Atmospheric CO2 levels and global temperatures are expected to rise in the next decades, and viticulture must face these changes. Within this context, exploiting the intra-varietal diversity of grapevine (Vitis vinifera L.) can be a useful tool for the adaptation of this crop to climate change. The aim of the present work was to study the effect of elevated temperature and elevated levels of atmospheric CO2, both individually and combined, on the growth, phenology and carbon partitioning of five clones of the cultivar Tempranillo (RJ43, CL306, T3, VN31 and 1084). The hypothesis that clones within the same variety that differ in their phenological development may respond in a different manner to the above mentioned environmental factors from a physiological point of view was tested. Grapevine fruit-bearing cuttings were grown from fruit set to maturity under two temperature regimes: ambient (T) vs elevated (ambient + 4 degrees C, T + 4), combined with two CO2 levels: ambient (ca. 400 ppm, ACO(2)) vs elevated (700 ppm, ECO2), in temperature-gradient greenhouses (TGGs). Considering all the clones, elevated temperature hastened grape development and increased vegetative growth, but reduced grape production, the later most likely associated with the heat waves recorded during the experiment. Plants in the elevated CO2 treatments showed a higher photosynthetic activity at veraison and an increased vegetative growth, but they showed signs of photosynthetic acclimation to ECO2 at maturity according to the C:N ratio, especially when combined with high temperature. The combination of ECO2 and T + 4, mimicking climate change environmental conditions, showed additive effects in some of the parameters analyzed. The clones showed differences in their phenological development, which conditioned some responses to elevated CO2 and temperature in terms of vegetative production and C partitioning into different organs. The work adds new knowledge on the use of different grapevine clones, that can be useful to improve the viticultural efficiency in future climate change scenarios.

Tempranillo is a grapevine (Vitis vinifera L.) variety extensively used for world wine production which is expected to be affected by environmental parameters modified by ongoing global climate changes, i.e., increases in average air temperature and rise of atmospheric CO2 levels. Apart from determining their effects on grape development and biochemical characteristics, this paper considers the intravarietal diversity of the cultivar Tempranillo as a tool to develop future adaptive strategies to face the impact of climate change on grapevine. Fruit-bearing cuttings of five clones (RJ43, CL306, T3, VN31, and 1084) were grown in temperature gradient greenhouses (TGGs), from fruit set to maturity, under two temperature regimes (ambient temperature vs. ambient temperature plus 4 degrees C) and two CO2 levels (ambient, ca. 400 ppm, vs. elevated, 700 ppm). Treatments were applied separately or in combination. The analyses carried out included berry phenological development, the evolution in the concentration of must compounds (organic acids, sugars, and amino acids), and total skin anthocyanins. Elevated temperature hastened berry ripening, sugar accumulation, and malic acid breakdown, especially when combined with high CO2. Climate change conditions reduced the amino acid content 2 weeks after mid-veraison and seemed to delay amino acidic maturity. Elevated CO2 reduced the decoupling effect of temperature on the anthocyanin to sugar ratio. The impact of these factors, taken individually or combined, was dependent on the clone analyzed, thus indicating certain intravarietal variability in the response of Tempranillo to these climate change-related factors.

Environmental stress factors caused by climate change affect plant growth and crop production, and pose a growing threat to sustainable agriculture, especially for tree crops. In this context, we sought to investigate the responses to climate change of two Prunus rootstocks (GF677 and Adesoto) budded with Catherina peach cultivar. Plants were grown in 15 L pots in temperature gradient greenhouses for an 18 days acclimation period after which six treatments were applied: [CO2 levels (400 versus 700 mol mol-1), temperature (ambient versus ambient + 4°C), and water availability (well irrigated versus drought)]. After 23 days, the effects of stress were evaluated as changes in physiological and biochemical traits, including expression of relevant genes. Stem water potential decreased under drought stress in plants grafted on GF677 and Adesoto rootstocks; however, elevated CO2 and temperature affected plant water content differently in both combinations. The photosynthetic rate of plants grafted on GF677 increased under high CO2, but decreased under high temperature and drought conditions. The photosynthetic rates of plants grafted onto Adesoto were only affected by drought treatment. Furthermore, in GF677-Catherina plants, elevated CO2 alleviated the effect of drought, whereas in those grafted onto Adesoto, the same condition produced acclimation in the rate.

