Abstract:
This invention describes the use of S-abscisic acid (S-ABA) to modify sensory characteristics such as aroma, bouquet, flavor, mouthfeel, astringency, balance, complexity or finish of red grapes and red wine.

Description:
FIELD OF THE INVENTION 
     The present invention is directed to the use of S-abscisic acid and its salts to modify the development of the sensory characteristics of red grapes and red wine, such as aroma, bouquet, flavor, mouthfeel, astringency, balance, complexity or finish. 
     BACKGROUND OF THE INVENTION 
     Abscisic acid (S-Abscisic acid, S-ABA, ABA) is a natural occurring plant hormone found in all higher plants (Cutler and Krochko. 1999. Trends in Plant Science. 4: 472-478. Finkelstein and Rock. 2002. The Arabidopsis Book. ASPB, Monona, Md., 1-52). S-ABA is involved in many major processes during plant growth and development including dormancy, germination, bud break, flowering, fruit set, general growth and development, stress tolerance, ripening, maturation, organ abscission, and senescence. S-ABA also plays an important role in plant tolerance to environmental stresses, such as drought, cold, and excessive salinity. 
     One key role of S-ABA in regulating physiological responses of plants is to act as a signal of reduced water availability to reduce water loss, inhibit growth and induce adaptive responses. All these functions are related to stomatal closure of plant leaves (Raschke and Hedrich, 1985, Planta, 163: 105-118). When stomata close, plants conserve water to survive in environmental stresses. However, stomatal closure also can result in the reduction of photosynthesis, and respiration and thus growth. Stomatal closure is a rapid response of plants to S-ABA. The mechanism of action of S-ABA-induced stomatal closure has been studied, and the effect has been shown to be due primarily to S-ABA&#39;s effect on guard cell ion channels. Specifically, S-ABA blocks H +  extrusion and K +  influx from guard cells and promotes K + , Cl − , and malate extrusion and Ca 2+  influx. The net effect of S-ABA is to reduce the total osmotica in the guard cells, which in turn decreases the water content in the cell. This causes the guard cells to lose their turgor and thus close the stomata (Assmann 2004 In:  Plant Hormones Biosynthesis, Signal Transduction, Action ! ed. Davies, p 391-412). The closing of stomata results in reduced transpiration of the plant leaf. In grapes, application of S-ABA has been reported to increase stomatal resistance in grapevines, thereby reducing the gas exchange and stomatal transpiration of the leaves (Düring and Broquedis, 1980, Sci. Hort., 13: 253-260). 
     The exogenous application of S-ABA to red grapes prior to harvest has been shown to increase the accumulation of anthocyanins and increase the red color of the grape berry skins (e.g. Han, D. H, S. M. Lee, and S. B. Kim. 1996, J. Kor. Soc. Hort. Sci. 37: 416-420; Lee, K. S., J. C. Less, Y. S. Hwang, and I. B. Hur, 1997, J. Kor. Soc. Hort. Sci. 38: 717-721; Kondo, S., Masuda, E. and Inoue, K., 1998, Acta Hort., 464: 35-40; Pepe, M. C., Fidelibus, M. W., Dokoozlian, N. 2006, HortScience, 41:1440-1445). 
     The sensory characteristics of wine such as aroma and flavor are complex and there is interest in altering wine grape berry and/or wine characteristics to produce more diverse or better wine, or wines with different balances of sensory characteristics. A patent application has been filed (Quaghebeur, K., 2005, US 2005/0198896 A1) claiming that ABA application enhances wine quality as a consequence of simulating drought in the grapevine leading to reduction in grape berry size in conjunction with increased sugar content. However, no mention is made of ABA application affecting sensory characteristics such as aroma, bouquet, flavor, mouthfeel, astringency, balance, complexity or finish. The literature reports that the effect of S-ABA application on grapes is to increase berry and cluster weight (Han, D. H, S. M. Lee, and S. B. Kim. 1996. J. Kor. Soc. Hort. Sci. 37: 416-420). While the effect of S-ABA to increase red color of red grapes has been studied and reported, there are no previous reports on the effect of and the use of S-ABA on red grapes to affect the various sensory characteristics of red grapes and/or the resulting wine. 
     SUMMARY OF THE INVENTION 
     The present invention is directed to the preharvest treatment of red wine grapes with S-ABA or its salts after fruit set. This treatment alters the development of red grape berry and wine flavor characteristics, allowing the berry and wine sensory characteristics, such as aroma, bouquet, flavor, mouthfeel, astringency, balance, complexity or finish to be manipulated to help achieve the wine style desired by the viticulturist and winemaker. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The present invention comprises the application of S-ABA or its salts to red grape grapevines to modify development of grape berry and wine sensory characteristics. S-ABA or a salt of S-ABA is applied to the grapevines as a foliar spray to the grape berries and leaves, by application to the roots of the grapevine through irrigation or fertigation methods, or by injection into the grapevine. 
