Carbon isotope discrimination (so-called δ 13 C) measured on grape juice is an accessible tool to monitor vine water status in production conditions

Assessment

Carbon isotope discrimination (so-called δ 13 C) measured on grape juice is an accessible tool to monitor vine water status in production conditions water deficit and, if so, its severity 2 .The ratio 13 C/ 12 C can be measured by isotope mass spectrometry with great precision.The δ 13 C is then obtained comparing the results against a standard with a known 13 C/ 12 C ratio.

Classification of water deficit stress levels
δ 13 C measured in grape berry juice sugars range between -28‰ from vines experiencing no water deficit, up to -20‰ from vines suffering severe water deficit stress.The classification of water deficit intensity across this range differs slightly across publications.and apply to regions with temperate climatic conditions.More precise site-and variety-specific threshold values can be set locally by combining δ 13 C and water potential measurements.Alternatively, a relative comparison can also be obtained: a variation of 1‰ in δ 13 C corresponds to a difference of ~0.2 MPa in midday stem water potential during the grape ripening period 5 .

Practical implementation of vine water status
assessment with δ 13 C δ 13 C measurement is carried out on samples of grape juice collected between three weeks after mid-veraison and harvest.Samples are sent to a laboratory equipped with an isotopic mass spectrometer.Some laboratories estimate δ 13 C using Fourier-transform infrared

Assessment of vine water status
Vine water status is a key parameter in grape growing, affecting both yield and berry composition.In turn, vine water status is driven by soil water availability (dependent on soil water holding capacity), climatic parameters (rainfall and evapotranspiration), training system (leaf area per hectare), plant material (rootstock and cultivar) and management practices (vineyard floor management and irrigation).Many methods for evaluating vine water status have been developed and can be grouped into: (1) soil-based measurements, (2) water balance modelling, and (3) plant-based measurements.Accuracy of soil-based and modelling approaches is limited by assumptions regarding the soil water holding capacity (SWHC) of the vine root zone, which is highly dependent on the rooting depth and root functioning, parameters that cannot be precisely assessed in field conditions.Plant-based indicators, however, naturally integrate the root zone SWHC of the vine and thus provide more reliable results.Among plant-based measurements, water potential by pressure chamber is widely used.Carbon isotope discrimination (δ 13 C) measured on berry juice is another plant-based indicator of vine water deficit with a huge potential for application in vineyard management.Although the first articles on this technique were published more than 20 years ago 1 , it is still not widely adopted by vineyard managers.

The principle of carbon isotope discrimination
Isotopes of an element have the same number of protons and electrons, but differing numbers of neutrons, and hence a differing atomic mass.There are three isotopes of carbon, but only 12 C and 13 C are stable in the environment, making them useful for the study of so-called isotope discriminating processes in nature. 12C is by far the most abundant carbon isotope in nature, with a proportion of approximately 99:1 compared to 13 C.During photosynthesis, there is discrimination in favor of CO 2 molecules containing 12 C isotopes, due to its higher reactivity with the enzymes in the photosynthetic reaction and a better diffusion through the stomata and the mesophyll.Hence, 13 C/ 12 C ratios in the sugars produced are lower when compared to atmospheric CO 2. When plants experience water deficit, these 13 C/ 12 C isotope ratios are further modified as stomata close and block diffusion of CO 2 into the intercellular space of the leaves.This causes the relative concentration of 13 CO 2 to 12 CO 2 to rise in the intercellular space, and hence in the sugars produced.During ripening, these sugars are accumulated in the berry juice, with the measured 13 C/ 12 C ratio in this juice providing an indication of whether the vines were subject to Limitations of water status monitoring with δ 13 C Because δ 13 C is measured at the end of the season, it is not useful for operational day-to-day irrigation management.For this purpose, the pressure chamber is preferred.Another limitation is that it represents vine water deficit during the period of sugar accumulation in grape berries.Hence, it does not provide a record of early vine water deficits, nor accounts for water deficits experienced after berry sugar loading is achieved.There are also variety specific differences in overall water use efficiency that must be considered when evaluating δ 13 C responses to water deficits across varieties 3 .The analysis of δ 13 C is currently offered by only a limited number of commercial laboratories, but will be more widely available as demand increases.

Conclusions
Being able to characterize vine water status is key to understanding how environmental factors and management practices may impact yield and wine quality potential.δ 13 C measurements on grape must or wine is an easily obtained and inexpensive indicator of vine water status during the berry ripening period and can be useful for managing both dry-farmed and irrigated vineyards. spectroscopy (FTIRS), although this method cannot be recommended because it is imprecise.A current limitation for the implementation of δ 13 C measurement is the small, but steadily increasing number of laboratories that offer the analysis.Only a small amount of juice (+/-5 μL) is needed for the analyses and can be obtained from samples taken for regular maturity control at the end of the season.These samples, however, need to be representative of the vines or vineyard block being investigated.The δ 13 C signature in grape sugar is conserved in wine ethanol, although fermentation induces a shift of -1.7‰ (Table 1 6 ).Hence, δ 13 C analysis can also be performed on wine in order to investigate the corresponding water status of the vines during berry ripening 7 8 .
Applications of water status monitoring with δ 13 C δ 13 C provides insight into the water status experienced by the vines during the grape berry ripening period, which extends approximately from one week prior to mid-veraison to three weeks after mid-veraison.This corresponds, in most cases, to the month of August on the Northern Hemisphere and February on the Southern Hemisphere, a key period during which vine water status can affect yield and wine quality potential.δ 13 C can be a useful tool for evaluating irrigation strategies at the end of the season with vine water status being affected by the amount and timing of applied water.In order to optimize wine quality for red wine making, growing vines under mild water deficit is desirable, with values of δ 13 C between -24‰ and -25‰ indicating vines were not over-irrigated.This can be important information when grapes are purchased from growers, as it provides a tool to check irrigation management post hoc.δ 13 C can also be used to easily evaluate the impact of any vineyard management practice (cover crop, tillage, leaf removal…) on vine water status.Because δ 13 C is a highly accessible indicator (easy to measure and not expensive), it can also be used to map vine water status over a vineyard block or a winegrowing estate (Figure 1A 2 ).δ 13 C can be a useful tool in terroir studies of dry-farmed vineyards, where vine water status depends largely on soil water holding capacity (figure 1B) and climate, both important terroir parameters 2,9 .Such maps of vine water status are very useful for fine-tuning plant material choices and management strategies.In dry areas of the estate, drought resistant rootstocks should be used to avoid severe water stress.In wet areas, wine quality may be impacted by excessive water availability, which can be improved by establishment of a competitive cover crop.At the intra-parcel level, mapping vine water status with δ 13 C is a tool for precision viticulture, such as explaining spatial variability in grape phenolics 5 .When measured on wine (see specific interpretation thresholds, table 1), δ 13 C allows to trace back the water status of the vines which produced the wine.This can be used to explain the effect of vine water status on quality related parameters, like aroma compounds 7 .
FIGURE 1.A -Vine water status measured on a dry-farmed wine-growing estate in Saint-Emilion (Bordeaux area, 2022).172 samples were taken over the 23 hectares.B -Soil water holding capacity map drawn from the soil map of the estate.
Thresholds in Table 1 are computed from van Leeuwen et al., 2009 2 and Santesteban et al., 2015

TABLE 1 .
Water potential and δ 13 C values with respect to vine water deficit thresholds.Water potential thresholds refer to the lowest values recorded during the grape ripening period.