Inmaculada Pascual

Plant availability of heavy metals in a soil amended with a high dose of sewage sludge under drought conditions

The objective of this research was to study the effect of water deficit on soil heavy metal availability and metal uptake by ryegrass (Lolium multiflorum Lam.) plants grown in a soil amended with a high dose of rural sewage sludge. Three fertility treatments were applied: sewage sludge (SS), mineral fertilizer (M), and control (C); unamended). The levels of irrigation were: well-watered (W) and water deficit (D). Microbial respiration decreased the total organic C (TOC) in sludge-treated soils, but this did not enhance soil DTPA-extractable heavy metal concentrations. Indeed, Zn, Cu, Mn and Ni availability decreased during the experiment. C- and M-treated soils showed either no changes or increases of some trace element concentrations during the incubation. In the plant experiment, ryegrass dry matter (DM) yield, relative water content (RWC) and leaf water potential (Ψw) decreased in drought conditions. Sludge addition increased metal concentrations in plants. However, in some instances, SS-treated plants showed either similar or lower transfer coefficient (Tc) values than did plants in the C and M treatments. Water deficit decreased the concentration and the Tc of some metals in roots of M and SS plants. Results indicate that sludge-borne heavy metals were maintained in chemical forms of low availability. The lower metal uptake by SS and M plants under dry conditions cannot be attributed to a lower availability of these elements in soil.

Ultraviolet-B radiation modifies the quantitative and qualitative profile of flavonoids and amino acids in grape berries.

Grapevine cv. Tempranillo fruit-bearing cuttings were exposed to supplemental ultraviolet-B (UV-B) radiation under controlled conditions, in order to study its effect on grape traits, ripening, amino acids and flavonoid profile. The plants were exposed to two doses of UV-B biologically effective (5.98 and 9.66kJm(-2)d(-1)), applied either from fruit set to ripeness or from the onset of veraison to ripeness. A 0kJm(-2)d(-1) treatment was included as a control. UV-B did not significantly modify grape berry size, but increased the relative mass of berry skin. Time to reach ripeness was not affected by UV-B, which may explain the lack of changes in technological maturity. The concentration of must extractable anthocyanins, colour density and skin flavonols were enhanced by UV-B, especially in plants exposed from fruit set. The quantitative and qualitative profile of grape skin flavonols were modified by UV-B radiation. Monosubstituted flavonols relative abundance increased proportionally to the accumulated UV-B doses. Furthermore, trisubstituted forms, which where predominant in non-exposed berries, were less abundant as UV-B exposure increased. Although total free amino acid content remained unaffected by the treatments, the increased levels of gamma-aminobutyric acid (GABA), as well as the decrease in threonine, isoleucine, methionine, serine and glycine, revealed a potential influence of UV-B on the GABA-mediated signalling and amino acid metabolism. UV-B had an overall positive impact on grape berry composition.

Short- and long-term physiological responses of grapevine leaves to UV-B radiation

The present study aimed at evaluating the short- and long-term effects of UV-B radiation on leaves of grapevine Vitis vinifera (cv. Tempranillo). Grapevine fruit-bearing cuttings were exposed to two doses of supplemental biologically effective UV-B radiation (UV-BBE) under glasshouse-controlled conditions: 5.98 and 9.66kJm(-2)d(-1). The treatments were applied either for 20d (from mid-veraison to ripeness) or 75d (from fruit set to ripeness). A 0kJm(-2)d(-1) UV-B treatment was included as control. The main effects of UV-B were observed after the short-term exposure (20d) to 9.66kJm(-2)d(-1). Significant decreases in net photosynthesis, stomatal conductance, sub-stomatal CO2 concentration, the actual photosystem II (PSII) efficiency, total soluble proteins and de-epoxidation state of the VAZ cycle were observed, whereas the activities of several antioxidant enzymes increased significantly. UV-B did not markedly affect dark respiration, photorespiration, the maximum potential PSII efficiency (Fv/Fm), non-photochemical quenching (NPQ), as well as the intrinsic PSII efficiency. However, after 75d of exposure to 5.98and 9.66kJm(-2)d(-1) UV-B most photosynthetic and biochemical variables were unaffected and there were no sign of oxidative damage in leaves. The results suggest a high long-term acclimation capacity of grapevine to high UV-B levels, associated with a high accumulation of UV-B absorbing compounds in leaves, whereas plants seemed to be tolerant to moderate doses of UV-B.

