Root transcriptomic provides insights on molecular mechanisms involved in the tolerance to water deficit in Pisum sativum inoculated with Pseudomonas sp.

MAIN CONCLUSION: Root transcriptomics and biochemical analyses in water-stressed Pisum sativum plants inoculated with Pseudomonas spp. suggested preservation of ABA-related pathway and ROS detoxification, resulting in an improved tolerance to stress. Drought already affects agriculture in large areas of the globe and, due to climate change, these areas are predicted to become increasingly unsuitable for agriculture. For several years, plant growth-promoting bacteria (PGPB) have been used to improve legume yields, but many aspects of this interaction are still unclear. To elucidate the mechanisms through which root-associated PGPB can promote plant growth in dry environments, we investigated the response of pea plants inoculated with a potentially beneficial Pseudomonas strain (PK6) and subjected to two different water regimes. Combined biometric, biochemical, and root RNA-seq analyses revealed that PK6 improved pea growth specifically under water deficit, as inoculated plants showed an increased biomass, larger leaves, and longer roots. Abscisic acid (ABA) and proline quantification, together with the transcriptome analysis, suggested that PK6-inoculated plant response to water deficit was more diversified compared to non-inoculated plants, involving alternative metabolic pathways for the detoxification of reactive oxygen species (ROS) and the preservation of the ABA stress signaling pathway. We suggest that the metabolic response of PK6-inoculated plants was more effective in their adaptation to water deprivation, leading to their improved biometric traits. Besides confirming the positive role that PGPB can have in the growth of a legume crop under adverse conditions, this study offers novel information on the mechanisms regulating plant-bacteria interaction under varying water availability. These mechanisms and the involved genes could be exploited in the future for the development of legume varieties, which can profitably grow in dry climates.

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

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

Unveiling resilience mechanisms of Quercus ilex seedlings to severe water stress: Changes in non-structural carbohydrates, xylem hydraulic functionality and wood anatomy.

Over the last few decades, extensive dieback and mortality episodes of Quercus ilex L. have been documented after severe drought events in many Mediterranean forests. However, the underlying physiological, anatomical, and biochemical mechanisms remain poorly understood. We investigated the physiological and biochemical processes linked to embolism formation and non-structural carbohydrates (NSCs) dynamics in Q. ilex seedlings exposed to severe water stress and rewatering. Measurements of leaf gas exchange, water relations, non-structural carbohydrates, drought-related gene expression, and anatomical changes in wood parenchyma were assessed. Under water stress, the midday stem water potential dropped below - 4.5 MPa corresponding to a ~ 50 % loss of hydraulic conductivity. A 70 % reduction in stomatal conductance led to a strong depletion of wood NSCs. Starch consumption, resulting from the upregulation of the ß-amylase gene BAM3, together with the downregulation of glucose (GPT1) and sucrose (SUC27) transport genes, suggests glucose utilization to sustain cellular metabolism in the wood parenchyma. After rewatering, the presence of residual xylem embolism led to an incomplete recovery of leaf gas exchanges. However, the partial restoration of photosynthesis allowed the accumulation of new starch reserves in the wood parenchyma and the production of new narrower vessels. In addition, changes in the cell wall composition of the wood parenchyma fibers were observed. Our findings indicate that thirty days of rewatering were sufficient to restore the NSCs reserves and growth rates of Q. ilex seedlings and that the carryover effects of water stress were primarily caused by hydraulic dysfunction.

Photoprotective Role of Photosynthetic and Non-Photosynthetic Pigments in Phillyrea latifolia: Is Their "Antioxidant" Function Prominent in Leaves Exposed to Severe Summer Drought?

