateral root development. The tomato (Solanum lycopersicum)Cyclophilin 1(SlCyp1) gene encodes a peptidyl-prolyl isomerase required for auxin response, lateral root development and gravitropic growth. The SlCyp1 protein is a phloem-mobile signal that moves from shoot to root to regulate lateral root development (Spiegelman et al., Plant J 83:853-863, 2015; J Exp Bot 68:953-964, 2017a). Here, we explored the mechanism of SlCyp1 movement by fusing it to the fluorescent protein mCherry. We found that, once trafficked to the root, SlCyp1 is unloaded from the phloem to the surrounding tissues, including the pericycle and lateral root primordia. Interestingly, SlCyp1 not only moves to the root system, but also to distant parts of the shoot. Grafting of theSlCyp1mutantdiageotropica(dgt) scions on VFN8 control rootstocks resulted in recovery ofdgtshoot gravitropism, which was associated with the restoration of auxin-response capacity. Application of the cyclophilin inhibitor cyclosporine A suppressed gravitropic recovery, indicating that SlCyp1 must be active in the target tissue to affect the gravitropic response. These results provide new insights on the mechanism of SlCyp1 transport and functioning as a long-distance signal regulating shoot gravitropism.

Cereals grains are the prime component of the human diet worldwide. To promote food security and sustainability, new approaches to non-chemical weed control are needed. Early vigor cultivars with enhanced weed-competitiveness ability are a potential tool, nonetheless, the introduction of such trait in breeding may be a long and labor-intensive process. Here, two image-driven plant phenotyping methods were evaluated to facilitate effective and accurate selection for early vigor in cereals. For that purpose, two triticale genotypes differentiating in vigor and growth rate early in the season were selected as model plants: X-1010 (high) and Triticale1 (low). Two modeling approaches, 2-D and 3-D, were applied on the plants offering an evaluation of various morphological growth parameters for the triticale canopy development, under controlled and field conditions. The morphological advantage of X-1010 was observed only at the initial growth stages, which was reflected by significantly higher growth parameter values compared to the Triticale1 genotype. Both modeling approaches were sensitive enough to detect phenotypic differences in growth as early as 21 days after sowing. All growth parameters indicated a faster early growth of X-1010. However, the 2-D related parameter [projected shoot area (PSA)] is the most available one that can be extracted via end user-friendly imaging equipment. PSA provided adequate indication for the triticale early growth under weed-competition conditions and for the improved weed-competition ability. The adequate phenotyping ability for early growth and competition was robust under controlled and field conditions. PSA can be extracted from close and remote sensing platforms, thus, facilitate high throughput screening. Overall, the results of this study may improve cereal breeding for early vigor and weed-competitiveness.

Given the potential health benefits (and adverse effects), of polyphenolic and steroidal glycoalkaloids in the diet there is a growing interest in fully elucidating the genetic control of their levels in foodstuffs. Here we carried out profiling of the specialized metabolites in the seeds of theSolanum pennelliiintrogression lines identifying 338 putative metabolite quantitative trait loci (mQTL) for flavonoids, steroidal glycoalkaloids and further specialized metabolites. Two putative mQTL for flavonols and one for steroidal glycoalkaloids were cross-validated by evaluation of the metabolite content of recombinants harboring smaller introgression in the corresponding QTL interval or by analysis of lines from an independently derived backcross inbred line population. The steroidal glycoalkaloid mQTL was localized to a chromosomal region spanning 14 genes, including a previously defined steroidal glycoalkaloid gene cluster. The flavonoid mQTL was further validated via the use of transient and stable overexpression of theSolyc12g098600andSolyc12g096870genes, which encode seed-specific uridine 5 `-diphosphate-glycosyltransferases. The results are discussed in the context of our understanding of the accumulation of polyphenols and steroidal glycoalkaloids, and how this knowledge may be incorporated into breeding strategies aimed at improving nutritional aspects of plants as well as in fortifying them against abiotic stress.

