Performance of resistant grape varieties (PIWI), 'Felicia', 'Calardis Blanc' and 'Helios' in two locations of Santa Catarina State (BR)

ABSTRACT Downy mildew (Plamopora viticola) is one of the major problems to produce wine grapes in Brazil, a main cause of yield losses in the Southern region of the country. An alternative to the use of pesticides is the use of resistant varieties (PIWI): wine grapes combining high oenological potential and resistance to diseases. The goal of this work was to assess the vegetative and productive performance and maturation of three PIWI varieties (‘Felicia’, ‘Calardis Blanc’ and ‘Helios’) produced at altitudes of 840m and 1250m in the 2020, 2021 and 2022 vintages. The study is focused on four phenological stages: budbreak (BBCH 7), full bloom (BBCH 65), veraison (BBCH 81) and maturity/harvest (BBCH 89). The criteria for measuring the productive performance were, the number of clusters, average cluster weight, production per plant, yield and Ravaz index. As parameters for technological and phenolic maturation, pH, soluble solids, total acidity and total polyphenols were evaluated. The budbreak occurred at the end of August until the second fortnight of September, the flowering occurred in the second week of October, the veraison occurred during the month of December, and the maturity/harvest occurred from the second fortnight of January until the beginning of February. The earliest budbreak occurred with ‘Felicia’ in the vineyard at 1250m and the latest occurred with ‘Calardis Blanc’. Regardless of the vineyard altitude, there was practically no difference in the flowering date between varieties and the vineyards altitude, the veraison and the maturity/harvest occurred earlier in the vineyard at 840m. The varieties produced at 840 m had a shorter cycle and proved to be more productive when compared to those produced at 1250 m. The studied varieties showed good phenological adaptation, high yield and maturation indices suitable for the production of quality still and sparkling white wines at both altitudes. From the results obtained, ‘Felicia’ and ‘Calardis Blanc’ seem to be the most promising varieties for cultivation in both altitude ranges.


Introduction
Brazil has recently stood out in grape production.The South region is the largest producer in the country, where most of grapes are used for processing (juices and wines) and the largest volume consists in 'common' grapes (V.labrusca and hybrids).Nonetheless nowadays, Santa Catarina State also stands out in processing wine grapes (V.vinifera), showing an increase in production and planted area in recent years (CALIARI, 2020).
Recently, the high altitude regions of Santa Catarina, characterized by altitudes ranging from 900 to 1400 meters, have shown to be a new frontier for wine production, producing grapes with distinct characteristics when compared to the other growing regions, enabling the production of high quality wines (FALCÃO et al., 2007;BORGHEZAN et al., 2011;MALINOVSKI et al., 2012, ALLEBRANDT et al., 2015;MACEDO et al., 2015;MARCON FILHO et al., 2015).

Performance of resistant grape varieties (PIWI), Felicia, Calardis
Blanc and Helios in two locations of Santa Catarina State (BR) 3 gens.It is therefore necessary to understand these climatic factors, in order to obtain the greater potential for vineyards in a given region (DELOIRE et al, 2005).Distinct characteristics can be attributed to the climatic particularities of these regions, since the high rate of rainfall combined with high temperatures observed in traditional producing regions, increases the incidence of fungal diseases interfering in fruit quality (BACK et al, 2013; EPAGRI/CIRAM, 2020).
Downy mildew and powdery mildew are considered the main vine fungal diseases in Brazil and around the world, causing losses up to 100% when not controlled (CARISSE;BEAULIOU 2018;SAIFERT et al. 2018).
In addition to phytosanitary treatments, the use of resistant varieties that are the result of interspecific crosses between Mediterranean (Vitis vinifera), North American and Asian species (Vitis spp) has been recognized as a way of overcoming the problem of fungal diseases.The use of marker-assisted selection, combined with backcrosses with vinifera varieties, allowed the development of genotypes resistant to fungal diseases, which combine disease resistance genes and a significant percentage (over 85%) of V. vinifera; these cultivars are known as PIWI (from the German "Pizwiderstandsfähige", meaning resistant to fungal diseases), accepted as vinifera varieties in European catalogs (SIVCEV et al., 2010;SAIFERT et al., 2018;VEZZULLI et al. 2018).