Carotenoids and phenylpropanoids play a dual role of limiting and countering photooxidative stress. We hypothesize that their "antioxidant" function is prominent in plants exposed to summer drought, when climatic conditions exacerbate the light stress. To test this, we conducted a field study on Phillyrea latifolia, a Mediterranean evergreen shrub, carrying out daily physiological and biochemical analyses in spring and summer. We also investigated the functional role of the major phenylpropanoids in different leaf tissues. Summer leaves underwent the most severe drought stress concomitantly with a reduction in radiation use efficiency upon being exposed to intense photooxidative stress, particularly during the central hours of the day. In parallel, a significant daily variation in both carotenoids and phenylpropanoids was observed. Our data suggest that the morning-to-midday increase in zeaxanthin derived from the hydroxylation of ß-carotene to sustain non-photochemical quenching and limit lipid peroxidation in thylakoid membranes. We observed substantial spring-to-summer and morning-to-midday increases in quercetin and luteolin derivatives, mostly in the leaf mesophyll. These findings highlight their importance as antioxidants, countering the drought-induced photooxidative stress. We concluded that seasonal and daily changes in photosynthetic and non-photosynthetic pigments may allow P. latifolia leaves to avoid irreversible photodamage and to cope successfully with the Mediterranean harsh climate.

An Improvement of SPME-Based Sampling Technique to Collect Volatile Organic Compounds from Quercus ilex at the Environmental Level.

Biogenic Volatile Organic Compounds (BVOCs) include many chemical compounds emitted by plants into the atmosphere. These compounds have a great effect on biosphere-atmosphere interactions and may affect the concentration of atmospheric pollutants, with further consequences on human health and forest ecosystems. Novel methods to measure and determine BVOCs in the atmosphere are of compelling importance considering the ongoing climate changes. In this study, we developed a fast and easy-to-handle analytical methodology to sample these compounds in field experiments using solid-phase microextraction (SPME) fibers at the atmospheric level. An improvement of BVOCs adsorption from SPME fibers was obtained by coupling the fibers with fans to create a dynamic sampling system. This innovative technique was tested sampling Q. ilex BVOCs in field conditions in comparison with the conventional static SPME sampling technique. The results showed a great potential of this dynamic sampling system to collect BVOCs at the atmosphere level, improving the efficiency and sensitivity of SPME fibers. Indeed, our novel device was able to reduce the sampling time, increase the amount of BVOCs collected through the fibers and add information regarding the emissions of these compounds at the environmental level.

Phenolic Compounds from Leaves and Flowers of Hibiscus roseus: Potential Skin Cosmetic Applications of an Under-Investigated Species.

The use of plant extracts in skin-care cosmetics is a modern trend due to their richness in polyphenols that act as anti-aging molecules. Hibiscus roseus is a perennial species naturalized in Italy, with beautiful soft pink flowers; its phenolic composition and biological activities have not been studied yet. The aim of this study was to characterize and quantify the phenolics and to evaluate the antioxidant, sun protection factor (SPF), and anti-collagenase activities of the ethanolic extracts of H. roseus leaves (HL) and flowers (HF). p-Coumaric, chlorogenic, and trans-ferulic acids derivatives as well as quercetin and kaempferol flavonoids were the main phenolic compounds detected. Catechin, epicatechin, kaempferol-3-O-rutinoside, kaempferol-3-O-glucoside, kaempferol-7-O-glucoside, tiliroside, oenin, and peonidin-3-O-glucoside were detected only in HF, while phloridzin was exclusive from HL, which also showed greater amounts of hydroxycinnamic acid derivatives. HF was richer in flavonoids and total phenolics, also exhibiting greater antioxidant capacity. The SPF and anti-collagenase activity of both extracts were similar and comparable to those of synthetic standards. The overall results demonstrate that H. roseus extracts are promising sources of bioactive phenolic compounds that could be potentially applied as anti-aging agents in skin-care cosmetics.

Ethyl acetate extract from Cistus x incanus L. leaves enriched in myricetin and quercetin derivatives, inhibits inflammatory mediators and activates Nrf2/HO-1 pathway in LPS-stimulated RAW 264.7 macrophages.