Wheat grain represents an important source of carbohydrates, proteins, lipids and minerals. Durum wheat is used mainly for the preparation of pasta, and in some Mediterranean areas is used for bread making. The atmospheric CO2 concentration influences wheat growth, yield and quality. The present work focuses on kernel quality under conditions of elevated [CO2] and subjected, or not, to water stress. The experiments were conducted with the durum wheat (Triticum durum Desf.) varieties cvv. Blanqueta, which is a historical Spanish landrace, and cvv. Sula, which is a modern variety. Sula demonstrated greater kernel weight (KW), insoluble protein (IP) content and amylose content, and also featured better potential test weight (TW) under projected future elevated [CO2] and drought conditions. Blanqueta exposed to drought conditions showed the highest C-13 isotopic composition (delta C-13) values indicating that, as a consequence of their higher biomass, they were subjected to a more severe stress. Under control conditions of ambient [CO2], the protein concentrations of both varieties were similar. This work provides data about the genetic diversity between a currently cultivated wheat cultivar derived from traditional breeding and another cultivated some decades ago.

The increase in the atmospheric CO2 concentration is predicted to influence wheat production and grain quality and nutritional properties. In the present study, durum wheat (Triticum durum Desf. cv. Sula) was grown under two different CO2 (400 versus 700 mu mol mol(-1)) concentrations to examine effects on the crop yield and grain quality at different phenological stages (from grain filling to maturity). Exposure to elevated CO2 significantly increased aboveground biomass and grain yield components. Growth at elevated CO2 diminished the elemental N content as well as protein and free amino acids, with a typical decrease in glutamine, which is the most represented amino acid in grain proteins. Such a general decrease in nitrogenous compounds was associated with altered kinetics of protein accumulation, N remobilization, and N partitioning. Our results highlight important modifications of grain metabolism that have implications for its nutritional quality.

Ammonia (NH3) emissions are linked to eutrophication, plant toxicity and ecosystem shifts from N to P limitation. Bryophytes are key components of terrestrial ecosystems, yet highly sensitive to N deposition. Hence, physiological responses of mosses may be indicative of NH3-related impacts, and thus useful to foresee future ecosystem damages and establish atmospheric Critical Levels (CLEs). In this work, samples of Hypnum cupressiforme Hedw. were seasonally collected along a well-defined NH3 concentration gradient in an oak woodland during a one-year period. We performed a comprehensive evaluation of tissue chemistry, stoichiometry, metabolic enzymes, antioxidant response, membrane damages, photosynthetic pigments, soluble protein content and N and C isotopic fractionation. Our results showed that all the physiological parameters studied (except P, K, Ca and C) responded to the NH3 gradient in predictable ways, although the magnitude and significance of the response were dependent on the sampling season, especially for enzymatic activities and pigments content. Nutritional imbalances, membrane damages and disturbance of cellular C and N metabolism were found as a consequence to NH3 exposure, being more affected the mosses more exposed to the barn atmosphere. These findings suggested significant implications of intensive farming for the correct functioning of oak woodlands and highlighted the importance of seasonal dynamics in the study of key physiological processes related to photosynthesis, mosses nutrition and responses to oxidative stress. Finally, tissue N showed the greatest potential for the identification of NH3-related ecological end points (estimated CLE = 3.5 mu g m(-3)), whereas highly scattered physiological responses, although highly sensitive, were not suitable to that end.

Foreseen climate change is expected to impact on grape composition, both sugar and pigment content. We tested the hypothesis that interactions between main factors associated with climate change (elevated CO2, elevated temperature, and water deficit) decouple sugars and anthocyanins, and explored the possible involvement of vegetative area, photosynthesis, and grape C uploading on the decoupling. Tempranillo grapevine fruit-bearing cuttings were exposed to CO2 (700 vs. 400ppm), temperature (ambient vs. +4 degrees C), and irrigation levels (partial vs. full) in temperature-gradient greenhouses. In a search for mechanistic insights into the underlying processes, experiments 1 and 2 were designed to maximize photosynthesis and enlarge leaf area range among treatments, whereas plant growth was manipulated in order to deliberately down-regulate photosynthesis and control vegetative area in experiments 3 and 4. Towards this aim, treatments were applied either from fruit set to maturity with free vegetation and fully irrigated or at 5-8% of pot capacity (experiments 1 and 2), or from veraison to maturity with controlled vegetation and fully irrigated or at 40% of pot capacity (experiments 3 and 4). Modification of air C-13 isotopic composition under elevated CO2 enabled the further characterization of whole C fixation period and C partitioning to grapes. Increases of the grape sugars-to-anthocyanins ratio were highly and positively correlated with photosynthesis and grape C-13 labeling, but not with vegetative area. Evidence is presented for photosynthesis, from fruit set to veraison, and grape C uploading, from veraison to maturity, as key processes involved in the establishment and development, respectively, of the grape sugars to anthocyanins decoupling.

