Grape Ripening Is Regulated by Deficit Irrigation/Elevated Temperatures According to Cluster Position in the Canopy.

The impact of water deficit on berry quality has been extensively investigated during the last decades. Nonetheless, there is a scarcity of knowledge on the performance of varieties exposed to a combination of high temperatures/water stress during the growing season and under vineyard conditions. The objective of this research was to investigate the effects of two irrigation regimes, sustained deficit irrigation (SDI, 30% ETc) and regulated deficit irrigation (RDI, 15% ETc) and of two cluster positions within the canopy (east- and west-exposed sides) on berry ripening in red Aragonez (Tempranillo) grapevines. The study was undertaken for two successive years in a commercial vineyard in South Portugal, monitoring the following parameters: pre-dawn leaf water potential, berry temperature, sugars, polyphenols, abscisic acid (ABA) and related metabolites. Additionally, expression patterns for different transcripts encoding for enzymes responsible for anthocyanin and ABA biosynthesis (VviUFGT, VvNCED1, VvßG1, VviHyd1, VviHyd2) were analyzed. In both years anthocyanin concentration was lower in RDI at the west side (RDIW- the hottest one) from véraison onwards, suggesting that the most severe water stress conditions exacerbated the negative impact of high temperature on anthocyanin. The down-regulation of VviUFGT expression revealed a repression of the anthocyanin synthesis in berries of RDIW, at early stages of berry ripening. At full-maturation, anthocyanin degradation products were detected, being highest at RDIW. This suggests that the negative impact of water stress and high temperature on anthocyanins results from the repression of biosynthesis at the onset of ripening and from degradation at later stages. On the other hand, berries grown under SDI displayed a higher content in phenolics than those under RDI, pointing out for the attenuation of the negative temperature effects under SDI. Irrigation regime and berry position had small effect on free-ABA concentration. However, ABA catabolism/conjugation process and ABA biosynthetic pathway were affected by water and heat stresses. This indicates the role of ABA-GE and catabolites in berry ABA homeostasis under abiotic stresses. Principal component analysis (PCA) showed that the strongest influence in berry ripening is the deficit irrigation regime, while temperature is an important variable determining the improvement or impairment of berry quality by the deficit irrigation regime. In summary, this work shows the interaction between irrigation regime and high temperature on the control of berry ripening.

Resilience of a semi-deciduous shrub, Cistus salvifolius, to severe summer drought and heat stress.

Shrubs often form the understorey in Mediterranean oak woodlands. These shrubs are exposed to recurrent water deficits, but how they will respond to predicted future exacerbation of drought is not yet understood. The ecophysiology of the shrub Cistus salvifolius L. was studied over the summer of 2005, which was during a heat-wave superimposed on the most severe drought in the Iberian Peninsula in the last 140 years. Branch water potential fell drastically during the summer, accompanied by stomatal closure and downregulation of PSII, with a concomitant loss of chlorophyll in the leaves. A parallel increase in the ratio of light-dissipating to light-capturing pigments and the proportion of xanthophyll cycle pigments in the de-epoxidated state, along with alterations in the structure of the light harvesting complex, may have reduced the potential for damage to leaves. Substantial increases in leaf tocopherol content during high radiation may have reduced damage from free radicals. Following autumn rains, leaves of the same shrubs showed physiological recovery, indicating the resilience of this Mediterranean species, for which an extremely dry hydrological year with 45% less rainfall than average, did not prevent healthy leaf functioning in response to renewed soil moisture availability.

Polyols in grape berry: transport and metabolic adjustments as a physiological strategy for water-deficit stress tolerance in grapevine.

