J Plnt Growth Regul (16) 35:245 255 DOI.07/s00344-015-9529-2 Responses of Vitis vinifer Pinot gris Grpevines to Exogenous Ascisic Acid (ABA): I. Yield, Fruit Qulity, Dormncy, nd Freezing Tolernce Shouxin Li 1 Imed E. Dmi 1 Received: 6 Ferury 15 / Accepted: 3 June 15 / Pulished online: 4 August 15 The Author(s) 15. This rticle is pulished with open ccess t Springerlink.com Astrct Economic losses due to cold wether events re mjor constrint to the expnsion of premium ut coldsensitive winegrpe cultivrs in colder regions, such s Ohio (USDA) Plnt Hrdiness Zones 6 5 (-17.8 to -26.1 C) (ARS-USDA 12). The purpose of this study ws to determine whether folir ppliction of scisic cid (ABA) could increse the freezing tolernce (FT) of field-grown Pinot gris grpevines (Vitis vinifer), nd whether the effectiveness of ABA tretments cn e influenced y the phenologicl timing of the ppliction. Mture Pinot gris grpevines were treted with folir ppliction of ABA t concentrtion of 0 mg L -1 (control), or 0 mg L -1 t vine phenologicl stges corresponding to vérison, post-vérison, nd posthrvest. ABA ppliction did not ffect yield components, fruit composition, or vegettive growth, ut cused erly lef senescence, lef scission, nd dvnced dormncy tht led to incresed FT of Pinot gris during the dormnt seson. The phenologicl timing of ppliction influenced ABA effectiveness, with spry pplictions mde t vérison nd post-vérison eing the most effective. It ws concluded tht folir ppliction of ABA incresed ud FT during the dormnt seson, nd thus, ABA is potentil culturl prctice tool for mitigting economic losses from cold injury in production regions with dmging cold events. Keywords Freezing tolernce Dormncy Therml nlysis Low-temperture exotherm Cold injury & Imed E. Dmi dmi.1@osu.edu 1 Deprtment of Horticulture nd Crop Science, Ohio Agriculturl Reserch nd Development Center, The Ohio Stte University, Wooster, OH 44691, USA Introduction Grpe production regions in the northern U.S. hve experienced sustntil economic losses in production due to cold wether events in 03, 04, 05, 09, 12, nd 14 (Zdl nd others 07; Dmi nd others 12, 14). The stte of Ohio hs experienced consecutive freezing-relted losses of grpe production over the pst 5 yers, with the gretest loss in 14 (Dmi nd others 14; Dmi nd Lewis 14). The recent wether events re of gret concern s they threten the expnsion of Vitis vinifer crege nd production nd thus their economic sustinility not only in Ohio (USDA Plnt Hrdiness Zones 6 5 (-17.8 to -26.1 C) (ARS-USDA 12) ut lso in other regions tht experience winter su-freezing events. To mitigte the thret of cold dmge in grpes, vrious protection methods hve een developed, with the purpose of either chnging the environmentl conditions in the vineyrd or improving the FT of grpevines. Active methods involve protection immeditely prior to or during the freeze event nd include the use of wind mchines, heters, nd over-vine wter sprinklers (Poling 08). Preventtive methods involve protection long prior to freeze event such s site nd vriety selection (Zdl nd others 07) nd the ppliction of chemicl protectnts (Dmi nd Bem 04). Among the chemicl protectnts, folir pplictions of the plnt growth regultor ABA hve een found to increse FT in vrious plnt species including Aridopsis (Mntyl nd others 1995), silver irch (Li nd others 03), rley (Brvo nd others 1998), rye (Churchill nd others 1998), whet (Dllire nd others 1994), chickpe (Kumr nd others 08), potto (Mor- Herrer nd Lopez-Delgelo 07), sugr mple (Bertrnd nd others 1997), nd pple (Guk nd Fuchigmi 01). Previous work y Dmi s group hs demonstrted tht
246 J Plnt Growth Regul (16) 35:245 255 ABA lso incresed FT of grpe species (Zhng nd Dmi 12, 12; Dmi nd others 15). The cultivr, Vitis vinifer, Pinot gris, is cold sensitive nd widely plnted in production regions where winter wether events commonly cuse cold injury (Zdl nd others 07). However, it is not known whether ABA would improve the FT of this economiclly importnt cultivr. Therefore, the purpose of this study ws to investigte whether folir ppliction of ABA nd the phenologicl timing of this ppliction would enhnce the FT of Pinot gris without dversely ffecting its reproductive or vegettive growth. Mterils nd Methods Plnt Mteril nd Tretments The field study ws conducted t the Ashtul Agriculturl Reserch Sttion (AARS), in Kingsville in northest Ohio (USDA Plnt Hrdiness Zone 6 (-17.8 to -.6 C) (ARS-USDA 12). Pinot gris grpevines (Clone 146, rootstock 1-14) were plnted in 1995 t AARS nd vines were spced 1.5 9 2.4 m 2 (vine 9 row). Grpevines were trined to ilterl low cordon