1. Introduction
Water is an essential production factor in agriculture, especially in semiarid areas where rainfall does not meet the demand for crops. Winegrowing is no exception.
Although the vine is well adapted to water shortages due to the depth and extent of its roots and to drought resistance mechanisms such as control of stomatal opening [
1] and osmotic adjustment [
2], increasing water availability for the plant means a radical change in its physiology [
3,
4]. These physiological changes decisively affect the grapes’ fruiting and ripening processes and the characteristics of the musts [
5,
6,
7,
8].
Knowing when and how much water to apply is essential to achieve, on the one hand, efficient management of this increasingly scarce and expensive natural resource and, on the other, to maintain farm profitability by balancing productivity and grape quality. Thus, correct water application requires precise knowledge of the vine’s response to water stress in its different phenological states to identify the periods of least sensitivity of the grape and define the best irrigation strategy to be applied. The period of greatest vegetative growth on the vine occurs from bud break to the cessation of shoot growth, which occurs near veraison [
8]. Water stress conditions during this period decrease shoot growth and leaf area in vine [
9]. This effect will be more pronounced the earlier the stress is present and used as a growth control tool in high vigorous plantations. On the other hand, after veraison, water stress has little or no effect on leaf area development [
10,
11,
12] as long as the vine stress is not severe, since high stress produces premature senescence of the leaves and a limitation in the translocation of photoassimilates to the bunches and reserve organs.
Vegetative growth decreases as berry growth and development become evident, so that the cluster becomes the main sink for photoassimilation to the detriment of the vegetative apices, which end up stopping their growth. The multiplication and elongation of berry cells decrease in response to early water stress, resulting in smaller berries [
13]. This smaller berry size has been associated with increases in must quality as a result of the increased skin/pulp ratio [
12]. However, [
14] reported that vine management throughout the cultivation cycle had a more significant influence on berry composition than on berry size.
Stress management through irrigation has also been used in hot climate areas as a tool to improve berry and wine composition [
15,
16,
17,
18,
19,
20,
21,
22,
23]. During pre-veraison, water stress produces metabolic changes in the berry, which are maintained even though later this stress decreases [
20,
24]. In contrast, the effects on berry of water stress during post-veraison are much more variable, generating greater discrepancy between the results obtained [
25,
26,
27,
28]. However, it can be generalized that moderate water stress improves crop quality by decreasing berry size and total acidity but increases the soluble solids content and the total concentration of anthocyanins and polyphenols [
29].
Intercepted solar radiation is the most crucial factor determining water consumption in vines [
30]. It is ultimately responsible for leaf photosynthetic activity, overall productivity, and vine quality through the relationship between exposed leaf area and fruit weight [
31,
32,
33,
34,
35].
Numerous studies have shown that a proper balance between vegetative growth and yield is necessary to improve berry quality [
31,
33]. This source–sink relationship is influenced by environmental factors and cultural practices to modify the amount of vegetation and illumination of the clusters [
36,
37] or the yield level, decreasing productivity [
12,
38]. In some fruit trees, fruit load adjustment has been proposed as a technique to improve the water status under conditions of limited water availability [
39,
40]. In the vineyard, [
41,
42] report that this load regulation does not affect vine water status, nor does it favor increases in leaf area, which is why cluster thinning is understood as a tool used to improve grape quality at the expense of a decrease in yield. However, studies conducted by [
43,
44] have shown that a reduction in yield is not necessarily associated with significant changes in grape quality and that the final effects on wine quality depend on the timing and intensity of harvest crop load regulation [
45]. In this regard, although some studies have reported that cluster thinning increases the concentration of sugars and anthocyanins [
46,
47], others concluded that this effect is slight or non-existent [
41,
44,
48]. However, other results obtained with cv. Tempranillo indicate that, at the same sugar concentration, cluster thinning reduces acidity and increases pH [
42]. This diversity of results suggests that cluster thinning effects depend on soil, climate, genetic factors, and possible variations in this practice’s implementation.
