Phytochemical and Antioxidant Profile of the Medicinal Plant Melia azedarach Subjected to Water Deficit Conditions.

Environmental stress triggered by climate change can alter the plant's metabolite profile, which affects its physiology and performance. This is particularly important in medicinal species because their economic value depends on the richness of their phytocompounds. We aimed to characterize how water deficit modulated the medicinal species Melia azedarach's lipophilic profile and antioxidant status. Young plants were exposed to water deficit for 20 days, and lipophilic metabolite profile and the antioxidant capacity were evaluated. Leaves of M. azedarach are rich in important fatty acids and oleamide. Water deficit increased the radical scavenging capacity, total phenol, flavonoids, and catechol pools, and the accumulation of ß-sitosterol, myo-inositol, succinic acid, sucrose, d-glucose and derivatives, d-psicofuranose, d-(+)-fructofuranose, and the fatty acids stearic, α-linolenic, linoleic and palmitic acids. These responses are relevant to protecting the plant against climate change-related stress and also increase the nutritional and antioxidant quality of M. azedarach leaves.

Plateau Waves of Intracranial Pressure: Methods for Automatic Detection and Prediction.

Plateau waves are recurrent phenomena observed in traumatic brain injury (TBI) patients, characterised by an increase in intracranial pressure (ICP) above 40 mmHg combined with an almost zero arterial blood pressure (ABP) variation and, hence, a decrease in cerebral perfusion pressure (CPP). A raised ICP for a long period of time, namely plateau waves, can lead to a secondary brain injury. Due to the impaired cerebral autoregulation mechanism these TBI patients present, they are admitted to neurocritical care units (NCCUs) to be under continuous multimodal monitoring, which allows a correct diagnosis for each patient. Plateau waves can end naturally by activating a vasoconstriction mechanism which decreases the amount of blood available in the brain. Alternatively, the phenomenon can end with therapeutic treatment.In this sense, the present study consists in the development of an algorithm capable of automatically detecting plateau waves using offline data, i.e. data already collected from patients. This creates an extra tool which allows for faster detection of events to assist their identification and final diagnosis. Despite the additional steps that can be included to improve the algorithm, the results show good performance, and thus it may be applied in NCCUs.

Olive Varieties under UV-B Stress Show Distinct Responses in Terms of Antioxidant Machinery and Isoform/Activity of RubisCO.

In recent decades, atmospheric pollution led to a progressive reduction of the ozone layer with a consequent increase in UV-B radiation. Despite the high adaptation of olive trees to the Mediterranean environment, the progressive increase of UV-B radiation is a risk factor for olive tree cultivation. It is therefore necessary to understand how high levels of UV-B radiation affect olive plants and to identify olive varieties which are better adapted. In this study we analyzed two Italian olive varieties subjected to chronic UV-B stress. We focused on the effects of UV-B radiation on RubisCO, in terms of quantity, enzymatic activity and isoform composition. In addition, we also analyzed changes in the activity of antioxidant enzymes (SOD, CAT, GPox) to get a comprehensive picture of the antioxidant system. We also evaluated the effects of UV-B on the enzyme sucrose synthase. The overall damage at biochemical level was also assessed by analyzing changes in Hsp70, a protein triggered under stress conditions. The results of this work indicate that the varieties (Giarraffa and Olivastra Seggianese) differ significantly in the use of specific antioxidant defense systems, as well as in the activity and isoform composition of RubisCO. Combined with a different use of sucrose synthase, the overall picture shows that Giarraffa optimized the use of GPox and opted for a targeted choice of RubisCO isoforms, in addition to managing the content of sucrose synthase, thereby saving energy during critical stress points.

Plant Flavonoids: Chemical Characteristics and Biological Activity.

