A synchronized, large-scale field experiment using Arabidopsis thaliana reveals the significance of the UV-B photoreceptor UVR8 under natural conditions.

This study determines the functional role of the plant ultraviolet-B radiation (UV-B) photoreceptor, UV RESISTANCE LOCUS 8 (UVR8) under natural conditions using a large-scale 'synchronized-genetic-perturbation-field-experiment'. Laboratory experiments have demonstrated a role for UVR8 in UV-B responses but do not reflect the complexity of outdoor conditions where 'genotype × environment' interactions can mask laboratory-observed responses. Arabidopsis thaliana knockout mutant, uvr8-7, and the corresponding Wassilewskija wild type, were sown outdoors on the same date at 21 locations across Europe, ranging from 39°N to 67°N latitude. Growth and climatic data were monitored until bolting. At the onset of bolting, rosette size, dry weight, and phenolics and glucosinolates were quantified. The uvr8-7 mutant developed a larger rosette and contained less kaempferol glycosides, quercetin glycosides and hydroxycinnamic acid derivatives than the wild type across all locations, demonstrating a role for UVR8 under field conditions. UV effects on rosette size and kaempferol glycoside content were UVR8 dependent, but independent of latitude. In contrast, differences between wild type and uvr8-7 in total quercetin glycosides, and the quercetin-to-kaempferol ratio decreased with increasing latitude, that is, a more variable UV response. Thus, the large-scale synchronized approach applied demonstrates a location-dependent functional role of UVR8 under natural conditions.

Litter decomposition of three halophytes in a Mediterranean salt marsh: Relevance of litter quality, microbial activity and microhabitat.

Studies of litter decomposition in salt marshes have been mainly focused on the measurement of decomposition rates, being litter quality, the type of microbial decomposers and their extracellular enzyme activity, rarely considered. Moreover, most of these studies have been conducted in Poaceae and Cyperaceae species, being scarce the literature on Chenopodiaceae species, which are abundant in Mediterranean salt marshes. Here we analyse the litter decomposition process of two Chenopodiaceae (Sarcocornia fruticosa and Halimione portulacoides) and one Poaceae (Elytrigia atherica) species, belonging S. fruticosa to a halophilous scrub habitat and the other two to a salt meadow habitat of a Mediterranean salt marsh. For each species, we analysed litter decomposition rates, litter quality, fungal and bacterial biomass and potential extracellular enzymes activities. In order to embrace the spatial heterogeneity, two zones were considered within each habitat. Litter of E. atherica decomposed 7- and 13-fold slower than those of S. fruticosa and H. portulacoides, respectively, suggesting that this species is the one that would favour most the carbon sequestration into the soil. The different decomposition rates would be explained by the higher initial lignin and cellulose content of E. atherica rather than by the initial carbon and nitrogen content and C/N ratio. Moreover, enzyme efficiency, compared to enzyme activity, better contributes to explain the different decomposition rates observed. Bacteria dominated throughout the litter decomposition process regardless the species, but fungi increased their relevance in the later stages, when the relative lignin litter content increased. Litter decomposition was affected by microhabitat spatial differences, although the responses depended on the species. Hence, flooding (in the habitat of S. fruticosa) or soil texture (in the habitat of E. atherica and H. portulacoides) might have modulated the decomposition process, being H. portulacoides the most sensitive species to the spatial differences of the salt meadow habitat.

Uptake, translocation and ligand of silver in Lactuca sativa exposed to silver nanoparticles of different size, coatings and concentration.