One of the advantages of these varieties is the reduction of the number of pesticide applications (FULLER et al. 2014), especially in warm humid areas such as southern Brazil.The use of PIWI cultivars can be considered an option for Brazilian growers, as a way to reduce the use of pesticides, consequently reducing costs and improving grape quality (DE BEM et al., 2016;BONIN et al., 2017).Also, more recent wine tastings have shown that it is possible to make wine from these varieties with a quality similar to that of traditional varieties (BASLER;PFENNINGER, 2003).
The fungal resistant grape are more vigorous and present a higher productivity when compared to V. vinifera in organic systems (REYNOLDS;VANDEN HEUVEL, 2009;SUN et al. 2011;ROSSEUAU et al., 2013;BARTHE, 2015).

Material and Methods
The  1 and Figure 2.   No irrigation was performed, plants were trained in VSP and pruned in a double spur cordon, leaving two buds per spur in a total of 50 buds per plant.Pruning was performed in August and early September in both areas.In the beginning of October was performed the shoot thining, leaving on average one shoot per sprouted bud.Leaf removal was performed in early December, after fruit set and before veraison.Vines were trimmed once shoots outgrew 20 cm above the top wire.Secondary shoot removal were performed at the end of spring until the beginning of summer, when vegetative growth was more intense.When nec-essary, mowing was carried out between the rows.Fungicide sprays were made at both locations, the first one for downy mildew at flowering time and the others according to specific needs identified during vineyard evaluations, the total number of phytosanitary treatments during the productions cycle were 50% less when compared to V. vinifera.
The experimental design was in completely randomized blocks, with five blocks and ten plants of each variety per plot.For every three plants per plot, the vine phenological stages were defined, based on the BBCH scale (LORENZ et al., 1995).The dates of occurrence of each phenological event were recorded between pruning and harvest.The four main phenological stages were budbreak, considering the date when 50% of the buds were at the green tip stage (BBCH 7); full bloom, when 50% (BBCH 65) of the inflorescences were open; beginning of ripening or veraison, when the berries have softened or changed color (BBCH 81); and maturity/ harvest, when physiological maturity was reached, that is, when the harvest point was reached according to chemical evaluation of the berries (BBCH 89).
Performance of resistant grape varieties (PIWI), Felicia, Calardis Blanc and Helios in two locations of Santa Catarina State (BR)

Giovanni et al. (2023)
Evaluations of productive variables, number and average weight of clusters (g), productivity per plant (Kg) and estimated yield (Ton) were carried out from three plants per plot.At the time of pruning, the Ravaz index was determined from the ratio between productivity per plant (kg) and the mass of pruned material from the same plant (Kg) (CUS, 2004).
After harvest, a sample of 50 berries per plot was sent to the Videira Experimental Station laboratory, where physical chemical analyzes were performed.Soluble solids content, expressed in °Brix, was determined by refractometer (Quimis®) reading; total acidity (mEq L -1 ) and pH obtained by automatic titrator Easy PlusTM (Metller Toledo) and total polyphenols obtained by the Folin-Ciocalteu colorimetric method in a spectrophotometer (RAY Leigh -UV 2601) reading at 760 nm (OIV 2019).
Phenology data were analyzed using descriptive statistics (mean and standard deviation).
The grape production and quality data were submitted to analysis of variance (ANOVA) and when treatment effects were detected, the average comparison test was carried out by the Tukey Test at 5% error probability, using the R software.A standardized principal component analysis (PCA) was used to describe the effect of varieties on the mean of the variables cluster weight, number of clusters, yield, Ravaz index, soluble solids, total acidity, pH and total polyphenols.