The effect of both irrigation strategy and cluster thinning depends mainly on the agro-climatic conditions. Although cv. Tempranillo is the variety with the largest planting area in Spain, there is still not enough information on its response to different irrigation strategies under high temperature conditions during ripening, nor on the combined effect of irrigation strategies and early cluster thinning. This work analyzes the impact of several deficit irrigation strategies combined with different cluster load levels, adjusted in setting, on vegetative development, production, and quality of Tempranillo grapes under southwestern Spain’s growing conditions.
2. Materials and Methods
2.1. Location and Vineyard Description
The trial was established for three years (2009, 2010, and 2011) in an experimental vineyard located in “Finca La Orden” (Badajoz, Spain). The vineyard is located in Las Vegas Bajas del Guadiana (38°51′ N, 6°40′ W, altitude 188 m). The area’s climate is Mediterranean with moderate Atlantic influence, with very hot and dry summers and mild winters, presenting irregular rainfall with an average of 450 mm per year. The soil is alluvial with a loamy to sandy texture, slightly acidic, and low organic matter content.
The experimental vineyard has an area of 1.7 ha of cv. Tempranillo on Richter 110 rootstock planted in 2001 on a trellis, formed in a double Royat cordon leaving six spurs per plant and two buds per spur, with a trunk height of 0.60 m, trellis height of 1.50 m, and a planting frame of 2.5 × 1.2 m (3333 plants/ha), with E-W row orientation. In spring, pruning was carried out to adjust to 12 shoots per vine, with several clippings, depending on the year, to contain the vegetation on the trellis following the area’s cultural practices.
A drip irrigation system was in place with two emitters of 4 L h−1 per plant, with the differential treatments carried out through programmed solenoid valves in the field.
2.2. Treatments and Irrigation Management
Four treatments were established in 2009: rainfed (T1), which did not receive any water input through irrigation; pre-veraison deficit (T2), which received 25% of crop evapotranspiration (ETc) from the beginning of irrigation until veraison, and 75% after veraison; post-veraison deficit (T3), which received 75% ETc from the beginning of irrigation until veraison and 25% after veraison, and control (T4), which received 100% ETc throughout the irrigation period. In the years 2010 and 2011, the percentages used in the regulated deficit irrigation (RDI) treatments, T2 and T3, were 56 and 19%, to increase the contrasts between treatments.
Two crop load levels were established for each irrigation treatment except T4: high load (A) (no cluster thinning) and low load (B) (with cluster thinning). Cluster thinning was carried out at fruit set, eliminating one cluster per shoot.
The experimental design was of randomized blocks with four repetitions per treatment for the irrigation treatments (16 subplots), and divided plots when considering irrigation x cluster thinning (24 subplots). The subplot comprised six rows of 18 vines each row (108 vines per subplot), considering the outer rows as borders, another two rows for destructive determinations, and another 2 for non-destructive determinations. The same vines were selected for all treatments during the 3 years of the study.
ET
c was determined in a weighing lysimeter of 2.67 × 2.25 m and installed to a depth of 1.5 m in an area of the experimental vineyard, which contained two vines irrigated to 100% of their ET
c and developed in the same conditions as the rest of the plot surrounding it [
49].
Irrigation was initiated when a threshold value of the stem water potential at midday (ψsmd) of −0.6 MPa was reached for optimal irrigation in pre-veraison (T3 and T4), and a ψsmd value of −1.0 MPa for the treatment of moderate water stress in pre-veraison (T2). After veraison, irrigation was cut in the RDI treatments and was not restarted until threshold ψsmd values of −0.8 and −1.2 MPa were reached for optimal irrigation (T2) and moderate water stress (T3), respectively.
Irrigation was applied five to six times per week, measuring the amount of water applied to each subplot through volumetric water meters and maintaining irrigation until the beginning or middle of October.
2.3. Climate and Phenology
Phenological monitoring was carried out according to [
50,
51] so that from mid-March, coinciding with the so-called “cotton bud” phenological stage, a weekly visual inspection was carried out. This phenological monitoring was carried out independently for each subplot, selecting ten control plants and noting on each day of observation the most delayed, the most advanced, and the most representative (in which there were at least 50% of vines) phenological states.
Meteorological data and reference evapotranspiration (ET
o) were obtained from an agrometeorological station and a weighing lysimeter located 100 m from the experimental vineyard, in a polyphytic meadow [
52].