Polyols are important metabolites that often function as carbon and energy sources and/or osmoprotective solutes in some plants. In grapevine, and in the grape berry in particular, the molecular aspects of polyol transport and metabolism and their physiological relevance are virtually unknown to date. Here, the biochemical function of a grapevine fruit mesocarp polyol transporter (VvPLT1) was characterized after its heterologous expression in yeast. This H(+)-dependent plasma membrane carrier transports mannitol (K m=5.4mM) and sorbitol (K m=9.5mM) over a broad range of polyols and monosaccharides. Water-deficit stress triggered an increase in the expression of VvPLT1 at the fully mature stage, allowing increased polyol uptake into pulp cells. Plant polyol dehydrogenases are oxireductases that reversibly oxidize polyols into monosaccharides. Mannitol catabolism in grape cells (K m=30.1mM mannitol) and mature berry mesocarps (K m=79mM) was, like sorbitol dehydrogenase activity, strongly inhibited (50-75%) by water-deficit stress. Simultaneously, fructose reduction into polyols via mannitol and sorbitol dehydrogenases was stimulated, contributing to their higher intracellular concentrations in water-deficit stress. Accordingly, the concentrations of mannitol, sorbitol, galactinol, myo-inositol, and dulcitol were significantly higher in berry mesocarps from water-deficit-stressed Tempranillo grapevines. Metabolomic profiling of the berry pulp by GC-TOF-MS also revealed many other changes in its composition induced by water deficit. The impact of polyols on grape berry composition and plant response to water deficit stress, via modifications in polyol transport and metabolism, was analysed by integrating metabolomics with transcriptional analysis and biochemical approaches.

Thermography to explore plant-environment interactions.

Stomatal regulation is a key determinant of plant photosynthesis and water relations, influencing plant survival, adaptation, and growth. Stomata sense the surrounding environment and respond rapidly to abiotic and biotic stresses. Stomatal conductance to water vapour (g s) and/or transpiration (E) are therefore valuable physiological parameters to be monitored in plant and agricultural sciences. However, leaf gas exchange measurements involve contact with leaves and often interfere with leaf functioning. Besides, they are time consuming and are limited by the sampling characteristics (e.g. sample size and/or the high number of samples required). Remote and rapid means to assess g s or E are thus particularly valuable for physiologists, agronomists, and ecologists. Transpiration influences the leaf energy balance and, consequently, leaf temperature (T leaf). As a result, thermal imaging makes it possible to estimate or quantify g s and E. Thermal imaging has been successfully used in a wide range of conditions and with diverse plant species. The technique can be applied at different scales (e.g. from single seedlings/leaves through whole trees or field crops to regions), providing great potential to study plant-environment interactions and specific phenomena such as abnormal stomatal closure, genotypic variation in stress tolerance, and the impact of different management strategies on crop water status. Nevertheless, environmental variability (e.g. in light intensity, temperature, relative humidity, wind speed) affects the accuracy of thermal imaging measurements. This review presents and discusses the advantages of thermal imaging applications to plant science, agriculture, and ecology, as well as its limitations and possible approaches to minimize them, by highlighting examples from previous and ongoing research.

Membrane transport, sensing and signaling in plant adaptation to environmental stress.

Plants are generally well adapted to a wide range of environmental conditions. Even though they have notably prospered in our planet, stressful conditions such as salinity, drought and cold or heat, which are increasingly being observed worldwide in the context of the ongoing climate changes, limit their growth and productivity. Behind the remarkable ability of plants to cope with these stresses and still thrive, sophisticated and efficient mechanisms to re-establish and maintain ion and cellular homeostasis are involved. Among the plant arsenal to maintain homeostasis are efficient stress sensing and signaling mechanisms, plant cell detoxification systems, compatible solute and osmoprotectant accumulation and a vital rearrangement of solute transport and compartmentation. The key role of solute transport systems and signaling proteins in cellular homeostasis is addressed in the present work. The full understanding of the plant cell complex defense mechanisms under stress may allow for the engineering of more tolerant plants or the optimization of cultivation practices to improve yield and productivity, which is crucial at the present time as food resources are progressively scarce.

Initial water deficit effects on Lupinus albus photosynthetic performance, carbon metabolism, and hormonal balance: metabolic reorganization prior to early stress responses.

