José Eduardo Eiras-Dias

Portuguese traditional grapevine cultivars and wild vines ( Vitis vinifera L.) share morphological and genetic traits

Portugal has a long tradition in viticulture and a great number of grapevine cultivars. To analyze the genetic relations among wild vines from Portuguese populations and old Portuguese grapevine cultivars we use morphological traits and chloroplastidial microsatellites from 53 accessions of four distinct populations of Vitis vinifera L. ssp. sylvestris (Gmelin) and 57 accessions of Vitis vinifera L. ssp. vinifera from the Portuguese National Ampelographic Collection. Principal coordinate analyses with the scores obtain from the descriptors of both the accessions of sylvestris and vinifera vines revealed two groups. One group is formed by the wild vine population of Alcácer do Sal and three vinifera accessions Rufete, Seara Nova and Trincadeira das Pratas and a second group includes all the other wild vines and grapevine cultivars. A total of four different chlorotypes (A, B, C and D) are present in the vinifera accessions and two in the sylvestris accessions (A, B). Chlorotype A is the most frequent in all the plants analyzed and correspond to 75.4% of the grapevine cultivars and 66% of the wild vines. The mixed distribution of chlorotypes in the Portuguese cultivars and the predominance of chlorotype A both in its wild populations and cultivars reinforced the hypothesis that West Europe was a domestication center for Vitis vinifera L. ssp. vinifera.

Characterization of Portuguese populations of Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi

Wild populations of Vitis vinifera L.␣have been located in Portugal. Morphological characterization was carried out in three populations located in Alcácer do Sal, Castelo Branco, and Montemor-o-Novo, and then compared using multivariate discriminant analysis. These populations were from three different hydrological basins, therefore cross-pollination was not possible. It was verified that in each population all plants were different. The data suggest that the frequency of female and male plants is rather variable in wild populations. The morphology of the adult leaf, from the Alcácer do Sal population, had particular features when compared with Castelo Branco and Montemor-o-Novo populations, which were more homogeneous. The length of teeth compared with width at the end of the base, and the density of prostrate hairs between and on main veins (lower side) were the variables which allowed the best discrimination among populations.

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

Modeling Phenology, Water Status, and Yield Components of Three Portuguese Grapevines Using the STICS Crop Model

Viticulture plays a key role in Portuguese economic growth. The possibility of obtaining early predictions of viticultural parameters, such as phenology, water status, yield, and potential wine characteristics, are of great value for growers and will influence the viticultural chain. The present study is a first approach to adapt the Simulateur mulTIdisciplinaire pour les Cultures Standard (STICS), a dynamic crop model, to Portuguese winegrowing conditions and grapevine varieties. Several site-specific parameters for climate, soil, and management practices were assessed. Varietal-specific parameters were measured from vineyards in the Douro and Lisboa winegrowing regions of Portugal. Three of the most important varieties in the country (Aragonez, Touriga-Franca, and Touriga-Nacional) were targeted for model calibration. Overall, the STICS crop model was successful in simulating yield, phenological stages, and water stress, but not alcohol content. The relatively high model accuracy will improve the performance of vineyard operations and winemaking practices, such as organizing harvests, vineyard intervention scheduling, irrigation procedures, and strategic planning of wineries. STICS may be feasible as a decision support tool for short- and long-term strategic planning in Portuguese viticulture, particularly taking into account the projected impacts of climate change. The efficiency gains obtained from the use of this crop model by wine industry stakeholders is expected to increase the competitiveness and sustainability of the wine sector in Portugal.

Examining the relationship between the Enhanced Vegetation Index and grapevine phenology

Abstract Monitoring the main grapevine phenological stages is a key procedure for optimizing vineyard activities and improving yield and quality attributes. Remote sensing may be an effective and practical monitoring tool, as data from on-board satellite sensors can measure vegetative growth. In the current study, a 12-year time series of four main phenophases (budburst, flowering, veraison and harvest) were obtained from an experimental vineyard located in Lisbon (Portugal). LANDSAT surface reflectances were used to calculate the Enhanced Vegetation Index (EVI) and derived metrics. Both time series were then analysed. Results show statistically significant relationships between vegetation metrics and phenological timings and intervals, such as the linkage between the peak greenness and flowering/veraison. The current study highlights the applicability of remote sensing to monitor grapevine phenology in both retrospective and real-time, bringing an added-value to the winemaking sector.

Molecular markers for assessing must varietal origin

Wine quality and market value greatly depend on the grapevine varietal composition, which may be characteristic of specific regions. In order to defend the distinct regions, Denominations of Origin were defined to protect against fraudulent practices. In this study, we evaluated the efficiency of two microsatellite-based systems (microsatellite (SSR) and intermicrosatellite (ISSR)) for must varietal composition determination and their potential role in certification purposes. Eleven Vitis vinifera L. varieties from leaf and monovarietal must DNA samples were screened with six SSR and 14 ISSR primers to discriminate polymorphisms. Principal coordinates analysis was performed with DCENTER on the resultant data using unweighted pair group mathematical average and revealed that ISSRs markers were not suitable for certification procedures, whereas nuclear SSR markers presented a complete correspondence between leaf and must samples, demonstrating that they were adequate for traceability purposes.