Results and Discussion
The average dates of occurrence of the main phenological events are presented in Table 1.'Felicia' and 'Helios' produced in Água Doce (1250 m) presented an anticipation of budbreak of 8 to 10 days when compared to Videira (840 m).For 'Calardis Blanc', no difference was observed in the average budbreak date in both evaluated sites (Table 1).The duration of budbreak-full bloom sub-period is 10 to 14 days shorter in the lower altitude vineyard (840 m).Among the evaluated varieties, Calardis B. has the shortest duration of budbreak-full bloom sub-period in both evaluated locations (Figure 3).In the full bloom-veraison sub-period, its duration was on average 14 days shorter in the lower altitude vineyard.There was little variation in the duration of this sub-period between the different varieties, but 'Helios' showed the shortest duration (Figure 3).
In the veraison-maturity sub-period, practically no differences were observed between the locations and the varieties 'Helios' and 'Felicia'.The variety Calardis B. grown in the vineyard located at higher altitude had an average duration of 54 days for this sub-period, 11 days longer than the vineyards at lower altitude (Figure 3).
Long maturation periods can increase the possibilities of problems such as cluster rot, and early maturation is a very advantageous feature in tropical and subtropical conditions due because of high precipitation during grape ripening period in these production sites (SCHAEFER, 2016).The results obtained in these works demonstrate a good adaptation of the studied cases in both studied regions.
When evaluating the same fungus resistant varieties in Videira (840m) in 2018 vintage, the same pattern of cycle duration was observed between the sub-periods evaluated (BRIGHENTI et al., 2019).Similar results were obtained for 'Felicia' and Calardis B. produced in São Joaquim (1100 m) (SOUZA et al., 2019).
As a result, the total cycle duration of the varieties was on average 25 days longer in the vineyard at the highest altitude (1250 m).Temperature helps to explain the differences between cycle lengths.Milder tempera-tures delay fruit development, prolonging the period of berry growth (KELLER, 2010).When a region has higher average temperatures, the grape growth cycle is shorter, due to the greater accumulation of heat that causes early maturation (MUNIZ et al. 2016).
The mean temperature over the cycle in the vineyard located at 840 m was 20.4°C, while the mean temperature in the vineyard located at 1250 m was 17.2°C.
The characterization of phenological sub-periods duration is important in the introduction of new grape varieties, as they allow the determination of vine development phases (vegetative growth, flowering, fructification, and fruit maturation), which occur seasonally, in relation to the climate, especially seasonal variations, in addition to being used to interpret how different regions interact with the plant (MATTAR, 2016, GRIS 2010, GIL;PSZCZÓLKOWISKI, 2007, BOLIANI, 1994).
Regarding the productive variables, it was observed that 'Calardis Blanc' stood out for the high number of clusters and, when produced at 840 m, it was the variety that presented the highest cluster number in all evaluated vintages.In all evaluated vintages and in both locations, 'Felicia' produced the lowest number of clusters.On the other hand, 'Felicia' cultivated at 840 m produced the heaviest clusters in all evaluated vintages (on average 186 g), while 'Calardis Blanc' cultivated at 1250 m produced the lowest weight clusters in all evaluated vintages (on average 94 g) (Table 2).Similar results were obtained by other authors when studying the same varieties in Santa Catarina.In the 2018 vintage, in the vineyard located at 840 m, an average of 28 clusters were observed for 'Helios' and 50 clusters for 'Calardis Blanc' and 'Felicia'.In the same experiment, 'Felicia' also presented the heaviest clusters (on average 176 g) and was followed by 'Calardis Blanc' (133 g) and 'Helios' (128 g) (BRIGHENTI et al., 2019).