Carbon balance, partitioning and photosynthetic acclimation in fruit-bearing grapevine (Vitis vinifera L. cv. Tempranillo) grown under simulated climate change (elevated CO2, elevated temperature and moderate drought) scenarios in temperature gradient greenhouses

Although plant performance under elevated CO2 has been extensively studied in the past little is known about photosynthetic performance changing simultaneously CO2, water availability and temperature conditions. Moreover, despite of its relevancy in crop responsiveness to elevated CO2 conditions, plant level C balance is a topic that, comparatively, has received little attention. In order to test responsiveness of grapevine photosynthetic apparatus to predicted climate change conditions, grapevine (Vitis vinifera L. cv. Tempranillo) fruit-bearing cuttings were exposed to different CO2 (elevated, 700ppm vs. ambient, ca. 400ppm), temperature (ambient vs. elevated, ambient +4°C) and irrigation levels (partial vs. full irrigation). Carbon balance was followed monitoring net photosynthesis (AN, C gain), respiration (RD) and photorespiration (RL) (C losses). Modification of environment (13)C isotopic composition (δ(13)C) under elevated CO2 (from -10.30 to -24.93‰) enabled the further characterization of C partitioning into roots, cuttings, shoots, petioles, leaves, rachides and berries. Irrespective of irrigation level and temperature, exposure to elevated CO2 induced photosynthetic acclimation of plants. C/N imbalance reflected the inability of plants grown at 700ppm CO2 to develop strong C sinks. Partitioning of labeled C to storage organs (main stem and roots) did not avoid accumulation of labeled photoassimilates in leaves, affecting negatively Rubisco carboxylation activity. The study also revealed that, after 20 days of treatment, no oxidative damage to chlorophylls or carotenoids was observed, suggesting a protective role of CO2 either at current or elevated temperatures against the adverse effect of water stress.

Methodological advances: using greenhouses to simulate climate change scenarios.

Human activities are increasing atmospheric CO2 concentration and temperature. Related to this global warming, periods of low water availability are also expected to increase. Thus, CO2 concentration, temperature and water availability are three of the main factors related to climate change that potentially may influence crops and ecosystems. In this report, we describe the use of growth chamber - greenhouses (GCG) and temperature gradient greenhouses (TGG) to simulate climate change scenarios and to investigate possible plant responses. In the GCG, CO2 concentration, temperature and water availability are set to act simultaneously, enabling comparison of a current situation with a future one. Other characteristics of the GCG are a relative large space of work, fine control of the relative humidity, plant fertirrigation and the possibility of light supplementation, within the photosynthetic active radiation (PAR) region and/or with ultraviolet-B (UV-B) light. In the TGG, the three above-mentioned factors can act independently or in interaction, enabling more mechanistic studies aimed to elucidate the limiting factor(s) responsible for a given plant response. Examples of experiments, including some aimed to study photosynthetic acclimation, a phenomenon that leads to decreased photosynthetic capacity under long-term exposures to elevated CO2, using GCG and TGG are reported.

Climate change conditions (elevated CO2 and temperature) and UV-B radiation affect grapevine (Vitis vinifera cv. Tempranillo) leaf carbon assimilation, altering fruit ripening rates

The increase in grape berry ripening rates associated to climate change is a growing concern for wine makers as it rises the alcohol content of the wine. The present work studied the combined effects of elevated CO2, temperature and UV-B radiation on leaf physiology and berry ripening rates. Three doses of UV-B: 0, 5.98, 9.66 kJm(-2)d(-1), and two CO2-temperature regimes: ambient CO2-24/14 °C (day/night) (current situation) and 700 ppm CO2-28/18 °C (climate change) were imposed to grapevine fruit-bearing cuttings from fruit set to maturity under greenhouse-controlled conditions. Photosynthetic performance was always higher under climate change conditions. High levels of UV-B radiation down regulated carbon fixation rates. A transient recovery took place at veraison, through the accumulation of flavonols and the increase of antioxidant enzyme activities. Interacting effects between UV-B and CO2-temperature regimes were observed for the lipid peroxidation, which suggests that UV-B may contribute to palliate the signs of oxidative damage induced under elevated CO2-temperature. Photosynthetic and ripening rates were correlated. Thereby, the hastening effect of climate change conditions on ripening, associated to higher rates of carbon fixation, was attenuated by UV-B radiation.

Growth, photosynthetic acclimation and yield quality in legumes under climate change simulations: An updated survey

Continued emissions of CO2, derived from human activities, increase atmospheric CO2 concentration. The CO2 rise stimulates plant growth and affects yield quality. Effects of elevated CO2 on legume quality depend on interactions with N2-fixing bacteria and mycorrhizal fungi. Growth at elevated CO2 increases photosynthesis under short-term exposures in C3 species. Under long-term exposures, however, plants generally acclimate to elevated CO2 decreasing their photosynthetic capacity. An updated survey of the literature indicates that a key factor, perhaps the most important, that characteristically influences this phenomenon, its occurrence and extent, is the plant source-sink balance. In legumes, the ability of exchanging C for N at nodule level with the N2-fixing symbionts creates an extra C sink that avoids the occurrence of photosynthetic acclimation. Arbuscular mycorrhizal fungi colonizing roots may also result in increased C sink, preventing photosynthetic acclimation. Defoliation (Anthyllis vulneraria, simulated grazing) or shoot cutting (alfalfa, usual management as forage) largely increases root/shoot ratio. During re-growth at elevated CO2, new shoots growth and nodule respiration function as strong C sinks that counteracts photosynthetic acclimation. In the presence of some limiting factor, the legumes response to elevated CO2 is weakened showing photosynthetic acclimation. This survey has identified limiting factors that include an insufficient N supply from bacterial strains, nutrient-poor soils, low P supply, excess temperature affecting photosynthesis and/or nodule activity, a genetically determined low nodulation capacity, an inability of species or varieties to increase growth (and therefore C sink) at elevated CO2 and a plant phenological state or season when plant growth is stopped.

