José Moutinho-Pereira

Physiological behaviour, oxidative damage and antioxidative protection of olive trees grown under different irrigation regimes

Irrigation effects were investigated on an 8-year-old olive (Olea europaea L., cv. Cobrançosa) commercial orchard located in northeast Portugal. Trees were subjected to a rainfed control (T0) and three treatments (T1, T2, T3) that received a seasonal water amount equivalent to 30%, 60% and 100% of the estimated local evaporative demand by a drip irrigation system. Irrigation increases the photosynthetic activity of olive trees, in association with increases in water status, and reduces the midday and afternoon depression in gas exchange. The closely association between photosynthetic rate (A) and stomatal conductance (gs) revealed that the decline in net photosynthesis over the course of the day was largely a consequence of stomatal limitation. However, the ratio of intercellular to atmospheric CO2 concentration increased markedly from morning to midday in non-irrigated plants, in spite of lower gs, suggesting that non-stomatal limitations of photosynthesis also occur when environmental conditions become more stressful. The occurrence of perturbations at chloroplastic level in rainfed plants was demonstrated by a lower maximum photochemical efficiency of photosystem II during the afternoon. Chlorophyll fluorescence measurements also revealed the occurrence of a dynamic photoinhibition in irrigated trees, mainly in T2 and T3, which seemed to be effective in protecting the photosynthetic apparatus from photodamage. Irrigation enhances antioxidant protection and decreases the oxidative damage at leaf level. Leaves grown under rainfed conditions revealed symptoms of oxidative stress, like the reduction (14%) in chlorophyll concentration and the increased levels (57%) of lipid peroxidation. We also found that the scavenging function of superoxide dismutase was impaired in rainfed plants. In contrast, the low thiobarbituric acid reactive substances concentration in T3 indicates that irrigation enhances the repairing mechanisms and decreases the oxidative damage by lipid peroxidation. Accordingly, leaves in T3 treatment had high levels of –SH compounds and the highest antioxidant potential. Meanwhile, the finding that guaiacol peroxidase activity increased in rainfed plants, associated with the appearance of oxidative damage, suggests that this enzyme has no major antioxidative function in olive.

Chromium (VI) induces toxicity at different photosynthetic levels in pea

In order to comprehensively characterize the effects of Cr (VI) on the photosynthetic performance of Pisum sativum, plants irrigated with Cr solutions (ranging from 20 to 2000 mg l(-1)) were evaluated using the following classical endpoints: gas exchange parameters, chlorophyll a (Chl a) fluorescence, leaf pigments, Rubisco activity, soluble sugars and starch content. Flow cytometry (FCM) was applied in an innovative approach to evaluate the morphological and fluorescence emission status of chloroplasts from plants exposed to Cr (VI). The parameters related to gas exchange, net CO(2) assimilation rate (A) and Rubisco activity were severally affected by Cr exposure, in some cases even at the lowest dosage used. While all biomarkers used to measure Chl a fluorescence indicated a decrease in fluorescence at the maximum dosage, pigment contents significantly increased in response to Cr (VI). The morphology of chloroplasts also was altered by Cr (VI) exposure, as a volume decrease was observed. Soluble sugars and starch showed an overall tendency to increase in Cr (VI) exposed plants, but sucrose and glucose decreased highly when exposed to 2000 mg l(-1). In conclusion, our results indicate that Cr (VI) affects photosynthesis at several levels, but the most Cr (VI)-sensitive endpoints were chloroplast morphology and biochemical processes; only at higher dosages the photochemical efficiency is compromised.

Future scenarios for viticultural zoning in Europe: ensemble projections and uncertainties

Optimum climate conditions for grapevine growth are limited geographically and may be further challenged by a changing climate. Due to the importance of the winemaking sector in Europe, the assessment of future scenarios for European viticulture is of foremost relevance. A 16-member ensemble of model transient experiments (generated by the ENSEMBLES project) under a greenhouse gas emission scenario and for two future periods (2011–2040 and 2041–2070) is used in assessing climate change projections for six viticultural zoning indices. After model data calibration/validation using an observational gridded daily dataset, changes in their ensemble means and inter-annual variability are discussed, also taking into account the model uncertainties. Over southern Europe, the projected warming combined with severe dryness in the growing season is expected to have detrimental impacts on the grapevine development and wine quality, requiring measures to cope with heat and water stress. Furthermore, the expected warming and the maintenance of moderately wet growing seasons over most of the central European winemaking regions may require a selection of new grapevine varieties, as well as an enhancement of pest/disease control. New winemaking regions may arise over northern Europe and high altitude areas, when considering climatic factors only. An enhanced inter-annual variability is also projected over most of Europe. All these future changes pose new challenges for the European winemaking sector.

