Touriga Nacional Research Articles

Effect of light intensity and CO 2 concentration on growth and the acquisition of in vivo characteristics during acclimatization of grapevine regenerated in vitro

This study reports the effects of light intensity and CO 2 concentration during the acclimatization of in vitro plantlets of grapevine (V itis vinifera L. cv. Touriga Nacional) on growth, chlorophyll and soluble carbohydrate contents, stomatal index and regulation of water loss. After in vitro phases at 45 µmol m -2 s -1 PFD, plantlets were transferred directly from in vitro to ex vitro acclimatization at different irradiation intensities (150 and 300 µmol m -2 s -1 ) and CO 2 concentrations (350 and 700 µl l -1 ). Growth, especially total biomass, was closer associated with light than with CO 2 . Ex vitro leaves, expanded at high light and high CO 2 , developed fully autotrophic characteristics, mainly with regard to specific leaf area, chlorophyll a/b ratio and down-regulation of sucrose accumulation. In all treatments stomatal indices of leaves were lower than those of in vitro leaves although regulation of water loss was reduced at high CO 2 .

Relating Water Deficiency to Berry Texture, Skin Cell Wall Composition, and Expression of Remodeling Genes in Two Vitis vinifera L. Varieties.

The cell wall (CW) is a dynamic structure that responds to stress. Water shortage (WS) impacts grapevine berry composition and its sensorial quality. In the present work, berry texture, skin CW composition, and expression of remodeling genes were investigated in two V. vinifera varieties, Touriga Nacional (TN) and Trincadeira (TR), under two water regimes, Full Irrigation (FI) and No Irrigation (NI). The global results allowed an evident separation between both varieties and the water treatments. WS resulted in increased anthocyanin contents in both varieties, reduced amounts in cellulose and lignin at maturation, but an increase in arabinose-containing polysaccharides more tightly bound to the CW in TR. In response to WS, the majority of the CW related genes were down-regulated in a variety dependent pattern. The results support the assumption that WS affects grape berries by stiffening the CW through alteration in pectin structure, supporting its involvement in responses to environmental conditions.

Molecular and oenological characterization of Touriga Nacional non-Saccharomyces yeasts.

The aim of this study was to evaluate non-Saccharomyces yeasts isolated from spontaneous fermenting musts of Touriga Nacional (TN), one of the most important Portuguese red grape variety, to improve and diversify TN wines. Two hundred and seventy nine isolates were assigned to 11 yeast species by conventional molecular and growth tests. Starmerella bacillaris was the most frequently detected yeast species, followed by Hanseniaspora guilliermondii and Hanseniaspora uvarum. Twenty-three isolates from 10 species were selected for oenological study, namely fermentation performance, physicochemical and quantitative descriptive sensory analysis of the wines produced. A significant species effect was observed for most of the variables evaluated, some species generating wines with quite interesting aromas. Candida diversa and S.bacillaris isolates produced wines with higher overall quality, higher balance and more intense and diverse aroma. Furthermore, S.bacillaris isolates generated wines with enhanced TN typical aroma, such as bergamot, violet and rock-rose, and were thus regarded as the most promising for improving TN wines. This study revealed the diversity of wine aroma profiles generated by non-Saccharomyces yeast isolates. This knowledge is particularly important given the growing trend from industry to use non-Saccharomyces yeasts as a tool for improving and diversifying the sensory characteristics of wine.

Heat stress in grapevine: the pros and cons of acclimation.

