CERTIFIED CLONE AND POWDERY MILDEW IMPACT ROTUNDONE IN DURAS WINES

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1 Olivier GEFFROY *, Olivier YOBRÉGAT, Thierry DUFOURCQ, Trcey SIEBERT,Éric SERRANO IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce IFV Pôle Sud-Ouest, Domine de Mons, 00 Cussens Frnce The Austrlin Wine Reserch Institute, PO Box 97, Glen Osmond SA Austrli *Corresponding uthor: Geffroy, tel , olivier.geffroy@vignevin.com CERTIFIED CLONE AND POWDERY MILDEW IMPACT ROTUNDONE IN DURAS WINES Few recent studies hve been investigting the effect of clone on rom compounds. The im of this reserch work ws to study the impct of certified clones from Vitis vinifer L. cv. Durs N - grpe vriety grown in the South West of Frnce - on grpe qulity nd rotundone, sesquiterpene responsible for peppery rom which hs been reported recently in red wines mde from this cultivr. u Experimentl site nd design: The experimentl site consisted of four consecutive rows, ech row plnted with one of the four certified clones of Durs N (554, 555, 67 nd 654). For ech clone, mesurements were replicted on three experimentl units of twelve continuous vines per row u Homogeneity control of the plot: Controls crried out on the plot using i) trunk circumference, vrible correlted to rotundone (), ii) δ C mesured on grpes t hrvest, nd iii) NDVI mpping confirmed the bsence of vribility, especilly between the rows u Mesurements on vines nd evlution of powdery mildew severity: At hrvest, number of clusters per vine nd yield (kg/vine) were monitored Severe powdery mildew (Erysiphe nector) infections were recorded on grpes in 04 t hrvest nd its severity ws ssessed by visully estimting the percentge of the cluster re colonized by the pthogen on 00 grpe clusters within ech experimentl unit u Anlyses: Conventionl enologicl prmeters were determined on helthy grpes nd rotundone ws mesured in wines prepred by microvinifiction techniques (-L Erlenmeyer flsks) u Sttisticl tretment: Anlyticl dt were subjected to n nlysis of vrince (ANOVA) tretment using Xlstt softwre (Addinsoft, Frnce). Results Clone number Trunk circumference (cm) Sugr concentrtion ( Brix). 9.9 c 0. b.0 Totl cidity (meq/l) 6 b c 58 c ph Amino cids (mg/l) NH4 + (mg/l) TPI c 77.7 b 77.9 b Anthocynin (mg/kg) b 99 b 95 b Number of clusters per vine Yield (kg/vine) δc Sugr concentrtion ( Brix) Totl cidity (meq/l) ph.8 b.88.8 b.8 b Amino cids (mg/l) NH4 + (mg/l) TPI Anthocynin (mg/kg) Number of clusters per vine Yield (kg/vine) δc Severity of powdery mildew (%) 0.0 c. d.4 b 4.5 Different letters indicte mens significntly different ccording to Fisher s test t P 0.05 u Effect of clone on TC, δ C, clssicl lbortory nlysis nd production ptitude While differences were observed in 0 between the four clones for most of the mesured vribles, few nlyticl dt llowed discrimintion of the clones in 04. In 04, the severity of powdery infections might hve brought some perturbtion nd vribility in the dt set. In 0, clone 554 showed better qulittive performnces (i.e. higher sugr concentrtion, TPI nd nthocynins) in comprison with the other clones nd especilly clone 555. Neither the number of clusters per vine nor yields (kg per vine) t hrvest were impcted in 0 nd 04 by clone type. As significnt differences in severity of PM on grpes t hrvest were observed between the clones in 04 nd PM infection leds to lighter bunch weight, differences in yields could hve been expected. To our knowledge, this is the first time tht clonl differences in powdery mildew susceptibility hve been observed on grpes. Indeed, the differences observed could reflect vritions in qulity of the phytosnitry tretment trgeted ginst PM in reltion with the sprying equipment. However, there ws no direct reltionship between the sequence of plnting of the four clones (554, 555, 67 nd 654) nd the severity of powdery mildew recorded on grpes in 04 (555 < 554 < 67 < 654). u Rotundone in wines Concentrtions of rotundone were significntly higher in wines from clones 554 nd 654 in comprison with clone 555, while clone 67 showed n intermedite level. Rotundone concentrtions observed in 04 (more thn 00 ng/l on verge for three of the studied clones), re well bove the rom threshold of this compound estblished t 6ng/L in red wine () nd re mong the highest ever reported which deserves further discussion. The 0 vintge ws cooler nd the two vintges were chrcterized by similr wter deficit s reflected by δ C mesurements. Therefore, in ccordnce with previous results (), higher rotundone concentrtions were expected in 0. Our results suggest tht nother mechnism is involved. On the experimentl site, correltion (r = 0.58) ws observed between the rotundone Different letters indicte mens significntly different ccording to Fisher s test t P 0.05 concentrtion in experimentl wine nd PM severity. As experimentl wines were mde from helthy berries, these results suggest tht infected berries could induce plnt defence mechnisms leding to synthesis of rotundone in the neighboring helthy berries. The difference of susceptibility to PM could hve been induced by i) constitutive fctors in reltion with physicl properties or brriers tht limits the penetrtion of the pthogen (i.e. cuticle) or ii) induced fctors in reltion with defence responses (i.e. phytolexin). As the correltion observed is positive, mechnisms involved in the clonl susceptibility re more likely to be relted to constitutive thn to induced fctors. Our results found differences in rotundone concentrtions in wines mde from the four Durs N certified clones. The results lso suggested tht grpevine defence response to PM could enhnce rotundone production in berries. Clonl