In recent decades, agricultural production is being affected by a sharp increase in atmospheric CO2 concentration. Due to the greenhouse effect gases, crops are impacted by enhanced temperatures and concomitantly by increased scarce water availability. All arid and semiarid areas, including Mediterranean viticulture, must face these three climate change-related factors: atmospheric CO2 concentration and temperature increases, and scarce water for irrigation. Scarce water is a problem even in irrigated viticulture, as irrigation is becoming more and more restricted. Within this context, the aim of this work was to investigate grapevine (Vitis vinifera L. cv. red and white Tempranillo) vegetative and reproductive growth. Fruit-bearing cuttings were grown under elevated CO2 (around 700 micromol mol-1 or ppm, versus 400), high temperature (ambient temperature +4°C, versus ambient) and water deficit (cyclic drought, versus full irrigated) in temperature gradient greenhouses for three consecutive growing seasons (years 2013, 2014 and 2015). Climate change impacted markedly vegetative growth. Within the abovementioned factors, vegetative growth (total vegetative mass) was significantly reduced by drought (consistent the three years) and was associated to a low substrate water status and low leaf stomatal conductance. Elevated CO2 stimulated total vegetative mass, whereas leaf area was not affected. When plants were grown under elevated CO2, the largest increases were observed ...

The intra-varietal genetic diversity of grapevine (Vitis vinifera L.) may be exploited to maintain grape quality under future warm conditions, which may alter grape berry development and composition. The present study assesses the effects of elevated temperature on the development of berry, grape composition and anthocyanins:sugars ratio of thirteen clones of V. vinifera. cv. Tempranillo that differed in length of the ripening period (time from veraison to berry total soluble solids, mainly sugars, of ca. 22 °Brix). Two temperature regimes (24 °C/14 °C or 28 °C/18 °C, day/night) were imposed to grapevine fruit-bearing cuttings from fruit set to maturity under greenhouse-controlled conditions. Elevated temperature hastened berry development, with a greater influence before the onset of ripening, and reduced anthocyanin concentration, colour intensity and titratable acidity. The clones significantly differed in the number of days that elapsed between fruit set and maturity. At the same concentration of total soluble solids, the anthocyanin concentration was lower at 28 °C/18 °C than 24 °C/14 °C, indicating a decoupling effect of elevated temperature during berry ripening. Thermal decoupling was explained by changes in the relative rate of response of anthocyanin and sugar build-up, rather than delayed onset of anthocyanin accumulation. Clones differed in the degree of thermal decoupling, but it was directly associated with differences neither in the length of their ripening ...

In the Mediterranean area, changes in crop production and quality are expected in the future, due to one or more stress factors associated with climate change. Among them, plant responses to atmospheric CO2 concentration increases, enhanced temperatures and scarce water availability are a matter that deserves further investigation. In this study, the effects of the three above-mentioned factors, acting individually and/or in interaction, on grapevine reproductive growth and berry quality were investigated in three consecutive growing seasons (2013, 2014, and 2015) in the cultivars red and white Tempranillo. Eight different treatments were applied from fruit set to maturity (2 CO2 levels (400 versus 700 mu mol mol(-1)) x 2 temperature treatments (ambient versus ambient +4 degrees C) x 2 water availability regimes (well irrigated versus cyclic drought)) in four temperature gradient greenhouses located at the University of Navarra (Pamplona, Spain). Yield was significantly reduced by drought and was year-dependent. Eventual heat shocks (above 35 degrees C) in the first week of July in 2015 induced berry burn, browning and loss of 50% of the berries. Regarding quality, simulated climate change scenarios affected to greater extent technological (primary metabolism) than phenolic (secondary metabolism) maturity. Indeed, high temperature and drought significantly and consistently increased must pH, due to decreases in malic acid. On the contrary, elevated CO2 decreased pH associated with significant increases in tartaric acid. Differences in the response of red and white Tempranillo were found. Acidity was lower (and pH higher) in white than in red Tempranillo, due to lower malic and tartaric acid concentrations. Also, total polyphenol index was lower in the white variety, in part due to the absence of anthocyanins. Fresh bunch weight and berry water content were higher in the white than in the red variety. Interactions found among variety, year, water availability, temperature and CO2 have been highlighted and discussed. (C) 2017 Elsevier B.V. All rights reserved.