Heat stress is a major limiting factor of grapevine production and quality. Acclimation and recovery are essential to ensure plant survival, and the recovery mechanisms can be independent of the heat response mechanisms. An experimental set up with and without acclimation to heat followed by recovery [stepwise acclimation and recovery (SAR) and stepwise recovery (SR), respectively] was applied to two grapevine varieties, Touriga Nacional (TN), and Trincadeira (TR), with different tolerance to abiotic stress. Major differences were found between leaves of SAR and SR, especially after recovery; in SAR, almost all parameters returned to basal levels while in SR they remained altered. Acclimation led to a swifter and short-term antioxidative response, affecting the plant to a lesser extent than SR. Significant differences were found among varieties: upon stress, TN significantly increased ascorbate and glutathione reduction levels, boosting the cell's redox-buffering capacity, while TR needed to synthesize both metabolites, its response being insufficient to keep the redox state at working levels. TR was affected by stress for a longer period and the up-regulation pattern of antioxidative stress genes was more obvious. In TN, heat shock proteins were significantly induced, but the canonical heat-stress gene signature was not evident probably because no shutdown of the housekeeping metabolism was needed.

Volatile components of vine leaves from two Portuguese grape varieties (Vitis vinifera L.), Touriga Nacional and Tinta Roriz, analysed by solid-phase microextraction

The purpose of this work was to study the volatile composition of vine leaves and vine leaf infusion prepared from vine leaves collected at 30 and 60 days after grape harvest of two Vitis vinifera L. species. Eighteen volatile compounds were identified by gas chromatography–mass spectrometry in vine leaves and in vine leaf infusions. It was observed that the volatile compounds present in vine leaves are dependent on the time of harvest, with benzaldehyde being the major volatile present in vine leaves collected at 30 days after harvesting. There are significant differences in the volatile composition of the leaves from the two grape cultivars, especially in the sample collected at 60 days after grape harvest. This is not reflected in the volatile composition of the vine leaf infusion made from this two cultivars, the more important being the harvesting date for the volatile profile of vine leaf infusion than the vine leaves grape cultivar.

Antioxidant capacity, radical scavenger activity, lipid oxidation protection analysis and antimicrobial activity of red grape extracts from different varieties cultivated in Portugal

The aim of this study was to investigate the antioxidant capacity, radical scavenger activity, lipid oxidation protection and antimicrobial activity of grape extracts from 12 different red grape varieties cultivated in Portugal. The mean values of total phenolic content quantified in grape extracts varied from 833.7 to 2005.6 mg/L gallic acid. Antioxidant capacity results showed different values for each grape variety ranging from 3.96 to 32.96 mm/L Fe(II). The scavenger activity values ranged from 15.99% to 54.82% for the superoxide radical and from 11.79% to 29.67% for the hydroxyl radical. The grape extracts with the highest antioxidant capacity had a positive effect on the lipid oxidation protection and induced low peroxide values in butter samples. Finally, concerning antimicrobial activity, grape extracts from Touriga Nacional and Tinta Roriz grape varieties had significant antimicrobial activity, especially notable for total mesophilic aerobics.

Assessment of (poly)phenols in grape (Vitis vinifera L.) stems by using food/pharma industry compatible solvents and Response Surface Methodology

A Box–Behnken design of Response Surface Methodology (RSM) was conducted to analyse the effect of time (10–30min), temperature (25–95°C), and solvents concentration (5–90%) on the extraction of total phenolics, flavonoids, ortho-diphenols, and anthocyanins as well as to assess the ABTS+ scavenging capacity, which were considered as response variables. Values coefficients of determination (R2), ranging from 0.903 to 0.996, fitted for describing efficient extraction of bioactive (poly)phenols and antioxidant activity. The recorded data allowed to establish the optimal extraction conditions at 23.0min, 95.0°C, and 57.9% of food-quality ethanol/water for Vitis vinifera L. var. ‘Viosinho’ (white variety), and 23.4min, 84.2°C, and 63.8% for var. ‘Touriga Nacional’ (red variety). The achievement of optimal extraction conditions of phenolics from grape stems using solvents compatible with further uses in food/pharma industries demonstrated that RSM constitutes a powerful tool for deriving optimal conditions for extraction of antioxidant phenolic compounds from grape stems.