differences in susceptibility to biotic stress, such s PM, might explin the differences observed in rotundone concentrtions between the four studied clones. Our results my ssist grpe growers to produce wines mde from Durs N with desired rom ttribute. One should consider plnting clone 554 in order to promote high levels of rotundone in wines mde from this cultivr. () Geffroy, O., Scholsch, T., Dufourcq, T., & Serrno, E. 05. Understnding nd mpping rotundone sptil vribility in Vitis vinifer L. cv Durs. In Proceedings of the 9th Interntionl Meeting GiESCO, pp () Wood, C., Siebert, T.E., Prker, M., Cpone, D.L., Elsey, G.M., Pollnitz, A.P., & Krmmer, G From wine to pepper: rotundone, n obscure sesquiterpene, is potent spicy rom compound. Journl of Agriculturl nd Food Chemistry, 56(0), () Cputi, L., Crlin, S., Ghiglieno, I., Stefnini, M., Vlenti, L., Vrhovsek, U., & Mttivi, F. 0. Reltionship of chnges in rotundone content during grpe ripening nd winemking to mnipultion of the peppery chrcter of wine. Journl of Agriculturl nd Food Chemistry, 59(0), Reserch works crried out with finncil support from FrnceAgriMer nd the Midi-Pyrénées region Copyright Institut Frnçis de l Vigne et du Vin June 06

2 Olivier Geffroy *, Cmille Buissière, Vlérie Lempereur, Bertrnd Chtelet IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce Fédértion viticole du Puy-de-Dôme, 670 Aubière - Frnce IFV Pôle Bourgogne - Beujolis - Jur - Svoie, 0 Boulevrd Victor Vermorel, CS 600, 6966 Villefrnche-sur-Sône cedex, Frnce *Corresponding uthor: Geffroy, tel , olivier.geffroy@vignevin.com A SENSORY, CHEMICAL AND CONSUMER STUDY OF THE PEPPERY TYPICALITY OF FRENCH GAMAY N WINES FROM COOL CLIMATE VINEYARDS In order to study the peppery typiclity of Gmy N wines from cool climte vineyrd study ws conducted on Gmy N wines from the 0 vintge nd from 4 French viticulturl res (Auvergne, Beujolis, Loire Vlley, nd South West). This reserch work contins sensory, chemicl nd consumer prt nd ws inspired by study conducted by Lund et l. () on Suvignon Blnc. u Wines: The wines selected for the study were from Auvergne (n = ), South West (n = ), Loire Vlley (n = ) nd Beujolis (n = 6). They were selected on the bsis of being predominntly from Gmy grpes (>85%) nd their retil price vried from 4.50 to 9.00 euros u Sensory Anlysis: 8 trined pnelists ssessed relevnt ttributes relted to rom nd tste on 5-point rting scle u Anlyses: Conventionl enologicl prmeters nd rotundone were determined u Consumer study: Four wines were chosen to prticipte in consumer study (n = 87). Rnking tests were performed nd the consumer pnelists hd to crry out n olfctory ssessment bsed on rom preferences. Wines were rnked from their fvourite to 4 the less pprecited u Sttisticl tretment: Sttisticl nlyses including liner regressions were conducted with Xlstt softwre (Addinsoft, Frnce). Olfctory dt were treted through PCA nd AHC. AHC ws used to select four wines with distinct sensory profiles for the consumer study. u Sensory nlysis The PCA plot shows tht four ttributes explin the min differences observed between the Gmy N wines of the study : intensity, fermenttive/lctic, fermenttive/mylic nd spicy/peppery. The scores discriminted group of wines from Auvergne with similr sensory chrcteristics (A-, A-, A-, A-5, A-6, A-7, A-0 nd A-) nd hevier peppery notes. S-, B-4 nd B-6 wines hd pronounced fermenttive/mylic notes, nd B- nd B-5 showed fermenttive/lctic rom sensory profile. The other wines of the study showed less distinctive rom profiles. u Rotundone in wines Wines from Auvergne show higher rotundone concentrtions which is 80 in ccordnce with the sensory observtions. They were produced using 60 trditionl winemking techniques wheres the other wines ll contined percentge of wine produced by semicrbonic 40 0 (whole berries) mcertion or prefermenttion het tretment of grpes. As rotundone is hydrophobic compound, these 00 higher levels my reflect rel terroir effect or rise from specific winemking techniques. Climtic indices clculted 80 for the winegrowing re showed tht Auvergne ws i) the coolest vineyrd over the whole wine-growing seson nd the 60 ripening period, nd ii) the wettest during the verison-hrvest period. Therefore in ccordnce with previous results (), 40 0 its «terroir» is the most likely fctor explining the differences observed between the wines from Auvergne nd the other 0 regions of the study. These differences my be further mplified by the winemking techniques. For the whole dt set, Auvergne Beujolis Loire South West significnt correltion ws observed between peppery rom scores nd rotundone concentrtion in wine (r = 0.66). There ws high vribility in rotundone content mong the wines from Auvergne. Wine rotundone concentrtions were mpped ccording to the plots from which the grpes were primrily sourced nd the rotundone distribution did not pper to be sptilly structured. The only prmeter tht ccounted for the vribility mong the wines ws lcohol content nd there ws significnt positive correltion between this lcohol content nd rotundone (r = 0.44). Rotundone concentrtion (ng/l) Results Wine / sensory profile S- B- A-6 A-8 Amylic Lctic Peppery Green Sum of the rnks Number of times rted s the fvorite 7 8 Number of times rted s the lest pprecited % of consumers who re femle when fvorite 8 9 % of consumers who re mngers nd professionls when fvorite % of consumers who re older thn 45 yers when fvorite % of consumers who re between 5 nd 45 yers old when fvorite % of consumers who re younger thn 5 yers when fvorite % of consumers who py more thn 5 when fvorite % of consumers who py more thn 0 when fvorite Number of times rted s the lest pprecited when A-6 (peppery) is