Leaf Gas Exchange and Water Relations of Grapevines Grown in Three Different Conditions

Diurnal and seasonal changes in the leaf water potential (Ψ), stomatal conductance (gs), net CO2 assimilation rate (PN), transpiration rate (E), internal CO2 concentration (Ci), and intrinsic water use efficiency (PN/gs) were studied in grapevines (Vitis vinifera L. cv. Touriga Nacional) growing in low, moderate, and severe summer stress at Vila Real (VR), Pinhão (PI), and Almendra (AL) experimental sites, respectively. In VR and PI site the limitation to photosynthesis was caused more by stomatal limitations, while in AL mesophyll limitations were also responsible for the summer decline in PN.

Effects of elevated CO2 on grapevine (Vitis vinifera L.): volatile composition, phenolic content, and in vitro antioxidant activity of red wine.

The impact of elevated carbon dioxide concentration ([CO2]) on the quality of berries, must, and red wine (with special reference to volatile composition, phenolic content, and antioxidant activity) made from Touriga Franca, a native grape variety of Vitis vinifera L. for Port and Douro wine manufacturing grown in the Demarcated Region of Douro, was investigated during 2005 and 2006. Grapevines were grown either in open-top chambers (OTC) with ambient (365 +/- 10 ppm) or elevated (500 +/-16 ppm) [CO2] or in an outside plot. In general, the increase of [CO2] did not affect berry characteristics, especially the total anthocyan and tannin concentrations. However, the total anthocyan and polyphenol concentrations of the red wine were inhibited under elevated [CO2]. The antioxidant capacity of the wines was determined by DPPH, ABTS, and TBARS assays and, despite the low concentrations of phenolics, the elevated [CO2] did not significantly change the total antioxidant capacity of the red wines. Thirty-five volatile compounds belonging to seven chemical groups were identified: C6 alcohols, higher alcohols, esters, terpenols, carbonyl compounds, acids, volatile phenols, and C13 norisoprenoids. Generally, the same volatile compounds were present in all of the wines, but the relative levels varied among the treatments. The effect of elevated [CO2] was significant because it was detected as an increase in ethyl 2-methylbutyrate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, butyric acid, and isovaleric acid concentrations and a decrease in ethyl acetate concentration when compared to wines produced in ambient [CO2] in 2005. In elevated [CO2], wines from 2006 had lower methionol, 1-octanol, and 4-ethylguaiacol and higher ethyl lactate and linalool concentrations. The increase in [CO2] did not significantly affect C6 alcohols, citronellol, carbonyl compounds, and beta-damascenone concentrations. This study showed that the predicted rise in [CO2] did not produce negative effects on the quality of grapes and red wine. Although some of the compounds were slightly affected, the red wine quality remained almost unaffected.

Physiological responses of different olive genotypes to drought conditions

Gas exchange rates, chlorophyll fluorescence, pressure–volume relationships, photosynthetic pigments, total soluble sugars, starch, soluble proteins and proline concentrations were investigated in five Olea europaea L. cultivars with different geographical origins (Arbequina, Blanqueta, Cobrançosa, Manzanilla and Negrinha) grown under Mediterranean field conditions. We found considerable genotypic differences among the cultivars. Comparing the diurnal gas exchange rates, we observed that Cobrançosa, Manzanilla and Negrinha had high photosynthetic rate than Arbequina and Blanqueta. The first group reveals to be better acclimated to drought conditions, and appears to employ a prodigal water-use strategy, whereas Blanqueta and Arbequina, with high water-use efficiency, appear to employ a conservative water-use strategy. The degree of midday depression in photosynthesis was genotype dependent, with a maximum in Arbequina and a minimum in Negrinha. The reductions in the photosynthetic rate were dependent from both stomatal and non-stomatal limitations. Elastic adjustment plays an important role as drought tolerance mechanism. The group of cultivars that employ a prodigal water-use strategy revealed high tissue elasticity, whereas Arbequina and Blanqueta revealed high tissue rigidity. We also identified the existence of drought tolerance mechanisms associated with soluble proteins accumulation in the foliage. The high levels of soluble proteins in Arbequina may represent an increased activity of oxidative stress defence enzymes and may also represent a reserve for post stress recovery. In all cultivars, especially in Manzanilla, free proline was accumulated in the foliage. The discussed aspects of drought stress metabolism may have an adaptative meaning, supporting the hypothesis that olive cultivars native to dry regions, such as Cobrançosa, Manzanilla and Negrinha, have more capability to acclimate to drought conditions than cultivars originated in regions with a more temperate climate, like Arbequina and Blanqueta.