In recent years, more attention has been paid to natural sources of antioxidants. Flavonoids are natural substances synthesized in several parts of plants that exhibit a high antioxidant capacity. They are a large family, presenting several classes based on their basic structure. Flavonoids have the ability to control the accumulation of reactive oxygen species (ROS) via scavenger ROS when they are formed. Therefore, these antioxidant compounds have an important role in plant stress tolerance and a high relevance in human health, mainly due to their anti-inflammatory and antimicrobial properties. In addition, flavonoids have several applications in the food industry as preservatives, pigments, and antioxidants, as well as in other industries such as cosmetics and pharmaceuticals. However, flavonoids application for industrial purposes implies extraction processes with high purity and quality. Several methodologies have been developed aimed at increasing flavonoid extraction yield and being environmentally friendly. This review presents the most abundant natural flavonoids, their structure and chemical characteristics, extraction methods, and biological activity.

Responses of olive plants exposed to different irrigation treatments in combination with heat shock: physiological and molecular mechanisms during exposure and recovery.

MAIN CONCLUSION: A water-deficit period, leading to stomatal control and overexpression of protective proteins (sHSP and DHN), contributes to olive´s tolerance to later imposed stress episodes. Aquaporins modulation is important in olive recovery. Olive is traditionally cultivated in dry farming or in high water demanding irrigated orchards. The impact of climate change on these orchards remains to unveil, as heat and drought episodes are increasing in the Mediterranean region. To understand how young plants face such stress episodes, olive plants growing in pots were exposed to well-irrigated and non-irrigated treatments. Subsequently, plants from each treatment were either exposed to 40 °C for 2 h or remained under control temperature. After treatments, all plants were allowed to grow under well-irrigated conditions (recovery). Leaves were compared for photosynthesis, relative water content, mineral status, pigments, carbohydrates, cell membrane permeability, lipid peroxidation and expression of the protective proteins' dehydrin (OeDHN1), heat-shock proteins (OeHSP18.3), and aquaporins (OePIP1.1 and OePIP2.1). Non-irrigation, whilst increasing carbohydrates, reduced some photosynthetic parameters to values below the ones of the well-irrigated plants. However, when both groups of plants were exposed to heat, well-irrigated plants suffered more drastic decreases of net CO2 assimilation rate and chlorophyll b than non-irrigated plants. Overall, OeDHN1 and OeHSP18.3 expression, which was increased in non-irrigated treatment, was potentiated by heat, possibly to counteract the increase of lipid peroxidation and loss of membrane integrity. Plants recovered similarly from both irrigation and temperature treatments, and recovery was associated with increased aquaporin expression in plants exposed to one type of stress (drought or heat). These data represent an important contribution for further understanding how dry-farming olive will cope with drought and heat episodes.

Water stress and recovery in the performance of two Eucalyptus globulus clones: physiological and biochemical profiles.

Eucalyptus plantations are among the most productive forest stands in Portugal and Spain, being mostly used for pulp production and, more recently, as an energy crop. However, the region's Mediterranean climate, with characteristic severe summer drought, negatively affects eucalypt growth and increases mortality. Although the physiological response to water shortage is well characterized for this species, evidence about the plants' recovery ability remains scarce. In order to assess the physiological and biochemical response of Eucalyptus globulus during the recovery phase, two genotypes (AL-18 and AL-10) were submitted to a 3-week water stress period at two different intensities (18 and 25% of field capacity), followed by 1 week of rewatering. Recovery was assessed 1 day and 1 week after rehydration. Drought reduced height, biomass, water potential, NPQ and gas exchange in both genotypes. Contrarily, the levels of pigments, chlorophyll fluorescence parameters (F(v) /F(m) and (φPSII)), MDA and ABA increased. During recovery, the physiological and biochemical profile of stressed plants showed a similar trend: they experienced reversion of altered traits (MDA, ABA, E, g(s), pigments), while other parameters did not recover ((φPSII), NPQ). Furthermore, an overcompensation of CO(2) assimilation was achieved 1 week after rehydration, which was accompanied by greater growth and re-establishment of oxidative balance. Both genotypes were tolerant to the tested conditions, although clonal differences were found. AL-10 was more productive and showed a more rapid and dynamic response to rehydration (namely in carotenoid content, (φPSII) and NPQ) compared to clone AL-18.