The broad use of silver nanoparticles (AgNPs) in daily life products enhances their possibilities to reach the environment. Therefore, it is important to understand the uptake, translocation and biotransformation in plants and the toxicological impacts derived from these biological processes. In this work, Lactuca sativa (lettuce) was exposed during 9 days to different coated (citrate, polyvinylpyrrolidone, polyethylene glycol) and sized (60, 75, 100 nm) AgNPs at different concentrations (1, 3, 5, 7, 10, 15 mg L-1). Total silver measurements in lettuce roots indicated that accumulation of AgNPs is influenced by size and concentration, but not by nanoparticle coating. On the other hand, nanosilver translocation to shoots was more pronounced for neutral charged and large sized NPs at higher NP concentrations. Single particle inductively coupled plasma mass spectrometry analysis, after an enzymatic digestion of lettuce tissues indicated the dissolution of some NPs. Ag K-edge X-ray absorption spectroscopy analysis corroborated the AgNPs dissolution due to the presence of less Ag-Ag bonds and appearance of Ag-O and/or Ag-S bonds in lettuce roots. Toxicological effects on lettuces were observed after exposure to nanosilver, especially for transpiration and stomatal conductance. These findings indicated that AgNPs can enter to edible plants, exerting toxicological effects on them.

Recent advances in the application of capillary electromigration methods for food analysis and Foodomics.

This review work presents and discusses the main applications of capillary electromigration methods in food analysis and Foodomics. Papers that were published during the period February 2015-February 2017 are included following the previous review by Acunha et al. (Electrophoresis 2016, 37, 111-141). The paper shows the large variety of food related molecules that have been analyzed by CE including amino acids, biogenic amines, carbohydrates, chiral compounds, contaminants, DNAs, food additives, heterocyclic amines, lipids, peptides, pesticides, phenols, pigments, polyphenols, proteins, residues, toxins, vitamins, small organic and inorganic compounds, as well as other minor compounds. This work describes the last results on food quality and safety, nutritional value, storage, bioactivity, as well as uses of CE for monitoring food interactions and food processing including recent microchips developments and new applications of CE in Foodomics.

Environmental plasticity of Pinot noir grapevine leaves: A trans-European study of morphological and biochemical changes along a 1,500-km latitudinal climatic gradient.

A 2-year study explored metabolic and phenotypic plasticity of sun-acclimated Vitis vinifera cv. Pinot noir leaves collected from 12 locations across a 36.69-49.98°N latitudinal gradient. Leaf morphological and biochemical parameters were analysed in the context of meteorological parameters and the latitudinal gradient. We found that leaf fresh weight and area were negatively correlated with both global and ultraviolet (UV) radiation, cumulated global radiation being a stronger correlator. Cumulative UV radiation (sumUVR) was the strongest correlator with most leaf metabolites and pigments. Leaf UV-absorbing pigments, total antioxidant capacities, and phenolic compounds increased with increasing sumUVR, whereas total carotenoids and xanthophylls decreased. Despite of this reallocation of metabolic resources from carotenoids to phenolics, an increase in xanthophyll-cycle pigments (the sum of the amounts of three xanthophylls: violaxanthin, antheraxanthin, and zeaxanthin) with increasing sumUVR indicates active, dynamic protection for the photosynthetic apparatus. In addition, increased amounts of flavonoids (quercetin glycosides) and constitutive ß-carotene and α-tocopherol pools provide antioxidant protection against reactive oxygen species. However, rather than a continuum of plant acclimation responses, principal component analysis indicates clusters of metabolic states across the explored 1,500-km-long latitudinal gradient. This study emphasizes the physiological component of plant responses to latitudinal gradients and reveals the physiological plasticity that may act to complement genetic adaptations.

UV-A radiation effects on higher plants: Exploring the known unknown.

Ultraviolet-A radiation (UV-A: 315-400nm) is a component of solar radiation that exerts a wide range of physiological responses in plants. Currently, field attenuation experiments are the most reliable source of information on the effects of UV-A. Common plant responses to UV-A include both inhibitory and stimulatory effects on biomass accumulation and morphology. UV-A effects on biomass accumulation can differ from those on root: shoot ratio, and distinct responses are described for different leaf tissues. Inhibitory and enhancing effects of UV-A on photosynthesis are also analysed, as well as activation of photoprotective responses, including UV-absorbing pigments. UV-A-induced leaf flavonoids are highly compound-specific and species-dependent. Many of the effects on growth and development exerted by UV-A are distinct to those triggered by UV-B and vary considerably in terms of the direction the response takes. Such differences may reflect diverse UV-perception mechanisms with multiple photoreceptors operating in the UV-A range and/or variations in the experimental approaches used. This review highlights a role that various photoreceptors (UVR8, phototropins, phytochromes and cryptochromes) may play in plant responses to UV-A when dose, wavelength and other conditions are taken into account.