rted s the fvorite Number of times rted s the lest pprecited when S- (mylic) is rted s the fvorite Different letters within the sme row indictes sums of the rnks significntly different ccording to the Nemenyi test t P Wines were rnked from their fvourite to 4 the less pprecited. u Consumer study Four wines showing distinct sensory profiles were selected for the consumer study. S- hd higher romtic intensity nd hevy fermenttive/mylic nd fermenttive/lctic notes. B- hd more complex profile mrked by significntly higher lctic notes. A-6 nd A-8 were chrcterized by peppery nd green notes. Regrding the olfctory ssessment of the wines, there were no significnt differences ccording to Friedmn s test between the four wines. A lck of consensus ws observed for wine S- nd A-8. The group of consumers who preferred the mylic wine frequently rted the peppery wine s the lest pprecited nd vice vers. This led us to think tht there is n opposition between the consumers who preferred the mylic nd the peppery wines. Consumers who rted the peppery wine s the most pprecited were mngers nd professionls who re willing to py more for bottle. These results tht scientificlly investigted the typiclity nd originlity of Gmy N wines from Auvergne should ssist winegrowers from this region in promoting their wines. They llow them to tke dvntge of the knowledge on rotundone obtined in nother wine-growing regions with the im to produce wines with desired level of peppery rom. The consumer study provides the key prmeters for developing the Côtes d Auvergne Gmy N wine rnge nd dpting the products to consumer profiles. () Lund, C.M., Thompson, M.K., Benkwitz, F., Wohler, M.W., Triggs, C.M., Grdner, R., & Nicolu, L New Zelnd Suvignon blnc distinct flvor chrcteristics: Sensory. chemicl. nd consumer spects. Americn Journl of Enology nd Viticulture. 60(). -. () Cputi, L., Crlin, S., Ghiglieno, I., Stefnini, M., Vlenti, L., Vrhovsek, U., & Mttivi, F. 0. Reltionship of chnges in rotundone content during grpe ripening nd winemking to mnipultion of the peppery chrcter of wine. Journl of Agriculturl nd Food Chemistry, 59(0), Reserch works crried out with finncil support from FrnceAgriMer nd the Auvergne region Copyright Institut Frnçis de l Vigne et du Vin June 06

3 Olivier GEFFROY *, Thibut SCHOLASCH, Thierry DUFOURCQ, Eric SERRANO IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce Fruition Sciences, MIBI, 67 rue du Ms de Verchnt, 4000 Montpellier - Frnce IFV Pôle Sud-Ouest, Domine de Mons, 00 Cussens Frnce *Corresponding uthor: Geffroy, tel , olivier.geffroy@vignevin.com DIFFERENTIAL VEST: SELECTING WITHIN- VINEYARD AREAS TO PRODUCE WINES WITH ENHANCED ROTUNDONE CONCENTRATION Recent reserch works highlighted lrge vribility in rotundone concentrtion - the min compound responsible for peppery rom in red wines - within vineyrd block ssocited with vrition in lnd underlying the vineyrd () nd vine wter deficit (). This study ims t determining prcticl wys nd setting up methodology to mp rotundone sptil vribility. u Experimentl vineyrd: The experiment ws crried out on 0.4 h Guyot trined Durs vineyrd from the South West of Frnce plnted in 999 u Climtic bckground: The vineyrd ws locted in n re under ocenic climtic influence with norml nnul rinfll nd men ir temperture of 68 mm nd.9 C, respectively. The 04 vintge ws chrcterized by n extremely riny summer (60mm of rin between bunch closure nd full verison) u Smpling optimiztion protocol: Vineyrd smpling ws optimized (). Vribility in vine rchitecture of the plot ws ssessed through 5 mesurements of trunk circumference (TC), vrible reflecting vine response to wter nd minerl deficits since plnttion (4). Dt were sptilized through the kriging method. On the bsis of the three clsses of TC obtined, six smrt points of 50 vines were positioned - with A nd B corresponding to high, C nd D to verge nd E nd F to low TC clsses - nd monitored independently (see mp on the right). The six smrt points represented 7.5% of the whole block re u Nitrogen uptke, wter sttus nd vine performnces: Nitrogen sttus ws chrcterized through i) petiole nlysis on verge smple of 50 bsl leves nd ii) 50 mesurements ( per vine) of Dulex, fluorescence bsed sensor which llows the clcultion of chlorophyll rtio nmed NBI. For ech smrt point, stem wter potentils (Ψstem) were mesured t 4 time points nd yields t hrvest (kg/vine) were monitored. On ech smrt point, pruning wood weights (PWW) were mesured through mnul weighing during winter dormncy. u Fruit composition nd rotundone: Conventionl enologicl prmeters nd δ C were determined t different time points. Rotundone determintions nd sensory nlysis on 5-point rting scle were conducted in wines prepred by minivinifiction techniques (60 L tnks) u Sttisticl tretment: Anlyticl dt were treted through PCA, ANOVA for those with replicted mesurements (Ψstem, NBI, yield nd PWW). u Results overview The PCA plot shows tht smrt points C nd D hve more similrities with smrt points E nd F nd should be merged with the low TC clss when considering selective hrvest. Liner regression models (n = 6) indicte tht rotundone is correlted with vribles relted directly or indirectly to wter deficit such s Ψstem t bunch closure (r = 0.75), t hrvest (r = 0.87), weight of 00 berries t hrvest (r = 0.85), mlic cid Smrt point t mid-verison (r = 0.76), δ C t mid-verison (r mesurement A B C D E F= 0.9) nd t hrvest (r = 0.8), which is consistent with previous observtions (). u Nitrogen uptke, wter sttus, vine performnce, fruit qulity, rotundone nd sensory nlysis Wter deficit nd differences between the smrt points were wek which is consistent with the hypothesis tht low wter deficit leds to low sptil