Organic Waste from the Management of the Invasive Oxalis pes-caprae as a Source of Nutrients for Small Horticultural Crops.

The management of invasive plants is a challenge when using traditional control methods, which are ineffective for large areas, leading to the abandonment of invaded areas and the subsequent worsening of the situation. Finding potential uses for waste resulting from invaders' management could motivate their control in the long-term, concurrently providing new bio-based resources with different applications. Oxalis pes-caprae is an invasive plant, widely distributed worldwide, which spreads aggressively through bulbils, creating a dense ground cover. This study was designed to assess the potential of Oxalis aboveground waste for use as fertilizer and in ameliorating deficit irrigation effects in growing crops. Diplotaxis tenuifolia (wild rocket) seedlings were planted in pots with soil mixed with Oxalis waste at 0, 2.2 and 4.3 kg m-2 or with commercial fertilizer, left to grow for 27 days and then irrigated at 100% or 50% field capacity for 14 days. The incorporation of the Oxalis waste improved the biomass, photosynthesis, sugars, total phenols and total antioxidant capacity in the crop, achieving commercial fertilization values, as well as increasing the phosphorus in soils. However, Oxalis waste seems not to directly affect plants' relative water contents. Our results support the use of Oxalis waste as fertilizer, which can encourage the long-term control of this invasive species.

Olive Tree Belowground Microbiota: Plant Growth-Promoting Bacteria and Fungi.

The olive tree is one of the most significant crops in the Mediterranean region. Its remarkable adaptability to various environments has facilitated olive cultivation across diverse regions and agricultural scenarios. The rising global demand for olive products, coupled with climate challenges, is driving changes in cultivation methods. These changes are altering the traditional landscape and may potentially reshape the structure and composition of orchard microbial communities, which can impact productivity and stress tolerance. Bacterial and fungal communities naturally associated with plants have long been recognized as crucial for plant growth and health, serving as a vital component of sustainable agriculture. In this review, we aim to highlight the significance of olive cultivation and the impact of abiotic stresses. We update the current knowledge on the profiles of rhizosphere and root fungal and bacterial communities in olive orchards and examine how (a)biotic factors influence these communities. Additionally, we explore the potential of plant growth-promoting bacteria and fungi in enhancing olive physiological performance and stress tolerance. We identify knowledge gaps and emphasize the need for implementing new strategies. A comprehensive understanding of olive-associated microbiota will aid in developing sustainable agronomic practices to address climatic challenges and meet the growing demand for olive products.

Olive Leaf Extracts from Three Italian Olive Cultivars Exposed to Drought Stress Differentially Protect Cells against Oxidative Stress.

Olive leaves are an abundant by-product of olive oil production. Olive leaf extracts (OLEs) are rich in polyphenols, which can be used for health benefits. As polyphenols are the main antioxidant molecules in plants, plants typically increase their polyphenol content when exposed to drought stress. However, the phenolic profile of OLEs can vary in relation to the origin and variety of the plant material. In this work, olive leaf extracts from three different Italian olive cultivars (Giarraffa, Leccino, and Maurino) both exposed and not exposed to drought stress were studied in terms of antioxidant properties and profile, intestinal permeation, and protection against oxidative stress of human umbilical vein endothelial cells (HUVECs), since HUVECs are considered a model to study a wide range of diseases. OLEs from stressed Maurino and Giarraffa plants showed the highest increase in antioxidant capacity compared to controls. The phenolic profile of Maurino' was mainly increased by water deficit, with a large increase in the compounds oleuropein and luteolin-7-O-rutinoside. All tested extracts exposed to a water deficit protected HUVECs against oxidative stress by reducing ROS production, and this effect was more pronounced in OLEs from Giarraffa and Maurino exposed to drought stress compared to all other extracts. Finally, OLE from the stressed Giarraffa group showed a higher apparent permeability of antioxidant molecules than that of Maurino.

Comparative metabolomics of leaves and stems of three Italian olive cultivars under drought stress.