trining system with verticlly shoot positioned (VSP), nd spur pruned to 15 uds per meter of cordon. Tretments were pplied on vines using rndomized complete lock with four locks nd four tretments ech consisting of 4-vine plots ordered y untreted uffer vines (2 4). The four tretments consisted of the following: (1) control (no ABA), (2) ABA pplied t % vérison ( % erry softening nd color chnge), (3) ABA pplied t postvérison (2 3 weeks fter % vérison), nd (4) ABA pplied t posthrvest (6 8 weeks fter % vérison). ABA nd control (no ABA) folir spry solutions were prepred using deionized wter nd the spreder-sticker Ltron B-1956 (Dow AgroScience, Midlnd, MI) t 0.05 % (v/v). The ABA solutions contining 0 mg L -1 were prepred using Protone SG (Vlent BioSciences Corportion, Liertyville, IL) which contins.0 % (w/ sderw) s-aba s the ctive ingredient. The sprys were pplied to oth sides of the vine cnopy (leves nd clusters) using ckpck spryer tht verged spry volume of 0.5 L per vine. The experiments were repeted for two growing sesons (12 nd 13). Lef Senescence nd Ascission In 12, lef senescence ws visully oserved, nd photos were tken of ll tretments. Lef scission ws determined y counting leves per shoot pre- nd post-aba ppliction nd computing lef scission percentge. In 13, lef senescence ws monitored y mesuring chlorophyll content using SPAD-5 chlorophyll meter (Spectrum Technologies, Inc. Est-Plinfield, IL). Four shoots (one shoot per vine) were rndomly tgged per tretment-replicte, nd chlorophyll mesurements were recorded i-weekly on the upper surfce of the sl four leves (node position 4, 5, 6, 7). Lef scission ws lso recorded t the sme time nd on the sme shoots s descried ove. Yield, Fruit Qulity, nd Vegettive Growth Yield components consisted of collecting crop weight per vine, cluster numer per vine, nd computing cluster weight (crop weight/cluster numer). A 0-erry smple from ech tretment-replicte ws collected t hrvest nd weighed using n electronic scle (Denver Instrument, Bohemi, NY), nd the erry numer per cluster ws clculted [(cluster weight/0-erry weight) 9 0]. The 0-erry smple ws crushed on the dy of smpling to ssess fruit composition y mesuring three sic prmeters: solule solids (SS), ph, nd titrtle cidity (TA). The relesed juice ws pressed through cheesecloth nd used to mesure SS (%) with digitl refrctometer (MISCO, Clevelnd, OH), ph with ph electrode (London Scientific, London, Cnd), nd TA y titrting 5 ml of juice with 0.1 N sodium hydroxide (NOH) to ph endpoint of 8.2 using n utomtic titrtor (PH/EP Titrtion Worksttion Model 3/352) with SAC80 Smple chnge (Denver Instrument, Bohemi, NY). The nnul vegettive growth ws estimted from the weight of one-yer-old cnes/vine, nd determined from vines left unpruned (not smpled for freezing tests) in ech tretment-replicte. The Rvz index, lso referred to s crop lod, ws clculted s the rtio of crop weight to pruning weight per vine (Rvz 1903). Bud Dormncy Bud smples were collected monthly from 31Aug 12 to 21 Mr 13 in the first yer of study nd from 6 Sept 13 to 21 Nov 13 in the second yer of study. All dormncy ssys were conducted on the sme dy of smple collection nd followed the method reported y Zhng nd others (11). Eighteen single-node cuttings were prepred from totl of 24 uds on three eight-node cnes. Cnes with visile periderm formtion were excised into one-node cuttings out 5-cm long, then inserted into 2.5 9 2.5 cm 2 fom medium (Smithers-Osis, Kent, OH) nd plced in 55 9 25 9 7cm 3 plstic trys (T.O. Plstics, Clerwter, MN) filled with deionized wter. Trys were then plced under forcing conditions in growth chmer (Conviron, Pemin, ND) with the following settings: 12-h photoperiod with 0 lmol m -2 s -1, 22 C, nd % reltive
J Plnt Growth Regul (16) 35:245 255 247 humidity (Zhng nd Dmi 12). Budurst ws recorded s Eichhorn-Lorenz (EL) stge 5 (Eichhorn nd Lorenz 1977) nd monitored three times per week for dys. Dormncy ws estimted s the numer of dys to % udurst (DBB), with higher vlues of DBB indicting more dormnt uds (Wke nd Fennell 00). Freezing Tolernce One-yer-old cnes with seven sl uds (node positions three to nine) were collected from ech tretment-replicte. The cnes were wrpped in plstic gs nd plced in cooler with ice pcks nd shipped overnight to the lortory in Wooster. Cnes were stored in refrigertor t 4 C, nd freezing tests were conducted within 48 h from time of collection. FT ws determined y therml nlysis descried riefly s follows. Buds were excised nd plced on thermoelectric modules (MELCOR, Trenton, NJ), which were plced in temperture-controlled chmer (Tenney ; Therml