contrst (5). Indictors used in routine nd well known by winegrowers such s yields t hrvest, fruit qulity, nitrogen uptke through petiole nlysis, pruning wood weights did not llow to discriminte the smrt points from n gronomicl stndpoint while better results were obtined with TC, Ψstem nd NBI. Ψstem especilly t bunch closure, llows perfect discrimintion of the smrt point ccording to TC clsses. Vribles mesured on fruit t hrvest were similr between the smrt points with the exception of Totl Acidity (TA), Yest Assimilble Nitrogen (YAN) nd rotundone. For the pepper rom compound, verge vlues of 94.5, 60.5 nd 6 ng/l were mesured in wines from the high, verge nd low TC re respectively. At tsting, significnt differences were noticed between the wines for the peppery score. Our results suggest tht it is possible under Atlntic conditions nd during vintge with low wter deficit to produce two wines from the sme vineyrd block, differing only by their rotundone concentrtion nd peppery tribute. Vrible Period of P-vlues Nitrogen sttus (NBI) FCFS b b - c Berry set < d 4.99 cd 5.4 bc 5. bc b Pe size b 4.87 b 5. b Bunch closure Verison Hrvest < c 5.86 c b 5.6 b 6.0 bc Totl N (% dry mtter) FCFS - b Berry set Pe size Bunch closure Verison Ψstem (Mp) Bunch closure < b b -0.6 b b Verison + 5 dys < b bc bc c Hrvest < c -0.7 b -0.8 bc -.07 d δ C Bunch closure Verison Hrvest PWW d (kg/vine) Winter dormncy Yields (kg/vine) Hrvest Sugr concentrtion ( Brix) Grpe t hrvest TA (g/l H SO 4 ) Grpe t hrvest ph Grpe t hrvest YAN (mg/l) Grpe t hrvest Anthocynins (mg/kg) Grpe t hrvest Totl Phenol Index (TPI) Grpe t hrvest Rotundone (ng/l) Finished wine Peppery score t tsting Finished wine b.44 b. b.8 b Acidity score t tsting Finished wine Astringency score t tsting Finished wine Flower cluster with floret seprtion. b Different letters within the sme row indictes mens significntly different t P c Not determined. d Pruning wood weights. The present works llowed to set up methodology for ssessing rotundone sptil distribution through the smrt point method. Our results suggest tht link exists between rotundone in wine nd plnt rchitecture (TC) nd tht this cost-effective indictor cn be used to pproch rotundone sptil distribution. Our study lso highlights the sensitivity of rotundone to fine vrition of grpevine wter sttus. () Scrlett, N.J., Brmley, R.G.V., & Siebert, T.E. (04). Within-vineyrd vrition in the pepper compound rotundone is sptilly structured nd relted to vrition in the lnd underlying the vineyrd. Austrlin Journl of Grpe nd Wine Reserch, 0(), 4-.. () Geffroy, O., Dufourcq, T., Crcenc, D., Siebert, T., Herderich, M., & Serrno, E. 04. Effect of ripeness nd viticulturl techniques on the rotundone concentrtion in red wine mde from Vitis vinifer L. cv. Durs. Austrlin Journl of Grpe nd Wine Reserch, 0(), () Meyers, J.M., Scks, G.L., Vn Es, H.M., & Vnden Heuvel, J.E., 0. Improving vineyrd smpling efficiency vi dynmic sptilly explicit optimistion. Austrlin Journl of Grpe nd Wine Reserch, 7, (4) Tisseyre, B., Ojed, H., Crillo, N., Deis, L., & Heywng, M Precision viticulture nd wter sttus, mpping the predwn wter potentil to define within-vineyrd zones. In Proceedings of the 4th GiESCO Congress, pp. -7. (5) Tylor J. A., Acevedo Opzo C., Ojed H., Tisseyre B., 00. Identifiction nd significnce of sources of sptil vrition in grpevine wter sttus. Austrlin Journl of Grpe nd Wine Reserch, 6, 8-6. Reserch works crried out with finncil support from FrnceAgriMer nd the Midi-Pyrénées region Copyright Institut Frnçis de l Vigne et du Vin - June 06

4 Olivier GEFFROY *, Trcey SIEBERT, Anthony SILVANO, Mrkus HERDERICH IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce The Austrlin Wine Reserch Institute, PO Box 97, Glen Osmond SA Austrli Lllemnd SAS, 9 rue des Briquetiers, BP 59, 70 Blgnc - Frnce *Corresponding uthor: Geffroy, tel , olivier.geffroy@vignevin.com DO DIFFERENT WINEMAKING TECHNIQUES AND PRESSING CONDITIONS IMPACT ROTUNDONE LEVELS IN RED WINE? Rotundone responsible for blck pepper rom in wine (), hs been only detected in the grpe berry exocrp (,) which suggests tht mcertion vribles re likely to impct its concentrtion in wine. Two seprte experiments were conducted to investigte the impct of i) key fermenttion prmeters nd winemking techniques t the lbortory scle nd ii) pressing conditions t semi-industril scle on the extrction of rotundone. u Lbortory scle experiment: 7 homogenous lots of 800 g destemmed berries were obtined in 05 from n irrigted Durs vineyrd locted in the South West of Frnce (PDO Gillc). Nine tretments were investigted in triplicte in L Erlenmeyer : control vinifiction t 5 C for 8 dys (Control), pectolytic enzyme ddition t 40 mg/kg (Enzyme), cold sok t 4 C for 7 hours (Cold), thermovinifiction t 70 C for hours followed by pressing, clrifiction nd fermenttion in liquid phse t 8 C (Thermo), vinifiction t 0 C for 8 dys (0 C), vinifiction t 5 C for 4 dys (4 dys), semi-crbonic mcertion t 0 C for 8 dys followed by pressing nd fermenttion in liquid phse t 8 C (Crbonic), rosé vinifiction fter 6-hour skin contct t 8 C (Rosé) nd fermenttion with Scchromyces uvrum (Uvrum) u Semi-industril scle experiment: tons of Syrh grpes from neighbouring dry-grown vineyrd were fermented t 5 C. After 0 dys of mcertion, grpes were pressed using pneumtic press. Smples were collected in the try of the press t different conditions of pressing : free run (Free run), dynmic in the press (De-juicing), pressing t 00mb (00mb), pressing t 400mb (400mb), pressing t 600mb (600mb) u Anlyses: Conventionl enologicl prmeters nd rotundone were determined in wines fter one week nd one month respectively u Sttisticl tretment: Anlyticl