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Can berry composition be explained by climatic indices? Comparing classical with new indices in the Portuguese Do region
Lopes1, CM; Egipto1, R.; Pedroso2, V; Braga3, R; Neto4, M; Pinto1, PA CBAA, Instituto Superior de Agronomia, Lisboa DRAPC/Centro de Estudos Vitivincolas do Do, Nelas, Portugal. 3 Escola Superior Agrria de Elvas, IPP, Elvas 4 ISEGI, Universidade Nova de Lisboa *Corresp. Author: Carlos M. Lopes, phone email:carlosmlopes@isa.utl.pt

+351213653450,

Keywords: Bioclimatic indices, statistical modeling, titratable acidity, total soluble solids, grapevine, Touriga Nacional. Abstract Climatic data collected between 1963 and 2010 in the Portuguese Do region were analyzed to evaluate the relationship between climatic indices, seasonal weather and the berry composition of the red variety Touriga Nacional. The trends over time for the classical temperature-based indices (Growing Season Temperature GST -, Growing Degree Days GDD -, Biologically Effective Degree Days BEDD -, Huglin Index HI -, Cool Night Index CI) were significantly positive and can be mostly attributed to the effects of climate change. The Dryness Index (DI) showed a negative trend although not significant. These indices were able to explain 9 and 45% of the variability in total soluble solids and titratable acidity, respectively, using a multiple stepwise regression analysis. The proportion of explained variability was much improved, to 52% for total soluble solids and 65% for titratable acidity, when new climatic indices were used. The new indices resulted from the generalization of the classical indices based upon chronological time specification as well as taking into consideration the phenological time instead. Our data shows that the classical climatic indices were not able to sufficiently explain the berry composition and that new climatic indices should be used for a better understanding of the climate drivers of berry quality. INTRODUCTION Climate affects all phases of the grapevine biological cycle being a key factor in determining the geographically suitable areas for viticulture. Within the meteorological parameters, temperature has a central role for viticulture production influencing the annual grapevine growth, yield, berry composition and wine style (Gladstones, 2011). The importance of temperature has resulted in the development of various bioclimatic indices (BI) based on temperature such as (Jones et al., 2010): Growing Season Temperature (GST), Growing Degree Days (GDD), Huglin Index (HI), Biologically Effective Degree Days (BEDD) and Cool Night Index (CI). Besides temperature, in warm and dry regions, water availability also plays an important role in viticulture mainly in warm and dry regions with Mediterranean climate. Several indices including water

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availability have been proposed (e.g. Branas, Bernon and Levadoux Hidrotermic Index, Constantinescu Index, Hidalgo Index, Dryness Index) being the Dryness Index (DI) the most used. The DI is based on a water balance in the theoretical budburst-harvest period (from 1 of April to 30 of September in the Northern Hemisphere) (Riou, 1994). The BI are mainly used to classify and compare regions in terms of climate suitability to producing grapes, to identify and characterize new viticultural regions and to determine the match between cultivar and region (Gladstones, 1992; Tonietto and Carbonneau, 2004).Some researchers have also used BI to explain the variability in berry composition and wine quality, however, in most cases, the resulting models can only explain a low proportion of berry composition variability at harvest (e.g. Riou, 1994; Giomo et al., 1996). One justification for the low robustness of BI models to explain berry composition variability might be the fact that many of the existing indices are only temperature based. Another important justification might be related to the accounted period (budburst-harvest) which does not take in consideration each phenological phase such as, for example, berry ripening. This work aims to test the relative performance of various classical and new climatic and bioclimatic indices, in explaining the variability of the berry composition of the Touriga Nacional variety in the Portuguese Do region during 47 seasons. MATERIALS AND METHODS Vineyard Site Viticultural data from 1963 to 2010 were collected in two experimental vineyards of the Do winegrowing region Research Centre (Centro de Estudos Vitivincolas do Do), located in Nelas, North East of Portugal (latitude 40 31 'N, longitude 7 51'W, elevation 440 m). The data from 1963 to 1987 were collected in a vineyard plot planted for a rootstocks/variety affinity study. In this work we used only data from one variety Touriga Nacional - grafted on seven rootstocks (420A, 161-49, 3309 Couderc, 110 Richter, 99 Richter, 420 A and 161-49). The vines were trained to a vertical shootpositioned trellis, pruned to a double Guyot system and spaced 1.1 m within and 1.8 m between rows. The data from 1995 to 2010 was collected in two more recent vineyard plots planted in the same research centre, with a slightly different row-spacing (1.8m x 1.0m) and spur pruned in a bilateral Royat cordon. The rootstocks used here were 1103P, SO4, 99R and 110R. The soil is from granitic origin, with a coarse texture and acid pH, with very good infiltration capacity, low organic matter content, medium content of phosphorus and high content in potassium. Climate, Phenological and Berry Composition Data Daily minimum and maximum temperatures and rainfall were collected from a weather station located within the Research Centre in the middle of the experimental vineyards. Budburst dates were noted when 50% of the buds reached Baggiolinis B stage. Flowering dates were noted when 50% of flowers were open and veraison when 50% of the berries changed color. Total soluble solids and titratable acidity were determined at harvest from juice samples made from each strain of rootstock.