The degree days (DD) were calculated using 10 °C as the base temperature [
53,
54].
2.4. Water Status
The ψ
smd was measured from leaves on the north side, in the lower zone of the vine, near the trunk and wrapped in aluminum foil 2 h before the time of measurement, with a pressure chamber (Soil Moisture Corp., Model 3500, Santa Barbara, CA, USA), according to the procedure described by [
55]. The measurement frequency was weekly, measuring one leaf per plant, in two plants per subplot.
2.5. Vegetative Growth
The fraction of photosynthetically active radiation intercepted by the vineyard (FiPAR) was determined by measuring the radiation at ground level with a 0.80 m PAR radiation ceptometer (Accupar LP-80 from Decagon Device) on eight vines per treatment (two per block), performing six transects per plant (2 measurements per transect), in high crop load treatments. The measurements were taken at noon on clear days since flowering, with a biweekly frequency. The linear sensor was placed horizontally at ground level, covering the planting frame and perpendicular to the vines. At the beginning and end of the data collection, two other measurements were performed in an open site and without interference from vegetation, serving as a reference of the total PAR radiation at ground level in the measurement session. The measures were weighted by their representativeness within the transect, adding those of the same transect to obtain a single value (
Scheme 1). Subsequently, the average value of each subplot’s 12 transects was obtained, thus obtaining the PAR radiation that arrives at ground level, which by difference with the reference measurement of the PAR radiation gives us the PAR radiation intercepted by the crop. The FiPAR is the result of dividing this difference by the reference of the PAR radiation.
2.6. Production of Biomass and Yield Components
Ten plants were selected by subplots (high load sub-plots), and the fresh weight and dry weight in the different interventions carried out were determined: clipping (removal of the tip of the shoots at different times of the herbaceous phase of the shoots); harvesting; pruning (removal of branches during dormancy excluding leaves fallen in senescence). A precision balance (Sartorius Mechatronics BP61S, Göttinger, GER) with a sensitivity of 0.01 g was used to determine the weight. A forced ventilation stove was used to dry the samples at 65 °C, and the samples were kept there until they reached a constant weight.
Harvesting was performed, on all sub-plots, when a concentration of soluble solids close to 23 °Brix (average value of each treatment) was reached. At harvest, all the clusters from the ten selected plants of each subplot were weighed and counted, both high and low loading, differentiating them. Berry weight was determined by sampling each subplot at harvest and weighing 100 berries.
2.7. Water Productivity
Water productivity (WP) was differentiated into two components, vegetative and productive. The vegetative component was determined as the ratio between the dry weight (g/plant) of clippings and pruning, except leaves in senescence, and the water received (L/plant) both through irrigation and total rainfall in the period from budbreak to harvest. Water supplied to each plant through irrigation was calculated by reading the volumetric meters installed in each subplot and divided by the number of vines in each subplot. In contrast, each plant’s rainfall was calculated from data obtained from a nearby agrometeorological station by multiplying this value by the area occupied by each vine. The efficiency attributed to the productive component was determined as the relationship between the fresh weight at harvest (g/plant) and the water received (L/plant) from irrigation plus precipitation for the same period.
2.8. Must Quality Parameters
At harvest, samples of grapes were taken from each subplot. The samples were crushed and blended, after manual removal of the pedicel, using a commercial blender (Taurus BAPI 850 INOX 916.251, Oliana, Spain) with speed control. The resulting must was separated into portions, measuring all the quality parameters under study.
The total soluble solids content (TSS) was determined using a digital refractometer (ATAGO PR32 Alpha, Tokyo, Japan) with a sensitivity of 0.2% and temperature compensation.
pH and total acidity expressed as g/L of tartaric acid were determined by titration with a 0.1 N NaOH solution to an endpoint of pH 8.2, with an automatic titrator (CRISON Micro TT), following the official methodology of the International Organisation of Vine and Wine [
56].
Tartaric acid (TH2, g/L) and malic acid (MH2, g/L) were analyzed by spectrophotometry, following the Rebelein [
57] and the enzymatic reaction methods, respectively, in an automatic analyzer (Chem Easy Plus, Systea R&D group, Anagni, Italy).