The early (2-4 d) effects of slowly imposed soil water deficit on Lupinus albus photosynthetic performance, carbon metabolism, and hormonal balance in different organs (leaf blade, stem stele, stem cortex, and root) were evaluated on 23-d-old plants (growth chamber assay). Our work shows that several metabolic adjustments occurred prior to alteration of the plant water status, implying that water deficit is perceived before the change in plant water status. The slow, progressive decline in soil water content started to be visible 3 d after withholding water (3 DAW). The earliest plant changes were associated with organ-specific metabolic responses (particularly in the leaves) and with leaf conductance and only later with plant water status and photosynthetic rate (4 DAW) or photosynthetic capacity (according to the Farquhar model; 6 DAW). Principal component analysis (PCA) of the physiological parameters, the carbohydrate and the hormone levels and their relative values, as well as leaf water-soluble metabolites full scan data (LC-MS/MS), showed separation of the different sampling dates. At 6 DAW classically described stress responses are observed, with plant water status, ABA level, and root hormonal balance contributing to the separation of these samples. Discrimination of earlier stress stages (3 and 4 DAW) is only achieved when the relative levels of indole-3-acetic acid (IAA), cytokinins (Cks), and carbon metabolism (glucose, sucrose, raffinose, and starch levels) are taken into account. Our working hypothesis is that, in addition to single responses (e.g. ABA increase), the combined alterations in hormone and carbohydrate levels play an important role in the stress response mechanism. Response to more advanced stress appears to be associated with a combination of cumulative changes, occurring in several plant organs. The carbohydrate and hormonal balance in the leaf (IAA to bioactive-Cks; soluble sugars to IAA and starch to IAA; relative abundances of the different soluble sugars) flag the initial responses to the slight decrease in soil water availability (10-15% decrease). Further alterations in sucrose to ABA and in raffinose to ABA relative values (in all organs) indicate that soil water availability continues to decrease. Such alterations when associated with changes in the root hormone balance indicate that the stress response is initiated. It is concluded that metabolic balance (e.g. IAA/bioactive Cks, carbohydrates/IAA, sucrose/ABA, raffinose/ABA, ABA/IAA) is relevant in triggering adjustment mechanisms.

The impact of drought on leaf physiology of Quercus suber L. trees: comparison of an extreme drought event with chronic rainfall reduction.

Understanding the responses of cork oak (Quercus suber L.) to actual and predicted summer conditions is essential to determine the future sustainability of cork oak woodlands in Iberia. Thermal imaging may provide a rapid method for monitoring the extent of stress. The ecophysiology of cork trees was studied over three years. Three treatments were applied by means of rainfall capture and irrigation, with plots receiving 120%, 100%, or 80% of natural precipitation. Despite stomatal closure, detected using both thermal imaging and porometry, leaf water potential fell during the summer, most drastically during the third year of accumulative stress. The quantum efficiency (ΦPSII) and the maximum efficiency Fv' /FM' of photosystem II also fell more intensely over the third summer, while non-photochemical quenching (NPQ) increased. The reduced precipitation treatment sporadically further reduced leaf water potential, stomatal conductance (gs), IG (an index of gs derived from thermal imaging), ΦPSII, and Fv' /FM', and increased leaf temperature and NPQ. It is concluded that these are very resilient trees since they were only severely affected in the third year of severe drought (the third year registering 45% less rainfall than average), and removing 20% of rainfall had a limited impact..

Dehydrins in Lupinus albus: pattern of protein accumulation in response to drought.

Dehydrins (DHNs) are proteins that accumulate abundantly in various plant tissues in response to environmental stresses and during seed maturation, possibly assisting cells in tolerating dehydration. White lupins (Lupinus albus L.) are able to withstand periods of severe water deficit (WD) and previous work suggested that the stem plays a central role as a survival structure. To investigate DHNs involvement in this strategy, we studied tissue specific protein accumulation of a RAB16-like DHN in lupin during a progressive WD and early recovery. Differences were found between leaves, stems and roots. In leaves and roots, the accumulation of the RAB16-like DHN was independent of the water status whereas in the stem (cortex and stele), DHNs were only detected under severe plant WD (stele relative water content, RWC, reduction of 6-7% and cortex RWC reduction of 20%). DHN mRNA analysis by RT-PCR, showed the presence of one DHN mRNA regardless of the tissue or the plant water status.

Hydraulic and chemical signalling in the regulation of stomatal conductance and plant water use in field grapevines growing under deficit irrigation.