Cistus x incanus L. is a Mediterranean evergreen shrub used in folk medicine for the treatment of inflammatory disorders but the underlying mechanisms are not fully understood. We therefore investigated the anti-inflammatory effects of an ethyl acetate fraction (EAF) from C. x incanus L. leaves on lipopolysaccharide (LPS) activated RAW 264.7 macrophages. HPLC analysis revealed myricetin and quercetin derivatives to be the major compounds in EAF; EAF up to 1 µM of total phenolic content, was not cytotoxic and inhibited the mRNA expression of interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) (p < 0.05) and the production of prostaglandins E2 (PGE2) (p < 0.05). Meanwhile, EAF triggered the mRNA expression of interleukin-10 (IL-10) and elicited the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), as well as the expression of its main target gene, heme oxygenase-1 (HO-1) (p < 0.05). These data indicate that EAF attenuates experimental inflammation via the inhibition of proinflammatory mediators and at least in part, by the activation of Nrf2/HO-1 pathway. These effects are likely due to myricetin and quercetin derivatives but the role of other, less abundant components cannot be excluded. Further studies to confirm the relevance of our findings in animal models and to highlight the relative contribution of each component to the anti-inflammatory activity of EAF should be conducted.

Are Flavonoids Effective Antioxidants in Plants? Twenty Years of Our Investigation.

Whether flavonoids play significant antioxidant roles in plants challenged by photooxidative stress of different origin has been largely debated over the last few decades. A critical review of the pertinent literature and our experimentation as well, based on a free-of-scale approach, support an important antioxidant function served by flavonoids in plants exposed to a wide range of environmental stressors, the significance of which increases with the severity of stress. On the other side, some questions need conclusive answers when the putative antioxidant functions of plant flavonoids are examined at the level of both the whole-cell and cellular organelles. This partly depends upon a conclusive, robust, and unbiased definition of "a plant antioxidant", which is still missing, and the need of considering the subcellular re-organization that occurs in plant cells in response to severe stress conditions. This likely makes our deterministic-based approach unsuitable to unveil the relevance of flavonoids as antioxidants in extremely complex biological systems, such as a plant cell exposed to an ever-changing stressful environment. This still poses open questions about how to measure the occurred antioxidant action of flavonoids. Our reasoning also evidences the need of contemporarily evaluating the changes in key primary and secondary components of the antioxidant defense network imposed by stress events of increasing severity to properly estimate the relevance of the antioxidant functions of flavonoids in an in planta situation. In turn, this calls for an in-depth analysis of the sub-cellular distribution of primary and secondary antioxidants to solve this still intricate matter.

Optimization of a Green Ultrasound-Assisted Extraction of Different Polyphenols from Pistacia lentiscus L. Leaves Using a Response Surface Methodology.

Pistacia lentiscus leaves are used in several applications, thanks to their polyphenolic abundance. Thiswork aimed to characterize the polyphenols and to optimize the extraction conditions to shorten the time, decrease the consumption of solvent, and to maximize the yield of different classes of phenolics, which have diverse industrial applications. The variables were optimized by applying a Box-Behnken design. Galloyl and myricetin derivatives were the most abundant compounds, and two new tetragalloyl derivatives were identified by LC-MS/MS. According to the models, the maximum yields of polyphenols (51.3 ± 1.8 mg g-1 DW) and tannins (40.2 ± 1.4 mg g-1 DW) were obtained using 0.12 L g-1 of 40% ethanol at 50 °C. The highest content of flavonoids (10.2 ± 0.8 mg g-1 DW) was obtained using 0.13 L g-1 of 50% ethanol at 50 °C, while 0.1 L g-1 of 30% ethanol at 30 °C resulted in higher amounts of myricitrin (2.6 ± 0.19 mg g-1 DW). Our optimized extraction decreased the ethanolic fraction by 25% and halved the time compared to other methods. These conditions can be applied differently to obtain P. lentiscus extracts richer in tannins or flavonoids, which might be employed for various purposes.

Improving Air Quality by Nitric Oxide Consumption of Climate-Resilient Trees Suitable for Urban Greening.