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Data Analysis For each season the following classical bioclimatic indices were calculated: growing season average temperature GST (Jones et al., 2010), Growing Degree Days GDD (Amerine &Winkler, 1944), Biologically Effective Degree-days - BEDD (Gladstones, 1992), Huglin Index HI (Huglin, 1978), Cool Night index - CI (Tonietto and Carbonneau, 2004) and Dryness index - DI (Riou, 1994).Using phenological and chronological periods a set of new climatic and bioclimatic indices were also calculated. A database with classical and new climatic indices, phenological and berry composition data (total soluble solids and titratable acidity) was built using available data. This database has the following missing values: Phenology: 1989, 1990, 1993 and 1998; total soluble solids: 1988 to 1994 and 1998; titratable acidity: 1988 to 1995 and 1999 to 2003; climate: 1973, 1975, 1992 to 1997. For each set of climatic indices a multiple regression analysis between berry composition data (dependent variables) and climatic indices (independent variables) was performed. Variable selection was done with a forward stepwise regression with 0.10 critical F statistic. For the set of the new climatic indices, as a result of the large number of independent variables and the resulting computational impossibility of equal consideration in a multiple regression variable selection algorithm, the selection of the best regression model followed a semi-automatic progressive process: (1) All independent variables significantly correlated to dependent variables were selected; (2) Amongst auto correlated independent variables, the most agronomic meaningful variable was chosen to represent each set; (3) Determination of the best (highest r2) 30 regression models using general multiple linear regression stepwise procedure. At this step only models with 4 to 6 variables were considered due to over fitting and collinearity issues. Finally, (4) the best model was refitted using multiple linear stepwise regression and variable collinearity was also re-checked. If collinearity persisted, the linear regression model was recalculated (after removing one of the collinear variables) and its r2 compared with the nest general regression model on the ranking list. RESULTS Classical Bioclimatic Indices The GDD values range from 1277.4 to 2051.6 C.day, with the average over the record period being 1601.3 147.1 C.day (Fig. 1A). The evolution between 1963 and 2010 shows a significant increase (6.3 C.day per year) over the recorded period resulting in an increase in the climate region classification proposed by Amerine and Winkler (1944). According to the established limits, the average GDD values result in a classification of the Do winegrowing region of Region II until 1985 and of Region III between 1985 and 2010. The GST average for the period was 17.4 0.7 C and present a similar trend to that of GDD (data not shown). The BEDD values range from 1224.4 to 1578.4 C.day, with the average of 1380.7 77.4 C.day (Fig. 1B). As observed with the GDD, the BEDD present an increase of 2.5C.day per year. Table 1 presents the average values for the three BI included in the Multicriteria Climatic Classification system proposed by Tonietto and Carbonneau (2004). According to this classification system, the Do winegrowing region is classified as having a temperate (Fig. 1C) and moderately dry (Fig. 1D) climate, with cool nights during the ripening period. Tonietto and Carbonneau (2004) stated that this climate class has no heliothermal constraint to ripen later varieties such as Cabernet Sauvignon, Ugni Blanc or