The Mediterranean will be one of the focal points of climate change. The predicted dry and hot summers will lead to water scarcity in agriculture, which may limit crop production and growth. The olive tree serves as a model woody plant for studying drought stress and improving water resource management; thus, it is critical to identify genotypes that are more drought tolerant and perform better under low irrigation or even rainfed conditions. In this study, the metabolomic approach was used to highlight variations in metabolites in stems and leaves of three Italian olive cultivars (previously characterized physiologically) under two and four weeks of drought stress. Phenolic and lipophilic profiles were obtained by gas chromatography-mass spectrometry and ultra-high performance liquid chromatography-mass spectrometry, respectively. The findings identified the leaf as the primary organ in which phenolic variations occurred. The Maurino cultivar exhibited a strong stress response in the form of phenolic compound accumulation, most likely to counteract oxidative stress. The phenolic compound content of 'Giarraffa' and 'Leccino' plants remained relatively stable whether they were exposed to drought or not. Variations in the lipid profile occurred in leaves and stems of all the cultivars. A high accumulation of compounds related to epicuticular wax components was observed in the leaf of 'Giarraffa', while a strong reduction of lipids and long-chain alkanes occurred in 'Maurino' when exposed to drought stress conditions.

Impact of Fe3O4-porphyrin hybrid nanoparticles on wheat: Physiological and metabolic advance.

Iron-magnetic nanoparticles (Fe-NMPs) are widely used in environmental remediation, while porphyrin-based hybrid materials anchored to silica-coated Fe3O4-nanoparticles (Fe3O4-NPs) have been used for water disinfection purposes. To assess their safety on plants, especially concerning potential environmental release, it was investigated for the first time, the impact on plants of a silica-coated Fe3O4-NPs bearing a porphyrinic formulation (FORM) - FORM@NMP. Additionally, FORM alone and the magnetic nanoparticles without FORM anchored (NH2@NMP) were used for comparison. Wheat (Triticum aestivum L.) was chosen as a model species and was subjected to three environmentally relevant doses during germination and tiller development through root application. Morphological, physiological, and metabolic parameters were assessed. Despite a modest biomass decrease and alterations in membrane properties, no major impairments in germination or seedling development were observed. During tiller phase, both Fe3O4-NPs increased leaf length, and photosynthesis exhibited varied impacts: both Fe3O4-NPs and FORM alone increased pigments; only Fe3O4-NPs promoted gas exchange; all treatments improved the photochemical phase. Regarding oxidative stress, lipid peroxidation decreased in FORM and FORM@NMP, yet with increased O2-• in FORM@NMP; total flavonoids decreased in NH2@NMP and antioxidant enzymes declined across all materials. Phenolic profiling revealed a generalized trend towards a decrease in flavones. In conclusion, these nanoparticles can modulate wheat physiology/metabolism without apparently inducing phytotoxicity at low doses and during short-time exposure. ENVIRONMENTAL IMPLICATION: Iron-magnetic nanoparticles are widely used in environmental remediation and fertilization, besides of new applications continuously being developed, making them emerging contaminants. Soil is a major sink for these nanoparticles and their fate and potential environmental risks in ecosystems must be addressed to achieve more sustainable environmental applications. Furthermore, as the reuse of treated wastewater for agricultural irrigation is being claimed, it is of major importance to disclose the impact on crops of the nanoparticles used for wastewater decontamination, such as those proposed in this work.

Impact of genome duplications in drought tolerance and distribution of the diploid-tetraploid Jasione maritima.