Characterization of the adaptive response of grapevine (cv. Tempranillo) to UV-B radiation under water deficit conditions

This work aims to characterize the physiological response of grapevine (Vitis vinifera L.) cv. Tempranillo to UV-B radiation under water deficit conditions. Grapevine fruit-bearing cuttings were exposed to three levels of supplemental biologically effective UV-B radiation (0, 5.98 and 9.66kJm(-2)day(-1)) and two water regimes (well watered and water deficit), in a factorial design, from fruit-set to maturity under glasshouse-controlled conditions. UV-B induced a transient decrease in net photosynthesis (Anet), actual and maximum potential efficiency of photosystem II, particularly on well watered plants. Methanol extractable UV-B absorbing compounds (MEUVAC) concentration and superoxide dismutase activity increased with UV-B. Water deficit effected decrease in Anet and stomatal conductance, and did not change non-photochemical quenching and the de-epoxidation state of xanthophylls, dark respiration and photorespiration being alternative ways to dissipate the excess of energy. Little interactive effects between UV-B and drought were detected on photosynthesis performance, where the impact of UV-B was overshadowed by the effects of water deficit. Grape berry ripening was strongly delayed when UV-B and water deficit were applied in combination. In summary, deficit irrigation did not modify the adaptive response of grapevine to UV-B, through the accumulation of MEUVAC. However, combined treatments caused additive effects on berry ripening.

Photosynthetic response of pepper plants to wilt induced by Verticillium dahliae and soil water deficit

Greenhouse experiments were conducted to compare stress effects caused by Verticillium dahliae and drought on gas exchange, chlorophyll (Chl) fluorescence and photosynthetic pigments of pepper plants. Three treatments were compared: Verticillium inoculated plants (+V), non-inoculated well-watered plants (-V) and non-inoculated plants subjected to progressive drought (D). Gas exchange, fluorescence and photosynthetic pigments were measured and represented along a gradient of relative water content (RWC) and stomatal conductance (g(s)). Net photosynthesis (A) and electron transport rate (ETR) decreased, as RWC and g(s) declined, similarly in both +V and D plants. However, dark respiration (R(D)) and photorespiration (R(L)) tended to increase in inoculated plants compared to those subjected to soil drought, as g(s) decreased. Photoinhibitory damage was not observed in infected or in droughted plants. Soil drought decreased intrinsic PSII efficiency (Phi(exc.)), which seemed to result in part from enhanced xanthophyll cycle- and/or lutein-related thermal energy dissipation. Nevertheless, the fact that 1-Phi(exc.) increased in D only at high values of the de-epoxidation state of the xanthophyll cycle (DPS) suggests that DeltapH could be the major factor controlling thermal energy dissipation in this treatment. By contrast, antheraxanthin, zeaxanthin and lutein, as well as Phi(exc.), were not markedly affected in +V. Water stress appeared to be the main limitation to photosynthesis in Verticillium infected plants, probably through stomatal closure, together with impaired mesophyll conductance (g(m)). However, our results indicate differential effects of V. dahliae on dark respiration, photorespiration, g(m) and on the capability of thermal energy dissipation under low g(s) values.

Sensitivity of Grapevine Phenology to Water Availability, Temperature and CO2 Concentration

In recent decades, mean global temperatures have increased in parallel with a sharp rise in atmospheric carbon dioxide (CO2) levels, with apparent implications for precipitation patterns. The aim of the present work is to assess the sensitivity of different phenological stages of grapevine to temperature and to study the influence of other factors related to climate change (water availability and CO2 concentration) on this relationship. Grapevine phenological records from 9 plantings between 42.75°N and 46.03°N consisting of dates for budburst, flowering and fruit maturity were used. In addition, we used phenological data collected from two years of experiments with grapevine fruit-bearing cuttings with two grapevine varieties under two levels of water availability, two temperature regimes and two levels of CO2. Dormancy breaking and flowering were strongly dependent on spring temperature, while neither variation in temperature during the chilling period nor precipitation significantly affected budburst date. The time needed to reach fruit maturity diminished with increasing temperature and decreasing precipitation. Experiments under semi-controlled conditions revealed great sensitivity of berry development to both temperature and CO2. Water availability had significant interactions with both temperature and CO2; however, in general, water deficit delayed maturity when combined with other factors. Sensitivities to temperature and CO2 varied widely, but higher sensitivities appeared in the coolest year, particularly for the late ripening variety, ‘White Tempranillo’. The knowledge gained in whole plant physiology and multi stress approaches is crucial to predict the effects of climate change and to design mitigation and adaptation strategies allowing viticulture to cope with climate change.