     Abscisic acid (S-ABA; ABA; S-(+)-abscisic acid; +-ABA, (+)-(S)-cis,trans-abscisic acid, (+)-(S)-cis,trans-ABA; S-ABA; (S)-5-(1-hydroxy-2,6,6,-trimethyl-4-oxo-2-cyclohexen-1-yl)-3-methyl-(2Z,4E)-pentadienoic acid; CAS registry no. [21293-29-8]) is available from Lomon BioTechnology Co., Ltd. (Shichuan, China). 
     Suitable salts of S-ABA include, but are not limited to sodium, potassium, calcium, ammonium, magnesium and amine salts. 
     S-ABA or its salts is applied to the grapevines after berry set to achieve the desired effect on the sensory characteristics of the grape berries and wine. The presently preferred timing of the S-ABA application is during the period of pre-veraison (approximately 3 weeks before veraison, where veraison is defined as berry softening) through the post-veraison period (when sugar levels in berries measure approximately 18-20° brix (10-11 Baume)). 
     Water is used as the carrier solvent for the applications. In the present invention, surfactants can be added to the chemical solution to improve the performance of the S-ABA or its salts, particularly for the foliar application. The water solution may contain between 0.01% to 0.5% v/v of a surfactant, such as Tween 20 (available from Sigma-Aldrich, St. Louis, Mo.). 
     The presently preferred surfactant for S-ABA or S-ABA salt performance is Brij 98 (polyoxyethylene (20) oleyl ether) available from Uniqema (Castle, Del.). Other surfactants are also useful in the present invention, including but not limited to, other surfactants in the Brij family (polyoxyethylene fatty alcohol ether) available from Uniqema (Castle, Del.), surfactants in the Tween family (Polyoxyethylene sorbitan ester) available from Uniqema (Castle, Del.), the Silwet family (Organosilicone) available from Momentive Performance Materials (Wilton, Conn.), the Triton family (Octylphenol ethoxylate) available from The Dow Chemical Company (Midland, Mich.), the Tomadol family (ethoxylated linear alcohol) available from Tomah3 Products, Inc. (Milton, Wis.), the Myrj family (Polyoxyethylene (POE) fatty acid ester) available from Uniqema (Castle, Del.), the Span family (Sorbitan ester) available from Uniqema (Castle, Del.), and the Trylox family (Ethoxylated Sorbitol and Ethoxylated Sorbitol Ester) available from Cognis Corporation (Cincinnati, Ohio) as well as commercial surfactants such as Latron B-1956 (77.0% modified phthalic/glycerol alkyl resin and 23.0% Butyl alcohol) available from Dow AgroSciences LLC (Indianapolis, Ind.), Capsil (Blend of Polyether-polymethylsiloxanecopolymer and nonionic surfactant) available from Aquatrols (Paulsboro, N.J.), Agral 90 (Nonyl phenol ethoxylate) available from Norac Concept. Inc. (Orleans, Ontario, Canada), Kinetic (99.00% proprietary blend of polyalkyleneoxide modified polydimethylsiloxane and nonionic surfactants) available from Setre Chemical Company (Memphis, Tenn.), and Regulaid (90.6% 2-butoxyethanol, poloxalene, monopropylene glycol) available from KALO, Inc. (Overland Park, Kans.). 
     Other additives are also useful in the present invention including, but not limited to, urea, nitrate salts such as ammonium nitrate, salts such as calcium chloride, humectants such as poly(ethylene glycol), and vegetable oils such as soybean oil, corn oil, cotton oil, and palm oil. 
     The effective concentration range of the active ingredient S-ABA varies depending on the water volume applied as well as other factors such as the plant variety, height, age, desired duration of effect and application method. The S-ABA concentration range is from about 1-about 10,000 ppm S-ABA. The preferred S-ABA concentration range for foliar applications is about 50-about 500 ppm. The preferred application volume for foliar applications is about 25-about 300 gallons/A. Foliar spray applications are directed at the grape bunches to achieve complete coverage of the grapes bunches and to maximize the effect of the S-ABA application. Soil applications are directed towards the rooting zone. Application methods include, but are not limited to application through irrigation/fertigation dripper line- or application of S-ABA formulations or solutions to the soil at the base of the vine, followed by application of water to the soil to transport the S-ABA to the roots of the vine. 
     EXAMPLES 
     The following examples illustrate that application of S-ABA to red grapes alters wine sensory characteristics. 
     Example 1 
     S-ABA (200 ppm) was applied by foliar spray using a standard vineyard airblast sprayer at 2000 L/ha to commercially grown Shiraz grapes approximately 3 weeks prior to initiation of veraison. The spray mix contained the surfactant/wetter Agral at 10 ml/100 L. Results reported are means of 4 replicates (grapes) and 3 replicates (wines) per treatment. Winemaking followed a standard small lot fermentation protocol with each replicate wine being made from 50 kg of harvested grapes. 
     Table 1 shows that S-ABA does not affect at-harvest grape sugar, pH, or total acidity or wine pH. However, at-harvest grape anthocyanin and wine color density and total phenolics are increased by S-ABA. 
     