Transcriptomic changes following the compatible interaction Vitis vinifera–Erysiphe necator. Paving the way towards an enantioselective role in plant defence modulation

The compatible interaction between Erysiphe necator and Vitis vinifera induces significant alterations in the host transcriptome, affecting essentially those genes involved in signalling and secondary metabolite biosynthetic pathways. The precise transcriptomic changes vary from the early events to later stages of infection. In the present work, suppressive subtraction hybridization (SSH) was used to identify several differentially expressed transcripts in symptomatic and asymptomatic leaves from powdery mildew infected grapevines following a long term interaction. The detected transcripts show little or no correlation with similar expression studies concerning the early stages of infection which suggests distinct host responses occur before and after the infection is established. The transcription level of thirteen genes was assessed through qRT-PCR using appropriately selected and validated normalization genes. With one exception, all these genes underwent moderate levels of differential transcription, with log2-fold change values ranging from-2.65 to 4.36. The exception, a dirigent-like (DIR) protein, was upregulated over 180 fold in symptomatic leaves, suggesting an important role for stereochemical selectivity in the compatible interaction E.necator-V.vinifera. DIR copy number was determined in the genome of three grapevine cultivars exhibiting high (Carignan), moderate (Fernão Pires) and low (Touriga Nacional) sensitivity to E.necator. It was found to be a two-copy gene in all cultivars analyzed. Further analysis involving DIR metabolic neighbourhood transcripts was performed. The possible physiological significance of the detected DIR upregulation is discussed.

Search for Transcriptional and Metabolic Markers of Grape Pre-Ripening and Ripening and Insights into Specific Aroma Development in Three Portuguese Cultivars

BackgroundGrapes (Vitis species) are economically the most important fruit crop worldwide. However, the complexity of molecular and biochemical events that lead to ripening of berries as well as how aroma is developed are not fully understood.Methodology/Principal FindingsIn an attempt to identify the common mechanisms associated with the onset of ripening independently of the cultivar, grapes of Portuguese elite cultivars, Trincadeira, Aragonês, and Touriga Nacional, were studied. The mRNA expression profiles corresponding to veraison (EL35) and mature berries (EL36) were compared. Across the three varieties, 9,8% (2255) probesets corresponding to 1915 unigenes were robustly differentially expressed at EL 36 compared to EL 35. Eleven functional categories were represented in this differential gene set. Information on gene expression related to primary and secondary metabolism was verified by RT-qPCR analysis of selected candidate genes at four developmental stages (EL32, EL35, EL36 and EL 38). Gene expression data were integrated with metabolic profiling data from GC-EI-TOF/MS and headspace GC-EI-MS platforms.Conclusions/SignificancePutative molecular and metabolic markers of grape pre-ripening and ripening related to primary and secondary metabolism were established and revealed a substantial developmental reprogramming of cellular metabolism. Altogether the results provide valuable new information on the main metabolic events leading to grape ripening. Furthermore, we provide first hints about how the development of a cultivar specific aroma is controlled at transcriptional level.

Study of polyamines during grape ripening indicate an important role of polyamine catabolism

Grapevine (Vitis species) is the most economically important fruit crop worldwide. Ripening of non-climacteric fruits such as grapes has been the subject of intense research. Despite this interest, little is known on the role played by polyamines in the onset of ripening of non-climacteric fruits. These growth regulators have been involved in plant development and stress responses. Molecular and biochemical studies were developed in three important Portuguese cultivars (Trincadeira, Touriga Nacional and Aragonês) during the year 2008 and in Trincadeira during 2007 in order to gather insights on the role of polyamines in grape ripening. Microarray and real-time qPCR studies revealed up-regulation of a gene coding for arginine decarboxylase (ADC) during grape ripening in all the varieties. This increase was not accompanied by an increase in free and conjugated polyamines that presented a strong decrease. Putrescine and Spermidine levels were higher at earlier stages of development, while Spermine level remained constant. Berries of Trincadeira cultivar presented the highest content in total free and conjugated polyamines at earlier stages of fruit development in particular in the year 2007. The decrease in polyamines content during grape ripening was accompanied by up-regulation of genes coding for diamine oxidase (CuAO) and polyamine oxidase (PAO), together with a significant increase in their enzymatic activity and in the hydrogen peroxide content. These results provide, for the first time, strong evidence of a role of polyamine catabolism in grape ripening possibly through interaction with other growth regulators.

