Efficient Use of Non-renewable Natural Resources for Quality Wine ...

Complimentary Contributor Copy

In: Agricultural Systems in the 21st Century Editor: Amir Raza

ISBN: 978-1-62808-992-9 © 2013 Nova Science Publishers, Inc.

Chapter 9

EFFICIENT USE OF NON-RENEWABLE NATURAL RESOURCES FOR QUALITY WINE THROUGH SUSTAINABLE VITICULTURE Sílvia Petronilho, António S. Barros, Manuel A. Coimbra and Sílvia M. Rocha QOPNA, Departamento de Química, Universidade de Aveiro, Aveiro, Portugal

ABSTRACT Sustainable viticulture appeared as a breakthrough approach aiming to enhance the quality of grape varieties as an efficient use of non-renewable natural resources. This leads to wine quality enhancing while maintaining an economically viable production. The detailed knowledge of the specificities of each region, namely environmental factors and agricultural practices, among others, is crucial for sustainability. Even within each Appellation, heterogeneity can be observed regarding the characteristics that may influence grape and wine composition and quality. The evaluation of the variety suitability regarding the region attributes should be considered a strategic issue for sustainable viticulture. An application of this concept was performed at Bairrada Appellation. With a perspective of sustainable viticulture, the wine makers are able to select the best suited binomial region specificities/variety to produce an optimized product and/or take advantage of the diversity and hence produce wines with different characteristics when using the same variety. Taking into account that a complex network of variables influences grape yield and quality into different extents, the development of multivariate models combining these variables with physical and chemical grapes compositions, are important tools for the establishment of relevant relationships between these natural parameters and grapes quality. This would allow the producers to know the variety suitability based on the region specificities. The concept of sustainable viticulture, although not yet globally established and accepted all over the world by the winemakers and consumers, tends to continue growing in the future.

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Corresponding author E-mail address: [email protected] (Sílvia M. Rocha), Tel. + 351 234401524; Fax. + 351 234370084.


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Sílvia Petronilho, António S. Barros, Manuel A. Coimbra et al.

Keywords: Sustainable viticulture, environmental factors, agricultural practices, binomial region specificities/variety, wine quality

1. SUSTAINABLE VITICULTURE: CONCEPT Sustainability is a visionary development paradigm that, at the beginning of the 21st century, is widely recognized by world leaders, and is a common topic of discussion by society all over the world (Zucca, Smith & Mitry, 2009; Lamastra, Fragkoulis, Trevisan & Capri, 2010). This term was established in 1987 by the Brundtland Commission that defined the sustainable development as a progress that "meets the needs of the present without compromising the ability of future generations to meet their own needs” (United, Nations General Assembly, 1987). This definition acknowledges that while development may be necessary to meet human needs and improve the quality of life, it must occur without depleting the capacity of the natural environment to meet present and future needs. The concept of sustainable development emerged as an attempt to bridge the gap between environmental concerns about the increasingly evident ecological consequences of human activities, and socio-political concerns about human development issues (Robinson, 2004). Sustainable development represents an actual concern of the society and influences the market trends and also some local and international policies. Thus, this concept was extended to several fields. The adaptation of this important concept to the agriculture sector defines sustainable agriculture as an integrated system of plant and animal production practices having a site-specific application that over the long-term will: i) satisfy human food and fibre needs, ii) enhance environmental quality and the natural resource based upon which the agriculture economy depends, iii) make the most efficient use of non-renewable and on-farm resources and integrate, where appropriate, natural biological cycles and controls, iv) sustain the economic viability of farm operations, and v) enhance the quality of life of the society as a whole (Lichtfouse et al., 2009; Lamastra et al., 2010). Permaculture is another area that adopts sustainable development concept. This offers a unique approach to the practice of sustainable farming, ranching, gardening and living. Permaculture integrates plants, animals, landscapes, structures and humans into symbiotic systems where the products of one element serve the needs of another, and once established, it can be maintained using a minimum of materials, energy and efforts. This system is designed to be diverse, so when one element fails, the system has enough stability and flexibility to prosper (Mollison, 1988). In line with the actual trends, sustainable viticulture emerged as a breakthrough approach for improving the quality of environmental and natural resources, namely grape varieties, based on an integrated and efficient use of non-renewable resources, integrating environmental, economic and social issues (Lamastra et al., 2010) (Figure 1). Thus, the wine quality enhancement is sought while maintaining an economically viable production. Sustainability is a particularly interesting challenge in wine sector, as it tends to grow up in an equilibrium between tradition and innovation (Jackson & Lombard, 1993).


Efficient Use of Non-Renewable Natural Resources for Quality Wine...

Economical

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Environmental

Social Figure 1. Graphical definition of sustainable viticulture (adapted from Lamastra et al., 2010).

The sustainable viticulture is not yet a global practice due to several factors. This concept and their advantages are not fully known by some winegrowers and wine producers (a broad topic that means many things to many people, because people often misunderstand the term ‘sustainable’). Also, economic pressures from several industries related to viticulture and winemaking, may also prevent its implementation. Global education and the definition of the limits and advantages of the sustainable viticulture should be done as a base for the development of an integrated platform of global sustainability. The wine producers of each region need to know the potentialities of their vineyards and varieties to go in front in this field. Achieving sustainable practices is viewed as a process requiring small, realistic, and step by step improvements. Several examples may be point out in different places around the world. Two examples were selected in two continents (America and Europe) to highlight the concept under discussion. For example, in California, sustainable viticulture began in the early 1990s, as a result of the efforts of growers and winemakers in the central valley of California around the town of Lodi. The Lodi–Winegrape Commission established a sustainable winegrowing program where a range of sustainable viticultural practices were implemented and tracked over time. This program involved work with a core group of 40 Lodi growers and about 15 pest control advisors in 60 different vineyards. Various sustainable viticultural practices were implemented in these vineyards, including pest monitoring and vineyard inputs such as water, fertilizer and pesticides, so that growers could see the effects of the sustainable practices. This sustainable winegrowing program has been adapted by several other regional winegrowing associations and regions in Californian (Zucca et al., 2009). Also, in Italy, a sustainable winegrowing program (SOStain) was developed. This program constituted a framework for viticultural and winemaking practices that protected the environment while efficiently and economically produced premium grapes and wines. In this program the assessment and the interpretation of the results occurred through the use of agro-environmental indicators, in this case the EIOVI (Environmental Impact of Organic Viticulture Indicator). EIOVI took into account the different agronomical practices used in viticulture (pest, disease, fertilizer, irrigation and soil management, and machinery used) and estimated the effect of vineyard


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management on soil organic matter and on the biodiversity. The indicator helped decisionmakers by informing them of the linkages between viticulture activities and environmental impacts and it could provide an early indication of potential changes in the state of the environment (Lamastra et al., 2010).

2. ENVIRONMENTAL FACTORS THAT INFLUENCE VITICULTURE Wine is produced around the world, under different climatic, soil and topographical conditions, using several hundreds of varieties selected in each region. Thus, it may be possible to find information about Vitis vinifera L. varieties produced in a wide range of conditions, even in extreme conditions. Indicatively, some ranges may be considered: temperatures (from < 0 °C to 38 °C), sunlight exposure (1200 to 2800 hours per year), precipitation (0.1 to 20 dm), soils (sandy, clayey, clay-calcareous, volcanic, loam, silt, siltloam, clay-loam), and topographical features (6 to 3000 m altitude and slope: terrace until 60 %). In this section influence of these parameters on viticulture and consequently on grape and wine composition are discussed and some examples are used to illustrate the topics under discussion. Grape components are produced by the plant itself, in leaves and in berries (sugars, acids, phenolics, volatiles, among others) during fruit development and ripening. Thus, the growth and the fructification of grapevines in the vineyard are of utmost importance to grapes, and consequently, to wine quality (Conde et al., 2007). The quality of grapes and wines is influenced by several environmental factors (Lila, 2006), namely type of soil, topography, agro-pedological features and edaphoclimatic conditions, among others (Smart, 2003; Turner & Creasy, 2003; Pozo-Bayón, Polo, Martín-Álvarez & Pueyo, 2004; Vaudour & Shaw, 2005; Coelho, Coimbra, Nogueira & Rocha, 2009). However, the dependence of grape berry attributes on the specific environmental conditions of many Appellations remains uncertain, although the specific and systematic knowledge is crucial for sustainability. Several physical and chemical parameters, such as berry weight, pH, acidity, volatile and phenolic composition are commonly considered in order to understand the effects of environment in specific grape variety, and consequently, in wine quality.