       
         
               
             
               
               
               
             
               
             
               
               
               
               
             
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Shiraz grape quality 
               
               
                 at harvest and wine quality. 
               
             
          
           
               
                   
                 Untreated 
                 S-ABA treated 
               
               
                   
                   
               
             
          
           
               
                 Grapes at harvest 
               
             
          
           
               
                   
                 Sugar (Be) 
                 13.6 
                 13.8 
               
               
                   
                 pH 
                 4.1 
                 4.1 
               
               
                   
                 Total acidity 
                 4.3 
                 4.2 
               
               
                   
                 Anthocyanins (mg/g) 
                 0.92 
                 1.02 
               
             
          
           
               
                 Wine 
               
             
          
           
               
                   
                 pH 
                 3.75 
                 3.76 
               
               
                   
                 Wine color density (a.u.) 
                 5.52 
                 7.23 
               
               
                   
                 Wine hue 
                 0.58 
                 0.56 
               
               
                   
                 Total phenolics 
                 23.84 
                 29.94 
               
               
                   
                   
               
             
          
         
       
     
     Table 2 shows the sensory profile as determined by 10 experienced sensory assessors. Shiraz wine made from S-ABA-treated fruit were described as have a more intense berry aroma and a more fruity, more acidic taste. Using a nine-point hedonic scale rating (dislike=1 to like=9) the sensors judged wine from the S-ABA treatment higher than the untreated (6.2 vs. 5.0, respectively) and thus a technically better wine. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Shiraz wine sensory profile. 
               