dt from the lbortory scle experiment were subjected to one-wy nlysis of vrince (ANOVA) tretment using Xlstt softwre (Addinsoft, Frnce). Different letters indicte mens significntly different ccording to Fisher s test t P 0.05 The dotted line represents the rom threshold of rotundone in red wine () u Fermenttion prmeters nd winemking techniques In comprison with the control vinifiction, the Enzyme nd Cold tretments hd wek impct on the mesured prmeters. For Enzyme, the pectin degrdtion mximized juice yield nd surprisingly, Totl Phenol Index (TPI) nd rotundone were not impcted. Cold sok hd no effect on nthocynins, TPI nd rotundone but induced gin in ph. The Thermo tretment hd lrge impct on the enologicl prmeters. In ccordnce with previous works (4), n increse in lcohol, ph, trtric cid, mlic cid nd VA ws noticed s consequence of lrger extrction of mino nd orgnic cids from the berry exocrp when pressing the grpes t hot temperture. As TPI vlues were incresed by the het tretment of grpes, gin in rotundone which lso hs hydrophobic behviour would hve been expected. Our results suggest tht rotundone is less het-extrctble thn pronthocynidins in queous phse. The impct of mcertion prmeters (time nd temperture of mcertion) on TPI nd especilly on rotundone reflected by the 0 C nd 4 dys tretments re unexpected s the literture usully supports the belief tht the extension of the time nd the increse in the temperture enhnce extrction of the skins nd seeds (5). Crbonic wines hd significnt smllest levels of lcohol due to n inversion of the Krebs cycle, nthocynins nd TA leding to higher ph. Even if the quntity of ethnol produced during the nerobic phse - typiclly bout to %. vol. - remins low t pressing, it llowed significnt extrction of rotundone. The rotundone in Rosé wines represented on verge % of the concentrtion found in the Control which indictes thn in most cses, rotundone is not likely to hve lrge sensory contribution to the rom of rosé wines. Rotundone, nthocynins nd ph were significntly impcted by the Uvrum tretment which must be the consequence of lesser extrction or lrger dsorption. Prmeter Control Enzyme Cold Thermo 0 C 4 dys Crbonic Rosé Uvrum Alcohol (% vol.).7 bc.6 bc.4 cd.4..8 b. d.9 b.5 bcd TA (g/l t. cid) 6.85 b b 6.8 c 6.46 c 6.55 bc 6.9 c 6.55 b 7.06 ph.57 d.5 e.60 c.7.65 b.67 b.70.4 f.6 c Trtric cid.76 de.7 de.9 bc.5.9 f.8 cd.95 b.0.69 e Mlic cid.74 b.76 b.86 b.9.7 b.77 b.7 b.4 c.75 b VA (g/l cetic cid) 0. b 0. b 0.0 b b 0. d 0.0 b 0.5 b 0.0 b Potssium b.56 Totl Phenol Index 5.6 bc 54.9 b 48.6 c c 46.0 c 56. b. d 46. c Anthocynins (mg/l) b 68 c 600 c 600 c 79 d 66 c Skin-to-juice rtio (%) 0.5 d 0.9 e 0.5 d 0.57 c 0. d 0. d 0.55 b d Different letters indicte mens significntly different ccording to Fisher s test t P 0.05 Tretment Alcohol content (% vol.) TA (g/l t. cid) ph Trtric cid Glucose / Fructose Voltile cidity K+ Totl Phenol Index Anthocynins (mg/l) Rotundone (ng/l) Free run Dynmic mb mb mb u Pressing conditions Pressing conditions hd limited impct on most of the mesured prmeters with the exception of glucose nd fructose. For rotundone, it remins difficult to drw firm conclusions,s concentrtions detected in wines were very low. The 05 vintge prticulrly dry nd hot in the South West of Frnce, ws not fvorble to rotundone ccumultion. None of the studied tretments resulted in enhnced rotundone concentrtions in wine compred to Control wine. Semi-crbonic mcertion, fermenttion with Scchromyces uvrum or longer skin contct resulted in wine with significnt reduction in rotundone, indicting prcticl opportunities for reducing pepper rom in wine. Pre-ferment removl of skins s typiclly used during thermovinifiction nd in rosé winemking resulted in the lowest wine rotundone concentrtions. Pressing conditions hd limited impct on rotundone in wine. () Wood C, Siebert TE, Prker M, Cpone DL, Elsey GM, Pollnitz AP, nd Krmmer, G From wine to pepper: rotundone, n obscure sesquiterpene, is potent spicy rom compound. J Agric Food Chem, 56(0), () Cputi L, Crlin S, Ghiglieno I, Stefnini M, Vlenti L, Vrhovsek U, nd Mttivi F. 0. Reltionship of chnges in rotundone content during grpe ripening nd winemking to mnipultion of the peppery chrcter of wine. J Agric Food Chem 59: () Siebert TE, nd Solomon MR. 0. Rotundone: development in the grpe nd extrction during fermenttion. In Proceedings of the 4th AWITC; Blir RJ et l. (eds.), pp AWITC Inc., Adelide, Austrli. (4) Geffroy O, Lopez R, Serrno E, Dufourcq T, Grci-Moreno E, Ccho J, nd Ferreir V. 05. Chnges in nlyticl nd voltile compositions of red wines induced by pre-fermenttion het tretment of grpes. Food Chem, 87, 4-5. (5) Scchi KL, Bisson LF, nd Adms DO A review of the effect of winemking techniques on phenolic extrction in red wines. Am J Enol Vitic, 56(), Reserch works crried out with finncil support from FrnceAgri- Mer, the Midi-Pyrénées region nd Wine Austrli, with mtching funds from the Austrlin government. Copyright Institut Frnçis de l Vigne et du Vin - June 06