Climate Conditions Growing any crop with quality and economically sustainable, anywhere in the world, is strongly dependent on climate (Anderson, Jones, Tait, Hall & Trought, 2012). Climate is a very complex, highly variable, and pervasive factor in our natural and human-based systems. It is widely recognized that climate has significant implications for the agricultural sector, including viticulture (Jones, White, Cooper & Storchmann, 2005). It is well known that climate have impact on grapes and wines quality, through the effect of both regional and local-scale climatic conditions during the growing season, which generates variations in grapevine growth, and then in berry composition (Jones et al., 2005; Soar, Sadras & Petrie, 2008). Climate can express its influence through several elements namely temperature,



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sunlight exposure, and precipitation, among others (Jones & Davis, 2000; Mateus, Marques, Gonçalves, Machado & Freitas, 2001a; Holland & Smit, 2010).

Temperature Among climate variables, temperature is recognized as having the greatest effect on physiological behaviour of the grapevine and on chemical changes in the berry during its formation and maturation affecting plant vitality, ripening rate and harvest date (Due, Morris, Pattison & Coombe, 1993). The effect of infield temperature on grapevine development may be reflected in a network that comprises several mechanisms, namely, photosynthesis, respiration and metabolism. Temperature influences photosynthesis, affecting all the biochemical reactions. A study comparing Trebbiano grapevines, grown in Italy, at 20, 27.5 and 35 °C in separated chambers, showed that the lowest photosynthetic rate was observed for the vines grown at the maximum temperature tested (Ferrini, Mattii & Nicese, 1995). The functional activities of the photosynthetic apparatus of two grapevines (Vitis vinifera L. cv. Müller-Thurgau and Lagrein) were investigated after low night temperature treatment (ca 5 ºC). During daylight, these plants were kept at ca 25 °C. The low night temperatures applied caused important reductions of the photosynthetic rate, limiting photosynthesis via inhibition of electron transport and photophosphorylation, via inhibition of key enzymes in sucrose, and starch biosynthesis (Bertamini, Zulini, K. & Nedunchezhian, 2007). A recent study carried out in Australia (Greer, 2012) showed that during the growing season of Vitis vinifera L. cv. Semillon, the higher photosynthetic rates was observed from 25 to 30 ºC. Temperature also influences the grape berry respiration, which is a key process in the grape ripening. The organic acids of grapes, namely malic acid, are formed in the combustion respiratory process of sugars. The organic acids formation represents intermediary steps of the respiratory process, releasing a part of the stored energy in sugars molecules (PopescuMitroi, Radua & Stoica, 2009). After their formation, malic acid diffuses from the interior to the periphery where it is metabolized (Staden, Volschenk, Vuuren & Viljoen-Bloom, 2005). If the berry temperature rises during ripening, simultaneous increase in malic acid respiration occurs, accounting for the low acidity of the sunburned berries. Such phenomenon implicitly suggests up-regulation of respiration as a temperature-sensitive malate metabolic pathway due to the involvement of malate in this pathway during ripening (Bondada & Keller, 2012). Thus, the influence of the temperature in respiration process regulates the organic acids and sugars content in the grapes. Temperature regulates the grapevine metabolism and the production and accumulation of metabolites, such as those responsible for aroma and colour. As an example, some studies showed the influence of temperature on the concentration of the grape components associated to colour properties (for example phenolic compounds) (Fregoni & Pezzutto, 2000; Montes, Perez-Quezada, Peña-Neira & Tonietto, 2012). It was found that cold days (15 ºC) during ripening improved colour development (due to anthocyanins increasing) in Cardinal, Pinot Noir, and Tokay berries, while hot days (35ºC) significantly reduced the formation of anthocyanins. A cold night temperature (10 or 15 ºC) does not reverse the effect of hot days influence on berry colour (Buttrose, Hale & Kleiwer, 1971; Kliewer & Torres, 1972). Similar results were obtained from Vitis vinifera cv. Cabernet-Sauvignon (Goto-Yamamoto, Mori, Numata, Koyama & Kitayama, 2009) and Merlot (Spayd, Tarara, Mee & Ferguson, 2002),


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suggesting that higher temperatures than 30 to 35 °C promote significant reductions on phenolic content, especially on anthocyanins.

Sunlight Exposure Berry temperature in the field is largely regulated by the flux density of absorbed radiation and convective heat loss and has been shown to increase linearly with incident radiation. The effects of light on grape composition are heavily dependent upon the extent to which berry temperature is elevated as a result of increased sunlight exposure (Bergqvist, 2001). This effect on grape berry development and composition has been investigated during the last few decades, showing that an increasing of sunlight grape exposure during ripening generally improves grape and wine composition (Price, Breen, Valladao & Watson, 1995; Bergqvist, 2001; Spayd et al., 2002), namely increasing the content of total soluble solids and phenolics, namely anthocyanins (Dokoozlian & Kliewer, 1996; Spayd et al., 2002). Otherwise, prolonged exposure to direct sunlight, rising temperatures in the field around 30 to 35°C, promoted the reduction on phenols content, namely on anthocyanins (Buttrose et al., 1971; Kliewer & Torres, 1972; Goto-Yamamoto et al., 2009). Sunlight-exposed grapes have also shown higher levels of sugar content and lower values of titratable acidity, malic acid content, and pH when compared to shaded grapes (Crippen & Morrison, 1986; Dokoozlian & Kliewer, 1996). Sunlight provides light energy for photosynthesis and other light-stimulated metabolic processes (namely the biosyntheses of phenolic compounds promoted by phenylalanine ammonia lyase), and provides heat, both by direct solar radiation on plant surfaces and by heating the surrounding air (Crippen & Morrison, 1986; Ribéreau-Gayon, Dubourdieu, Doneche & Lonvand, 2000). Heat from sunlight can influence reaction rates of metabolic processes and can also cause stress, either by direct temperature stress or by dehydration (Crippen & Morrison, 1986). The environment surrounding the vines, namely the presence of trees, the abundance of vegetation, and excessive leaf areas, may affect the sunlight exposure, reducing the levels of sun and light penetration and also reducing the air flow (Smithyman, Howell & Miller, 1997). Grapes less exposed to the sunlight contain less sugar content, lower pH and higher concentrations of malic acid than the grapes more exposed to the sunlight (Macaulay & Morris, 1993; Ribéreau-Gayon et al., 2000). The effect of surrounding vegetation height on Touriga Nacional grapes, from Douro Appellation, in Portugal, was evaluated. The results showed that grapes grown in vineyards with higher vegetation height (100 cm) had higher carotenoid levels, while grapes grown in vineyards with lower vegetation height (60 cm), less protected from sunlight exposure, had higher weight and sugar content. Furthermore, during the maturation period, a lower decrease in carotenoid degradation was observed in vineyards surrounded with higher vegetation height, explaining their higher carotenoid content (Oliveira, Ferreira, Costa, Guerra & Guedes de Pinho, 2004). A study carried out on Gewürztraminer grape berries composition showed that the levels of glycosylated monoterpenoids were much higher on the sun-exposed grapes when compared with grapes that remained in the partial or total shadow (Reynolds & Wardle, 1989b). The sunlight also promoted the biosynthesis of carotenoids from the first stage of berry formation until véraison, decreasing between véraison and maturity, giving rise to the glycosylated C13 norisoprenoids and other compounds (Baumes, Wirth, Bureau, Gunata & Razungles, 2002). Furthermore, modifications on grape environment such as the hedging and basal leaf removal, or crop level reduction, increased the level of both free and glycosylated terpenoids (Reynolds



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& Wardle, 1989a; Reynolds, Wardle & Dever, 1996; Reynolds, Wardle & Naylor, 1996; Belancic et al., 1997).