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, 700 ppm vs. ambient, ca. 400 ppm), temperature (ambient vs. elevated, ambient +4 degrees C) and irrigation levels (partial vs. full irrigation). Carbon balance was followed monitoring net photosynthesis (A(N), C gain), respiration (R-D) and photorespiration (R-L) (C losses). Modification of environment C-13 isotopic composition (delta C-13) 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 700 ppm 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.

C sink/source balance and N assimilation have been identified as target processes conditioning crop responsiveness to elevated CO2 . However, little is known about phenology-driven modifications of C and N primary metabolism at elevated CO2 in cereals such as wheat. Here, we examined the differential effect of elevated CO2 at two development stages (onset of flowering, onset of grain filling) in durum wheat (Triticum durum, var. Sula) using physiological measurements (photosynthesis, isotopes), metabolomics, proteomics and (15) N labelling. Our results show that growth at elevated CO2 was accompanied by photosynthetic acclimation through a lower internal (mesophyll) conductance but no significant effect on Rubisco content, maximal carboxylation or electron transfer. Growth at elevated CO2 altered photosynthate export and tended to accelerate leaf N remobilization, which was visible for several proteins and amino acids, as well as lysine degradation metabolism. However, grain biomass produced at elevated CO2 was larger and less N rich, suggesting that nitrogen use efficiency rather than photosynthesis is an important target for improvement, even in good CO2 -responsive cultivars.

In the Mediterranean area, climate change is associated with atmospheric CO2 concentration increases, enhanced temperatures and scarce water availability, limiting seriously crop yield and decreasing quality. The aim of this study was to investigate the effects of elevated CO2, elevated temperature and water deficit, acting individually and/or interacting, on vegetative and reproductive growth, substrate and plant water status, and must quality in fruit-bearing cuttings of two grapevine (Vitis vinifera L.) cultivars (red and white Tempranillo). In four temperature gradient greenhouses, eight treatments were applied, from fruit set to maturity: CO2 level (400 versus 700 ¿mol mol¿1), temperature (ambient versus ambient +4 °C), and water availability (full irrigation versus cyclic drought). Effects of climate change on grape yield and quality were cultivar dependent. Generally, red Tempranillo had more vegetative growth and grape yield than the white cultivar. Also, grape yield was less affected by the treatments than vegetative growth. Drought, especially under elevated temperature, drastically reduced vegetative growth, bunch fresh and dry weights in both cultivars. Interestingly, elevated CO2 attenuated these negative effects of drought. The effects of climatic factors on yield were not associated with a worse water status of the vegetative or reproductive organs. In red Tempranillo, the combination of elevated CO2, elevated temperature and drought reduced total polyphenol index (TPI), malic acid and increased color density, but did not modify anthocyanin concentration. In white Tempranillo, the combined action of the three factors associated with climate change modified only tartaric acid. In this latter cultivar, drought increased TPI under ambient temperature, regardless of CO2 level, when compared with full-irrigated plants. In conclusion, climate change-related factors (elevated CO2, elevated temperature and water deficit) individually (especially drought) and/or interacting affected to different extent red and white Tempranillo vegetative growth and yield. Drought combined with elevated temperatures reduced grapevine performance, and elevated CO2 mitigated such deleterious effect.

The natural N-15/N-14 isotope composition (delta N-15) of a tissue is a consequence of its N source and N physiological mechanisms in response to the environment. It could potentially be used as a tracer of N metabolism in plants under changing environmental conditions, where primary N metabolism may be complex, and losses and gains of N fluctuate over time. In order to test the utility of delta N-15 as an indicator of plant N status in N-2-fixing plants grown under various environmental conditions, alfalfa (Medicago sativa L.) plants were subjected to distinct conditions of [CO2] (400 vs. 700 mu mol mol(-1)), temperature (ambient vs. ambient +4 degrees C) and water availability (fully watered vs. water deficiency-WD). As expected, increased [CO2] and temperature stimulated photosynthetic rates and plant growth, whereas these parameters were negatively affected by WD. The determination of delta N-15 in leaves, stems, roots, and nodules showed that leaves were the most representative organs of the plant response to increased [CO2] and WD. Depletion of heavier N isotopes in plants grown under higher [CO2] and WD conditions reflected decreased transpiration rates, but could also be related to a higher N demand in leaves, as suggested by the decreased leaf N and total soluble protein (TSP) contents detected at 700 mu mol mol(-1) [CO2] and WD conditions. In summary, leaf delta N-15 provides relevant information integrating parameters which condition plant responsiveness (e.g., photosynthesis, TSP, N demand, and water transpiration) to environmental conditions.