             
          
           
               
                 Character 
                 Untreated 
                 S-ABA treated 
               
               
                   
               
               
                 Aroma 
                 Berry, vegetal 
                 More intense berry 
               
               
                 Taste 
                 Dark fruit, soft tannins 
                 More fruity, more acidic 
               
               
                 Color 
                 Red, purple hues 
                 Increased intensity 
               
               
                 Hedonic scale 
                 5.0 
                 6.2 
               
               
                 (1-dislike, 9 like) 
               
               
                   
               
             
          
         
       
     
     Example 2 
     S-ABA (200 ppm) was applied by foliar spray to Cabernet Sauvignon grapes when sugar levels were approximately 11 Be using a standard vineyard airblast sprayer at approximately 2000 L/ha. The spray mix contained the surfactant/wetter Agral at 10 ml/100 L. Wine was prepared from the treated and untreated grapes. Table 3 shows that S-ABA did not affect sugar, pH, total acidity, and anthocyanins of grapes at harvest and did not affect the alcohol, pH, wine color density, wine hue, and total phenolics of the wine. 
     
       
         
               
             
               
               
               
             
               
             
               
               
               
               
             
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Cabernet Sauvignon 
               
               
                 grape quality at harvest and wine quality. 
               
             
          
           
               
                   
                 Untreated 
                 S-ABA treated 
               
               
                   
                   
               
             
          
           
               
                 Grapes at harvest 
               
             
          
           
               
                   
                 Sugar (Be) 
                 12.9 
                 13.1 
               
               
                   
                 pH 
                 3.94 
                 3.97 
               
               
                   
                 Total acidity 
                 4.9 
                 4.9 
               
               
                   
                 Anthocyanins (mg/g) 
                 0.91 
                 0.93 
               
             
          
           
               
                 Wine 
               
             
          
           
               
                   
                 Alcohol (%) 
                 13.3 
                 13.2 
               
               
                   
                 pH 
                 3.49 
                 3.52 
               
               
                   
                 Wine color density (a.u.) 
                 7.0 
                 7.1 
               
               
                   
                 Wine hue 
                 0.6 
                 0.6 
               
               
                   
                 Total phenolics 
                 26.3 
                 25.7 
               
               
                   
                   
               
             
          
         
       
     
     Although S-ABA did not affect sugar, total acidity, and anthocyanin/color of grapes at harvest or wine, an experienced sensory panel judged Cabernet Sauvignon wine from the S-ABA treatment as having a longer finish, increased aroma, and greater color intensity than the untreated wine (Table 4). 
     
       
         
               
             
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Cabernet Sauvignon 
               
               
                 wine quality (1-5 scale; 1 = low/short; 5 = high/long). 
               
             
          
           
               
                   
                 Untreated 
                 S-ABA treated 
               
               
                   
                   
               
             
          
           
               
                   
                 Finish 
                 1.70 
                 2.3 
               
               
                   
                 Flavor intensity 
                 3.0 
                 2.7 
               
               
                   
                 Body/fullness 
                 2.0 
                 1.7 
               
               
                   
                 Aroma intensity 
                 2.3 
                 3.7 
               
               
                   
                 Color intensity 
                 3.0 
                 4.0 
               
               
                   
                   
               
             
          
         
       
     
     Example 3 
     S-ABA (0.4 grams per vine) was applied to commercially grown Shiraz grapes through the drip irrigation system at approximately 3 weeks prior to initiation of veraison. No surfactant was added to the irrigation water. Results reported are means of 3 replicates. Winemaking followed a standard small lot fermentation protocol with each replicate wine being made from 50 kg of harvested grapes. 
     Table 5 illustrates that S-ABA increased anthocyanin and phenolics of grape/juice at crush and wine color density and total phenolics and reduced wine hue. S-ABA had no effect on sugar, pH, or total acidity of grapes/juice at crush or on alcohol or pH of wine. Wine made from S-ABA treated Shiraz berries produced a wine that had more intense fruit and was less acidic based on the judging of 6 experienced wine sensory assessors. 
     