Precipitation The period and precipitation level and the moment when it occurs influence vine water status. The effect of precipitation during initial vegetative phase, flowering, berry development and prior to harvest are considered, in this chapter. The grapevine vegetative phase is influenced by water availability (Acevedo-Opazo, Ortega-Farias & Fuentes, 2010) and, depending on the intensity and period of water stress, different effects may be observed such as shoot growth stopping (Hsiao & Xu, 2000; van Leeuwen et al., 2009) and high reduction of leaves size and number, which increases the risks of berry sunburn during its development (Bondada & Keller, 2012). In these cases irrigation can increase the shoot growth rates and the leaf area. However, a regulation of the water administration needs to be done, as overdone water amounts can originate denser canopies that decrease the radiation levels and airing inside the canopy. Thus, lately, this can lead to a deficient berry development and an increase of the risk of diseases, having a negative effect in the berry quality. Water deficit between anthesis (flowering period) and véraison decreases flowers formation which lead to the diminishing of berry formation and also of its size, and this is often irreversible even if there is no water shortage after the beginning of ripening (McCarthy, 1997). Cell division of pericarp occurs during the first growth phase of the berries. Early water stress reduces the rate of cell division, which explains the inability of berries to recover in size after an early water deficit (during the flowering period). Furthermore, organic acids (tartaric and hydroxycinnamic acids), phenolics (tannins) and several other compounds such as minerals, micronutrients, and aroma compounds are accumulated during the first phase of berry growth (Cardoso, Carvalheira, Coimbra & Rocha, 2005). Thus, an early water stress reduces the accumulation of these components, affecting grapes quality (Conde et al., 2007). Water stress during the development phase of the berry may also decrease berry weight, but in this case the reduction is related to reduce cell volume or diminished solutes (sugar) in the cells. Nutrient deficiencies and other disorders that reduce photosynthesis may also reduce berry growth or slow ripening by decreasing the supply of sugars to the berries (Ojeda, Deloire & Carbonneau, 2001). The water status during berry development influences the berry sugar content, which is yield-dependent. For low yields, vine water deficit enhances berry sugar content (Trégoat, van Leeuwen, Choné & Gaudillère, 2002; van Leeuwen et al., 2009). However, extreme water stress, due to low precipitation, is harmful to the development of the berries, and may lead to yield and quality losses (Ojeda, Andary, Kraeva, Carbonneau & Deloire, 2002; van Leeuwen et al., 2009). In this case, the irrigation should be an adequate option. Precipitation just prior to harvest can affect grape sugar content, usually expressed as ºBrix, by diluting the sugar and causing ºBrix to drop, thus it is expecting the production of wines with low alcohol content (van Leeuwen & Seguin, 2006). Excessive precipitation at this period can also increase berry size and promote the decreasing of organic acids, anthocyanins, and tannins content (Keller, Smith & Bondada, 2006). Several vine diseases and grapes rotting can also be observed. Otherwise, vines that experience low amount of water have been found to produce fewer and smaller grapes, but with higher sugar, phenolic and volatile content (Jackson & Lombard, 1993; van Leeuwen & Seguin, 2006). In these cases, it is expected to produce wines with high alcohol content, and


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more intense aroma and colour. This fact is particularly interesting in red wine production due to the fact that controlled water stress of the vine increases berry colour, specially due to the increasing of anthocyanins and tannin content (van Leeuwen & Seguin, 1994; Koundouras, Marinos, Gkoulioti, Kotseridis & van Leeuwen, 2006).

Soil Soil has also an important role for viticulture sustainability, since several environmental factors affecting the vine growing and grape and wine composition are related to the soil properties. Soil acts as a regulator of the climate elements, because soil may affect water and nutrient availability to the plant by its retaining capacity, may affect the microclimate by its heat-retaining and light reflecting capacity, and may affect the root growth by its penetrability (Jackson & Lombard, 1993; Martinez, Ascacibar, Espinoza & Lorza, 2011). Soil types may be defined according to different criteria, namely, taxonomy, morphology, genesis, and texture, among others (USDA, 1999; Gerrard, 2000). Considering the texture features, soil is usually classified as clay, clay-calcareous and sandy. The soil type is highly related to the water status through its water-holding capacity (Oliveira et al., 2003; van Leeuwen et al., 2004). The restriction of water supply plays a significant role in vine behaviour and berry composition. A limitation in vine water uptake reduces shoot growth, berry weight and yield and increases berry anthocyanin and tannin content (van Leeuwen & Seguin, 1994; Choné, Leeuwen, Chery & Ribereau-Gayon, 2001; Koundouras et al., 2006), which, if not excessive, are favourable to grape quality potential (Kennedy, Matthews & Waterhouse, 2002; Roby, Harbertson, Adams & Matthews, 2004; van Leeuwen et al., 2009). For example, water stress imposed by some types of soils (namely siltclay) have been shown to increase the oenological potential of Agiorgitiko red grape variety by: i) accelerating sugar accumulation and malic acid breakdown in the juice, ii) promoting the concentration of anthocyanins and total phenolics in berry skins, and iii) increasing the amount of glyco conjugates of the main aroma components of grapes (Koundouras et al., 2006). The influence of soil regarding its texture, depth, chemical composition, and water availability on the characteristics of wines has been evaluated (van Leeuwen et al., 2004; Prado et al., 2007). The soil type was found to influence significantly the volatile composition of sparkling wines obtained from Fernão-Pires and Baga varieties from Bairrada Appellation, in Portugal. Sparkling wines produced from the clay-calcareous soil presented the highest content of volatiles related to aroma properties when compared to those obtained from grapes produced in clay and sandy soils (Coelho et al., 2009). According to these data, clay-related soils seems to improve wine quality (namely increasing its phenolic and volatile contents), when compared to other kind of soils. However, the outcome observed results from a network of other natural factors, intrinsically related to each region, thus its extension to other vineyards and Appellations is not possible. These studies are helpful and needed for each specific wine region, which has specific environments, agricultural practices, varieties, and other inherently natural parameters.



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Topographical Features Altitude and slope can strongly regulate the climatic conditions since they are directly associated to the resulting temperature, precipitation, humidity, vineyards surrounding vegetation height, sunlight exposure and shadow of vineyards, vineyards orientation and trellising, influencing grapevine vigour and grape maturation (Jackson & Lombard, 1993). Scarce information is available about the influence of these topographical features on viticulture. The effect of altitude and its related climatic conditions on phenolic composition (procyanidin and anthocyanin components) of grapes and wines of Touriga Nacional and Touriga Francesa red varieties, from Douro Appellation, have been reported (Mateus et al., 2001a; Mateus, Proença, Ribeiro, Machado & Freitas, 2001b; Mateus, Machado & Freitas, 2002). At berry maturation, low altitude (100 to 150 m above the sea level) was shown to be an important factor favouring the biosynthesis of higher concentrations of grape-skin catechin monomers ((+)-catechin, (-)-epicatechin gallate), procyanidin dimers, trimer C1, as well as total extractable proanthocyanidins, when compared to higher altitudes (250 to 350 m above the sea level), contributing to improve wine quality (Mateus et al., 2001a). Higher altitudes are associated with lower temperatures and high humidity, which affect grape maturation, diminishing its polyphenolic composition (Mateus et al., 2001a; Mateus et al., 2001b; Mateus et al., 2002). Similar trend was observed in Cabernet Sauvignon wines produced at different altitudes (909 m and 1280 m) in Loess Plateau (China). The content of phenolic compounds (flavonoids and flavanols) and antioxidant activity of wine from lower altitude vineyard (909 m) were relatively higher than those from higher altitude vineyard (1280 m) (Jiang, Zhang & Zhang, 2011). Altitude effect was also studied using grapes from two varieties harvested in different terraces from Douro Appellation (Touriga Franca: 85, 145 and 180 m, and Touriga Nacional: 90, 155 and 210 m) (Oliveira et al., 2004). Touriga Franca grapes grown at lower altitude (85 m) had the lowest carotenoid concentrations. Conversely, for this variety, grapes grown at higher altitudes (145 and 180 m) had higher carotenoid levels. High altitude, which presented lower temperature and higher humidity, is associated with a lower berry growth, decreasing the carotenoid degradation during the maturation period, and this could explain the higher carotenoid values in the high-elevation sites in Touriga Franca variety. This observation was not so evident in Touriga Nacional grapes. At maturity, the carotenoids content of these grapes produced at 155 m altitude were significantly higher than at 90 and 210 m, suggesting that moderate temperatures on moderate slope positions with good sunlight exposure are ideal for colour development and also for carotenoid accumulation. The latter, as precursors of aroma compounds, may also contribute positively to the wine sensorial properties. It was also reported that aroma potential, given by monoterpene and norisoprenoid components, showed higher content for Vitis vinifera cv. Veneto grapes grown at lower altitude (Tomasi et al., 2000), being expected to obtain wines with higher aroma quality. According to the selected examples, in the same vineyard, grapevines located at lower altitudes produced grapes with better oenological potential, specially related to phenolic and volatile composition. The slope of a vineyard has also high influence on the temperature and soil drainage, both of which are critical for the growing of grapevines. The manner by which the vines are trellising is the one that best overcomes the restraints imposed upon them by climate, soil, plant needs, and production goals. Vineyards can be planted on very steep slopes (until 60 %)


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or on terraces. A slope can reduce exposure to heat or cold depending on its aspect (is the term used to describe the direction that slope faces). The aspect of the slope is important for sunlight interception: more sunlight intercepted lead to warm the vineyard. The contribution of slope and its aspect depends on the region; however, it is common to consider that Southern aspects and the steepest slopes are preferred due to its highest temperature and higher sunlight interception, being great for grapes maturation. The slope of a vineyard lead to changes in several parameters, namely in grapevine vegetative height and also in grapes chemical composition.