Although climate scenarios have predicted an increase in [CO2] and temperature conditions, to date few experiments have focused on the interaction of [CO2] and temperature effects in wheat development. Recent evidence suggests that photosynthetic acclimation is linked to the photorespiration and N assimilation inhibition of plants exposed to elevated CO2. The main goal of this study was to analyze the effect of interacting [CO2] and temperature on leaf photorespiration, C/N metabolism and N transport in wheat plants exposed to elevated [CO2] and temperature conditions. For this purpose, wheat plants were exposed to elevated [CO2] (400 vs 700 mu molmol(-1)) and temperature (ambient vs ambient+4 degrees C) in CO2 gradient greenhouses during the entire life cycle. Although at the agronomic level, elevated temperature had no effect on plant biomass, physiological analyses revealed that combined elevated [CO2] and temperature negatively affected photosynthetic performance. The limited energy levels resulting from the reduced respiratory and photorespiration rates of such plants were apparently inadequate to sustain nitrate reductase activity. Inhibited N assimilation was associated with a strong reduction in amino acid content, conditioned leaf soluble protein content and constrained leaf N status. Therefore, the plant response to elevated [CO2] and elevated temperature resulted in photosynthetic acclimation. The reduction in transpiration rates induced limitations in nutrient transport in leaves of plants exposed to elevated [CO2] and temperature, led to mineral depletion and therefore contributed to the inhibition of photosynthetic activity.

Although climate scenarios have predicted an increase in [CO2] and temperature conditions, to date few experiments have focused on the interaction of [CO2] and temperature effects in wheat development. Recent evidence suggests that photosynthetic acclimation is linked to the photorespiration and N assimilation inhibition of plants exposed to elevated CO2. The main goal of this study was to analyze the effect of interacting [CO2] and temperature on leaf photorespiration, C/N metabolism and N transport in wheat plants exposed to elevated [CO2] and temperature conditions. For this purpose, wheat plants were exposed to elevated [CO2] (400 vs 700 mu molmol(-1)) and temperature (ambient vs ambient+4 degrees C) in CO2 gradient greenhouses during the entire life cycle. Although at the agronomic level, elevated temperature had no effect on plant biomass, physiological analyses revealed that combined elevated [CO2] and temperature negatively affected photosynthetic performance. The limited energy levels resulting from the reduced respiratory and photorespiration rates of such plants were apparently inadequate to sustain nitrate reductase activity. Inhibited N assimilation was associated with a strong reduction in amino acid content, conditioned leaf soluble protein content and constrained leaf N status. Therefore, the plant response to elevated [CO2] and elevated temperature resulted in photosynthetic acclimation. The reduction in transpiration rates induced limitations in nutrient ...

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.

Despite its relevance, few studies to date have analysed the role of harvest index (HI) in the responsiveness of wheat (Triticum spp.) to elevated CO2 concentration ([CO2]) under limited water availability. The goal of the present work was to characterise the role of HI in the physiological responsiveness of durum wheat (Triticum durum Desf.) exposed to elevated [CO2] and terminal (i.e. during grain filling) water stress. For this purpose, the performance of wheat plants with high versus low HI (cvv. Sula and Blanqueta, respectively) was assessed under elevated [CO2] (700¿¿mol¿mol¿1 vs 400¿¿mol¿mol¿1 CO2) and terminal water stress (imposed after ear emergence) in CO2 greenhouses. Leaf carbohydrate build-up combined with limitations in CO2 diffusion (in droughted plants) limited the responsiveness to elevated [CO2] in both cultivars. Elevated [CO2] only increased wheat yield in fully watered Sula plants, where its larger HI prevented an elevated accumulation of total nonstructural carbohydrates. It is likely that the putative shortened grain filling period in plants exposed to water stress also limited the responsiveness of plants to elevated [CO2]. In summary, our study showed that even under optimal water availability conditions, only plants with a high HI responded to elevated [CO2] with increased plant growth, and that terminal drought constrained the responsiveness of wheat plants to elevated [CO2].