Still in the 2018 vintage, but in a vineyard located at 1100 m, an average of 50 clusters were observed for 'Calardis Blanc' and 'Felicia'.In this experiment 'Felicia' also presented the heaviest clusters (on average 177 g) and was followed by 'Calardis Blanc' (134 g) (SOUZA et al., 2019).In a recent work that evaluated the adaptation of resistant varieties in northern Italy, it was found for 'Soreli' values of 3.5 kg per plant, 30 clusters per plant and an average of 120 g per cluster; for 'Sauvignon Kretos' it was found 2.4 kg per plant, 25 clusters per plant and an average of 107 g per cluster; and for 'Sauvignon Rytos' it was found 2.4 kg per plant, 29 clusters per plant and an average of 85 g per cluster (FRIONI et al., 2021).
When evaluating the productivity per plant and estimated yield per hectare, it is possible to affirm that 'Felicia' and 'Calardis Blanc' produced at 840 m have the highest productive potential, with an average productivity of 6.0 kg per plant or 18 tons per hectare.With the exception of 2020 vintage, 'Helios' produced at 840 m also showed high yields, with an average of 5.0 kg per plant or 13 tons per hectare (Table 3).and Cabernet Sauvignon produced in São Joaquim (1400 m) and Campo Belo do Sul (900 m), regions with altitudes similar to Videira and Água Doce, the productive rates of resistant varieties were much higher than those obtained by Muniz et al (2015).When studying the Chardonnay variety in the 2018 and 2019 vintages in São Joaquim (1200 m), other authors found productivity values between 1.7 and 8.4 tons per hectare (PANCERI et al., 2018;WÜRZ et al., 2021).
Nowadays, remunerative productivity is a necessary requirement for new genotypes, and it should be associated with desired grape quality and low to moderate susceptibility to biotic and abiotic stress.Moreover, high basal bud fruitfulness allows the implementation of spur-pruning systems prone to the mechanization of winter operations (PONI et al., 2018).The Ravaz index is a parameter used to establish the balance and vigor of the plants, the ideal is that these indexes are between 4 and 10.Values below 4 indicate an excess of vigor or low grape production, while values a from 7 onwards, may indicate excess of production and risk of plant depletion (carbohydrates and nutrients) (ZALAMENA, et al. 2013;KLIEWER;DOKOOZLIAN, 2005YUSTE, 2005).The Ravaz index values found in this work indicate that the plants are in balance, or with a tendency towards excess production, especially in the vineyard located at 840 m (Table 3).Overproduction did not affect the quality of the grapes and can be resolved by bud number adjustment during pruning or cluster thinning.
The highest pH values were found in the 'Felicia', the average of the three evaluated vintages was 3.61 to 840 m and 3.52 to 1250 m. 'Calardis Blanc' showed intermediate pH values, on average 3.48 at 840 m and 3.36 at 1250 m.And 'Helios' had the lowest pH values in 2020 and 2021 vintages, on average the values obtained in this experiment were 3.37 at 840 m and 3.33 at 1250 m.In general, regardless of the variety, the grapes produced in the lower altitude vineyard had higher pH values (Table 4).4).The values obtained in this study ranged from 1134.9 mg L -1 for 'Felicia' to 221.6 mg L -1 for 'Calardis Blanc'.When evaluating the concentration of total polyphenols in white grape varieties produced in São Joaquim (1400 m), values of 995 mg L -1 were found for Trebbiano Toscano, 648 mg L -1 for Chardonnay, 506 mg L -1 for Moscato Giallo and 283 mg L -1 for Riesling Renano were found (BRIGHENTI et al., 2017).
Many authors have reported that polyphenol composition is due not only to the type of cultivar but also to the location where the grapes are grown and to environmental and management practices, as well as to the growing season (JONES; DAVIS, 2000;MATEUS et al., 2002).Climatic variables such as precipitation and solar radiation play an important role in the accumulation of phenolic compounds in grapes.
Precipitation is an important climatic factor in terms of the formation of phenolic compounds in the skin of the grapes (JONES; DAVIS, 2000).Polyphenols have the function of protecting plants from physical (such as the sun UV radiation) and biological (fungi, viruses, bacteria) attacks.Typically, in places where there are high rainfall levels, there is stress from the plant to combat, mainly, fungal diseases, inducing the production of phenolic compounds (DOWNEY et al., 2006;DAI et al., 2011).