3. AGRICULTURAL FACTORS THAT INFLUENCE VITICULTURE The detailed knowledge about all the factors that influence grapes and wines quality is crucial for sustainability in viticulture sector. In addition to environmental conditions, which are not controllable, the grapes and wines quality also depends on several agricultural practices, which the winemakers can act on. All over the world, several kinds of agricultural practices are employed in order to improve vineyards performance, in particular, to improve grape yield and quality. However, these are dependent on local environment factors (soil, topography and climate, among others), that are well known for their influence on grapes and wines quality. Thus, it is essential to adapt appropriate viticulture management practices, namely irrigation, thinning, trellising, and mulching, among others, to the specificities of each vineyards and/or Appellation. The influence of several viticultural management operations on viticulture and, consequently, on grape and wine composition, is discussed in this section.

Irrigation Vine water status may be regulated by irrigation and its management is crucial to grape quality. Excessive irrigation, which is common due to poor assessment of vine water requirements, can promote grape and wine quality losses caused by yield increase, vigorous canopy growth and berry dilution. Full irrigation increases yield production however is detrimental for grapes quality, diminishing its phenolic and volatile contents (van Leeuwen et al., 2009). Vines that suffer severe water stress due to precipitation deficit, begin to defoliate, exposing more the berries that had been shaded by foliage, thus may not attaining their full size, and be prone to sunburn and shrivelling. The severity of water stress can affect vines either reversibly or irreversibly. Furthermore, severe water stress tends to decrease vigour but also the sugar and acid content since photosynthetic activity may be compromised (Bahar, Carbonneau & Korkutal, 2011). However, when a severe water stress is observed, deficit irrigation strategies can be developed in order to produce high quality wines, with higher phenolic content and aroma components, while limiting yield losses (Dry, Loveys, Mccarthy & Stoll, 2001). Berry sugar content was greatest when water deficit was moderate (van Leeuwen et al., 2009). Thus, a moderate water stress is the ideal, where the positive effects outweigh the negative ones (Zarrouk et al., 2012), promoting the production of grapes containing higher concentrations of sugar, anthocyanin and tannins (van Leeuwen & Seguin, 1994). However, the level of moderate water deficits, in order to produce quality grapes and



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consequently quality wines, in irrigated vines, is difficult to achieve. For this type of finelytuned irrigation management to yield positive results, accurate methods for assessing vine water status are needed. A large range of methods for assessing vine water uptake conditions have been developed over the years: i) soil water monitoring by means of tensiometers (Nadal & Arola, 1995; Hanson, Douglas & Orloff, 2000), (ii) water balance modelling (Lebon, Dumas, Pieri P. & Schultz, 2003; Pellegrino, Gozé, Lebon & Wery, 2006), or (iii) the use of physiological indicators as stem water potential (Choné, van Leeuwen, Dubourdieu & Gaudillère, 2001) and carbon isotope discrimination, measured on grape sugar at ripeness (Gaudillère, van Leeuwen & Ollat, 2002). From these, stem water potential was considered the most accurate tool for irrigation management (van Leeuwen et al., 2009) because it represents whole vine water status during the day, even if soil water content is heterogeneous (the case of irrigated vineyards). It can also be used in dry-farmed vineyards for measuring residual water deficits after rainfall (Choné et al., 2000). Nowadays, several irrigation technologies are available, and their selection is ruled by the specific conditions of the vineyard and its price as well.

Thinning In a vineyard, the balance between shoot growth and berry development is important to determine the production quality and quantity (Reynolds, Price, Wardle & Watson, 1994). It is known that vines with too much load have shorter central roots, less knots and smaller leafs (Edson, Howell & Flore, 1993). In order to achieve a correct balance, winemakers have available several viticultural management operations such as thinning, which is described as the suppression of leaves, flowers or clusters before full maturation (Pastore et al., 2011). Thinning is a very controversial operation, and there are many inconsistencies between several studies. Some authors support that the environmental conditions, such as temperature and soil composition, are more relevant to grapes quality than thinning. They minimize the hypothesis that thinning accelerates ripening and improves grapes composition (Ridomi, Pezza, Intrieri & Silvestroni, 1995; Keller, Mills, Wample & Spayd, 2005). On the other hand, other authors demonstrate that thinning is an essential ripening and quality tool (Prajitna et al., 2007) used to correct over cropping or improve fruit composition (Reynolds et al., 2007), namely in terms of soluble solids, phenolics and aroma compounds. The process of thinning has a direct effect on photosynthesis, due to the fact that with less amount of fruits, the photosynthetic assimilation is redistributed and improved, increasing soluble solids, pH, anthocyanins and must colour (Reynolds et al., 1994). It directly influences the yield/quality ratio (Naor & Gal, 2002; Kurtural, Dami & Taylor, 2006; Petrie & Clingeleffer, 2006; Reynolds et al., 2007). A study conducted through 5 years (2000 to 2004) to evaluate the effects of 3 thinning levels (10, 20 and 30 clusters per vine) on yield and grape composition (in terms of soluble solids, pH and titratable acidity) of Chambourcin, grown in Ohio, concluded that thinning reduced the yield per vine and crop load but increased the cluster and berry weights. Thinning also improved grapes composition by increasing soluble solids and pH but not acidity. Furthermore, under the climatic conditions of the study, thinning to 10 clusters per vine produced the lowest yield but the highest vine size with the most optimum fruit composition. Therefore, vines from the 10 thinning treatment were considered the most balanced in the 5 years study (Dami, Ferree, Prajitna & Scurlock, 2006). Furthermore, it was


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also shown that thinning increased the total anthocyanins, phenolics (including resveratrol), and antioxidant capacity of wines produced from Chambourcin grapes (Prajitna et al., 2007). Thinning has also been considered a viticulture process that improves canopy sanitary conditions as it allows more illumination and fresh air penetration in the vegetation and grapes (Smithyman, Howell & Miller, 1998). The thinning time should be established as it influences the grape composition. Thinning between berries establishment and cluster closure has been shown to increase berry size and weight (Prajitna et al., 2007) and to increase the number of berries per cluster (Smithyman et al., 1998). Soluble solids increase with thinning severity due to the fact that the vine has less fruit to ripen, and this occurs independently of when the thinning is performed, after berry set or at berry development phase (Jackson & Lombard, 1993; Reynolds et al., 2007). Recently, the effects of different thinning times at different stages of berry development (pre-véraison and post-véraison) on grape quality and yield components of Sauvignon Blanc grapevines, from 2008, were investigated. Yield and grape composition characteristics were variously influenced by thinning times. It was found that pre-véraison thinning treatments led to enhancements in grape quality, namely improving grape composition in terms of free and potentially volatile terpenoids, total soluble solids, total acidity, and pH, and particularly improving the monoterpene levels (Kok, 2011). Thinning can help growers to avoid delays in ripening and improve grape quality however it reduces the amount of harvestable fruits. Thus, for a grower to consider thinning as a management tool, the loss in income from lower overall yields and increased labour costs must be compensated by a sufficient increase in the fruit price (Preszler, Schmit & Heuvel, 2010). Furthermore, pre-véraison thinning treatments seems to be more useful in order to improve grapes quality, than during berries development, improving volatiles and phenolics content, pH, acidity, and soluble solids. In addition, thinning processes are highly dependent on environmental conditions, mainly with sunlight exposures. Thus, in order to adopt accurate thinning treatments with a correct balance between sun exposed berries and the level of thinning treatments, it is essential to take into consideration each local environmental condition.