Contribution for a better understanding of grapevine fungal trunk diseases in the Portuguese Dão wine region

Summary. Esca and Petri disease, two of the most important fungal trunk diseases of grapevine, are responsible for significant losses by causing premature decline and dieback in vineyards worldwide. The Portuguese Dao wine region is no exception. Local winegrowers’ knowledge on Grapevine Trunk Diseases (GTD) in general and of esca and Petri disease in particular is incomplete. The real scope of those problems has been based largely on individual perceptions rather than on a methodical evaluation of the situation. In order to get a full picture of the diseases impact, a leaflet with color pictures was produced and issued to winegrowers, accompanied by a simple questionnaire. The results of this survey represent a first indication of the extent of grapevine trunk diseases in the Dao wine region, specifically its economic impact and relevance to the local wine industry. In conjunction with the survey, several samples of wood collected from esca and Petri disease symptomatic vines, identified during the survey throughout the entire region, were processed and a collection of isolates of Phaeomoniella chlamydospora obtained. To determine the intra-specific variability among these isolates, morphological, cultural and molecular characteristics were evaluated. A protocol to study the pathogenicity with P. chlamydospora was conducted, consisting on the inoculation of cv. Touriga Nacional’s spurs with a previously studied P. chlamydospora isolate.

Polynomial relation between mean shoot area and mean shoot weight for vigour calculation on cv. Touriga Nacional

<p style="text-align: justify;"><strong>Aim</strong>: To develop a new methodology for determining vine vigour based on a polynomial relation between mean shoot area and mean shoot weight.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Thirty grapevines (cv. Touriga Nacional) characterized by low, medium and high vigour were selected for a threeyear study. Vine vigour was determined each year by using the traditional “mean shoot weight” methodology. At the same time, the mean shoot area of each vine was determined by using digital image processing, where RGB images (Red Green Blue) were converted to HSV images (Hue Saturation Value) and then to binary images based on the saturation values with a threshold set to 0.15. Finally, we established a polynomial relation between mean shoot area and mean shoot weight, given by the equation <em>f(x)</em> = 0.0047<em>x</em><sup>2</sup> + 0.2254<em>x</em> + 0.0474, where <em>f(x)</em> corresponds to the mean shoot weight (vine vigour estimation) and <em>x</em> corresponds to the mean shoot area.</p><p style="text-align: justify;"><strong>Conclusions</strong>: The proposed method for calculating vine vigour based on a digital image of a vine and a polynomial relation between mean shoot area and mean shoot weight provides a highly reliable estimate of the actual vigour. Correlation for the 30 grapevines with three-year average of 99.8%.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: With this new methodology, it is unnecessary to determine the pruning weight and the number of shoots per plant in the vineyard. Its implementation is simple, based on a digital photography of the vine under study, and independent of the plot pruning date.</p>

Chloroplast Genome Diversity in Portuguese Grapevine (Vitis vinifera L.) Cultivars