Potassium was determined using an atomic absorption spectrophotometer (AA 240 FS, Varian, CA, USA), according to the official methodology [
56].
Total polyphenols and anthocyanins were determined following the procedure proposed by the Australian Wine Research Institute and described in [
58].
2.9. Statistical Analysis
The results were subjected to a one-way and two-way (irrigation x load) analysis of variance (ANOVA) for each year, using the general linear model, considering irrigation as the main factor and load as a subplot of the design-divided plots. The comparison of means for the four irrigation treatments at high load was done by the Tukey test with a significance level p < 0.05. To analyze the effect of load on each irrigation treatment, the year and treatment to be analyzed were selected (except T4), comparing the means using a T-test for independent samples. The interaction between the factors considered was done through multivariate analysis.
The analyses were carried out using SPSS version 20 (IBM, Armonk, NY, USA).
4. Discussions
In the cultivation of vineyards for winemaking, the value of the harvest is determined by the production/quality binomial so that, as in other fruit production, lower yield production systems or costly cultivation practices can be adopted to enhance aspects that give greater value to the final product. However, this quality is marked by a complex set of factors that can interact with each other. Agro-climatic conditions are decisive for berry composition, as are cultural practices. It is this complicated situation that causes the disparity of results that can be found in the literature when the effect of practices such as irrigation or cluster thinning on the production and quality of a vineyard is analyzed, and that justifies the need to adapt cultural practices to specific growing conditions with a medium-term vision. This work aimed to analyze the response of a cv. Tempranillo vineyard to different irrigation strategies and cluster loading in the Guadiana Valley’s growing conditions, in southwestern Spain.
A determining aspect of the results obtained was the year’s characteristics, both in terms of weather conditions and the “memory” of the vineyard. Under these conditions, the response of the water status of the vines to the irrigation treatments was variable, with irrigation dose interacting with the characteristics of the year despite establishing the irrigation criteria (start of irrigation and changes in veraison) according to ψsmd levels and calculating the doses according to water consumption (ETc). It was possible to establish different levels of water stress in the pre-veraison and post-veraison periods in RDI treatments.
The average water needs of the vineyard obtained in this study were 568 mm, allowing the RDI strategies to save irrigation water between 70 and 62%, while this same vineyard, in the period 2005–2008 [
42], presented very variable water needs between years, with an average value of 500 mm and promoting the management of sustained deficit irrigation, water reductions according to the percentages applied.
The production cycle duration was variable between years, faster in the driest year with less accumulation of DD from budbreak to harvest, coinciding with lower crop load. These results also show an acceleration of ripening due to cluster thinning, which is in agreement with the findings of authors such as [
21,
47].
In this study, vine water status was not affected by crop load. A similar result was found for cv. Tempranillo by [
42] and for cv. Cabernet Sauvignon by [
41], although cluster thinning was carried out in veraison in both cases. However, [
60] found that reducing crop load on Thomson Seedless improved water status, so there seems to be a lack of consensus regarding the effect of cluster thinning on water status.
Figure 2 shows a strong parallelism between the seasonal evolution of ψ
smd in the dry treatment and ET
o over the three years of the trial, indicating a high degree of coupling between the two. These results question the isohydric behavior in vines [
3,
61,
62], which was put forward as an argument to question the validity of the ψ
smd measure as an indicator of water status in this crop [
63,
64]. This controversy regarding isohydric or anisohydric behavior in the vine may be due to the influence of multiple factors as found by several authors [
4,
62,
65,
66,
67,
68].
A positive response was found in terms of the relationship between dry matter production and PAR radiation intercepted by the crop. As the interception of PAR radiation increased, an increase in dry matter production was observed (
Figure 4), obtaining low coefficients of determination, both for total dry matter and productive dry matter, in line with the results obtained for cv. Tempranillo by [
69]. The correlation with vegetative organ biomass was higher, coinciding with the results of [
70] in which it was reported that several factors affect the relationship between dry matter production and intercepted PAR radiation. In this study, shoot clipping carried out to maintain the vegetation in a vertical position and facilitate the passage of machinery contained the vegetation and reduced the intercepted PAR radiation with greater intensity in the treatments with less water stress. This could distort the correlation between dry matter and intercepted PAR radiation. Given these results, it is clear that the use of irrigation strategies requires adapted cultivation techniques that do not limit their potential, such as forming plants with free vegetation or increases in the height of the vegetation, which have demonstrated a greater capacity for intercepting radiation [
71].