Trellising Grapevine orientation in space through the trellising system has significant effects, particularly on light distribution through the vines. In all wine regions, trellising needs to be adapted to the local climatic conditions in order to improve grape ripeness and productivity and also grape composition and quality (Ferree et al., 2002). Trellising is done in order to develop a structure that: i) optimizes the utilization of sunlight and promotes productivity, ii) adapts to the characteristics of the grape cultivar, iii) promotes efficient and sustainable vineyard management practices, and iv) is economically feasible to establish and to maintain (Palliotti, 2012). The production of quality grapes relies on two basic characteristics of trellising systems: a) the adequate functional leaf area, which is the source of the soluble solids that are transported to the fruit, therefore, a characteristic of a good vine trellising system is the ability to display a large amount of leaf area in a way that all leaves are well exposed to the sunlight, and b) the exposure of fruit to the sunlight. This is most important in a cool to moderate climate because the temperature of the fruit during the period of its



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ripening directly influences the reduction of acid levels and increases the specific aroma profile in the fruit (Trioli & Hofmann, 2009). Several trellising systems have been used in order to better match trellis configuration to anticipate vine vigour. Several of these systems incorporate canopy division (either horizontal or vertical) and/or shoot positioning (Gladstone & Dokoozlian, 2003) in order to increase the exposed leaf area, to optimize the canopy microclimate (increasing solar interception by the canopy surface and sunlight penetration into the canopy interior), and to improve grape quality (Gladstone & Dokoozlian, 2003; Palliotti, 2012). Different trellising systems may be considered: i) Australian (head-trained vines with eight or ten bud canes and two bud-renewal spurs), ii) Lenz Moser (cordon-cane systems with eight or ten bud canes and two bud-renewal spurs), iii) bilateral cordon (two cordons are fastened to the top wire at 1.6 m and three to five-bud canes plus spurs totalling 20 buds are left on each side, and after cluster thinning the shoots are positioned), iv) bilateral cordon sylvos (seven to nine-bud canes are arched over a wire at 1.2 m and tied to a lower wire (0.8 m), v) 2 types of upright cordons (two equal cordons in the shape of a “U” with shoots developing in the bottom of the “U” (0.8 m) removed and no shoot positioning or summer tipping are received): upright cordon-spur pruned (20 buds are left on each side in two-to-three bud spurs), and upright cordon-cane pruned (three 5-bud canes are left on each side plus spurs to equal 40 buds), vi) Vertical Shoot Position (VSP) (vine spacing is 2.70 × 1.50 m, trained on a bilateral Royat Cordon at 1.0 m aboveground with shoots positioned upwards in three foliage wires, 1.0 m of surface area) and vii) modified Geneva Double Curtain (GDC) (vine spacing is 3.0 × 1.5 m with trunk height at 1.90 m aboveground with shoots horizontally divided and trained downwards). Little attention was paid to environmental factors that influence vine vigour when selecting trellising systems. Thus, the improper utilization of trellising systems can result in excessive fruit zone shading under vigorous conditions and/or inefficient vineyard design in low-vigour situations (Gladstone & Dokoozlian, 2003). The effect of trellising on the Round Seedless grapes was evaluated for several quality characteristics (cluster and berry weight, length and width, total soluble solids, pH, acidity and total soluble solids/acidity), conducted in 2004/2005 and 2005/2006, in the Mediterranean region of Turkey, which has a subtropical climate. The vines were trained with the Australian and Lenz Moser trellising systems. For this variety and environmental conditions, no significant differences were found between the trellising systems used, where the cluster and berry properties (acidy and pH) were similar for both trellising systems (Kamiloğlu, 2012). The Seyval Blanc vines were evaluated over 5 years in 4 trellising systems: bilateral cordon, bilateral cordon sylvos, upright cordon-spur pruned and upright cordon-cane pruned. All systems were dormant pruned annually to leave a total of 40 count buds. In this study, cumulative yields over 5 years of the upright cordon systems were higher than vines in the sylvos system. The upright cordon system tended to increase vegetative growth and yield. On the other hand, the sylvos system reduced growth and resulted in a more open canopy with less Botrytis bunch rot and higher total soluble solids than other systems. (Ferree et al., 2002). Syrah grapes, grown in Minas Gerais State (Brazil) using VSP and GDC trellising systems, present similar pH, berry size and weight, and total phenolic contents. However, the GDC system produced grapes with the highest °Brix and lowest titratable acidity. On the other hand, berries from the VSP system presented lower anthocyanin content than those from the GDC system. GDC wines were characterized by high anthocyanin content and red colour,


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resulting in wines with high colour intensity. These data suggest that in the tropical region of Minas Gerais State, with high temperature and high sunlight intensity, the GDC trellising system, which protects bunches against excessive radiation, should be chosen (Mota, Amorim, Favero, Purgatto & Regina, 2011). Trellising systems are one of the most important factors for controlling vine growth and development and is essential to improve grape ripeness and productivity and also grape composition and quality (Falcão et al., 2008). As the overall physiological effects of a trellising system depend on specific environment conditions, it is not possible to extrapolate results to other growing areas, being important to adapt the trellising system to each vineyard and local conditions.

Oporto

Atlantic Ocean

Bairrada Appellation Spain

Lisbon

Figure 2. Location of Bairrada Appellation (Portugal).

Mulching To improve viticulture sustainability, it has been suggested that vineyard and winery wastes could be incorporated into mulches (described as any bulk material placed on the soil surface to control weeds and/or preserve moisture) for use in vineyards (Mundy & Agnew, 2002). The potential benefits from using mulches include weed control (Frederikson, Skinkis & Peachey, 2011), minimization of water loss and improved soil infiltration (Varga & Májer, 2004), increasing soil biological activity (Thomson & Hoffmann, 2007), and vine health status (Mundy & Agnew, 2002; Jacometti, Wratten & Walter, 2010). However, an increased incidence of fungal diseases due to increased soil water content can be a concern in mulched vineyards (Varga & Májer, 2004).



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Some studies carried out in New Zealand tested various mulch materials, namely vineyard pruning and thinning materials, green waste, pine bark, and grape pomaces, in several vineyards and indicated that Botrytis bunch rot did not increase (Mundy & Agnew, 2002; Jacometti, Wratten & Walter, 2007). However, the effect of mulching on vineyard pest control is not yet clear. Ideally, an economic assessment of the potential benefits of having a high number of natural pests in a vineyard is needed to develop a cost/benefit analysis of mulching applications (Thomson & Hoffmann, 2007). However, possible direct environmental benefits associated with a reduced herbicide load on the vineyard should not be ignored (Thomson, Sharley & Hoffmann, 2007). Mulching also has a great impact in grapes composition. It was shown that mulches composed of fresh plant residues increased grape tartaric acid and soluble solids (Varga & Májer, 2004). Furthermore, mulches composed of wastewater sludge in combination with bark, reduced the need for chemical weed control, without reducing vigour, yields, or grapes quality, and may improve grape yield, berry skin strength, and resistance (Jacometti et al., 2007). Thus, this was a suitable alternative to fertilizers for the sustainable production of grapes (Guerra & Steenwerth, 2012).

4. VARIETY ADEQUACY AS A STRATEGY FOR THE SUSTAINABLE VITICULTURE: A CASE STUDY The evaluation of the variety adequacy regarding the Appellation characteristics should be considered as a strategy for sustainable viticulture. As the environment is determinant for grapes characteristics, the winemakers can choose the best suited binomial environment/variety to produce a specific product and/or it is also possible to take advantage of this diversity and produce products with different characteristics, obtained from the same variety. With this perspective on sustainable viticulture it is possible to maximize the potential of each variety, minimizing further interventions in the winemaking process, which should contribute to the production of high quality wines, reducing costs and environmental impact. To demonstrate this concept, Bairrada, a Portuguese Appellation legally established in 1979 (Portaria nº 709-A/79, December 28), was selected as a case study. Bairrada Appellation is located in the Beiras region, in northwest of Portugal (Figure 2), located between Vouga and Mondego rivers, at east of Caramulo and Bussaco hills and at west of the Atlantic Ocean (Salvador, 1993). The total surface area of this region extends over 108.000 hectares, of which only 12.000 hectares are planted with grapes for wine production. For each Portuguese Appellation there are specific recommended and authorized grape varieties. According to Decreto-Lei nº 301/2003, for the Bairrada Appellation there are a list of 10 white and 16 red Vitis vinifera L. varieties, from these Bical and Castelão were studied in this chapter. Bairrada Appellation presents some natural environmental heterogeneity at several levels, namely soil type, climate, sunlight exposure, and altitude. The vineyards of Bairrada Appellation were planted mostly in soils from inferior and medium Jurassic, which are claycalcareous soils. Therefore, the soil in this region has some heterogeneity in its texture and was usually classified into 3 types: i) clayey; ii) clay-calcareous; and iii) sandy. This Appellation presents a wave relief with ca. 50 to 150 m of altitude. Furthermore, its climate is


210


essentially Atlantic and it may present some Mediterranean characteristics with maximum temperatures ranging between ca 25 and 35 ºC during summer (June-September). Further, in this Appellation of remarkable maritime influence, it is important to point out the large temperature range between day and night at the time the grapes ripe (reaching a range of 20 º C), which contributes to maintain the acidity of the grapes and consequently freshness of the wines. The occurrence of precipitation in Spring is common until the middle of April or, rarely, until the first days of May, conditioning mainly the early harvesting varieties. In some years, the abundance of rain in the second half of September may cause rotten whose extent depends on the varieties.