Grapevine chloroplast (cp) DNA diversity was analysed for the first time through amplification and digestion of fragments of the large single copy (LSC) region by polymerase chain reaction-restriction fragment length polymorphism methodology and also by amplification of three microsatellite loci, previously described as polymorphic in grapevine. Thirty-eight grapevine cultivars collected mainly in the North of Portugal, including some neglected cultivars, four international cultivars (Chasselas, Muscat of Alexandria, Muscat of Hamburg and Pinot) and Vitis riparia and Vitis rupestris, were used in this study with the main goal of finding out their cpDNA diversity and compare the obtained results with previously published data on cultivars from other regions to ascertain their possible origin. Two different alleles were found in each of the three cpSSR loci. Allele variants of the three loci combined in a total of three different haplotypes (A, B and D). The most frequent haplotype, A, was previously reported as the most frequent in Iberian Peninsula and Occidental Europe. Haplotype B was unique to Rabigato, Muscat of Alexandria, V. riparia and V. rupestris. This haplotype was previously proposed to be an ancestral haplotype. Twenty-seven fragments of the LSC region of Vitis vinifera cpDNA were amplified and then digested with HinfI and TaqI restriction enzymes. Polymorphisms were found in the trnT-psbC (TC) and orf184-petA (OA) fragments. In the TC fragment, the polymorphism corresponds to a point mutation in a restriction site of TaqI and is only present in all cultivars with cpSSR haplotype D. In the OA fragment, a short deletion exclusive to the Rabigato cultivar was found. In this case, one sequence tagged site-based marker was developed and will be very useful in future phylogenetic and fingerprinting studies in a broader number of cultivars and in wild grapevine populations. Inference about the progenitors of the Touriga Franca cultivar is done. The present work supports and completes its origin as a descendent of the female and male parents, Marufo and Touriga Nacional.

Towards comprehensive utilization of winemaking residues: Characterization of grape skins from red grape pomaces of variety Touriga Nacional

The chemical composition and the structural features of macromolecular components from red grape skins (variety Touriga Nacional) have been evaluated. The results of chemical analyses revealed the following contents of the main constituents: cellulose (20.8%), hemicelluloses (12.5%), proteins (18.8%), tannins (13.8%), extractives soluble in dichloromethane (5.0%) and ash (7.8%). Cellulose of grape skins is typical cellulose I polymorph with a degree of crystallinity of 66.1% as assessed by X-ray scattering. The hemicelluloses were analyzed by neutral sugars analysis, methylation analysis and 1D/2D 1H NMR. The water-soluble compounds (26.4%) are composed mainly of monomeric sugars (glucose and fructose) and a complex mixture of hemicelluloses, the most abundant being pectin and acetylated glucomannan. The major part of structural polysaccharides (cellulose, xylan, xyloglucan, etc.) is embedded in cuticular layer and is poorly accessible to the acid hydrolysis. The cuticular substances contributed to ca. 8.5% of grape skins and their chemical structure has been evaluated by methanolysis. The possibility of comprehensive utilization of grape skins has been discussed.

Influence of soil water content and atmospheric conditions on leaf water potential in cv. “Touriga Nacional” deep-rooted vineyards

In this study, the influence of soil and atmosphere conditions on noon and basal leaf water potential of vines “Touriga Nacional” in the Dão region submitted to different irrigation treatments is analysed. Both indicators showed to be dependent on environmental conditions at the time of measurement. Leaf water potential at noon of fully watered plants was linearly related with atmospheric conditions, with values registered when vapour pressure deficit (VPD) was higher than approximately 3 kPa being no different from the values registered in stressed plants. Therefore, this indicator cannot be reliably used to distinguish different plant water stress levels when atmospheric conditions induce high evaporative demands. The basal leaf water potential (ψb) was also influenced by VPD at the time of measurement for all soil water conditions. In well irrigated plants, it was even possible to establish a baseline that can therefore be used to identify nonwater stressed conditions (ψb (MPa) = −0.062–0.0972 VPD (kPa), r 2 = 0.78). A good correlation was found between soil humidity and ψb. However, more than the average value of the whole thickness of soil monitored, the ψb values were dependent on the distribution of soil humidity, with the plants responding to the presence of wet layers.