Nitrogen oxides (NOx), mainly a mixture of nitric oxide (NO) and nitrogen dioxide (NO2), are formed by the reaction of nitrogen and oxygen compounds in the air as a result of combustion processes and traffic. Both deposit into leaves via stomata, which on the one hand benefits air quality and on the other hand provides an additional source of nitrogen for plants. In this study, we first determined the NO and NO2 specific deposition velocities based on projected leaf area (sV d) using a branch enclosure system. We studied four tree species that are regarded as suitable to be planted under predicted future urban climate conditions: Carpinus betulus, Fraxinus ornus, Fraxinus pennsylvanica and Ostrya carpinifolia. The NO and NO2 sVd were found similar in all tree species. Second, in order to confirm NO metabolization, we fumigated plants with 15NO and quantified the incorporation of 15N in leaf materials of these trees and four additional urban tree species (Celtis australis, Alnus spaethii, Alnus glutinosa, and Tilia henryana) under controlled environmental conditions. Based on these 15N-labeling experiments, A. glutinosa showed the most effective incorporation of 15NO. Third, we tried to elucidate the mechanism of metabolization. Therefore, we generated transgenic poplars overexpressing Arabidopsis thaliana phytoglobin 1 or 2. Phytoglobins are known to metabolize NO to nitrate in the presence of oxygen. The 15N uptake in phytoglobin-overexpressing poplars was significantly increased compared to wild-type trees, demonstrating that the NO uptake is enzymatically controlled besides stomatal dependence. In order to upscale the results and to investigate if a trade-off exists between air pollution removal and survival probability under future climate conditions, we have additionally carried out a modeling exercise of NO and NO2 deposition for the area of central Berlin. If the actually dominant deciduous tree species (Acer platanoides, Tilia cordata, Fagus sylvatica, Quercus robur) would be replaced by the species suggested for future conditions, the total annual NO and NO2 deposition in the modeled urban area would hardly change, indicating that the service of air pollution removal would not be degraded. These results may help selecting urban tree species in future greening programs.

Short-Term Pre-Harvest UV-B Supplement Enhances the Polyphenol Content and Antioxidant Capacity of Ocimum basilicum Leaves during Storage.

Ocimum basilicum (basil) leaves are rich in polyphenols, conferring them a high antioxidant activity. The application of UV-B can be used to maintain the post-harvest nutraceutical quality of basil leaves. We aimed to investigate the effects of pre-harvest UV-B application on polyphenolic and pigment contents, antioxidant capacity, and the visual quality of basil stored leaves. We also evaluated the applicability of the non-invasive Dualex® for monitoring the accumulation of leaf epidermal phenolics (Flav Index). After exposing plants to white light (control) and to supplemental UV-B radiation for 4 d, the leaves were harvested and stored for 7d (TS7). The UV-B leaves showed both a higher phenolic content and antioxidant capacity than the controls at TS7. In addition, the correlations between the Flav Index and phenolic content demonstrated that Dualex® can reliably assess the content of epidermal phenolics, thus confirming its promising utilization as a non-destructive method for monitoring the phytochemical quality of O. basilicum leaves. In conclusion, a pre-harvesting UV-B application may be a tool for enhancing the content of polyphenols and the antioxidant potential of basil stored leaves without detrimental effects on their visual quality. These results are important considering the nutraceutical value of this plant and its wide commercial distribution.

Phenotypic plasticity of two M. oleifera ecotypes from different climatic zones under water stress and re-watering.

Moringa oleifera is a fast-growing hygrophilic tree native to a humid sub-tropical region of India, now widely planted in many regions of the Southern Hemisphere characterized by low soil water availability. The widespread cultivation of this plant worldwide may have led to populations with different physiological and biochemical traits. In this work, the impact of water stress on the physiology and biochemistry of two M. oleifera populations, one from Chaco Paraguayo (PY) and one from Indian Andhra Pradesh (IA) region, was studied in a screenhouse experiment where the water stress treatment was followed by re-watering. Through transcriptome sequencing, 2201 potential genic simple sequence repeats were identified and used to confirm the genetic differentiation of the two populations. Both populations of M. oleifera reduced photosynthesis, water potential, relative water content and growth under drought, compared to control well-watered plants. A complete recovery of photosynthesis after re-watering was observed in both populations, but growth parameters recovered better in PY than in IA plants. During water stress, PY plants accumulated more secondary metabolites, especially ß-carotene and phenylpropanoids, than IA plants, but IA plants invested more into xanthophylls and showed a higher de-epoxidation state of xanthophylls cycle that contributed to protect the photosynthetic apparatus. M. oleifera demonstrated a high genetic variability and phenotypic plasticity, which are key factors for adaptation to dry environments. A higher plasticity (e.g. in PY plants adapted to wet environments) will be a useful trait to endure recurrent but brief water stress episodes, whereas long-term investment of resources into secondary metabolism (e.g. in IA plants adapted to drier environments) will be a successful strategy to cope with prolonged periods of drought. This makes M. oleifera an important resource for agro-forestry in a climate change scenario.