Vineyards under Study Using Bairrada Appellation as a case study to assess the viticulture sustainability, a quite thorough and up-to-date approach to assess the suitability of Castelão and Bical varieties (Figure 3) will be briefly presented. The varieties under study were harvested in 3 environments: 3 types of soils (clayey, clay-calcareous, and sandy), each one corresponding to different sunlight exposure, and topographical characteristics. Similar agricultural conditions were observed for all the environments under study: the vineyards were not irrigated, thinning was not performed, and also no kind of mulches was added to the soil. Furthermore the bilateral cordon trellising system was used in all the vineyards. Variety Castelão

Bical

Figure 3. Clusters and leaves of Vitis vinifera L. cv. Castelão and Bical.

Figure 4 shows the location and the environmental conditions of the vineyards, where the varieties under study were harvested. A high heterogeneity in the environments may be observed: Castelão and Bical were harvested at clayey, clay-calcareous and sandy soils. From each type of soil, each variety is cultivated at different altitude. For example, in sandy soil, Castelão is cultivated at 50 m and Bical at 150 m. Despite the fact that all the vineyards were implemented at low altitudes, the slight differences in altitude may influence the sunlight and wind exposures and the resulting vines temperature, which may influence grapes maturity and



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composition. Further, Bical vineyard with clayey soil (Figure 4 - 1 C) was near to pine trees, thus this vineyard can be influenced by its shadow, and the presence of trees can also protect this vineyard from winds. In addition, all the other vineyards under study were at an open space, which promoted higher sunlight exposure. Ancas

São Mateus Altitude (m)

2 CC

Anadia 1 S

Altitude (m) 100 2 S

50

6 Km

2 C

1 C 50

1 CC

100

São Lourenço do Bairro

Legend: 1 – Bical 2 – Castelão

Vale de Azar

Figure 4. Vineyards under study are located in Manuel dos Santos Campolargo, Herdeiros company, in Bairrada Appellation. Varieties (Vitis vinifera L cv. Bical and Castelão), altitude (50 to 200 m) and soil type (C – clayey soil; CC – clay-calcareous soil, and S – sandy soil) are indicated.

Varieties Characterization The classical physical-chemical parameters (berry weight, sugar content and acidity), volatile composition, phenolic content, and antioxidant activity were evaluated for the two varieties under study. Healthy-state grapes from Vitis vinifera L. cv. Castelão and Bical were collected from véraison to post-maturity, from the 2010 harvest. In order to obtain random samples and avoid picking grapes from other environments, every vine in the vineyards was previously marked. According to Table 1, at maturity similar berry weight and titratable acidity were observed for Castelão grapes obtained from clayey (ca. 50 m), clay-calcareous (ca. 125 m) and sandy (ca. 50 m) soils. However, differences may be noticed related to sugar content, which decreased in the following order: clayey, clay-calcareous and sandy grapes. Opposite trend was observed for white variety, as sandy grapes exhibited the higher sugar content. For Bical variety, similar results were observed for grapes obtained from clayey and claycalcareous soils both at ca. 100 m, whereas for grapes obtained from sandy soil (at ca. 150 m), higher berry weight and lesser titratable acidity were observed. It is crucial that wine producers can obtain this kind of information about their vines and varieties, which represent a useful tool to support winemaking decisions. According to specific desirable wine characteristics, the binomial variety/environment should be selected, minimizing further interventions in the winemaking process.


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Table 1. Weight, sugar and titratable acidity of Castelão and Bical berries in the sampling period (three independent aliquots were considered for each parameter, prepared from 200 grape berries harvested for each variety for each environment) Clayey Sugar (g L-1)

Titratable acidity (g L-1)

Véraison (day 0) Maturity (day 28) Postmaturity (day 35)

0.84 (2.3)* 1.85 (3.4)

51.0 (1.1) 241.4 (0.7)

29.3 (0.5) 5.2 (2.2)

1.81 (2.5)

255.0 (0.8)

4.3 (1.0)

1.91 (1.5)

227.8 (1.1)

Véraison (day 0) Maturity (day 35) Postmaturity (day 42)

1.09 (1.0) 1.38 (1.0)

102.0 (1.6) 173.4 (0.5)

16.0 (0.3) 5.7 (0.8)

0.98 (1.3) 1.18 (2.3)

1.30 (4.3)

170.0 (1.3)

5.7 (1.5)

1.20 (0.9)

Samplin g time

*

Clay-calcareous Berry Titratable Sugar weight acidity (g L-1) (g) (g L-1) Castelão 0.95 44.2 29.7 (1.7) (0.7) (0.7) 1.84 221.0 5.0 (1.8) (1.2) (1.9)

Berry weight (g)

Sandy Berry weight (g)

Sugar (g L-1)

Titratable acidity (g L-1)

1.16 (1.6) 1.88 (1.4)

64.6 (1.0) 217.6 (0.9)

28.1 (0.5) 5.5 (2.1)

4.6 (2.5)

_

_

_**

Bical 115.6 (1.7) 170.0 (0.6)

15.5 (0.3) 6.1 (1.4)

0.99 (1.1) 1.44 (3.4)

119.0 (1.4) 187.0 (0.5)

12.8 (0.7) 4.2 (2.8)

171.7 (1.3)

5.4 (1.6)

1.33 (1.4)

183.6 (1.4)

4.4 (3.4)

Relative standard deviation (RSD, % in parentheses). grapes not available.

**

To go further in the field of studying the suitability of the varieties under study to the different environmental conditions, beyond these classical parameters, other molecules such as volatile and phenolic compounds, were also considered. Varietal aroma is directly associated to the grape variety, biosynthesized during grape development, being dependent on climate, soil type, and grape maturity. The terpenic compounds and C13 norisoprenoids are associated to the varietal aroma (Rocha et al., 2000). The pre-fermentative aromas result from several mechanic or technological operations (transport, crushing, maceration and clarification) performed before the beginning of the fermentation process, and this aroma results mainly from C6 alcohols and aldehydes (Coelho, Rocha, Barros, Delgadillo & Coimbra, 2007). The varietal and pre-fermentative volatile components were studied by headspace-solid phase micro extraction combined with comprehensive two dimensional gas chromatography with time-of-flight mass spectrometry (HS-SPME/GC × GC-ToFMS), a high sensitive and throughput methodology, previously used to characterized Vitis vinifera L. related products (Perestrelo, Barros, Câmara & Rocha, 2011). Figure 5 shows a scheme of the methodology used for volatile profile analysis. Figures 6 and 7 show the screening of volatile components during ripening for Castelão and Bical varieties, respectively, and the results obtained were distributed over the chemical families of C6 alcohols and aldehydes, aromatic alcohols, monoterpenoids, C13 norisoprenoids, sesquiterpenoids, and diterpenoid. According to both Figures 6 and 7, the content of varietal and pre-fermentative volatile components increased in the first or second weeks of analysis and then tends to stabilize, or continues to increase at maturity, and then a decreasing was observed. The exception was observed for C13 norisoprenoids that increase in the first week of analysis, and then a continuous decrease was observed. This could be explained by the fact that between véraison



213

and maturity occurs the glycosylation of norisoprenoids, and this leads to a decrease of these compounds in the free form (Baumes et al., 2002; Coelho, Rocha, Delgadillo & Coimbra, 2006). Despite the observed variability during Castelão grapes ripening, it was observed that grapes from clay-calcareous soil tend to exhibit the higher volatile content, especially on terpenoids. On the other hand, grapes from sandy soil tend to present the lower content (Figure 6). Besides, cultivation altitudes of the 3 vineyards were also different (Figure 4): the higher volatile content was found for the vineyard at ca. 125 m of altitude (clay-calcareous soils) followed by vineyards at ca. 50 m (clayey and sandy ones). Furthermore, in general, clayey soils have better water retention capacity and volumetric water content than sandy soils. In opposition, the drainage is larger in sand than in clay. Thus, the clay-calcareous and clay soils that have good water-holding and drainage capacities should allow obtaining wines richer in volatiles than sandy soils (Coelho et al., 2009).