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Syrah, and have a ripening period that occurs under conditions where the maximum threshold of favorable ripening night temperature is not exceeded for any variety. The same authors also stated that the values of the DI indicate a moderately dry condition where the vines will potentially face a certain level of dryness, conducing to a significant stomatal regulation usually favorable to maturation. However, the DI values present a very high variability ranging from -168.7 to +194.9 mm (Fig. 1D). Using the classical BI as independent variables, a forward stepwise regression analysis was performed with berry total soluble solids (TSS) and titratable acidity (TAc) as dependent variables. The best model for TSS explained only 9% of total soluble solids (TSS) variability (Eq.1) with DI and HI as the variables with higher and lower contribution, respectively. For TAc only two variables were selected to the model. HI was the first one, explaining 33% of the variability and DI the last one with a lower contribution (12%). The final model explained 45% of the titratable acidity variability (Eq. 2). TSS (Brix) = 14.700 + 0.002 * DI + 0.164 * CI 0.002 * HI (Eq. 1) R2 = 0.09 (p < 0.01); RMSE= 1.2; n = 127 TAc (g tart. ac./L) = 21.5 0.007 *HI 0.007 * DI (Eq. 2) R2 = 0.45 (p < 0.001); RMSE= 1.0; n = 122 New Climatic and Bioclimatic Indices In order to test the use of the new climatic and bioclimatic indices as berry composition drivers, the classical BI were replaced in the stepwise regression by a subset of new indices selected according to the statistical procedures reported in the chapter 2. For TSS six variables entered into the model (Table 2), being the average temperature of July (TJul) the first one selected, explaining 9.7% of the variability, and the total rainfall in the veraison-harvest period (RVH) the last one with a 4.4% contribution. The final model explained 52% of total soluble solids variability (Eq. 3). TSS(Brix) = -47.413+0.542 * TJul + 0.137 * Hdate+0.220 * TbH + 0.403 * TSep 0.017 * DSep - 0.004 * RVH (Eq. 3) R2 = 0.52 (p< 0.001); RMSE= 0.8; n = 127(For legend see Table 2) For titratable acidity six variables entered into the model, being ET0Mar-Sep the first one, explaining 29.4% of the variability and ET0VH was the last one selected, explaining 2.8%. The final model explained 65% of the TAc variability (Eq. 4). TAc(g tart. ac./L) = 27.217 0.022 * ET0Mar-Sep + 0.412 * DTRAug 0.025 * ETRJul 0.025 * RAug-Sep 0.026 * DSep -0.007 * ET0VH (Eq. 4) R2 = 0.65 (p<0.001); RMSE= 1.9; n =132(For legend see Table 2)

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DISCUSSION AND CONCLUSIONS The trends over time of the classical temperature based indices indicate an increase in the air temperature. These results are consistent with Portuguese climate change scenarios for which higher climatic variability and extreme events probability, rainfall concentration in the coolest months, and the increase in the air temperature are anticipated (Pinto et al., 2006). When we used the new climatic and bioclimatic indices that include temperature and rainfall, calculated in months (or phenological phases) with large influence on the vegetative growth and fruit ripening, a higher berry quality components variability were described by the models. The obtained model for TSS integrates the positive effect of temperature in the month before veraison and during ripening, as well as the date of harvest, in sugar accumulation. The negative effects on sugar accumulation of rainfall at the veraison-harvest period and/or soil water deficit in September may be explained by a diluting effect (Williams& Matthews, 1990). The titratable acidity model integrates the negative effects of ET0 (accumulated during all vegetative cycle and in summer months), rainfall and soil water deficit during ripening, revealing the known effects of soil water availability on must acidity, mainly through the effect on malic acid (Williams& Matthews, 1990; Esteban et al., 1999). Our results show that the classical bioclimatic indices, based upon air temperature, were not sufficiently robust to explain berry composition. New indices encompassing phenological, climatic and soil water balance variables related to the ripening period should be preferred for a better understanding of the climate drivers of berry quality. ACKNOWLEDGMENTS The research leading to these results has received funding from the Fundao para a Cincia e Tecnologia (FCT), under the grant agreement PTDC/AAC-MB/105024/2008, project SIAMVITI. Literature Cited Amerine, M.A. andWinkler, A.J. 1944. Composition and quality of musts and wines of California grapes. Hilgardia, 15:493-675. Esteban, M.A.; Villanueva, M.J. and Lissarrague, J.R. 1999. Effect of irrigation on changes in berry composition of Tempranillo during maturation. Sugars, organique acids and mineral elements. Am. J. Enol. Vitic., 50:418433. Giomo, A.; Borsetta, P. and Zirono, R. 1996. Grape quality: research on the relationships between grape composition and climatic variables. Acta Horticulturae, 427:227-285. Gladstones, J. 1992. Viticulture and environment. Winetitles, Adelaide, Australia. Gladstones, J. 2011. Wine, Terroir and Climate Change. Wakefield Press. Adelaide, SA. Huglin, P. 1978. Nouveau mode dvaluation des possibilits hliothermiques dun milieau viticole. C.R. Acad. Agr. France, 64: 1117-1126. Jones, G.V.; Duff, A.A.; Hall, A. andMyers, J.W. (2010). Spatial analysis of climate in winegrape growing regions in the Western United States. Am.J.Enol.Vitic., 61:313326. Pinto, P.A.; Braga, R. and Brando, A.P. 2006. Agricultura. In: Santos, F.D.; Miranda,P. (eds). Alteraes climticas em Portugal. Cenrios, Impactos e Medidas de Adaptao. Projecto SIAM II, pp. 209-231. Gradiva, Lisboa.