Environmental Factors Correlated with the Metabolite Profile of Vitis vinifera cv. Pinot Noir Berry Skins along a European Latitudinal Gradient.

Mature berries of Pinot Noir grapevines were sampled across a latitudinal gradient in Europe, from southern Spain to central Germany. Our aim was to study the influence of latitude-dependent environmental factors on the metabolite composition (mainly phenolic compounds) of berry skins. Solar radiation variables were positively correlated with flavonols and flavanonols and, to a lesser extent, with stilbenes and cinnamic acids. The daily means of global and erythematic UV solar radiation over long periods (bud break-veraison, bud break-harvest, and veraison-harvest), and the doses and daily means in shorter development periods (5-10 days before veraison and harvest) were the variables best correlated with the phenolic profile. The ratio between trihydroxylated and monohydroxylated flavonols, which was positively correlated with antioxidant capacity, was the berry skin variable best correlated with those radiation variables. Total flavanols and total anthocyanins did not show any correlation with radiation variables. Air temperature, degree days, rainfall, and aridity indices showed fewer correlations with metabolite contents than radiation. Moreover, the latter correlations were restricted to the period veraison-harvest, where radiation, temperature, and water availability variables were correlated, making it difficult to separate the possible individual effects of each type of variable. The data show that managing environmental factors, in particular global and UV radiation, through cultural practices during specific development periods, can be useful to promote the synthesis of valuable nutraceuticals and metabolites that influence wine quality.

Physiological adjustments of a Mediterranean shrub to long-term experimental warming and drought treatments.

Warmer temperatures and extended drought in the Mediterranean Basin are becoming increasingly important in determining plant physiological processes and affecting the regional carbon budget. The responses of plant physiological variables such as shoot water potential (Ψ), carbon-assimilation rates (A), stomatal conductance (gs) and intrinsic water-use efficiency (iWUE) to these climatic regimes, however, are not well understood. We conducted long-term (16 years) field experiments with mild nocturnal warming (+0.6°C) and drought (-20% soil moisture) in a Mediterranean early-successional shrubland. Warming treatment moderately influenced Ψ, A and gs throughout the sampling periods, whereas drought treatment strongly influenced these variables, especially during the summer. The combination of a natural drought in summer 2003 and the treatments significantly decreased A and iWUE. Foliar δ13C increased in the treatments relative to control, but not significantly. The values of Ψ, A and gs were correlated negatively with vapor-pressure deficit (VPD) and positively with soil moisture and tended to be more dependent on the availability of soil water. The plant, however, also improved the acclimation to drier and hotter conditions by physiological adjustments (gs and iWUE). Understanding these physiological processes in Mediterranean shrubs is crucial for assessing further climate change impacts on ecosystemic functions and services.

Interactive effects of UV radiation and reduced precipitation on the seasonal leaf phenolic content/composition and the antioxidant activity of naturally growing Arbutus unedo plants.

The effects of UV radiation and rainfall reduction on the seasonal leaf phenolic content/composition and antioxidant activity of the Mediterranean shrub Arbutus unedo were studied. Naturally growing plants of A. unedo were submitted to 97% UV-B reduction (UVA), 95% UV-A+UV-B reduction (UV0) or near-ambient UV levels (UVBA) under two precipitation regimes (natural rainfall or 10-30% rainfall reduction). Total phenol, flavonol and flavanol contents, levels of eight phenols and antioxidant activity [DPPH(●) radical scavenging and Cu (II) reducing capacity] were measured in sun-exposed leaves at the end of four consecutive seasons. Results showed a significant seasonal variation in the leaf content of phenols of A. unedo, with the lowest values found in spring and the highest in autumn and/or winter. Leaf ontogenetic development and/or a possible effect of low temperatures in autumn/winter may account for such findings. Regardless of the watering regime and the sampling date, plant exposure to UV-B radiation decreased the total flavanol content of leaves, while it increased the leaf content in quercitrin (the most abundant quercetin derivative identified). By contrast, UV-A radiation increased the leaf content of theogallin, a gallic acid derivative. Other phenolic compounds (two quercetin derivatives, one of them being avicularin, and one kaempferol derivative, juglanin), as well as the antioxidant activity of the leaves, showed different responses to UV radiation depending on the precipitation regime. Surprisingly, reduced rainfall significantly decreased the total amount of quantified quercetin derivatives as well as the DPPH scavenging activity in A. unedo leaves. To conclude, present findings indicate that leaves of A. unedo can be a good source of antioxidants throughout the year, but especially in autumn and winter.