5 min. in a 5 min. in a thermostatic thermostatic bath at 60.0 ºC bath at 60.0 ºC 4 g of grapes 42ggof ofgrapes NaCl 2 g of 5 mL ofNaCl H2O 5 mL of H2O

Injector Injector T = 250 ºC T = 250 ºC

Extraction time: 20 min Extraction time: 20 60min ºC0.1 ºC Extraction temperature: 60.0 Extraction temperature: 60.0 0.1 ºC

N2 N2 Injector

ModulationInjector time: 6 s Modulation time: 6 s Modulator Modulator

100 spectra/s 100 spectra/s EI:70 eV EI:70 eV 33 - 300 m/z Modulator control 33 - 300 m/z

Modulator control Time, temperature Time, temperature

Column 1: Non-polar Column 1: Non-polar

1st column HP-5 1st column HP-5 30 m 0.32 mm I.D., 0.25 mm 30 m 0.32 mm I.D., 0.25 mm

TOFMS TOFMS detector detector

Column 2: Polar Column 2: Polar Detector: ToFMS Detector: ToFMS

2nd column DB-FFAP 2nd column DB-FFAP 1 m 0.25 mm I.D., 0.25 mm 1 m 0.25 mm I.D., 0.25 mm nd Dimension 2nd 2Dimension (s) (s)

Vitis vinifera L. Vitis grapes vinifera L. grapes 1/β = 0.5 1/β = 0.5

1st Dimension (s) Data analysis 1st Dimension (s)

Data analysis

Data Data Processing Processing

Figure 5. Scheme of HS-SPME/GC × GC-TOFMS used for grape volatiles analysis.

For Bical variety, grapes from clay-calcareous and sandy vineyards exhibit the higher volatile content comparatively to grapes from clay soils (Figure 7). The Bical vineyards present slightly differences in altitudes and also in sunlight exposure (Figure 4): the vineyard with clayey soil (ca. 100 m) is influenced by the shadow of pine trees, presenting lower sunlight exposure, which may modulate grapes volatile composition. The higher level of


214


volatile components of Bical grapes from clay-calcareous (ca. 100 m) and sandy soils (ca. 150 m) can also be related to the higher sunlight exposure (Bureau, Baumes & A., 2000; Baumes et al., 2002), explained by its localization in open spaces. The level of phenolic compounds and antioxidant activity is modulated by environmental conditions under which the grapes are cultivated. Thus, in order to study these parameters, the total phenolic content was determined using the Folin–Ciocalteu method, and the antioxidant activity was determined using DPPH• (1,1-diphenyl-2-picril-hidrazil) radical scavenging assay (Paixão, Perestrelo, Marques & Câmara, 2007). Similarly to the tendency observed for Castelão volatile components, at harvesting day, grapes from clay-calcareous soil, at ca. 125 m of altitude, presented higher phenolic content, as well as higher antioxidant activity, followed by grapes from the clayey and sandy soils, both at ca. 50 m (Figure 8). Aromatic alcohols

Total chromatographic area x106 (a.u.)

C6 compounds 250.0

10.0

200.0

8.0 *

150.0

*

4.0

50.0

2.0

0.0

0

7

Clayey


6.0

100.0

14

21

28

Clay-calcareous

35

0.0

0

Monoterpenoids

7 Clayey

Sandy

21

28

Clay-calcareous

17.5

60.0

14.0

45.0

35 Sandy

C13 norisoprenoids

75.0

10.5 *

30.0

7.0

15.0

3.5

*

0.0 0

7

14

21

28

35

0.0

0

7

Sesquiterpenoids Total chromatographic area x106 (a.u.)

14

14

21

28

35

Diterpenoid

15.0

2.0

12.0

1.6

9.0

1.2 *

*

6.0

0.8

3.0

0.4 0.0

0.0 0

7

Clayey

14

21

Days Clay-calcareous

28

35

Sandy

0

Clayey

7

14

21


28

35

Sandy

Figure 6. Varietal and pre-fermentative volatile components evolution during Vitis vinifera L. cv. Castelão ripening, obtained from different environments. The harvesting day is indicated with a dash rectangle. The areas are expressed as arbitrary units (a.u.). * Grapes from sandy soil were not available for the last harvest.


Efficient Use of Non-Renewable Natural Resources for Quality Wine... Aromatic alcohols


C6 compounds 300.0

10.0

250.0

8.0

200.0

6.0

150.0

4.0

100.0

2.0

50.0

0.0 0

7

14

21

28

35

42

0

7


14

21

28

35

42

28

35

42

28

35

42

C13 Norisoprenoids

Monoterpenoids 50.0

25.0

40.0

20.0

30.0

15.0

20.0

10.0

10.0

5.0 0.0

0.0 0


215

7

14

21

28

35

0

42

Sesquiterpenoids 12.5

1.0

10.0

0.8

7.5

0.6

5.0

0.4

2.5

0.2

7

14

21

Diterpenoid

0.0

0.0 0

Clayey

7

14

21


28

35

42

Sandy

0

Clayey

7

14

21


Sandy

Figure 7. Varietal and pre-fermentative volatile components evolution during Vitis vinifera L. cv. Bical ripening, obtained from different environments. The harvesting day is indicated with a dash rectangle. The areas are expressed as arbitrary units (a.u.).

At harvesting day, Bical grapes obtained from sandy soil (ca. 125 m) followed by grapes from clay-calcareous (ca. 100 m) and clayey soils (ca. 100 m), presented higher phenolic content and antioxidant activity (Figure 9). As grapes from sandy and clay-calcareous exhibited the higher volatile content, thus, it is possible to conclude that Bical grapes obtained from sandy soil seems to be related to higher phenolic content and antioxidant activity, and also volatile components. Table 2 summarizes the results for volatile components, phenolic content and antioxidant activity of Castelão and Bical varieties. Considering that the same viticultural practices were used for both varieties, the environmental conditions seem to modulate the grape composition take into account the parameters under study. The behaviour of each variety is different in the same environment, i.e., sandy related environment seems to favour Bical white grape composition, but the opposite trend was observed for Castelão red variety. Thus, this suggests that it is possible to take advantage of the natural resources and produce products with different characteristics obtained from the same variety, minimizing costs during the


216


winemaking process, increasing the sustainability in this sector. Furthermore, these results also showed that this type of study includes the evaluation of a network of several parameters, such as soil, altitude, sunlight exposure, water availability, on grape composition. Using these natural systems, representing uncontrollable environments, it is not possible to study one parameter at a time. Thus, as these environmental conditions are very complex and influence the grapes characteristics at several levels, in order to access the sustainability of viticulture, it is important in future studies to develop multivariate models to evaluate and predict relevant relationships between these natural parameters and grape quality in order to increase the understanding of the variety suitability regarding the Appellation characteristics. A

1800

Galic acid (mg/L)

1400

1000 *

600

200 0

7

14

21

28

35

B

% DPPH remaining

100

80 *

60

40

20 0

7 Clayey

14

Days

Clay-calcareous

21

28

35

Sandy

Figure 8. Total phenolic components (A) and antioxidant activity (B) evolution of V. vinifera L. cv. Castelão, during ripening. The harvesting day is indicated with a dash rectangle.* Grapes from sandy soil were not available for the last harvest.



217

A

Galic acid (mg/L)

350

300

250

200

% DPPH remaining

B

0

7

14

0

7

14

21

28

35

42

21

28

35

42

100

80

60

40

Days Clayey

Clay-calcareous

Sandy

Figure 9. Total phenolic components (A) and antioxidant activity (B) evolution of Vitis vinifera L. cv. Bical during ripening. The harvesting day is indicated with a dash rectangle.