Gas exchange and water relations of three Vitis vinifera L. cultivars growing under Mediterranean climate

Optical characteristics, contents of photosynthetic pigments, total soluble sugars, and starch, rates of gas exchange, chlorophyll (Chl) a fluorescence, and leaf water relations were analysed in three Vitis vinifera L. cultivars, Tinto Cão (TC), Touriga Nacional (TN), and Tinta Roriz (TR), grown in Mediterranean climate. Chl content was significantly lower in TC than in TN and TR leaves, while the Chl a/b ratio was higher. TR had the lowest net photosynthetic rate, stomatal conductance, and contents of soluble sugars and starch than TN and TC. In spite of low Chl content, TC showed the lowest photon absorbance and the highest photochemical efficiency of photosystem 2. TC had the lowest predawn and midday leaf water potential. The capability for osmotic adjustment was similar among cultivars and the calculated modulus of elasticity was higher in TC leaves. The typical lighter green leaves of TC seemed to be an adaptive strategy to high irradiance and air temperature associated to water stress.

Integrated Analysis of Climate, Soil, Topography and Vegetative Growth in Iberian Viticultural Regions

The Iberian viticultural regions are convened according to the Denomination of Origin (DO) and present different climates, soils, topography and management practices. All these elements influence the vegetative growth of different varieties throughout the peninsula, and are tied to grape quality and wine type. In the current study, an integrated analysis of climate, soil, topography and vegetative growth was performed for the Iberian DO regions, using state-of-the-art datasets. For climatic assessment, a categorized index, accounting for phenological/thermal development, water availability and grape ripening conditions was computed. Soil textural classes were established to distinguish soil types. Elevation and aspect (orientation) were also taken into account, as the leading topographic elements. A spectral vegetation index was used to assess grapevine vegetative growth and an integrated analysis of all variables was performed. The results showed that the integrated climate-soil-topography influence on vine performance is evident. Most Iberian vineyards are grown in temperate dry climates with loamy soils, presenting low vegetative growth. Vineyards in temperate humid conditions tend to show higher vegetative growth. Conversely, in cooler/warmer climates, lower vigour vineyards prevail and other factors, such as soil type and precipitation acquire more important roles in driving vigour. Vines in prevailing loamy soils are grown over a wide climatic diversity, suggesting that precipitation is the primary factor influencing vigour. The present assessment of terroir characteristics allows direct comparison among wine regions and may have great value to viticulturists, particularly under a changing climate.

Photosynthetic parameters of Ulmus minor plantlets affected by irradiance during acclimatization

In order to set up large-scale acclimatization protocols of micropropagated plants, an in-depth knowledge of their physiological responses during in vitro to ex vitro transfer is required. This work describes the photosynthetic performance of Ulmus minor micropropagated plants during acclimatization at high irradiance (HI; 200 ± 20 μmol m−2 s−1 or low irradiance (LI; 100 ± 20 μmol m−2 s−1). During this experiment, leaf pigment content, chlorophyll a fluorescence, gas exchange, stomata morphology, the activity of the Calvin cycle enzymes and saccharides were measured in persistent and new leaves. The results indicated that HI induces a higher photosynthetic performance compared to LI. Therefore, plants acclimatized under HI are likely to survive better after field transfer.

Statistical modelling of grapevine phenology in Portuguese wine regions: observed trends and climate change projections

Phenological models are considered key tools for the short-term planning of viticultural activities and long-term impact assessment of climate change. In the present study, statistical phenological models were developed for budburst (BUD), flowering (FLO) and veraison (VER) of 16 grapevine varieties (autochthonous and international) from the Portuguese wine-making regions of Douro, Lisbon and Vinhos Verdes. For model calibration, monthly averages of daily minimum (Tmin), maximum (Tmax) and mean (Tmean) temperatures were selected as potential regressors by a stepwise methodology. Significant predictors included Tmin in January–February–March for BUD, Tmax in March–April for FLO, and Tmin, Tmax and Tmean in March–July for VER. Developed models showed a high degree of accuracy after validation, representing 0·71 of total variance for BUD, 0·83 for FLO and 0·78 for VER. Model errors were in most cases