The results obtained confirm that water deficit reduces vineyard productivity in terms of both grape fresh weight (
Table 5), as well as the production of vegetative and reproductive biomass (
Table 7 and
Figure 3), which is in accordance with previous works by other authors [
8,
72]. The effect was more evident in the vegetative organs, since, as pointed out in [
73,
74], water stress primarily affects cell expansion and, therefore, vegetative growth, and, at more severe levels, cell multiplication and photosynthesis. The RDI strategies tested resulted in yield losses, unlike the results obtained in other woody crops under similar agro-climatic conditions [
75]. On the other hand, the strategy with the best yields was that which in each year managed to maintain the best vine water status throughout the crop (
Figure 5). Evidence has been obtained that early stress after budbreak was detrimental to production, both in the current and the following year (
Figure 5) [
42,
76,
77,
78]. This is due to the decrease in bud fertility (
Figure 5A) caused by the stress of the previous year at the time of fruit bud initiation and differentiation [
73] and a decrease in berry set due to stress in the current year (
Figure 5B), thus reducing cluster weight [
10,
79]. Irrigation treatment did not modify berry size (
Table 5), unlike the results obtained in other studies [
26,
80], probably because in no year did the berry reach a sufficiently severe stress level during the three weeks after flowering, which according to [
81], is when the potential size of the berry is determined.
Early cluster thinning decreased yield (
Table 5), although this was slightly compensated for by increased cluster weight due to a higher number of berries per cluster, as individual berry weight was not significantly modified.
The effect of irrigation treatment on grape quality was very variable between years (
Table 6), as also previously noted [
26], no clear effect was found in this study (2009–2011), nor in previous studies carried out on the same vineyard (2005–2008) [
42,
47]. Considering that the greatest differences between irrigation treatments were found in 2009 (
Table 6), it is evident that it is necessary to generate notable differences in vine water status to affect the characteristics measured at harvest.
The establishment of different water statuses in the different phases of the crop, favored by irrigation management, did not imply a significant decrease in the concentration of soluble solids (
Table 9). This concurs with the results obtained for cv. Tempranillo by authors such as [
82,
83] but differs from those obtained by [
84] who found a relationship between water status and TSS, which increased with the water stress integral. These results are explained by the fact that no stress levels were reached in this study that compromised berry size, and so it was not possible to establish differential soluble solids concentrations [
85].
As for must total acidity, the improvement observed in water status was observed to be correlated with the increase in total acidity, with a good correlation being obtained for the average of the whole period but which was more conditioned by the water status in the pre-veraison period (
Table 9). When breaking down this total acidity into its main components (malic and tartaric acid), it can be observed that this increase was more influenced by an increase in malic acid, mostly affected by the supported pre-veraison water status (
Table 9). This result, also reported by other authors [
82,
83], may be due to irrigation increasing vegetative growth [
86,
87] resulting in a more unfavorable microclimate for malic acid combustion, with vegetative growth closely related to must malic acid content [
26]. For its part, tartaric acid content remained very stable, regardless of water status, which is in agreement with results obtained for cv. Tempranillo by other authors [
82,
83]. The above leads to a decrease in the tartrate–malate ratio as the water status improves [
83]; a relationship that can have a major impact on the final pH of the wine due to the greater presence of a weak acid (the malic acid) against a strong acid (the tartaric acid).
As for the effect of irrigation on pH, the results support the variability obtained by other authors. An apparent effect was observed in 2009, but there was no effect of irrigation in 2010, as previously observed [
42], or there was a decrease as the result of irrigation in 2011 (
Table 6), as also observed by some authors [
5,
88]. Similarly, no conclusive result could be obtained when analyzing the effect of cluster thinning on this parameter.