Interactive effects of UV radiation and water availability on seedlings of six woody Mediterranean species.

To assess the effects of UV radiation and its interaction with water availability on Mediterranean plants, we performed an experiment with seedlings of six Mediterranean species (three mesophytes vs three xerophytes) grown in a glasshouse from May to October under three UV conditions (without UV, with UVA and with UVA+UVB) and two irrigation levels (watered to saturation and low watered). Morphological, physiological and biochemical measures were taken. Exposure to UVA+UVB increased the overall leaf mass per area (LMA) and the leaf carotenoids/chlorophyll a + b ratio of plants in relation to plants grown without UV or with UVA, respectively. In contrast, we did not find a general effect of UV on the leaf content of phenols or UVB-absorbing compounds of the studied species. Regarding plant growth, UV inhibited the above-ground biomass production of well-watered plants of Pistacia lentiscus. Conversely, under low irrigation, UVA tended to abolish the reduction in growth experienced by P. lentiscus plants growing in a UV-free environment, in accordance with UVA-enhanced apparent electron transport rate (ETR) values under drought in this species. UVA also induced an overall increase in root biomass when plants of the studied species were grown under a low water supply. In conclusion, while plant exposition to UVA favored root growth under water shortage, UVB addition only gave rise to photoprotective responses, such as the increase in LMA or in the leaf carotenoids/chlorophyll a + b ratio of plants. Species-specific responses to UV were not related with the xerophytic or mesophytic character of the studied species.

UV responses of Lolium perenne raised along a latitudinal gradient across Europe: a filtration study.

Lolium perenne (cv. AberDart) was grown at 14 locations along a latitudinal gradient across Europe (37-68°N) to study the impact of ultraviolet radiation (UV) and climate on aboveground growth and foliar UV-B absorbing compounds. At each location, plants were grown outdoors for 5 weeks in a replicated UV-B filtration experiment consisting of open, UV-B transparent (cellulose diacetate) and UV-B opaque (polyester) environments. Fourier transform-infrared spectroscopy was used to compare plant metabolite profiles in relation to treatment and location. UV radiation and climatic parameters were determined for each location from online sources and the data were assessed using a combination of anova and multiple regression analyses. Most of the variation in growth between the locations was attributable to the combination of climatic parameters, with minimum temperature identified as an important growth constraint. However, no single environmental parameter could consistently account for the variability in plant growth. Concentrations of foliar UV-B absorbing compounds showed a positive trend with solar UV across the latitudinal gradient; however, this relationship was not consistent in all treatments. The most striking experimental outcome from this study was the effect of presence or absence of filtration frames on UV-absorbing compounds. Overall, the study demonstrates the value of an European approach in studying the impacts of natural UV across a large latitudinal gradient. We have shown the feasibility of coordinated UV filtration at multiple sites but have also highlighted the need for open controls and careful interpretation of plant responses.

Water-use responses of 'living fossil' conifers to CO2 enrichment in a simulated Cretaceous polar environment.