Quantification of Stilbenes in Vitis Genotypes with Different Levels of Resistance to Plasmopara viticola Infection

Stilbenic phytoalexins have been associated with disease resistance. In Vitis spp., stilbene synthesis can be induced by UV irradiation, treatment with a variety of substance extracts, and inoculation with Botrytis cinerea or Plasmopara viticola . The aim of the present work was to examine the relationship between stilbene production and the level of resistance of different Vitis genotypes to P. viticola . The ability of different grapevine genotypes ( Vitis vinifera L. cvs. Tempranillo, Touriga Nacional, Pinot noir, and Cabernet Sauvignon and Vitis riparia cv. Gloire de Montpellier) to resist Plasmopara viticola infection was assessed via their potential to accumulate toxic stilbenic phytoalexins. Leaf discs taken from plants belonging to these genotypes were inoculated with P. viticola sporangia and disease severity was determined five days later. Stilbene production was quantified in similar leaf material at 6, 24, 48, and 72 hr postinoculation. After P. viticola infection, the resistant genotype V. riparia showed high production of the phytotoxic stilbenes ɛ- and δ-viniferin, which limited the development of the pathogen and prevented it from producing spores. Indeed, this genotype was associated with the lowest sporulation values. No relationship was observed, however, between resistance and these compounds in the V. vinifera genotypes. Further study is required to define the role of stilbenic phytoalexins in resistance to P. viticola.

Multivariate statistical approaches for wine classification based on low molecular weight phenolic compounds

Background and Aims: Phenolic compounds influence colour, flavour and astringency of wines. Technology and grape variety are the main factor affecting the phenolic content of wines. Different multivariate statistical approaches were used to investigate the relationships between the profile of phenolic compounds and grape variety and also the impact of malolactic fermentation (MLF). Methods and Results: A reversed phase liquid chromatography/diode array detection method was used for the analysis of major non-flavonoid phenolic compounds in wines from Trincadeira, Aragonez, Cabernet Sauvignon, Alfrocheiro, Casteão and Touriga Nacional varieties before and after MLF. The impact of MLF and grape variety on phenolic profile was evaluated by principal component analysis (PCA), variation partitioning analysis (VPA) and artificial neural network (ANN). PCA explained 86.5% of the total variance among samples. ANN showed a significant clustering of samples according to grape variety and confirmed that MLF has a minor effect on wine phenolic profile. VPA enabled more information to be extracted from the data by identifying explanatory variables responsible for variability among samples. Conclusions: Compared with PCA and ANN, VPA provides more information concerning the variability on the sample system. Also, grape varieties have a more effective impact on wine low molecular weight phenolic compounds than MLF. Significance of the Study: Each one of the three multivariate statistical approaches showed ways of analysing large chemistry experimental datasets. VPA is a step forward in data analysis, providing more solid and complete assessment of sample system variability, not possible by PCA and ANN.

NMR spectroscopy and chemometrics as a tool for anti-TNFα activity screening in crude extracts of grapes and other berries

The identification of active ingredients in crude plant extracts offers great advantages. In this study, nuclear magnetic resonance and chemometrics were used for the screening of in vitro anti-TNFα activity in different berry types. Solid phase extraction was applied and the resulting water, methanol–water (1:1), and methanol fractions were tested for the activity. The methanol–water fraction contained most of the phenolics and showed significantly higher activity than the other two fractions. In the second phase of this study, grapes from ‘Trincadeira’, ‘Touriga Nacional’, and ‘Aragonês’, at four developmental stages were metabolically classified and tested for the TNFα inhibition. The initial stages of grape development, green and veraison, were found more active against TNFα production as compared to the later ripe and harvest stages. Among the cultivars, ‘Touriga Nacional’ was found to be the most potent inhibitor. Different multivariate data analyses algorithms based on projections to latent structures were applied to correlate the NMR and TNFα inhibition data. The variable importance in the projections plot showed that phenolics like quercetin, myricetin, (+)-catechin, (−)-epicatechin, caftarate, and coutarate, were positively correlated with high activity. This work demonstrates the great potential of NMR spectroscopy in combination with chemometrics for the screening of large set of crude extracts, to study the effects of different variables on the activity, and identifying active compounds in complex mixtures like plant extracts.Electronic supplementary materialThe online version of this article (doi:10.1007/s11306-012-0406-8) contains supplementary material, which is available to authorized users.