5 Olivier GEFFROY *, Juliette DESCOTES, Christin DEBORD, Thierry DUFOURCQ IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce IFV - Pôle Bordeux Aquitine, 9 rue Michel Montigne, 90 Blnquefort - Frnce IFV - Pôle Sud-Ouest, Domine de Mons, 00 Cussens - Frnce *Corresponding uthor: Geffroy, tel , olivier.geffroy@vignevin.com MODELLING IMPACTS OF VITICULTURAL AND ENVIRONMENTAL FACTORS ON ROTUNDONE IN DURAS WINES It hs been shown tht rotundone - the rom compound responsible for peppery rom in red wines - ws impcted by temperture nd wter sttus within vineyrd (,). A study ws conducted to determine the key environmentl nd viticulturl vribles ffecting the rotundone concentrtion in Durs wines grown in the South West of Frnce (PDO Gillc) nd mde from 0 different vineyrd blocks in 0 nd in 04. u Vineyrds blocks : 0 vineyrd blocks plnted with Durs from the PDO Gillc (400 h) were used for this study. At ech site, 0-vine pnel from consecutive rows ws selected for dt collection nd smpling. No lef removl ws performed. u Climtic dt : Antilope dt (T C, rinflls, evpotrnspirtion, hygrometry) were provided by MeteoFrnce t kilometer scle. Solr rdition dt were obtined from HelioClim-, stellite-bsed surfce solr irrdition dtbse u Site, phenology, vine nd crop dt : 4 vribles were collected or mesured t ech experimentl site. Stem wter potentils were clculted using the WLIS wter blnce () u Rotundone : Rotundone ws quntified in wine (L Erlenmeyer) mde from helthy grpes hrvested 44 dys fter verison u Sttisticl tretment : Prtil lest squres regression (PLSR) ws conducted with Xlstt (Addinsoft, Pris, Frnce) to model rotundone. Site Phenology Vine dt Crop Climte Metric Metric Metric Period(s) of mesurement Metric Period(s) of mesurement Metric Period(s) of mesurement yer of plnttion 5d before VER pruning wood weights WIN yield men ir T C VER- slope onset of VER exposble lef re VER cluster weight minimum ir T C VER- clone VER (50%) height-to-width rtio VER 00 berry weight, VER mximum ir T C VER- rootstock full VER lef-to-fruit rtio soluble solids men ir hygrometry VER- trining system b ψstem c 5d before VER totl cidity men dily irrdition VER- row orienttion durtion of VER onset of VER ph sunshine hours VER- % of cover crop VER mlic cid men evpotrnspirtion VER- vine density / h full VER trtric cid cumultive rinflls April-0 September ltitude mino cids 5d before VER-VER trunk circonference mmonium VER- verison b hrvest c stem wter potentil TPI nthocynins δ C d gluconic cid PM severity skin to juice rtio, VER u Overview of the 0 nd 04 vintges The 0 vintge ws chrcterized by cold spring, regulr nd lrge rinfll events over the vine vegettive cycle which induced dely of lmost weeks t hrvest nd ripening difficulties. In 04, winter ws riny nd wrm; summer ws extremely riny with 60 mm of rin recorded between mid- July nd the end of August nd conditions during ripening were dry nd hot. Severe bunch rot (Botrytis cinere) ws recorded on bunches in 0. Men ir tempertures over the VER- period were lower in 0 (8.5 C ± 0.6) thn in 04 (9.5 ± 0.4). For the sme period, levels of wter deficit reflected by mesurements of δ C were equivlent for the two vintges (-6.4 ±. in 0; -6.7 ± 0.6 in 04). u Rotundone concentrtions Rotundone concentrtions were higher in the cooler nd wetter 0 growing seson in ccordnce with previous results (4). Surprisingly, some high rotundone vineyrds in 0 (i.e. blocks, 4 nd 8) showed low to moderte levels of concentrtions in 04. In the sme wy, some blocks hving high concentrtions in 04 (i.e. blocks, nd 5) showed low rotundone in 0. This led us to think tht fixed vribles such s yer of plnttion, ltitude, trining system, clone or rootstock don t hve lrge contribution to the rotundone model. Dotted line represent men rotundone vlues found in wines in 0 nd 04 u Multisite PLSR models for rotundone in wine During the 0 vintge with ripening difficulties, precocity of verison, gluconic cid - secondry metbolite of B. cinere - nd cumultive rinflls over the VER- period hd negtive regression coefficients. B. cinere hs the bility to withstnd the toxic effects of plnt such s sesquiterpenes by its lccse through oxidtion nd detoxifiction (5). Therefore, our results suggest tht the fungi cn induce the degrdtion of rotundone. For the 04 nd 0-04 models, rotundone ws best predicted by cumultive rinflls over the st of April-0th of September nd by stem wter potentils 5 dys before verison in 04, which is in greement with previous results (6,). Vrible PLSR models : bsics sttistics nd regression coefficients for the best rotundone model Men vlues nd stndrd devitions Metric Cumultive rinflls VER- (mm) ± 8 6 ± Cumultive rinflls st April-0th September (mm) ± 58 ± 9 Onset of VER (Julin dy) ± 4 4 ± ψstem5d before VER (MP) ± ± 0.7 Gluconic cid (mg/l) ± 69 4 ± R RMSE indictes vribles not in the model Importnt vribles identified for modelling rotundone in wine were those ssocited with cumultive rinflls nd B. cinere dmges. Our results suggest tht mesoclimte is the key fctor to explin the differences in rotundone observed mong the sites. () Zhng, P. et l. (05). Within-vineyrd, within-vine, nd within-bunch vribility of the rotundone concentrtion in berries of Vitis vinifer L. cv. Shirz. Journl of Agriculturl nd Food Chemistry, 6(7), () Geffroy, O. et l. (04). Effect of ripeness nd viticulturl techniques on the rotundone concentrtion in red wine mde from Vitis vinifer L. cv. Durs. Austrlin Journl of Grpe nd Wine Reserch, 0(), () Dufourcq, T. et l. (00). Use of the wter blnce model WLIS to predict stem wter potentil on Colombrd vriety in Gscony vineyrd. Ciênc e Técnic Vitivinicol, 8: (4) Cputi, L. et l. (0). Reltionship of chnges in rotundone content during grpe ripening nd winemking to mnipultion of the peppery chrcter of wine. Journl of Agriculturl nd Food Chemistry 59: (5)Myer, A.M., nd Stples, R. C. (00). Lccse: new functions for n old enzyme. Phytochemistry, 60(6), (6) Zhng, P.et l. (05). Environmentl fctors nd sesonlity ffect the concentrtion of rotundone in Vitis vinifer L. cv. Shirz wine. PloS one, 0(7), e07. Works crried out with finncil support from FrnceAgriMer nd the Midi-Pyrénées region Institut Frnçis de l Vigne et du Vin June 06