5. MULTIVARIATE REGRESSION MODELS Over many years research has been aimed at developing a simple model or method that could define and predict grape and wine quality. In fact, in the last years, different types of models constructed by application of different multivariate methods were developed (Due et al., 1993; Marais, Calitz & Haasbroek, 2001; Valdés-Gómez, Celette, García de CortázarAtauri, Jara-Rojas & Gary, 2009; Santos, Malheiro, Karremann & Pinto, 2011; Urhausen, Brienen, Kapala & Simmer, 2011; Santos, Wample, Sachidhanantham & Kaye, 2012). Multivariate analysis, machine learning and pattern recognition techniques play an important role in the assessment of the relationships that may occur between several factors such as climate, viticulture techniques, vineyard ecosystem, and grape composition, relevant for the definition of the wine quality. The general approach could include, on the one hand, unsupervised studies of the major sources of variability, using Principal Component Analysis


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(PCA) and Hierarchical Cluster Analysis (HCA). On the other hand, the use of factors driven methods such as APCA (ANOVA PCA), Partial Least Squares-Discriminant Analysis (PLSDA), and Canonical Correlation Analysis (CCA), could bring out important relationships among measured factors as well as to assess the influence of confounding factors that are pervasive in the analysis of environmental uncontrolled measures. Some models to predict wine quality based on chemical, climatic and agricultural data are available in literature. A discussion about these examples should be important to highlight their relevance and drawbacks. A model to predict wine quality that correlates microclimatic data (temperature and radiation) with volatile component concentrations and wine sensory parameters of Sauvignon Blanc variety, was developed (Marais et al., 2001). In this model, seventy-two independent data sets consisting of 3 harvests (1997 to 1999), 3 climatically different regions in South Africa, 2 canopy treatments (canopy is a function of different climatic and viticultural factors, which determines the effects of temperature and radiation), and 4 ripening stages, were developed. The microclimatic data within the 2 canopies were recorded as independent variables while the grape and wine measurements such as volatiles (monoterpenes and norisoprenoids) and sensory data (fruitiness and vegetative/ asparagus/ green pepper intensity) were recorded as dependent variables. Pearson´s correlation coefficients were calculated between the above mentioned independent and dependent variables (Marais et al., 2001). The model utilises above- and within-canopy radiation and can explain 68.8% of the variation in the cultivar-typical vegetative/asparagus/green pepper intensity of Sauvignon Blanc wine. Other selected example is a study carried out from 2005 to 2008 to calibrate and apply near infrared spectroscopy to assess the spatial behaviour of 3 grape varieties (Cabernet Sauvignon, Syrah and Merlot) quality parameters along the vineyards of Sao Joaquin Valley (USA), and promote differential mechanical harvesting, according to quality zone delineation, was developed (Santos et al., 2012). The quality indicators (anthocyanin content, pH, titratable acidity and soluble solids) were subject to geospatial modelling, and calibrations were developed using PLS. Subsequently, the data set was utilized to delineate “within-field” grape quality zone and to determine the harvest time. The approach for field prediction of grape quality parameters and zone delineation allowed to distinguish two wines based on their different chemical composition, principally on anthocyanin content (Santos et al., 2012). The studies reported above and others available on the literature are restricted in time, in space and in the number of parameters evaluated, consequently, the prediction power of the models is very limited. As the ultimate goal of the multivariate analysis in the context of sustainability should be to combine all the different measurements to provide predictive/classification models that increases the knowledge about the relationships that arise between the many different measured parameters, one should envisage a data fusion approach. Each set of different parameters could be collected into separate data blocks which can then be more effectively analyzed using multi-block techniques such as "Common Component and Specific Weights Analysis" - CCSWA. The objective of CCSWA is to find the directions describing common distributions of the samples in the spaces defined by the different data blocks. This approach will allow to better understand which parameters are really relevant to the vineyard ecosystem and also to recognize which one of those parameters has significant influence onto the wine quality.



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Table 2. Castelão and Bical grapes composition in distinct environments Variety Castelão

Bical

Altitude (m)

Soil type

50 125 50 100 100 150

Clayey Clay-calcareous Sandy Clayey Clay-calcareous Sandy

Climate • Temperature • Sunlight exposure • Precipitation

Volatile components Middle Higher Lower Lower Higher

Topography • Altitude • Slope

Phenolic content Middle Higher Lower Lower Middle Higher

Antioxidant activity Middle Higher Lower Lower Middle Higher

Soil • Composition • Water retain capacity • Heat retain capacity

Water status Viticultural practice • Irrigation •Thinning • Trellising • Mulching

Vine behaviour

Grapes Composition

Yield production

Harvesting moment

Wine making process Wine quality

Future? Appellation / Variety Figure 10. Environmental factors and wine making process that influence wine composition and quality.


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CONCLUSION AND FUTURE TRENDS The future of the viticulture should incorporate the sustainability concept as a strategy of innovation in equilibrium with tradition, highly associated and valorised in this sector. This concept may be explored in different areas, namely i) in the adaptation of agricultural practices and winemaking technologies to each variety and region, reducing costs and environmental impacts, ii) in the implementation of more eco-friendly agriculture with respect to environment, iii) in the development of new products or improvement of quality of the wine usually produced taking advantages from variety potential, iv) in the development of tools that may support winemakers and local/national/international agencies decisions and policies, v) in the development of marketing and communication with the consumers. The grapes characteristics and composition, and wine quality depend on a complex network of variables known to influence viticulture, including environmental conditions, and agricultural practices (Figure 10). Thus, the detailed knowledge of these specificities for each region is crucial for the sustainability in this sector. With this perspective on sustainable viticulture it is possible to maximize the potential of the varieties, minimizing further interventions during winemaking process, which should contribute to the production of high quality wines, reducing costs and also environment impact. To go further into the sustainable viticulture, several other approaches should also be considered. The genetic diversity of the varieties and the exploitation of endophyte-plant interactions (defined as fungi and bacteria that colonize the internal tissue of plants, some even reside inside the plant cells, showing no external sign of infection or negative effect on their host) that can benefits plant growth and control diseases, are two lines that should be more explored. Genetic diversity is helpful for vine improvement (This, Lacombe & Thomas, 2006). Understanding the entire variability existent within a variety and its distribution across the different regions (so the different environmental conditions and the agricultural practices) where it is grown, is very important allowing a more efficient recognition and preservation of genetic resources, as well as higher genetic gains through selection (Gonçalves, St.Aubyn & Martins, 2010). This helps to have a more realistic knowledge about the influence of the local conditions in the variety suitability. Furthermore, endophytes can promote plant growth and yield, can act as biocontrol agents, have potential to remove soil contaminants by enhancing phyto remediation and may play a role in soil fertility. An understanding of the mechanisms enabling these endophytes to interact with plants will be essential to fully achieve the biotechnological potential of efficient plant-endophytes partnerships for a range of applications (Ryan, Germaine, Franks, Ryan & Dowling, 2008). Until now, this concept was not properly valued in the context of the viticulture; however, several applications were also explored in other areas of the agriculture and forestry. The use of endophytes to control plantpathogenic bacteria and fungi is receiving increasing attention as a sustainable alternative to synthetic pesticides. In order to reduce the input of pesticides and fertilizers and to make ecofriendly agriculture, endophytes were used to promote the sustainable production of biopesticides and biofertilizers in conjunction with biomass and bio energy crops (Germaine et al., 2006; Ryan et al., 2007). To understand the variety suitability regarding the region characteristics, and mainly considering the uncontrollable environment conditions, the development of models in order to obtain fast and reliable information that helps the winemaker decision is crucial. Robust



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models including long term data may represent a key tool for the analysis of emerging policies, price sets, and to reduce the environmental impact on wine quality, by implementing adequate winemaking technologies. This concept was already successful applied in dairy farming systems (Doole, Romera & Adler, 2012; Doole, Marsh & Ramilan, 2013). Finally, it is important to point out that the concept of sustainable viticulture is not yet globally implemented. So far, wine industry is doing its own self assessment, and therefore, many government regulatory agencies have not become involved. Clearly, sustainability in viticulture sector is going to continue to grow, step by step, in the future. As the local environmental and agriculture conditions influence the quality of each variety, in order to establish a viable and useful concept, it must remain a local program and be modified to meet local needs. Thus, in order to implement sustainable viticulture programs in new areas, the wine producers of each region need to know the potentialities of their vineyards and varieties to go further in this field. With the implementation of these sustainable programs the consumer awareness and acceptance of sustainable winegrowing and winemaking practices grows, thus the market will see growing acceptance and demand for wines produced from sustainably farmed grapes and made in sustainable vineyards. In order to help the consumers identifying and distinguishing these products from others, the wine producers need to cooperatively develop and support appropriate marketing programs.

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