Polyploidy has important ecological effects, including ploidy-mediated effects on morphology, breeding system and ecological tolerances. However, there is still little comprehensive research available to test its adaptive significance and its role in driving distributional patterns. This work aimed to assess the contribution of genome duplications to ecological divergence using an experimental approach with the diploid-tetraploid Jasione maritima polyploid complex. We explored if individuals with different ploidy differ in their tolerance to water deficit and if this may contribute to explaining the distribution patterns along a latitudinal gradient in the northwest Iberian Peninsula. For that, we used three cytogenetic entities: diploids and established tetraploids collected in natural populations along a latitudinal gradient, and neotetraploids synthesized from diploid populations after treatments with colchicine. Thirty plants from each of the nine populations were grown under controlled conditions with half randomly assigned to the water deficit treatment, and half used as control. We determined experimental plants' response by measuring fitness-related parameters, such as above and belowground biomass, plant water status, photosynthetic efficiency and pigments, membrane stability, antioxidant capacity and sugars content. Our data shows that biomass, chlorophyll content, photochemical quenching (qP) and non-photochemical quenching (NPQ) in neotetraploids and established tetraploids were significantly higher than in diploids and that these differences could be attributed to genome duplications. In response to the water deficit, diploids seem to use a strategy of avoidance, whereas tetraploids seem to employ the strategy of tolerance to overcome water deficit stress, which appears equally efficient. Additionally, we did not observe a response pattern along the latitudinal gradient of the distributional range of the J. maritima complex. The results indicate that the response to water deficit is population dependent. Further studies are necessary to understand the role of ploidy in explaining the distribution patterns of the J. maritima complex.

Functional Groups Response to Water Deficit in Mediterranean Ecosystems.

Enhanced drought, more frequent rainfall events and increased inter-annual variability of precipitation are the main trends of climate expected for the Mediterranean. Drought is one of the most important stressors for plants and significantly impacts plant communities causing changes in plant composition and species dominance. Through an experiment under controlled conditions, we assessed the response of Mediterranean species from different functional groups (annual grass, annual forb, annual legume, and perennial shrub) to moderate and severe water deficit. Changes in plant traits (leaf dry matter), biomass and physiology (water status, photosynthesis, pigments, and carbohydrate) were evaluated. The studied species differed in their response to water deficit. Ornithopus compressus, the legume, showed the strongest response, particularly under severe conditions, decreasing leaf relative water content (RWC), pigments and carbohydrates. The grass, Agrostis pourreti and the forb, Tolpis barbata, maintained RWC, indicating a higher ability to cope with water deficit. Finally, the shrub, Cistus salviifolius, had the lowest response to stress, showing a higher ability to cope with water deficit. Despite different responses, plant biomass was negatively affected by severe water deficit in all species. These data provide background for predicting plant diversity and species composition of Mediterranean grasslands and Montado under climate change conditions.

Metabolomics as a Tool to Understand Nano-Plant Interactions: The Case Study of Metal-Based Nanoparticles.

Metabolomics is a powerful tool in diverse research areas, enabling an understanding of the response of organisms, such as plants, to external factors, their resistance and tolerance mechanisms against stressors, the biochemical changes and signals during plant development, and the role of specialized metabolites. Despite its advantages, metabolomics is still underused in areas such as nano-plant interactions. Nanoparticles (NPs) are all around us and have a great potential to improve and revolutionize the agri-food sector and modernize agriculture. They can drive precision and sustainability in agriculture as they can act as fertilizers, improve plant performance, protect or defend, mitigate environmental stresses, and/or remediate soil contaminants. Given their high applicability, an in-depth understanding of NPs' impact on plants and their mechanistic action is crucial. Being aware that, in nano-plant interaction work, metabolomics is much less addressed than physiology, and that it is lacking a comprehensive review focusing on metabolomics, this review gathers the information available concerning the metabolomic tools used in studies focused on NP-plant interactions, highlighting the impact of metal-based NPs on plant metabolome, metabolite reconfiguration, and the reprogramming of metabolic pathways.

Cadmium-induced cyto- and genotoxicity are organ-dependent in lettuce.