BACKGROUND AND AIMS: During the Mesozoic, the polar regions supported coniferous forests that experienced warm climates, a CO(2)-rich atmosphere and extreme seasonal variations in daylight. How the interaction between the last two factors might have influenced water use of these conifers was investigated. An experimental approach was used to test the following hypotheses: (1) the expected beneficial effects of elevated [CO(2)] on water-use efficiency (WUE) are reduced or lost during the 24-h light of the high-latitude summer; and (2) elevated [CO(2)] reduces plant water use over the growing season. METHODS: Measurements of leaf and whole-plant gas exchange, and leaf-stable carbon isotope composition were made on one evergreen (Sequoia sempervirens) and two deciduous (Metasequoia glyptostroboides and Taxodium distichum) 'living fossil' coniferous species after 3 years' growth in controlled-environment simulated Cretaceous Arctic (69 degrees N) conditions at either ambient (400 micromol mol(-1)) or elevated (800 micromol mol(-1)) [CO(2)]. KEY RESULTS: Stimulation of whole-plant WUE (WUE(P)) by CO(2) enrichment was maintained over the growing season for the three studied species but this pattern was not reflected in patterns of WUE inferred from leaf-scale gas exchange measurements (iWUE(L)) and delta(13)C of foliage (tWUE(L)). This response was driven largely by increased rates of carbon uptake, because there was no overall CO(2) effect on daily whole-plant transpiration or whole-plant water loss integrated over the study period. Seasonal patterns of tWUE(L) differed from those measured for iWUE(L). The results suggest caution against over simplistic interpretations of WUE(P) based on leaf isotopic composition. CONCLUSIONS: The data suggest that the efficiency of whole-tree water use may be improved by CO(2) enrichment in a simulated high-latitude environment, but that transpiration is relatively insensitive to atmospheric CO(2) in the living fossil species investigated.

Contrasting growth changes in two dominant species of a Mediterranean shrubland submitted to experimental drought and warming.

BACKGROUND AND AIMS: Climate projections predict drier and warmer conditions in the Mediterranean basin in the next decades. The possibility of such climatic changes modifying the growth of two Mediterranean species, Erica multiflora and Globularia alypum, which are common components of Mediterranean shrublands, was assessed. METHODS: A field experiment was performed from March 1999 to March 2002 to prolong the drought period and to increase the night-time temperature in a Mediterranean shrubland, where E. multiflora and G. alypum are the dominant species. Annual growth in stem diameter and length of both species was measured and annual stem biomass production was estimated for 1999, 2000 and 2001. Plant seasonal growth was also assessed. KEY RESULTS: On average, drought treatment reduced soil moisture 22 %, and warming increased temperature by 0.7-1.6 degrees C. Erica multiflora plants in the drought treatment showed a 46 % lower annual stem elongation than controls. The decrease in water availability also reduced by 31 % the annual stem diameter increment and by 43 % the annual stem elongation of G. alypum plants. New shoot growth of G. alypum was also strongly reduced. Allometrically estimated biomass production was decreased by drought in both species. Warming treatment produced contrasting effects on the growth patterns of these species. Warmer conditions increased, on average, the stem basal diameter growth of E. multiflora plants by 35 %, raising also their estimated stem biomass production. On the contrary, plants of G. alypum in the warming treatment showed a 14 % lower annual stem growth in basal diameter and shorter new shoots in spring compared with controls. CONCLUSIONS: The results indicate changes in the annual productivity of these Mediterranean shrubs under near future drier and warmer conditions. They also point to alterations in their competitive abilities, which could lead to changes in the species composition of these ecosystems in the long term.

Complex spatiotemporal phenological shifts as a response to rainfall changes.

• Climatic warming produces significant gradual alterations in the timing of life-cycle events, and here we study the phenological effects of rainfall-pattern changes. • We conducted ecosystem field experiments that partially excluded rain and runoff during the growing season in a Mediterranean forest and in a mediterranean shrubland. Studies of time-series of leaf-unfolding, flowering and fruiting over the last 50 yr in central Catalonia were carried out, and greenup onset in the Iberian Peninsula was monitored by satellite images. • Experimental, historical and geographical changes in rainfall produced significant, complex and strongly species-specific, as well as spatially and temporally variable, phenological effects. Among these changes, it was found that in the Iberian Peninsula, greenup onset changes from spring (triggered by rising temperatures) in the northern cool-wet regions to autumn (triggered by the arrival of autumn rainfalls) in the southern warm-dry regions. Even in the mesic Mediterranean central Catalonia (NE of the peninsula) rainfall had a stronger relative influence than temperature on fruiting phenology. • The results show that changes in rainfall and water availability, an important driver of climate change, can cause complex phenological changes with likely far-reaching consequences for ecosystem and biosphere functioning and structure. The seasonal shift in the Iberian Peninsula further highlights this importance and indicates that vegetation may respond to climate change not only with gradual, but also with abrupt temporal and spatial, changes in the timing of greenup onset.