The incidence of solar radiation in grape clusters also contributes to the activation of metabolism and the formation of substances responsible for wine quality, such as phenolic compounds, especially flavonols (DOWNEY et al., 2006).This difference can give the wines produced in high altitude regions of Santa Catarina State more intense coloration, more complex aroma, greater structure and persistence (GRIS et al., 2010;MALINOVSKI et al., 2012).
For soluble solids, 'Felicia' grown at 840 m showed high values in all evaluated vintages.For the other varieties there was a marked effect of the vintage, in 2020, 'Felicia' (840 m) presented values of 20.7 °Brix, while the lowest values were obtained for 'Calardis Blanc' (1250 m) and 'Helios' (1250 m), with 15.6 ° Brix and 17.4 °Brix respectively.In the vintage 2021, 'Helios' (1250 m) presented the lowest values of soluble solids, with 16.5 °Brix, while the other varieties did not differ.In the vintage 2022, in both locations, 'Calardis Blanc' produced grapes with the lowest concentration of soluble solids, 16.9 °Brix at 840 m and 18.3 °Brix at 1250 m, the other varieties did not differ from each other (Table 5).Among two the production sites, the total acidity of grapes produced in Videira was lower than in Água Doce, which can be explained by the shorter maturation cycle due to lower altitude and higher temperatures.High altitude regions tend to produce grapes with higher levels of acidity and aromas, which favors the production of white and sparkling wines (MANSOUR et al. 2022).
It is important to point out that the final quality of a wine is related to the grape stage of maturation, especially the concentration of soluble solids and total acidity, in the case of the production of sparkling wines, a medium sugar content and the presence of acidity result in products with high market acceptance (CALIARI et al, 2014;MANSOUR et al, 2022).Thus, based on the results obtained for soluble solids and acidity, it is possible to infer that still wines and quality sparkling wines can be produced from grapes produced in the evaluated locations.In Figure 4 it is possible to observe the PCA of the PIWI varieties grown at different altitudes, in this case factor 1 (PC1) and factor 2 (PC2) explain, respectively, 55.12 and 34.09% of the data variation; together, they explain 89.21% of the total variability.The dispersion of treatments can be observed from their distribution in the coordinate system.

Conclusions
The varieties produced at 840 m had a shorter cycle and proved to be more productive when compared to those produced at 1250 m.The Calardis Blanc variety produced at both altitudes, sprouted later, and it can be considered an advantage to avoid the late frosts, which eventually occur in these regions.
The studied varieties showed good phenological adaptation, high yield, and maturation indices suitable for the production of quality still and sparkling white wines at both altitudes.'Felicia' and 'Calardis Blanc' proved to be the most promising varieties for cultivation in both altitude ranges.
Performance of resistant grape varieties (PIWI), Felicia, Calardis Blanc and Helios in two locations of Santa Catarina State (BR)
Oscillations in soluble solids values in different vintages can be associated with high variations in precipitation during the grape ripening period, as can be seen in Figure2.The soluble solids contents verified in this study were like those found byBrighenti et al. (2019)  in São Joaquim for the 'Felicia','Calardis Blanc'  and 'Helios'.For Helios, in Trento -Italy,  Pedò et al. (2019)  observed a high amount of soluble solids in 'Helios' (21, 3°Brix) and Tuchshmid et al (2006) observed 21,6°Brix for Helios and 19,0°Brix for 'Bronner'.When studying the adaptation of resistant varieties in northern Italy, Frioni et al. (2021) found values above 22° Brix for Soreli, Sauvignon Rytos and Sauvignon Kretos.The lowest values of total acidity, approximately 67 mEq L -1 , were obtained in 'Calardis Blanc' (840 m) in 2020 and 2021 vintages.Low acidity values were also obtained in 'Felicia' (in both locations) in 2020 and 2021 and in 'Calardis Blanc' (1250 m) in 2021.In the vintage 2022 there was no difference between the different varieties.The results obtained by Brighenti et al. (2019) in São Joaquim (1100 m) were higher when compared to those produced in Videira and Água Doce for both varieties.These values are lower than those obtained by other authors, who obtained 130.5 mEq L -1 and 127.3 mEq L -1 evaluating the variety Chardonnay (PANCERI et al., 2018; WÜRZ et al., 2020).Tuchshmid et al (2006) noted total acidity in Swizerland for 'Helios' 113,5 mEq/L.Pedò et al. 2006 observed the values of 71,9 mEq/L for 'Helios' in Trento -Italy; and Baietto (2022) in Canelones -Uruguai observed values of 81,17 for 'Fleurtai'.