       
         
               
             
               
               
               
             
               
             
               
               
               
             
               
             
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Shiraz grape quality 
               
               
                 at harvest and wine quality. 
               
             
          
           
               
                   
                 Untreated 
                 S-ABA treated 
               
               
                   
                   
               
             
          
           
               
                 Grapes/Juice at crush 
               
             
          
           
               
                 Sugar (Be) 
                 13.7 
                 13.6 
               
               
                 pH 
                 3.78 
                 3.75 
               
               
                 Total acidity 
                 4.3 
                 4.3 
               
               
                 Anthocyanins (mg/g) 
                 0.88 
                 1.06 
               
               
                 Phenolics (mg/g) 
                 0.99 
                 1.06 
               
             
          
           
               
                 Wine 
               
             
          
           
               
                 Alcohol (%) 
                 13.7 
                 13.9 
               
               
                 pH 
                 3.66 
                 3.65 
               
               
                 Wine color density (a.u.) 
                 6.66 
                 7.87 
               
               
                 Wine hue 
                 0.73 
                 0.68 
               
               
                 Total phenolics 
                 34.33 
                 37.41 
               
               
                 Sensory panel of 6 assessors 
                 — 
                 More intense fruit and less 
               
               
                   
                   
                 acidic 
               
               
                   
               
             
          
         
       
     
     Example 4 
     S-ABA (200 ppm) was applied by foliar spray at veraison to commercially grown Shiraz grapes through standard agricultural spray equipment on 12 Jan. 2007. Kendeen 20 (Tween 20) was added to the spray mix at 50 ml/100 L. Reported results are means of 3 replicates. Winemaking followed a standard small lot fermentation protocol with each replicate wine being made from 50 kg of harvested grapes. 
     Table 6 shows that S-ABA did not affect sugar, pH, total acidity, anthocyanins, or phenolics of grapes/juice at crush. S-ABA increased color density and total phenolics, but did not affect alcohol, pH, or wine hue. However, wine made from S-ABA treated Shiraz berries produced a wine that had more intense fruit, spice and herbaceous flavors and more tannin based on the judging of 6 experienced wine sensory assessors. 
     
       
         
               
             
               
               
               
             
               
             
               
               
               
             
               
             
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Effect of abscisic acid (S-ABA) on Shiraz grape quality 
               
               
                 at harvest and wine quality. 
               
             
          
           
               
                   
                 Untreated 
                 S-ABA treated 
               
               
                   
                   
               
             
          
           
               
                 Grapes/Juice at crush 
               
             
          
           
               
                 Sugar (Be) 
                 13.9 
                 13.5 
               
               
                 pH 
                 3.84 
                 3.85 
               
               
                 Total acidity 
                 3.8 
                 3.5 
               
               
                 Anthocyanins (mg/g) 
                 0.93 
                 0.92 
               
               
                 Phenolics (mg/g) 
                 1.04 
                 1.04 
               
             
          
           
               
                 Wine 
               
             
          
           
               
                 Alcohol (%) 
                 13.7 
                 13.8 
               
               
                 pH 
                 3.71 
                 3.72 
               
               
                 Wine color density (a.u.) 
                 9.37 
                 10.63 
               
               
                 Wine hue 
                 0.55 
                 0.53 
               
               
                 Total phenolics 
                 40.1 
                 43.4 
               
               
                 Sensory panel of 6 assessors 
                 — 
                 More intense fruit, spice and 
               
               
                   
                   
                 herbaceous. More tannin 
               
               
                   
               
             
          
         
       
     
     Thus, regardless of the effect of S-ABA on sugar, acidity, anthocyanin/color or phenolics of red grapes at harvest or the subsequent wine made from these berries, these examples illustrate that S-ABA consistently alters the sensory characteristics of the wine such as aroma and flavor.