Medicago sativa L. (alfalfa) can exhibit photosynthetic down-regulation when grown in greenhouse conditions under elevated atmospheric CO2. This forage legume can establish a double symbiosis with nitrogen fixing bacteria and arbuscular mycorrhizal fungi (AMF), which may increase the carbon sink effect of roots. Our aim was to assess whether the association of alfalfa with AMF can avoid, diminish or delay the photosynthetic acclimation observed in previous studies performed with nodulated plants. The results, however, showed that mycorrhizal (M) alfalfa at the end of their vegetative period had lower carbon (C) discrimination than non-mycorrhizal (NM) controls, indicating photosynthetic acclimation under ECO2 in plants associated with AMF. Decreased C discrimination was due to the acclimation of conductance, since the amount of Rubisco and the expression of genes codifying both large and small subunits of Rubisco were similar or slightly higher in M than in NM plants. Moreover, M alfalfa accumulated a greater amount of soluble sugars in leaves than NM plants, thus favoring a down-regulation effect on photosynthetic rates. The enhanced contents of sugars in leaves coincided with a reduced percentage of arbuscules in roots, suggesting decreased sink of carbohydrates from shoots to roots in M plants. The shorter life cycle of alfalfa associated with AMF in comparison with the NM controls may also be related to the accelerated photosynthetic acclimation in M plants. Further research is needed to clarify to what extent this behavior could be extrapolated to alfalfa cultivated in the field and subjected to periodic cutting of shoots under climatic 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.

Although responsiveness of N2-fixing plants to elevated CO2 conditions have been analyzed in previous studies, important uncertainties remain in relation to the effect enhanced CO2 in nodule proteomic profile and its implication in leaf responsiveness. The aim of our study was to deepen our understanding of the relationship between leaf and nodule metabolism of N2-fixing alfalfa plants after long-term exposure to elevated CO2. After 30-day exposure to elevated CO2, plants showed photosynthetic down-regulation with reductions in the light-saturated rate of CO2 assimilation (Asat) and the maximum rate of rubisco carboxylation (Vcmax). Under elevated CO2 conditions, the rubisco availability limited potential photosynthesis by around 12 %, which represented the majority of the observed fall in Vcmax. Photosynthetic down-regulation has been associated with decreased N availability even if those plants are capable to assimilate N2. Diminishment in shoot N demand (as reflected by the lower rubisco and leaf N content) suggests that the lower aboveground N requirements affected negatively nodule performance. In this condition, specific nodule activity was reduced due to an effect on nodule metabolism that manifested as a lower amount of nitrogenase reductase. Moreover, the nodule proteomic approach also revealed that nodule functioning was altered simultaneously in various enzyme quantity apart from nitrogenase. At elevated CO2, the tricarboxylic acid cycle was also altered with a red

The expansion of the worlds population requires the development of high production agriculture. For this purpose, it is essential to identify target points conditioning crop responsiveness to predicted [CO2]. The aim of this study was to determine the relevance of ear sink strength in leaf protein and metabolomic profiles and its implications in photosynthetic activity and yield of durum wheat plants exposed to elevated [CO2]. For this purpose, a genotype with high harvest index (HI) (Triticum durum var. Sula) and another with low HI (Triticum durum var. Blanqueta) were exposed to elevated [CO2] (700 mol mol(1) versus 400 mol mol(1) CO2) in CO2 greenhouses. The obtained data highlighted that elevated [CO2] only increased plant growth in the genotype with the largest HI; Sula. Gas exchange analyses revealed that although exposure to 700 mol mol(1) depleted Rubisco content, Sula was capable of increasing the light-saturated rate of CO2 assimilation (A(sat)) whereas, in Blanqueta, the carbohydrate imbalance induced the down-regulation of A(sat). The specific depletion of Rubisco in both genotypes under elevated [CO2], together with the enhancement of other proteins in the Calvin cycle, revealed that there was a redistribution of N from Rubisco towards RuBP regeneration. Moreover, the down-regulation of N, NO3, amino acid, and organic acid content, together with the depletion of proteins involved in amino acid synthesis that was detected in Blanqueta grown at 700 mol mol(1) CO2, revealed that inhibition of N assimilation was involved in the carbohydrate imbalance and consequently with the down-regulation of photosynthesis and growth in these plants.