Changes in abscisic acid content during and after drought are related to carbohydrate mobilization and hydraulic recovery in poplar stems.

Drought compromises plant's ability to replace transpired water vapor with water absorbed from the soil, leading to extensive xylem dysfunction and causing plant desiccation and death. Short-term plant responses to drought rely on stomatal closure, and on the plant's ability to recover hydraulic functioning after drought relief. We hypothesize a key role for abscisic acid (ABA) not only in the control of stomatal aperture, but also in hydraulic recovery. Young plants of Populus nigra L. were used to investigate possible relationships among ABA, non-structural carbohydrates (NSC) and xylem hydraulic function under drought and after re-watering. In Populus nigra L. plants subjected to drought, water transport efficiency and hydraulic recovery after re-watering were monitored by measuring the percentage loss of hydraulic conductivity (PLC) and stem specific hydraulic conductivity (Kstem). In the same plants ABA and NSC were quantified in wood and bark. Drought severely reduced stomatal conductance (gL) and markedly increased the PLC. Leaf and stem water potential, and stem hydraulic efficiency fully recovered within 24 h after re-watering, but gL values remained low. After re-watering, we found significant correlations between changes in ABA content and hexoses concentration both in wood and bark. Our findings suggest a role for ABA in the regulation of stem carbohydrate metabolism and starch mobilization upon drought relief, possibly promoting the restoration of xylem transport capacity.

Seasonal and Diurnal Variation in Leaf Phenolics of Three Medicinal Mediterranean Wild Species: What Is the Best Harvesting Moment to Obtain the Richest and the Most Antioxidant Extracts?

Mediterranean plants biosynthesize high amounts of polyphenols, which are important health-promoting compounds. Leaf polyphenolic composition changes according to environmental conditions. Therefore, it is crucial to know the temporal variation in their production. This study aimed to: i) evaluate the monthly and daily changes in polyphenols of Phyllirea latifolia, Cistus incanus, and Pistacia lentiscus to identify their best harvesting moment, ii) verify the possible correlations between phenolic production and temperature and irradiation, iii) evaluate their antioxidant capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical (OH)scavenging assays. The extracts of leaves harvested at 8:00, 13:00 and 18:00, in May, July, and October for two years were analysed by HPLC-DAD. Both "month" and "time of the day" affected the polyphenolic content in all species. July at 13:00 was the best harvesting moment for all polyphenolic classes of P. latifolia and only for some classes of C. incanus and P. lentiscus. Environmental parameters positively correlated with the polyphenols of C. incanus and P. latifolia, while the antioxidant capacity only varied in this last species, reaching the highest value in July. Results of the study allow to determine the balsamic time for each species. Moreover, the relationship between polyphenols and environmental data can be useful for the cultivation of these plants under controlled conditions.

Phellem Cell-Wall Components Are Discriminants of Cork Quality in Quercus suber.