Figure 4 :
Figure 4: Projection of the principal component analysis of 8 variables used to evaluate the performance of vine fungal resistant varieties (PIWI), during the 2020, 2021 and 2022 vintages in Videira (840 m) and Água Doce (1250 m).

Table 1 .
Dates Pacifico et al. (2013)vents of 'Felicia','Calardis Blanc' and 'Helios' cultivatedin two altitude ranges of Santa Catarina.Average of 2020, 2021 and 2022 vintages.The average date of veraison also occurred earlier in the lower altitude vineyard (840 m), between December 2 and 16.In the highest altitude vineyard (1250 m) the average date of veraison occurred between December 19th and January 4. 'Felicia' presented the veraison on December 2 (840 m) and December 19 (1250 m).'Helios' presented the veraison on December 8 (840 m) and January 4 (1250 m).And Calardis B. showed the veraison on December 16 (840 m) and January 1 (1250 m) (Table1).Maturity/harvest occurred on average 16 days earlier in the lower altitude vineyard.In the lower altitude vineyard, the harvest took place between January 14th and 21st, while in the higher altitude vineyards, the It seems that resistant varieties have lower requirements in terms of heat accumulation to shift from vegetative activity to a prioritized reproductive activity.The early veraison renders these varieties more suitable for cool climates or areas located at higher altitudes(FRIONI et al., 2020).In a previous study, evaluating 19 resistant varieties,Pacifico et al. (2013)verified that on average, the blooming, veraison and harvest dates for resistant varieties in northern Italy occurred later than observed with "Pinot gris".
* Averages followed by the same letter, in the column, belong to the same group by Tukey test at 5% probability.Performance
* Averages followed by the same letter, in the column, belong to the same group by Tukey test at 5% probability.ns = not significant by analysis of variance (ANOVA) at 5% probability.
* Averages followed by the same letter, in the column, belong to the same group by Tukey test at 5% probability.ns = not significant by analysis of variance (ANOVA) at 5% probability.

Giovanni et al. (2023) Rev. Bras. Frutic. 2023; 45: e-001 Performance of resistant grape varieties (PIWI), Felicia, Calardis Blanc and Helios in two locations of Santa Catarina State (BR) 13
According to the PCA, it was possible to separate the varieties into 4 groups according to similarity standards.In group A are the varieties 'Helios' and 'Calardis Blanc' grown at 1250 m, they are associated with higher levels of total acidity, higher number of bunches, lower cluster weight, lower levels of total polyphenols, lower yield, lower levels of solids soluble and lower Ravaz index.In group B are 'Helios' cultivated at 840 m and 'Felicia' cultivated at 1250 m, these varieties are associated with a smaller number of clusters per plant and intermediate values of soluble solids, pH, yield and cluster weight.In group C is the 'Calardis Blanc' variety grown at 840 m, it is associated with a high Ravaz index, high number of clusters, high yield and low acidity.And in group D is the variety 'Felicia' grown at 840 m, which is associated with higher values of soluble solids, cluster weight, total polyphenols, pH and higher yield.