Cadmium is a priority pollutant. Its mechanisms and effects within different plant organs remain unclear. Here, cyto-genotoxicity biomarkers were evaluated in roots and leaves after Cd exposure (0, 1, 10, and 50 µM) of the model crop Lactuca sativa L. (cv. "Reine de Mai"). Overall, superoxide dismutase (SOD) and catalase (CAT) activities were stimulated in leaves, where Cd accumulation was lower in comparison to that in roots. In roots, SOD and peroxidase (POX, APX) activities were stimulated. Moreover, in both organs glutathione reductase (GR) was not affected by Cd. Overall, the H(2)O(2) content increased in both organs, while the total antioxidant capacity decreased in leaves and increased in roots with Cd concentrations. In both organs, lipid and protein oxidation rose with consequent increase of membrane permeability. Simultaneously, the comet assay showed that tail moment, tail length, and % tail DNA were maximum for 1 µM. For 10 µM, shorter tails were found suggesting induced Cd-DNA adducts that lead to DNA-DNA/DNA-protein cross-links, and/or formation of longer DNA fragments, and/or impairment of DNA repair mechanisms, while at 50 µM, nucleoids sensitivity to the technique was evident. This result was consistent with the maximum micronuclei frequency found for the 10 µM Cd dose in roots, suggesting that the surviving cells in this organ had an increase of mitotic catastrophe and that DNA repair systems for blocking cell cycle were dysfunctional. In lower Cd concentrations, root cells might have developed strategies to repair damaged DNA by blocking the cell cycle at specific checkpoints, thus avoiding mitotic catastrophe. Roots at 1 µM showed a cell cycle blockage trend at the G(2) checkpoint, while those at higher concentrations presented S phase delay. We finally discuss a general model of Cd-organ interaction covering these cyto- and genotoxic effects and the potential use of this cultivar in phytoremediation strategies.

Titanium and Zinc Based Nanomaterials in Agriculture: A Promising Approach to Deal with (A)biotic Stresses?

Abiotic stresses, such as those induced by climatic factors or contaminants, and biotic stresses prompted by phytopathogens and pests inflict tremendous losses in agriculture and are major threats to worldwide food security. In addition, climate changes will exacerbate these factors as well as their negative impact on crops. Drought, salinity, heavy metals, pesticides, and drugs are major environmental problems that need deep attention, and effective and sustainable strategies to mitigate their effects on the environment need to be developed. Besides, sustainable solutions for agrocontrol must be developed as alternatives to conventional agrochemicals. In this sense, nanotechnology offers promising solutions to mitigate environmental stress effects on plants, increasing plant tolerance to the stressor, for the remediation of environmental contaminants, and to protect plants against pathogens. In this review, nano-sized TiO2 (nTiO2) and ZnO (nZnO) are scrutinized, and their potential to ameliorate drought, salinity, and xenobiotics effects in plants are emphasized, in addition to their antimicrobial potential for plant disease management. Understanding the level of stress alleviation in plants by these nanomaterials (NM) and relating them with the application conditions/methods is imperative to define the most sustainable and effective approaches to be adopted. Although broad-spectrum reviews exist, this article provides focused information on nTiO2 and nZnO for improving our understanding of the ameliorative potential that these NM show, addressing the gaps in the literature.

Strategies and prospects for biostimulants to alleviate abiotic stress in plants.

Global climate change-induced abiotic stresses (e.g., drought, salinity, extreme temperatures, heavy metals, and UV radiation) have destabilized the fragile agroecosystems and impaired plant performance and thereby reducing crop productivity and quality. Biostimulants, as a promising and eco-friendly approach, are widely used to address environmental concerns and fulfill the need for developing sustainable/modern agriculture. Current knowledge revealed that plant and animal derived stimulants (e.g., seaweeds and phytoextracts, humic substances, and protein hydrolysate) as well as microbial stimulants (e.g., plant beneficial bacteria or fungi) have great potential to elicit plant tolerance to various abiotic stresses and thus enhancing plant growth and performance-related parameters (such as root growth/diameter, flowering, nutrient use efficiency/translocation, soil water holding capacity, and microbial activity). However, to successfully implement biostimulant-based agriculture in the field under changing climate, the understanding of agricultural functions and action mechanism of biostimulants coping with various abiotic stresses at physicochemical, metabolic, and molecular levels is needed. Therefore, this review attempts to unravel the underlying mechanisms of action mediated by diverse biostimulants in relation to abiotic stress alleviation as well as to discuss the current challenges in their commercialization and implementation in agriculture under changing climate conditions.

Chemical Profiling of Two Italian Olea europaea (L.) Varieties Subjected to UV-B Stress.