Effects of irrigation strategies on stomata and plant water use were studied in field-grown grapevines (Vitis vinifera L.). We assessed the importance of root-derived chemical signals vs. hydraulic signalling in stomatal regulation. The experiment included two treatments with the same water added to the soil (50% ETc) applied either to the whole root system (DI) or to half of the roots, alternating irrigation side every 15 days (PRD). Well-watered plants (FI) (100% ETc) and non-irrigated grapevines (NI) were also studied. Partial stomata closure occurred in both PRD and DI plants. [ABA] of xylem sap remained constant during the day and was maintained throughout the season, with higher values in NI plants. Xylem sap pH was not affected by soil water availability. A positive correlation between ψpd and maximum g s was found, indicating that grapevine stomata strongly respond to plant water status. In contrast, ABA did not explain stomatal control at veraison. At mid-ripening g s was significantly correlated with ABA, apparently interacting with the rise in xylem sap pH. Therefore, our data suggest that hydraulic feedback and feed-forward root-to-shoot chemical signalling mechanisms might be involved in the control of stomata in response to decreased soil water availability, hydraulic signals playing the dominant role.

Responses to chilling of two Eucalyptus globulus clones with contrasting drought resistance.

The effect of chilling on growth and plant hydraulic properties in a drought-resistant clone (CN5) and a drought-sensitive clone (ST51) of Eucalyptus globulus Labill. was evaluated. Chilling (10/5°C, day/night) led to a general decrease in growth of both clones and significant reductions in root hydraulic conductivity, rate of photosynthesis and stomatal conductance in comparison to plants grown at control temperature (24/16°C). The drought-resistant CN5 clone maintained higher root growth and lower leaf-to-root-area ratio than the drought-sensitive ST51 clone, in both temperature treatments. Conversely, ST51 exhibited greater carbon allocation to the foliage and higher hydraulic conductance than clone CN5 at both temperatures. Plants of both clones, when acclimated to chilling, maintained a higher hydraulic conductivity than control plants exposed to chilling temperatures without acclimation. Under chilling, the main differences between clones were a higher water status and anthocyanin concentration in CN5 plants, and a stronger inhibition of root growth in ST51 plants. Except for roots, the hypothesis of a lower depression of growth rate in the drought-resistant clone under chilling was not verified. However, higher root growth under low temperatures, as observed in CN5, can be an advantageous trait in Mediterranean-type environments, protecting trees against summer water-stress.

Exploring thermal imaging variables for the detection of stress responses in grapevine under different irrigation regimes.

Temperatures of leaves or canopies can be used as indicators of stomatal closure in response to soil water deficit. In 2 years of field experiments with grapevines (Vitis vinifera L., cvs Castelão and Aragonês), it was found that thermal imaging can distinguish between irrigated and non-irrigated canopies, and even between deficit irrigation treatments. Average canopy temperature was inversely correlated with stomatal conductance measured with a porometer. Variation of the distribution of temperatures within canopies was not found to be a reliable indicator of stress. A large degree of variation between images was found in reference 'wet' and 'dry' leaves used in the first year for the calculation of an index proportional to stomatal conductance. In the second year, fully irrigated (FI) (100% Et(c)) and non-irrigated (NI) canopies were used as alternatives to wet and dry leaves. A crop water stress index utilizing these FI and NI 'references', where stressed canopies have the highest values and non-stressed canopies have the lowest values, was found to be a suitable measure for detecting stress. It is suggested that the average temperatures of areas of canopies containing several leaves may be more useful for distinguishing between irrigation treatments than the temperatures of individual leaves. Average temperatures over several leaves per canopy may be expected to reduce the impact of variation in leaf angles. The results are discussed in relation to the application of thermal imaging to irrigation scheduling and monitoring crop performance.

Impact of deficit irrigation on water use efficiency and carbon isotope composition (delta13C) of field-grown grapevines under Mediterranean climate.