Cork is a renewable, non-wood high valued forest product, with relevant ecological and economic impact in the Mediterranean-type ecosystems. Currently, cork is ranked according to its commercial quality. The most valuable planks are chosen for cork stoppers production. Cork planks with adequate thickness and porosity are classified as stoppable quality cork (SQC). The chemical composition of cork is known, but the regulation of metabolic pathways responsible of cork production and composition, hence of cork quality, is largely unknown. Here, we tested the hypothesis that post-genomic events may be responsible for the development of SQC and N-SQC (non-stoppable quality cork). Here, we show that combined proteomics and targeted metabolomics (namely soluble and cell wall bound phenolics) analyzed on recently formed phellem allows discriminate cork planks of different quality. Phellem cells of SQC and N-SQC displayed different reducing capacity, with consequential impact on both enzymatic pathways (e.g., glycolysis) and other cellular functions, including cell wall assembly and suberization. Glycolysis and respiration related proteins were abundant in both cork quality groups, whereas the level of several proteins associated to mitochondrial metabolism was higher in N-SQC. The soluble and cell wall-bound phenolics in recently formed phellem clearly discriminated SQC from N-SCQ. In our study, SQC was characterized by a high incorporation of aromatic components of the phenylpropanoid pathway in the cell wall, together with a lower content of hydrolysable tannins. Here, we propose that the level of hydrolysable tannins may represent a valuable diagnostic tool for screening recently formed phellem, and used as a proxy for the quality grade of cork plank produced by each tree.

Dissecting Adaptation Mechanisms to Contrasting Solar Irradiance in the Mediterranean Shrub Cistus incanus.

Molecular mechanisms that are the base of the strategies adopted by Mediterranean plants to cope with the challenges imposed by limited or excessive solar radiation during the summer season have received limited attention. In our study, conducted on C. incanus plants growing in the shade or in full sunlight, we performed measurements of relevant physiological traits, such as leaf water potential, gas exchange and PSII photochemistry, RNA-Seq with de-novo assembly, and the analysis of differentially expressed genes. We also identified and quantified photosynthetic pigments, abscisic acid, and flavonoids. Here, we show major mechanisms regulating light perception and signaling which, in turn, sustain the shade avoidance syndrome displayed by the 'sun loving' C. incanus. We offer clear evidence of the detrimental effects of excessive light on both the assembly and the stability of PSII, and the activation of a suite of both repair and effective antioxidant mechanisms in sun-adapted leaves. For instance, our study supports the view of major antioxidant functions of zeaxanthin in sunny plants concomitantly challenged by severe drought stress. Finally, our study confirms the multiple functions served by flavonoids, both flavonols and flavanols, in the adaptive mechanisms of plants to the environmental pressures associated to Mediterranean climate.

Aspergillus flavus as a Model System to Test the Biological Activity of Botanicals: An Example on Citrullus colocynthis L. Schrad. Organic Extracts.

Citrullus colocynthis L. Schrader is an annual plant belonging to the Cucurbitaceae family, widely distributed in the desert areas of the Mediterranean basin. Many pharmacological properties (anti-inflammatory, anti-diabetic, analgesic, anti-epileptic) are ascribed to different organs of this plant; extracts and derivatives of C. colocynthis are used in folk Berber medicine for the treatment of numerous diseases-such as rheumatism arthritis, hypertension bronchitis, mastitis, and even cancer. Clinical studies aimed at confirming the chemical and biological bases of pharmacological activity assigned to many plant/herb extracts used in folk medicine often rely on results obtained from laboratory preliminary tests. We investigated the biological activity of some C. colocynthis stem, leaf, and root extracts on the mycotoxigenic and phytopathogenic fungus Aspergillus flavus, testing a possible correlation between the inhibitory effect on aflatoxin biosynthesis, the phytochemical composition of extracts, and their in vitro antioxidant capacities.

Dynamic changes in ABA content in water-stressed Populus nigra: effects on carbon fixation and soluble carbohydrates.