Elevated CO2 leads to a decrease in potential net photosynthesis in long-term experiments and thus to a reduction in potential growth. This process is known as photosynthetic downregulation. There is no agreement on the definition of which parameters are the most sensitive for detecting CO2 acclimation. In order to investigate the most sensitive photosynthetic and molecular markers of CO2 acclimation, the effects of elevated CO2, and associated elevated temperature were analyzed in alfalfa plants inoculated with different Sinorhizobium meliloti strains. Plants (Medicago sativa L. cv. Aragn) were grown in summer or autumn in temperature gradient greenhouses (TGG). At the end of the experiment, all plants showed acclimation in both seasons, especially under elevated summer temperatures. This was probably due to the lower nitrogen (N) availability caused by decreased N-2-fixation under higher temperatures. Photosynthesis measured at growth CO2 concentration, rubisco in vitro activity and maximum rate of carboxylation were the most sensitive parameters for detecting downregulation. Severe acclimation was also related with decreases in leaf nitrogen content associated with declines in rubisco content (large and small subunits) and activity that resulted in a drop in photosynthesis. Despite the sensitivity of rubisco content as a marker of acclimation, it was not coordinated with gene expression, possibly due to a lag between gene transcription and protein translation.

An understanding of the mechanisms that determine plant response to reduced water availability is essential to improve water-use efficiency (WUE) of stone fruit crops. The physiological, biochemical and molecular drought responses of four Prunus rootstocks (GF 677, Cadaman, ROOTPAC 20 and ROOTPAC(®) R) budded with 'Catherina' peach cultivar were studied. Trees were grown in 15-l containers and subjected to a progressive water stress for 26 days, monitoring soil moisture content by time domain reflectometry. Photosynthetic and gas exchange parameters were determined. Root and leaf soluble sugars and proline content were also measured. At the end of the experiment, stressed plants showed lower net photosynthesis rate, stomatal conductance and transpiration rate, and higher intrinsic leaf WUE (AN/gs). Soluble sugars and proline concentration changes were observed, in both root and leaf tissues, especially in an advanced state of stress. The accumulation of proline in roots and leaves with drought stress was related to the decrease in osmotic potential and increase in WUE, whereas the accumulation of sorbitol in leaves, raffinose in roots and proline in both tissues was related only to the increase in the WUE. Owing to the putative role of raffinose and proline as antioxidants and their low concentration, they could be ameliorating deleterious effects of drought-induced oxidative stress by protecting membranes and enzymes rather than acting as active osmolytes. Higher expression of P5SC gene in roots was also consistent with proline accumulation in the tolerant genotype GF 677. These results indicate that accumulation of sorbitol, raffinose and proline in different tissues and/or the increase in P5SC expression could be used as markers of drought tolerance in peach cultivars grafted on Prunus rootstocks

Elevated CO2 may decrease alfalfa forage quality and in vitro digestibility through a drop in crude protein and an enhancement of fibre content. The aim of the present study was to analyse the effect of elevated CO2, elevated temperature and Sinorhizobium

Photosynthesis in C3 plants is CO2 limited and therefore any increase in Rubisco carboxylation substrate may increase net CO2 fixation, unless plants experience acclimation or other limitations. These aspects are largely unexplored in grapevine. Photosynthesis analysis was used to assess the stomatal, mesophyll, photochemical and biochemical contributions to the decreasing photosynthesis observed in Tempranillo grapevines (Vitis vinifera) from veraison to ripeness, modulated by CO2, temperature and water availability. Photosynthesis and photosystem II photochemistry decreased from veraison to ripeness. The elevated CO2 and temperature increased photosynthesis, but transiently, in both well irrigated (WI) and water-stressed plants. Photosynthetic rates were maxima 1 week after the start of elevated CO2 and temperature treatments, but differences with treatments of ambient conditions disappeared with time. There were not marked changes in leaf water status, leaf chlorophyll or leaf protein that could limit photosynthesis at ripeness. Leaf total soluble sugars remained at ripeness as high as 2 weeks after the start of treatments. On the other hand, and as expected, CO2 diffusional limitations impaired photosynthesis in grapevine plants grown under water scarcity, stomatal and mesophyll conductances to CO2 decreased and in turn low chloroplastic CO2 concentrations limited photosynthetic CO2 fixation. In summary, photochemistry and photosynthesis from veraison to ripeness in Tempranillo grapevine were dominated by a developmental-related decreasing trend that was only transiently influenced by elevated CO2 concentrations.