Water stress can reduce potassium absorption due to reduced root activity and growth [
89]. This is reflected in
Table 9, as it can be seen that, as other authors have also found [
83,
90], as vine water status improves, there is an increase in must potassium content, although this correlation was only significant in this study for water status during the ripening period (post-veraison).
Color very much conditions the quality of the wine, and so the incidence that different irrigation managements can have on this parameter is of great importance. The different studies on this subject sometimes obtain contradictory results [
83,
84,
87,
91]. The results of this study could not establish a clear correlation between the water status supported by the vines and TPI, nor with anthocyanin content (
Table 9). This somewhat demystifies the idea that irrigation and quality are opposing concepts and contradicts what has been established by other authors [
83,
84] who did see a clear effect of irrigation on the reduction in these parameters. The small difference in quality induced by the different water regimes may be because no differences were established in berry size (
Figure 5C), a parameter that is inversely correlated with polyphenolic load [
84].
Cluster thinning is an expensive practice that reduces the production of the vineyard and can only be justified by a significant improvement in grape characteristics. In this sense, this work shows an increase in parameters such as acidity, soluble solids, and color compounds, although these differences were, once again, dependent on the year. Cluster thinning increased TSS when the availability of assimilates was lower, that is to say, in the treatments with higher water stress in 2009 and 2010, and even decreased in T3 in 2011 probably due to the considerable increase in cluster size caused by cluster thinning increasing the competition for assimilates between berries (
Table 5). Cluster thinning increased anthocyanins and TPI, confirming other authors’ results on this same cultivar [
47,
90,
92]. As previously mentioned, when differences in yield occurred due to irrigation management, no conclusive results were obtained on must concentrations of polyphenols and anthocyanins, with a different effect to load regulation by cluster elimination, as also observed by [
91,
93]. This highlights the importance in the synthesis of compounds responsible for color of other factors such as the effective illumination of leaves and bunches [
94] and the volumes of water used [
95].
An important aspect of this work is the comparison of the effect produced by the different dates of cluster thinning on the same vineyard. In this study, there was an improvement in the compounds responsible for wine color in the year in which the highest production was recorded (
Table 5) and the lowest vegetative development (
Table 4), which can be added to the improvement found for late cluster thinning on the same vineyard by [
42]. This contributed to improving the source–supply ratio due to cluster thinning, an effect that was more noticeable in early than in late cluster thinning, as also noted in [
93]. On the other hand, late cluster thinning has been reported to reduce must acidity and increase pH [
42], while, in this study, the same cluster thinning carried out earlier improved acidity in two of the years studied and decreased pH in one of them (
Table 6). Another aspect to highlight with respect to cluster thinning was the advance in ripening that was produced, which was more marked in early thinning (13 days in this study) than was observed for late thinning (4 days) for this same vineyard by [
42].
5. Conclusions
As can be seen in our study, vine water status is influenced by irrigation management and conditioned by the characteristics of the year, being the evolution of the water status of rainfed vines influences by the annual evolution of ETo, which gives an idea of isohydric behavior in the vine. In this sense, it could be seen as pre-veraison water status is more decisive in vine productivity, as it affects the berries’ setting in the current year and bud fertility the following year. On the other hand, it was noted that crop load level, adjusted by early cluster thinning, does not affect vine water status.
Water deficit reduces vineyard yield in terms of both fresh grape weight and vegetative and reproductive biomass production. However, this water deficit due to regulated deficit irrigation is an excellent tool to increase water productivity, as this paper has shown.
About the quality of the musts, our study can conclude that the improvement in vine water status in pre-veraison increases must total acidity, mainly due to the increase in malic acid, while this improvement in vine water status after veraison leads to an increase in must potassium content.
Another point addressed in this study was the regulation of the crop load by early cluster thinning, which results in a reduction in yield, although with a compensatory effect by increasing cluster size, also producing an acceleration in ripening. Improving this crop load regulation, the source–sink ratio, especially in high production years, and improving also limiting factors for wine quality in semiarid areas, such as acidity and must color.
Given the vine’s variable response to the irrigation treatments proposed in this and previous works on the same experimental vineyard, it seems more interesting not so much to talk about irrigation management but rather about the management of water stress in specific periods of the phenological cycle of the crop.