The objective of this study was to evaluate the effect of deficit irrigation on intrinsic water use efficiency (A/g(s)) and carbon isotope composition (delta13C) of two grapevine cultivars (Moscatel and Castelão), growing in a commercial vineyard in SW Portugal. The study was done in two consecutive years (2001 and 2002). The treatments were full irrigation (FI), corresponding to 100% of crop evapotranspiration (ETc), rain-fed (no irrigation, NI), and two types of deficit irrigation (50% ETc): (i) by supplying the water either to one side of the root system or to the other, which is partial rootzone drying (PRD), or (ii) dividing the same amount of water by the two sides of the root system, the normal deficit irrigation (DI). The water supplied to the PRD treatment alternated sides approximately every 15 d. The values of predawn leaf water potential (Psi(pd)) and the cumulative integral of Psi(pd) (S(Psi)) during the season were lower in 2001 than in the 2002 growing season. Whereas differences in Psi(pd) and S(Psi) between PRD and DI were not significantly different in 2001, in 2002 (a dryer year) both cultivars showed lower values of S(Psi) in the PRD treatment as compared with the DI treatment. This suggests that partial rootzone drying may have a positive effect on water use under dryer conditions, either as a result of better stomatal control and/or reduced vigour. The effects of the water treatments on delta13C were more pronounced in whole grape berries and pulp than in leaves. The delta13C of pulp showed the best correlation with intrinsic water use efficiency (A/g(s)) as well as with S(Psi). In spite of the better water status observed in PRD compared with DI in the two cultivars in 2002, no statistical differences between the two treatments were observed in A/g(s) and delta13C. On the other hand, they showed a higher delta13C compared with FI. In conclusion, it is apparent that the response to deficit irrigation varies with the environmental conditions of the particular year, the driest conditions exacerbating the differences among treatments. The highest values of delta13C found in the pulp of NI vines in Castelão compared with Moscatel suggest different sensitivities to water deficits in the two cultivars, as was empirically observed.

Partial rootzone drying: regulation of stomatal aperture and carbon assimilation in field-grown grapevines (Vitis vinifera cv. Moscatel).

The effects of 'partial rootzone drying' (PRD) irrigation compared with other irrigation systems, namely non-irrigated (NI), full irrigation (FI) and deficit irrigation (DI), on stomatal conductance and carbon assimilation were evaluated in field-grown grapevines (Vitis vinifera L. cv. Moscatel). At the end of the growing season, pre-dawn leaf water potential was highest in FI (-0.18 ± 0.01 MPa; mean ± s.e.), intermediate in PRD (-0.30± 0.01 MPa) and DI (-0.36 ± 0.02 MPa), and lowest in NI vines (-0.64 ± 0.03 MPa). Stomatal conductance measured under controlled conditions of light and temperature was reduced in NI (ca 60%) and PRD (ca 30%) vines compared with DI and FI vines. Under ambient conditions, NI vines had lower rates of stomatal conductance (ca26%), net CO2 assimilation (ca 28%) and light-adapted PSII quantum yields (ca 47%) than PRD, DI and FI vines. No significant differences were found among the three irrigated treatments. Both maximum electron transport rate (Jmax; ca 30%) and triose-phosphate utilization rates (TPU; ca 20%) were significantly lower in NI and PRD vines than in DI and FI vines. Carbon isotope composition (δ13C) of grape berries was highest in NI vines (-24.3), followed by PRD (-25.4) and DI (-25.8) and lowest in FI (-26.4) vines, suggesting a long-term increase in the efficiency of leaf gas exchange in NI compared with PRD, DI and FI vines. Sap-flow data and estimates of relative stomatal limitation are in accordance with the observed stomatal closure in PRD vines. In this study, we show that PRD irrigation was able to maintain a vine water status closed to FI, but with double water use efficiency, which was due to a reduction of stomatal conductance with no significant decrease in carbon assimilation.

Partial rootzone drying: effects on growth and fruit quality of field-grown grapevines (Vitis vinifera).