Impacts of leafroll‐associated viruses (GLRaV‐1 and ‐3) on the physiology of the Portuguese grapevine cultivar ‘Touriga Nacional’ growing under field conditions

AbstractThe impact of mixed infection of grapevine leafroll‐associated virus 1 and 3 (GLRaV‐1&‐3) on physiological performance of the Portuguese grapevine variety ‘Touriga Nacional’ was evaluated during 3 years with the main purpose of understanding the drastic reduction in yield. Overall, gas exchange was negatively affected in leaves with these leafroll virus infections. Particularly at ripeness stage, the reduction in stomatal conductance (gs) was higher than in net CO2 assimilation rate (A), leading to higher intrinsic water use efficiency (A/gs) in infected leaves. However, the decrease in gs and A were not a consequence of the decrease in bulk water potential, as the water index/normalised difference vegetation index ratio suggested similar magnitude for both treatments. The maximum quantum efficiency of photosystem II was unaffected by GLRaV‐1&‐3, whereas quantum effective efficiency of PSII, apparent electron transport rate and photochemical quenching significantly decreased in infected leaves and these was paralleled by a significant increase of non‐photochemical quenching. Relative to carbon metabolism, the analyses of the net CO2 assimilation rate/photosynthetic photon flux density (A/PPFD) and net CO2 assimilation rate/internal CO2 concentration (A/Ci) curves revealed that virus infection had a negative effect on light saturated rate of CO2 fixation at high irradiances and carboxylation efficiency but, in contrast, apparent quantum yield of CO2 fixation was significantly higher. Meanwhile, the presence of GLRaV‐1&‐3 resulted in a marked decrease in photosynthetic pigments, soluble sugars and soluble proteins contents, while starch and anthocyanins were significantly improved. N, P, Ca, S and Fe leaf concentrations significantly decreased, while K, Mg, B, Cu, Zn and Mn were unaffected by these two leafroll virus species. Infected plants showed a significant decrease in yield, mainly due to a lower cluster weight. These results emphasised the important role of GLRaV‐1&‐3 as a biotic stress for the grapevine physiology and consequently to yield attributes.

Grapevine varieties exhibiting differences in stomatal response to water deficit.

Knowledge on variety traits and physiological responses to stress is still scarce in Vitis vinifera L., limiting the optimisation of irrigation and breeding for high water use efficiency. We have characterised five grapevine varieties using thermal imaging, leaf gas exchange, leaf morphology and carbon isotope composition. Plants of the varieties Aragonez, Trincadeira, Cabernet Sauvignon, Syrah and Touriga Nacional were grown in field conditions. Two experiments were performed. In Experiment I (2006), vines of Aragonez and Trincadeira were either well irrigated (WI, 80% ETc), non-irrigated but rain fed (NI) or subjected to regulated deficit irrigation (RDI, 40% ETc) and studied along the summer season. In Experiment II (2006 and 2007), vines of the five varieties were subjected to RDI (30-40% ETc) and studied at veraison. In Experiment I, leaf temperature (Tleaf) correlated negatively with stomatal conductance (gs) and leaf water potential (Ψpd). The inverse relationship between gs and Tleaf was highly significant in the afternoon. In Experiment II, the different genotypes showed different Tleaf for similar Ψpd. Stomatal density did not correlate with gs suggesting that varieties have different stomatal control. Our results show that combined measurements of canopy temperature and Ψpd can aid in better understanding of stomatal regulation in different grapevine varieties. Such variation in stomatal regulation should be taken into account in determining irrigation strategies.