The use of auxins to improve the vase life of cut flowers is very limited. Previous studies demonstrated that a pulse treatment of Red Cestrum (Cestrum elegansSchlecht.) cut flowers with 2,4-dichlorophenoxyacetic acid (2,4-D) significantly reduced floret bud abscission, whereas 1-naphthaleneacetic acid (NAA) was ineffective. This difference resulted, at least in part, from the higher acropetal transport capability of 2,4-D compared to that of NAA. The present research focused on examining the factors affecting the acropetal transport, and hence the efficacy of the two auxins in reducing floret bud abscission of Red Cestrum cut flowers. We assumed that the differential acropetal transport capability of the two auxins results from the difference in their dissociation constants (pKa), with values of 2.75 and 4.23 for 2,4-D and NAA, respectively, which affects their pH-dependent physicochemical properties. Thus, increasing the pH of the pulsing solution above the pKa of both auxins might improve their acropetal movement. Indeed, the results of the present research show that raising the pH of the pulsing solution to pH 7.0 and above improved the efficacy of the two auxins in reducing floret bud abscission, with a higher effect on 2,4-D than that on NAA. Raising the pH of the pulsing solution decreased the adsorption and/or uptake of the two auxins by the cells adjacent to the xylem vessels, leading to an increase in their acropetal transport. The high pH of the pulsing solution increased the dissociation and hence decreased the lipophilicity of the auxin molecules, leading to improved acropetal movement. This effect was corroborated by the significant reduction in their 1-octanol/water partition coefficient (K-OW) values with the increase in the pH. A significant increase in theCeIAA1transcript level was obtained in response to 2,4-D pulsing at pH 7.0 and 8.25 and to NAA pulsing at pH 8.25, indicating that the acropetally transported auxins were taken up by the cells under these conditions. Our data suggest that raising the pH of the pulsing solution would significantly contribute to the increased efficacy of auxins in improving the vase life of cut flowers.

Aerial organs of plants, being highly prone to local injuries, require tissue restoration to ensure their survival. However, knowledge of the underlying mechanism is sparse. In this study, we mimicked natural injuries in growing leaves and stems to study the reunion between mechanically disconnected tissues. We show that PLETHORA (PLT) and AINTEGUMENTA (ANT) genes, which encode stem cell-promoting factors, are activated and contribute to vascular regeneration in response to these injuries. PLT proteins bind to and activate the CUC2 promoter. PLT proteins and CUC2 regulate the transcription of the local auxin biosynthesis gene YUC4 in a coherent feed-forward loop, and this process is necessary to drive vascular regeneration. In the absence of this PLT-mediated regeneration response, leaf ground tissue cells can neither acquire the early vascular identity marker ATHB8, nor properly polarise auxin transporters to specify new venation paths. The PLT-CUC2 module is required for vascular regeneration, but is dispensable for midvein formation in leaves. We reveal the mechanisms of vascular regeneration in plants and distinguish between the wound-repair ability of the tissue and its formation during normal development.

In anticipation of a drier climate and to project future changes in forest dynamics, it is imperative to understand species-specific differences in drought resistance. The objectives of this study were to form a comprehensive understanding of the drought resistance strategies adopted by Eastern Mediterranean woodland species, and to elaborate specific ecophysiological traits that can explain the observed variation in survival among these species. We examined leaf water potential (Psi), gas exchange and stem hydraulics during 2-3 years in mature individuals of the key woody species Phillyrea latifolia L., Pistacia lentiscus L. and Quercus calliprinos Webb that co-exist in a dry woodland experiencing similar to 6 rainless summer months. As compared with the other two similarly functioning species, Phillyrea displayed considerably lower Psi (minimum Psi of -8.7 MPa in Phillyrea vs -4.2 MPa in Pistacia and Quercus), lower Psi at stomatal closure and lower leaf turgor loss point (Psi(TLP)), but reduced hydraulic vulnerability and wider safety margins. Notably, Phillyrea allowed Psi to drop below Psi(TLP) under severe drought, whereas the other two species maintained positive turgor. These results indicate that Phillyrea adopted a more anisohydric drought resistance strategy, while Pistacia and Quercus exhibited a more isohydric strategy and probably relied on deeper water reserves. Unlike the two relatively isohydric species, Phillyrea reached complete stomatal closure at the end of the dry summer. Despite assessing a large number of physiological traits, none of them could be directly related to tree mortality. Higher mortality was observed for Quercus than for the other two species, which may result from higher water consumption due to its 2.5-10 times larger crown volume. The observed patterns suggest that similar levels of drought resistance in terms of survival can be achieved via different drought resistance strategies. Conversely, similar resistance strategies in terms of isohydricity can lead to different levels of vulnerability to extreme drought.