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Riou, C. 1994. Le dterminisme climatique de la maturation du raisin: application au zonage de la teneur en sucre dans la communaut europenne. Luxembourg, Office des Publications Officielles des Communauts Europennes. Tonietto, J. and Carbonneau, A. 2004. A multicriteria climatic classification system for grape-growing regions worldwide Agricultural and Forest Meteorology. 124: 81-97 Williams, L.E. and Matthews, M.A. 1990. Grapevine. In: Stewart, B.A., Nielsen, D.R. (Eds.), Irrigation of Agricultural Crops. Series of Agronomy, vol. 30. American Society of Agronomy, Madison, Wisconsin, USA, pp. 10191055.

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Tables Table 1 - Bioclimatic indices integrating the Multicriteria Climatic Classification (MCC) system calculated for Nelas (Do), Portugal. HI - Huglin index; CI - Cool Night index; DI - Dryness index.
PERIOD 1963-2010 MCC classification HI (C.day) 2026.6 172.3 HI - 1 CI (C) 12.6 1.3 CI + 1 DI (mm) -5.8 87.2 DI + 1

232 233 234 235 Table 2 Summary of stepwise regression analysis between berry total soluble solids and titratable acidity (dependent variables) and new climate indices (independent variables) (n=127 for TSS; n=132 for TAc).
VARIABLE TJul Hdate TSS (Brix) TbH TSep DSep RVH ET0Mar-Sep DTRAug TAc (g tart. ac./L) ETRJul RAug-Sep DSep Step +in/-out 1 2 3 4 5 6 1 2 3 4 5 R-square change 0,093 0,100 0,139 0,056 0,088 0,044 0,294 0,067 0,085 0,076 0,105 Multiple R-square 0,093 0,193 0,332 0,387 0,475 0,519 0,294 0,361 0,446 0,522 0,627 p-level <0,001 <0,001 <0,001 <0,01 <0,001 <0,01 <0,001 <0,001 <0,001 <0,001 <0,001

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0,028 0,655 ET0VH 6 <0,01 Legend: TJul- average temperature in July; Hdate- harvest date; TbH-average temperature in the 15 days before harvest; TSep-average temperature of September; DSep-soil water deficit in September(Thornthwaite-Matter); RVH- total rainfall in the veraison-harvest period; ET0Mar-Sep- Reference Evapotranspiration MarchSeptember; DTRAug- daily temperature range in August; ETRJul- actual Evapotranspiration in July; RAug-Seprainfall August-September; ET0VH- Reference Evapotranspiration in the veraison-harvest period.

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Figures

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Figure 1 Evolution of Growing Degree Days (A), Biologically Effective Degree Days (B), Huglin Index (C) and Dryness Index (D) over 39 years (1963-2010 2010). Dotted lines represent the class limits of each index.