Identification by SNP Analysis of a Major Role for Cayetana Blanca in the Genetic Network of Iberian Peninsula Grapevine Varieties

Cayetana Blanca is a grapevine variety widely distributed in the Iberian Peninsula (Spain and Portugal). Its numerous synonyms and its presence in northern Africa point to a long history of cultivation. The goal here was to identify the genetic relationships of Cayetana Blanca with other Iberian and Mediterranean cultivars using a panel of SNP markers. A total of 243 single nucleotide polymorphisms (SNPs) and four chloroplast microsatellite loci allowed for identifying synonyms and trio (parents and offspring) and duo (parent-offspring) relationships. Six trios including Cayetana Blanca as a parent were found and supported by very high LODs. Five of the trios had the variety Alfrocheiro Preto as the other parent and gave rise to cultivars used in Portugal: Cornifesto, Camarate, Mouratón, Malvasia Preta, and Periquita. The Spanish cultivar Jaén Tinto is confirmed to be an offspring of Cayetana Blanca and Listán Prieto. Additionally, synonyms and duos were detected that, in some cases, were not anticipated. The parents of Cayetana Blanca remain unknown, while the geographical origin of Cayetana Blanca could be a border region between Portugal and Spain. Results show that this cultivar has had a relevant impact in the viticulture of that area. As in other European regions, viticulture in the Iberian Peninsula is also marked by the impact of certain varieties that have worked as progenitors of many present cultivars. The results also show the interconnection between Portuguese and Spanish viticulture. Finally, the study demonstrates that SNP is a powerful tool for parentage inference in grapevine.

High Resolution Melting (HRM) applied to wine authenticity

Wine authenticity methods are in increasing demand mainly in Denomination of Origin designations. The DNA-based methodologies are a reliable means of tracking food/wine varietal composition. The main aim of this work was the study of High Resolution Melting (HRM) application as a screening method for must and wine authenticity. Three sample types (leaf, must and wine) were used to validate the three developed HRM assays (Vv1-705bp; Vv2-375bp; and Vv3-119bp). The Vv1 HRM assay was only successful when applied to leaf and must samples. The Vv2 HRM assay successfully amplified all sample types, allowing genotype discrimination based on melting temperature values. The smallest amplicon, Vv3, produced a coincident melting curve shape in all sample types (leaf and wine) with corresponding genotypes. This study presents sensitive, rapid and efficient HRM assays applied for the first time to wine samples suitable for wine authenticity purposes.

Phenolic Compounds Involved in Grafting Incompatibility of Vitis spp: Development and Validation of an Analytical Method for their Quantification

INTRODUCTION Graft incompatibility of Vitis spp is an unresolved worldwide problem with important economic consequences. Grafting comprises a complex set of morphological and physiological alterations, in which the phenolic compounds seem to be strongly involved. Therefore, a detailed analysis and recognition of structural phenolic compounds diversity in the two partners of a Vitis graft is of great importance to evaluate their role as markers of graft establishment. OBJECTIVE To optimise a sample extraction method, and to develop and validate a high-performance liquid chromatography (HPLC) method for the simultaneous determination of phenolic acids and flavonols in the graft union so as to understand their behaviour in the metabolism of the scion-rootstock system, using compatible and incompatible combinations of a Syrah cultivar and two rootstocks (R110 and SO4). METHODS Sixty extracts of Vitis grafting tissues were prepared and analysed by HPLC for the qualitative and quantitative determination of their phenolic profile. RESULTS Among the phenolic compounds identified in the samples, one benzoic acid (gallic acid), three cinnamic acids (caffeic acid, ferulic acid and sinapic acid) and two flavonols (catechin and epicatechin) are potentially suitable as markers of graft incompatibility. CONCLUSION The method developed presents good performance and lends itself readily for application in routine analysis of the phenolic composition of Vitis grafting tissues to distinguish compatible and incompatible combinations in the graft callusing stage.

Portuguese Vitis vinifera L. Germplasm: Accessing Its Diversity and Strategies for Conservation

Grapevine (Vitis vinifera L.) is the most widely cultivated and economically important fruit crop in the world. In the different Portuguese agro-ecosystems, grapevine plays an impor‐ tant role either as a border culture or as an extensive crop. The surface area used by vine‐ yards amounts to 4.9 % of the arable land [1], representing 240,000 ha, being the 7th largest area in the world and the 4th in the European Union [2]. In 2011 Portugal produced 5.9 mil‐ lion hectoliters of which 2.9 million hectoliters were exported, making the country the 12th world wine producer [2]. There are fourteen wine regions with Protected Geographical Indi‐ cation (Figure 1) and 31 wine areas with Designation of Origin status including Porto, estab‐ lished since 1756, the oldest legally established wine production region in the world. Each one of the wine regions has a particular set of grapevine cultivars adapted to its specific ter‐ roirs. Officially there are 343 cultivars allowed to be use in wine production in Portugal [3].