A study to assess the effects of the Partial Rootzone Drying (PRD) irrigation strategy in comparison to other irrigation systems was carried out in southern Portugal in two field-grown grapevines varieties, Moscatel and Castelão. We addressed the question of whether by regulating growth and plant water use, the PRD system would enable an equilibrated vegetative development, leading to a favourable capture of solar radiation for photoassimilate production and, at the same time to provide an optimum environment for fruit maturation. Three irrigation schemes were applied in addition to the non-irrigated (NI) vines: partial root drying (PRD), 50% of crop evapotranspiration (ETc), supplied to only one side of the root system while the other one was allowed to dry, alternating sides every 15 days; deficit irrigated (DI), 50% ETc supplied, half to each side of the root system and full irrigated (FI, 100% ETc). During the whole season FI plants of both varieties exhibited a high leaf predawn water potential (ψpd , ca-0.2MPa) while a progressive decline was observed in NI plants, reaching ψpd values near -0.7 MPa at the end of August. PRD and DI presented intermediate values. PRD vines exhibited a stronger control over vegetative growth as compared with DI and FI plants. This was expressed by lower values of total leaf area at harvest, leaf layer number, canopy wideness and water shoots number, allowing a higher light interception at the cluster zone that induced an improvement in some berry quality characteristics. Watering had no significant effects on sugar accumulation in the berries but led to a favourable increase in the must titratable acidity, mainly in Castelão. Whereas in DI and FI treatments berry skin anthocyanins and phenols content were always lower than in NI, in PRD there was either no reduction or the reduction was much lower than in the other irrigation treatments. Water use efficiency (WUE) was increased by about 80% in PRD and DI when compared with FI, as a result of almost similar yields in the three treatments. Yield gains of irrigated plants in relation to NI were modest, explained by the rainy spring in both years.

Use of infrared thermography for monitoring stomatal closure in the field: application to grapevine.

This paper reviews and discusses strategies for the use of thermal imaging for studies of stomatal conductance in the field and compares techniques for image collection and analysis. Measurements were taken under a range of environmental conditions and on sunlit and shaded canopies to illustrate the variability of temperatures and derived stress indices. A simple procedure is presented for correcting for calibration drift within the images from the low-cost thermal imager used (SnapShot 225, Infrared Solutions, Inc.). The use of wet and dry reference surfaces as thresholds to eliminate the inclusion of non-leaf material in the analysis of canopy temperature is discussed. An index that is proportional to stomatal conductance was compared with stomatal measurements with a porometer. The advantages and disadvantages of a possible new approach to the use of thermal imagery for the detection of stomatal closure in grapevine canopies, based on an analysis of the temperature of shaded leaves, rather than sunlit leaves, are discussed. Evidence is presented that the temperature of reference surfaces exposed within the canopy can be affected by the canopy water status.

Limitations to leaf photosynthesis in field-grown grapevine under drought - metabolic and modelling approaches.

The effects of a slowly-imposed drought stress on gas-exchange, chlorophyll a fluorescence, biochemical and physiological parameters of Vitis vinifera L. leaves (cv. Aragonez, syn. Tempranillo) growing in a commercial vineyard (South Portugal) were evaluated. Relative to well-watered plants (predawn water potential, ΨPD = -0.13 ± 0.01 MPa), drought-stressed plants (ΨPD = -0.97 ± 0.01 MPa) had lower photosynthetic rates (ca 70%), stomatal conductance, and PSII activity (associated with a higher reduction of the quinone A pool and lower efficiency of PSII open centres). Stomatal limitation to photosynthesis was increased in drought-stressed plants relative to well-watered plants by ca 44%. Modelled responses of net photosynthesis to internal CO2 indicated that drought-stressed plants had significant reductions in maximum Rubisco carboxylation activity (ca 32%), ribulose-1,5-bisphosphate regeneration (ca 27%), and triose phosphate (triose-P) utilization rates (ca 37%) relative to well-watered plants. There was good agreement between the effects of drought on modelled biochemical parameters, and in vitro activities of key enzymes of carbon metabolism, namely Rubisco, glyceraldehyde-3-phosphate dehydrogenase, ribulose-5-phosphate kinase and fructose-1,6-bisphosphate phosphatase. Quantum yields measured under both ambient (35 Pa) and saturating CO2 (100 Pa) for drought-stressed plants were decreased relative to well-watered plants, as well as maximum photosynthetic rates measured at light and CO2 saturating conditions (three times ambient CO2 levels). Although stomatal closure was a strong limitation to CO2 assimilation under drought, comparable reductions in electron transport, CO2 carboxylation, and utilization of triose-P capacities were also adaptations of the photosynthetic machinery to dehydration that slowly developed under field conditions. Results presented in this study confirm that modelling photosynthetic responses based on gas-exchange data can be successfully used to predict metabolic limitations to photosynthesis.