Diurnal and seasonal variations in the photosynthetic performance and water relations of two co-occurring Mediterranean shrubs, Erica multiflora and Globularia alypum.

Diurnal and seasonal fluctuations in the photosynthetic performance and water relations of two co-occurring Mediterranean shrubs, Erica multiflora and Globularia alypum were monitored throughout two consecutive years at Garraf Natural Park in north-east Spain. Leaf gas exchange rates, chlorophyll fluorescence and shoot water potentials were measured once each season. Leaf nitrogen and carbon concentrations, leaf delta13C and delta15N and specific leaf area (SLA) were also measured once a year (August) on well developed mature leaves. Globularia alypum experienced seasonal fluctuations in their water potential, with the lowest values recorded in summer, whereas E. multiflora did not show significant differences in water potential among seasons. Moreover, lower water potentials were found in G. alypum than in E. multiflora throughout the entire study, suggesting that the latter behaved as a drought-avoiding species, whereas the former tolerated lower water potentials. In both species, maximum leaf gas exchange rates were observed in autumn and secondarily in spring; in contrast, photosynthetic and transpiration rates reached absolute minima in summer. The stronger fluctuations in water potential and leaf gas exchange rates found in G. alypum compared to E. multiflora, suggest that G. alypum is, sensu Levitt (1980), a water spender, whereas E. multiflora is a water conservative. This hypothesis is further supported by a higher integrated water-use efficiency (higher delta13C values) and a higher degree of sclerophylly (lower SLA) in E. multiflora in comparison with G. alypum. Globularia alypum showed higher leaf gas exchange rates and higher predawn potential photochemical efficiency (Fv/Fm) than E. multiflora during most of the study. In spring and autumn, predawn Fv/Fm values were within the optimal range, whereas chronic photoinhibition in summer and winter was detected in both species. However, whereas both species could maintain positive photosynthetic rates in winter, frequent negative values were found in summer, suggesting higher levels of stress during the drought period. These results together with the high correlations that were found between the net photosynthetic rates and several parameters of water availability (accumulated rainfall, soil moisture or midday water potential) provided further evidence of the key role of water availability in the regulation of the photosynthetic rates in these Mediterranean species. Warmer and drier conditions in future decades, as a consequence of climate change, may alter the present, slight competitive advantage of G. alypum and the fitness of both shrub species within semi-arid Mediterranean environments.

Variations in Quercus ilex chloroplast pigment content during summer stress: involvement in photoprotection according to principal component analysis.

We examined chloroplast pigment variation in holm oak (Quercus ilex L.) leaves for two periods under two climatic conditions, at midday during summer. We compared variation between control (unburned) plants and plants burned the preceding summer, since post-fire resprouts show higher photosynthetic rates and lower thermal energy dissipation. Principal component (PC) analysis was performed on nine pigment-content variables for the two periods separately. Two PC factors (PC1 and PC2) explained 83 and 84% of the variance of the data for each period. In both periods, PC1 was marked by positive loading of pigments associated with light absorption or structural function namely neoxanthin, lutein, ß-carotene, chlorophyll a, and chlorophyll b. These pigments were only affected by leaf age. In contrast, PC2 was marked by high loadings of xanthophyll-cycle pigments (associated with photoprotection), and lutein-5,6-epoxide. Leaf content of these pigments was affected by climatic conditions. In the situations considered in PC analysis (leaf types, periods), the lutein-5,6-epoxide content presented a variation pattern similar to that of violaxanthin, and was significantly correlated with thermal dissipation of excess energy (represented by non-photochemical quenching or NPQ). These results suggest a relationship of lutein and lutein-5,6-epoxide with photoprotection.