6 Olivier GEFFROY, Frédéric CRIER, Pscl POUPAULT, Rémi SCHNEIDER, Ricrdo LOPEZ 4, Lure GONTIER, Thierry DUFOURCQ * IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn Frnce IFV - Pôle Vl de Loire, Châteu de l Frémoire Vertou - Frnce IFV - Pôle Rhône-Méditerrnée, Domine de Pech Rouge, 40 Gruissn - Frnce 4 LAAE, Universidd Zrgoz, Zrgoz - Spin *Corresponding uthor: Dufourcq, tel , thierry.dufourcq@vignevin.com FOLIAR NITROGEN AND SULFUR SPRAYING ALLOW TO PRODUCE WINES WITH ENHANCED CONCENTRATION IN VARIETAL THIOLS Vrietl thiols, especilly -mercptohexnol (MH) nd -mercptohexylcette (MHA), re desirble rom compounds identified in white, rosé nd red wines mde from severl cultivrs. These fruity compounds re linked to the plnt nitrogen (N) uptke () nd sulfur (S) metbolism by its S-cyteine nd S-glutthione conjugtes described s rom precuror (). Between 005 nd 0, severl experiments were conducted to ssess the impct of folir pplictions of nitrogen (formulted forms of ure, orgnic fertilizers) combined or not with sulfur on severl cultivrs. u Conditions of sprying : To llow sufficient dilution of the fertilizer nd void toxicity, folir spryings were crried out t 400L/h nd divided into two pplictions (onset of verison nd full verison) u st experiment on white nd rosé wines : A formulted form of ure (Folur, Trdecorp, Belgium) ws pplied through folir wy t the rte of 0, 5 nd 0kg N/h. These pplictions were combined or not with elementl soluble sulfur (Microthiol, Cerexgri, Frnce) t reduced doses (from 5 to 0kg S/h). Spryings were tested on Colombrd B, Gros Mnseng B, Négrette N, Suvignon B, Melon B nd Suvignon B between 005 nd 008 u nd experiment with orgnic fertilizers : commercil products obtined from enzymtic digestion of niml (Aminovitl, Biof, Germny), vegetl (Diglutin, Biof, Germny) nd mrin (Liquoplnt B6, Plntin, Frnce) proteins were tested t 0kg N/h in 00 on Suvignon B grown in the South West of Frnce u rd experiment on red wines : A formulted form of ure (Azofol SR, Agronutrition, Frnce) ws pplied in 0 t 0 kg N/h without sulfur due to the risk of developing reductive off-flvours, on Fer N nd Crignn N u Winemking nd nlyzes : For ll the experiments, wines were elborted t pilot scle (0L). Grpes were processed under strict non-oxidtive protocol for white nd rosé wines. For red wines, fermenttion nd mcertion took plce t 5 C for 8 dys. Nitrogen in must ws determined s the sum of mmoni nd mino-cid concentrtions. Wines were nlyzed for their rom composition, especilly for their concentrtion of vrietl thiols (MH nd MHA). The molr sum of these two compounds ws used s n indictor of the vrietl thiol potentil. u Cn the technique be dpted to orgnic viticulture? u A lrge impct on the nitrogen content of the must nd the rom composition of white nd rosé wines The gin in nitrogen induced by the folir ppliction ws liner: 50% nd 00% increses were noticed for 0 nd 0kg N/h pplictions respectively. In comprison with controls, nitrogen nd sulfur spryings induced three to four-fold gin in vrietl thiols. This increse ws observed even for control wines with high concentrtions in thiols (from 0 to 40 nm/l). When performing sensory nlysis, wines from the N nd S tretment were judged more intense nd presented higher scores for grpefruit nd tropicl fruit ttributes. No undesirble sulfur/reductive notes were perceived t tsting. Our results showed tht orgnic fertilizers cn be ssimilted by grpevine through folir wy. The gin obtined for Aminovitl reching 48%, ws equivlent to the increse expected for ure-bsed ppliction (50%). Concentrtions in vrietl thiols mesured in wines were very wek nd folir spryings hd deprecitive impct on the quntity of molecules. No differences were found between the four wines t tsting. Severe burn dmges, prticulrly mrked for the Liquoplnt B6 nd Diglutin tretments, were noticed on the folige t hrvest.this phytotoxicity cn be minly explined by the concentrtion in nitrogen of the orgnic fertilizers which doesn t llow sufficient dilution. The cost of this product lso poses serious thret to the development of the technique. Tretment Nitrogen in must Content (mg/l) Gin in comprison with the control (%) MH (ng/l) MHA (ng/l) Molr sum MH +MHA (nm/l) Control Liquoplnt B6 6 8% 8.0 Diglutin 5 % Aminovitl 70 48% u Is the technique lso suitble for the production of red wines? Red wines from the folir tretment hd lower Totl Phenol Index (TPI) which is not completely unexpected s it hs been shown tht biosynthesis pthwys of pronthocynidins were down-regulted by n excessive nitrogen uptke (). Surprisingly, despite verge gins of 5% nd 0% in glutthione nd cysteine precursors of MH respectively, concentrtions in MH were not impcted in finished wines. Red winemking conditions (higher temperture nd turbidity) re less fvourble to the relese of MH (4) nd the bsence of nitrogen/sulfur combintion in this experiment might lso hve plyed role (5). From sensory point of view, wines from the nitrogen tretment hd mrked reductive off-fvours which cn be explined by the higher concentrtion in hydrogen sulphide found in wines. The ppliction of sulfur nd nitrogen through folir wy t verison is powerful viticulturl technique to over-express the vrietl chrcter of white nd rosé wines without undesirble side effects. The technique cnnot be yet dpted to orgnic viticulture nd its interest is limited when mking red wines. () des Gchons et l. (005). Influence of wter nd nitrogen deficit on fruit ripening nd rom potentil of Vitis vinifer L cv Suvignon blnc in field conditions. Journl of the Science of Food nd Agriculture, 85: () Toming, T. et l. (998). A New Type of Flvor Precursors in Vitis v inifer L. cv. Suvignon Blnc: S-Cysteine Conjugtes. Journl of Agriculturl nd Food Chemistry, 46: () Lillo, C. et l. (008). Nutrient depletion s key fctor for mnipulting gene expression nd product formtion in different brnches of the flvonoid pthwy. Plnt Cell Environnement, : (4) Msneuf-Pomrède, I. et l. (006). Influence of fermenttion temperture on voltile thiols concentrtions in Suvignon blnc wines. Interntionl Journl of Food Microbiology., 08: (5) Lcroux, F. et l. (008). Effect of folir nitrogen nd sulphur ppliction on romtic expression of Vitis vinifer L. cv. Suvignon blnc. Journl Interntionl des Sciences de l Vigne et du Vin, 4: 5. These studies were crried out with finncil support from FrnceAgriMer, the Midi-Pyrénées region nd the Europen Community through the POCTEFA. Copyright IFV - June 06