BACKGROUND AND AIMS: Hydraulic and chemical signals operate in tandem to regulate systemic plant responses to drought. Transport of abscisic acid (ABA) through the xylem and phloem from the root to shoot has been suggested to serve as the main signal of water deficit. There is evidence that ABA and its ABA-glycosyl-ester (ABA-GE) are also formed in leaves and stems through the chloroplastic 2-C-methylerythritol-5-phosphate (MEP) pathway. This study aimed to evaluate how hormonal and hydraulic signals contribute to optimize stomatal (gs), mesophyll (gm) and leaf hydraulic (Kleaf) conductance under well-watered and water-stressed conditions in Populus nigra (black poplar) plants. In addition, we assessed possible relationships between ABA and soluble carbohydrates within the leaf and stem. METHODS: Plants were subjected to three water treatments: well-watered (WW), moderate stress (WS1) and severe stress (WS2). This experimental set-up enabled a time-course analysis of the response to water deficit at the physiological [leaf gas exchange, plant water relations, (Kleaf)], biochemical (ABA and its metabolite/catabolite quantification in xylem sap, leaves, wood, bark and roots) and molecular (gene expression of ABA biosynthesis) levels. KEY RESULTS: Our results showed strong coordination between gs, gm and Kleaf under water stress, which reduced transpiration and increased intrinsic water use efficiency (WUEint). Analysis of gene expression of 9-cis-epoxycarotenoid dioxygenase (NCED) and ABA content in different tissues showed a general up-regulation of the biosynthesis of this hormone and its finely-tuned catabolism in response to water stress. Significant linear relationships were found between soluble carbohydrates and ABA contents in both leaves and stems, suggesting a putative function for this hormone in carbohydrate mobilization under severe water stress. CONCLUSIONS: This study demonstrates the tight regulation of the photosynthetic machinery by levels of ABA in different plants organs on a daily basis in both well-watered and water stress conditions to optimize WUEint and coordinate whole plant acclimation responses to drought.

Seasonal and daily variations in primary and secondary metabolism of three maquis shrubs unveil different adaptive responses to Mediterranean climate.

Maquis species play a central role in the maintenance of coastal ecosystems thanks to anatomical, physiological and biochemical features evolved to cope with severe stress conditions. Because the seasonal and daily dynamics of physiological and biochemical traits of maquis species are not fully addressed, we performed a field study on three coexisting Mediterranean shrubs (Pistacia lentiscus L. and Phillyrea latifolia L., evergreen schlerophylls, and Cistus incanus L., semi-deciduous) aiming at detecting the main adaptive differences, on a seasonal and daily basis, in primary and secondary metabolism along with the principal climatic determinants. These species differed in their physiological and biochemical responses especially on a seasonal level. In P. latifolia, a great investment in antioxidant phenylpropanoids contributed to maintain high photosynthetic rates throughout the whole growing season. In C. incanus, high carotenoid content associated with chlorophyll (Chl) regulation alleviated oxidative damage during the hot and dry summers and help recover photosynthesis in autumn. In P. lentiscus, high abscisic acid levels allowed a strict control of stomata, while fine Chla/Chlb regulation concurred to avoid photoinhibition in summer. Temperature resulted the most important climatic factor controlling the physiological and biochemical status of these coexisting shrubs and, thus, in determining plant performances in this Mediterranean coastal habitat.

Metabolic plasticity in the hygrophyte Moringa oleifera exposed to water stress.

Over the past decades, introduction of many fast-growing hygrophilic, and economically valuable plants into xeric environments has occurred. However, production and even survival of these species may be threatened by harsh climatic conditions unless an effective physiological and metabolic plasticity is available. Moringa oleifera Lam., a multipurpose tree originating from humid sub-tropical regions of India, is widely cultivated in many arid countries because of its multiple uses. We tested whether M. oleifera can adjust primary and secondary metabolism to efficiently cope with increasing water stress. It is shown that M. oleifera possesses an effective isohydric behavior. Water stress induced a quick and strong stomatal closure, driven by abscisic acid (ABA) accumulation, and leading to photosynthesis inhibition with consequent negative effects on biomass production. However, photochemistry was not impaired and maximal fluorescence and saturating photosynthesis remained unaffected in stressed leaves. We report for the first time that M. oleifera produces isoprene, and show that isoprene emission increased three-fold during stress progression. It is proposed that higher isoprene biosynthesis helps leaves cope with water stress through its antioxidant or membrane stabilizing action, and also indicates a general MEP (methylerythritol 4-phosphate) pathway activation that further helps protect photosynthesis under water stress. Increased concentrations of antioxidant flavonoids were also observed in water stressed leaves, and probably cooperate in limiting irreversible effects of the stress in M. oleifera leaves. The observed metabolic and phenotypic plasticity may facilitate the establishment of M. oleifera in xeric environments, sustaining the economic and environmental value of this plant.