The depletion of the stratospheric ozone layer due to natural and/or anthropogenic causes decreases the amount of UV-B radiation filtered, and consequently increases the risk of potential damage to organisms. In the Mediterranean region, high UV-B indices are frequent. Even for species typical of this region, such as the olive tree, the progressive increase in UV-B radiation represents a threat. This work aimed to understand how high UV-B radiation modulates the phenolic and lipophilic profile of olive varieties, and identify metabolites that enhance olive stress tolerance. Two Italian olive varieties were subjected to chronic UV-B stress, and leaves were analyzed by gas and liquid chromatography. The results indicated that the most representative phenolic and lipophilic compounds of Giarraffa and Olivastra Seggianese were readjusted in response to UV-B stress. The Giarraffa variety seemed better suited to prolonged UV-B stress, possibly due to the higher availability of flavonoids that could help control oxidative damage, and the accumulation of hydroxycinnamic acid derivatives that could provide strong UV-B shield protection. In addition, this variety contained higher levels of fatty acids (e.g., palmitic, α-linolenic, and stearic acids), which can help to maintain membrane integrity and accumulate more sorbitol (which may serve as an osmoprotectant or act a free-radical scavenger), terpenes, and long-chain alkanes, providing higher protection against UV-B stress.

Quercus suber Roots Activate Antioxidant and Membrane Protective Processes in Response to High Salinity.

Cork oak (Quercus suber) is a species native to Mediterranean areas and its adaptation to the increasingly prevalent abiotic stresses, such as soil salinization, remain unknown. In sequence with recent studies on salt stress response in the leaf, it is fundamental to uncover the plasticity of roots directly exposed to high salinity to better understand how Q. suber copes with salt stress. In the present study we aimed to unveil the antioxidants and key-genes involved in the stress-responses (early vs. later responses) of Q. suber roots exposed to high salinity. Two-month-old Q. suber plants were watered with 300 mM NaCl solution and enzymatic and non-enzymatic antioxidants, lipid peroxidation and the relative expression of genes related to stress response were analysed 8 h and 6 days after salt treatment. After an 8 h of exposure, roots activated the expression of QsLTI30 and QsFAD7 genes involved in stress membrane protection, and QsRAV1 and QsCZF1 genes involved in tolerance and adaptation. As a result of the continued salinity stress (6 days), lipid peroxidation increased, which was associated with an upregulation of QsLTI30 gene. Moreover, other protective mechanisms were activated, such as the upregulation of genes related to antioxidant status, QsCSD1 and QsAPX2, and the increase of the antioxidant enzyme activities of superoxide dismutase, catalase, and ascorbate peroxidase, concomitantly with total antioxidant activity and phenols. These data suggest a response dependent on the time of salinity exposure, leading Q. suber roots to adopt protective complementary strategies to deal with salt stress.

Spatiotemporal Variation in Pollination Deficits in an Insect-Pollinated Dioecious Crop.

Inadequate quantity and quality of pollen reaching the stigmas decreases the sexual reproductive output of plants, compromising yield. Still, the current extent of pollen limitation affecting yield (i.e., pollination deficits) is poorly quantified. This study is aimed at quantifying pollination deficits in kiwifruit orchards, a dioecious plant with a fruit caliber and market value largely dependent on pollination services. For that, we set up a pollination experiment and quantified services and yield provided by current pollination vectors, and under optimal pollination, over two years in a total of twenty-three orchards covering the kiwifruit production range in Portugal. We characterized nine fruit traits and used: (1) fruit weight to calculate pollination deficits and relate them with pollinator diversity and abundance, and environmental variables; and (2) production values, fruit caliber, and market values to calculate economic impact of pollination deficits. Results showed that pollination deficits were variable in time and space and were significantly and negatively correlated with pollinator abundance, while the opposite pattern was obtained for production, supporting the notion that a higher pollinator's abundance is related to lower pollination deficits and higher yields. Understanding the factors affecting pollination deficits is crucial to depict the need for nature-based solutions promoting pollinators and to resort to management practices assisting pollination.