7 Olivier GEFFROY, Fnny PREZMAN*, Ginni TRIOLI, Mrijn BUBOLA, Cristin CARLOS4, Andres KORTEKAMP5, Eric SERRANO IFV Pôle Sud-Ouest, V innopôle, BP, 80 Lisle sur Trn - Frnce; VINIDEA srl, 908 Ponte dell Olio - Itly; Institute of Agriculture nd Tourism, 5440 Poreč - Croti; 4 ADVID, Vil Rel - Portugl; 5 DLR Rheinplz, 6745 Neustdt - Germny *Corresponding uthor: Fnny Prezmn: tel , fnny.prezmn@vignevin.com WINETWORK : Identifiction nd dissemintion of innovtive control strtegies for Flvescence Dorée nd Grpevine Trunk Diseses mngement WINETWORK is Europen collbortive project for the exchnge nd trnsfer of innovtive knowledge between Europen wine-growing regions to increse the productivity nd sustinbility of the sector. For yers, prtners of 7 Europen countries will exchnge knowledge on two importnt diseses in vineyrd: Grpevine Trunk Diseses nd Flvesence dorée. These diseses re well-known in mny vineyrds nd hve been extending for severl yers in different Europen countries, nd hving big economic importnce for the Europen wine industry. As mny winegrowers re testing innovtive nd sustinble pproches to fight ginst these diseses, it is very beneficil to cpture these ides nd to shre them between Europen countries. AN INNOVATIVE METHODOLOGY Project WINETWORK is developing the innovtion-driven methodology, which promotes the exchnge of knowledge between science nd prctice. This network is implemented in ten winegrowing regions, within seven Europen countries. The methodology is built on network of winegrowers nd regionl experts gthered in regionl Technicl Working Groups, Scientific Working Groups (one for ech disese), nd ten regionl Fcilittor Agents tht stimulte their interction. Bottom-up pproch pplied in WINETWORK includes surveys, conducted nd synthesised by Fcilittor Agents, to identify winegrower s innovtive prctices, while scientists revise scientific dt nd dpt it to dissemintion nd prcticl use. This prticiptory pproch will llow trnsferring results from science nd prcticl knowledge to mterils dpted to end-users. This network will promote interctions between scientists nd prctitioners to gther nd shre experiences nd knowledge of different ctors from the min wine producing Europen regions. THE KNOWLEDGE RESERVOIR Collected knowledge will be gthered in the knowledge reservoir, nd will serve s bsis for Fcilittor Agents in collbortion with Technicl Working Groups nd Scientific Working Groups to co-crete orignil mteril dpted to innovtion support services nd to winegrowers. The knowledge reservoir is prticiptive tool hving the mbition to host ll existing knowledge on the topics, both developed by reserch or derived from prcticl experience. Those stkeholders-such s scientists, dvisors or winegrowers-who would like to shre their documented knowledge cn contribute to this webrchive by uploding videos, imges nd documents. All the creted mteril will be vilble for the Europen wine sector nd will llow, for exmple, winegrowers to recognize diseses symptoms nd to better understnd their mngement. Flvescence Dorée is qurntine disese cused by phytoplsm ffecting grpevine. This pthology is trnsmited from vine to nother by n insect vector, Scphoideus titnus. Once fected grpevine cn express symptoms s n bsence of lignifiction of green shoots which become rubbery nd dngling, dditionnlly yellow or red discolortion nd rolling down of leves is observed, berry setting nd wilting of clusters re lso reduced. EXAMPLES OF INNOVATIVE CONTROL STRATEGIES Grpevine Trunk Diseses gther three min diseses: Esc, Botryospheri diebck nd Eutyp diebck. These diseses re widely spred nd cuse lrge dmges in Europen vineyrds. Depending on grpe vriety, symptoms re expressed with vrible intensity. Folir symptoms nd internl symptoms re typicl of grpevine trunk diseses. By visiting regionl vineyrds, nd reviewing scientific literture nd reserch results, severl control strtegies hve been identified. For Flvescence Dorée, control strtegies re bsed on vector monitoring, insecticide ppliction nd removl of infected vines nd secondry hosts. Hot wter tretment (HWT) is technique pplied in nursery to suppress potentil phytoplsm from plnting mteril. In field, insecticides tretments re used by winegrowers to limit Scphoideus titnus popultion. Alterntives in orgnic viticulture re existing s bsed nturl pyrethrin product nd sprying of cholinite or ornge oil/extrct on folige to reduce plnt ttrctiveness. Control strtegies for Grpevine Trunk diseses re bsed on preventive techniques which minimize news infections. Scientific Working Group identified the most efficient chemicls nd bio-control gents regrding one or two diseses. Then, they provided recomendtions for nursery nd vineyrd mngement. In field, most Europen winegrowers re using pruning techniques nd protection of pruning wounds to limit pthogens infection (Guyot Poussrd, Simonit nd Sirch, bio-control gent sprying), trunk clening nd combintion of prctices insuring good vineyrd helth sttus. This project hs received funding from the Europen Union s Horizon 00 reserch nd innovtion progrmme under grnt greement No 6560