Products Solutions, New Columi, PA). The chmer temperture ws held t -2 C for min nd then lowered to - C t the rte of 4 C h -1. The FT of uds ws determined y identifying the medin or men low-temperture exotherm (MLTE), which corresponds to the lethl temperture tht kills % of the popultion, or LT (Wolf nd Pool 1987). Cnes for freezing tests were collected t the sme times s those used to determine ud dormncy from 31 Aug 12 to 21 Mr 13 in the first yer of study nd from 6 Sept 13 to 21 Nov 13 in the second yer of study. On 7 Jn 14, the vineyrd site experienced sufreezing events with minimum low temperture of -25.3 C. This temperture ws lower thn the LT of Pinot gris (-21.3 C) mesured on 25 Nov 13. As result, cnes were collected to ssess the extent of cold dmge. On 13 Jn 14, ten cnes with ten uds (node position one to ten) per cne were collected from ech tretment-repliction comintion nd stored for 24 h t room temperture. Buds were excised using rzor ldes to visully ssess the viility of primry, secondry, nd tertiry uds s well s cne phloem tissue nd ctegorized s ded or live sed upon the presence or sence of rown tissue (Dmi nd others 12). Winter injury ws expressed s percentge y dividing the numer of dmged uds or cnes with ded tissue y the smple size nd multiplying y 0. Field Budurst After pruning in 13, the totl numer of uds per vine ws counted nd recorded. Budurst ws quntified y recording the percent of uds per vine t the EL (Eichhorn nd Lorenz 1977) stge 5 nd monitored three times per week until 80 % or more of uds urst. Budurst ws not evluted in 14 ecuse of winterkill. Wether Dt nd Sttisticl Anlysis Dily minimum nd mximum tempertures were collected nd recorded y the wether sttion locted t the AARS reserch vineyrd in Kingsville (OARDC Wether Sttion System 14). Cumultive growing-degree dys (GDD), which indicte the growing seson het, were computed s cumultive dily men tempertures with se C from 1 Apr. to 31 Oct. The dtes of first fll frost events, which correspond to the dte of the first occurrence of temperture elow 0 C in the fll, were lso recorded. Frost-free dys (FFD), which indicte the growing seson durtion, were computed s the successive numer of dys with minimum dily temperture t or ove 0 C. Dys fter udurst (DAB) indicte the numer of dys from udurst to the corresponding vine phenologicl stge (Tle 1). All dt were sujected to nlysis of vrince using SAS sttisticl softwre (version 9.3; SAS Institute, Cry, NC). When tretments were sttisticlly different, n LSD test ws used for men comprison (P B 0.05). Results ABA Appliction Dtes nd Corresponding Wether nd Phenology In oth yers, ABA pplictions t vérison, post-vérison, nd posthrvest corresponded to the following GDD rnges: 943 1138, 1292 1296, nd 17 1493, respectively (Tle 1). ABA pplictions t the three phenologicl stges lso corresponded to the following DAB rnges 5 7, 1 134, nd 148 158, respectively. In yer 1, grpes were hrvested on 19 Sept 12, nd the cumultive GDD ws 1637 with 191 FFD. In yer 2, grpes were hrvested on 24 Sept 13, nd the cumultive GDD ws 1438 with 199 FFD (Tle 1). Lef Senescence nd Ascission In erly Octoer of 12, ABA-treted Pinot gris grpevines showed color chnge of leves erlier thn the untreted (control) vines with incresed intensity of yellowing from vérison to posthrvest tretment (Fig. 1). During the sme period of time, no differences of lef scission were oserved mong tretments (* %) except for posthrvest tretment (*80 %), which exhiited the erliest defolition (Figs. 1, 2). In 13, lef senescence ws monitored sed on chlorophyll content, which ws stedy in Septemer then declined in Octoer in oth
248 J Plnt Growth Regul (16) 35:245 255 Tle 1 Ascisic cid (ABA) ppliction dtes (t ve rison, post-ve rison, nd posthrvest) nd hrvest dtes nd corresponding growing degree dys (GDD) nd dys fter udurst (DAB) reltive to the phenology of Pinot gris Ve rison Dte Post-ve rison GDD Hrvest Posthrvest GDDc FFDd DAB Dte GDD DAB Dte GDD DAB Dte GDD DAB 5 31Aug 1296 1 19 Sept 1464 139 28 Sept 1493 148 1637 191 7 17 Sept 1292 134 24 Sept 1225 141 11 Oct 17 158 1438 199 Yer 1 (12) 16 Aug. 1138 Yer 2 (13) 21Aug 943 GDD cumultive dily men tempertures with se C from 1 Apr. through dte of corresponding stge of development DAB dys fter udurst c GDD cumultive dily men tempertures with se C from 1 Apr. through 31 Oct d FFD successive dys with minimum dily tempertures t or ove 0 C Fig. 1 Effect of scisic cid (ABA) on lef senescence nd scission of Pinot gris grpevines grown in Kingsville, OH. Photos were tken on 09 Oct 12 nd 17 Oct 12. Grpevines were spryed with 0 mg L-1 (control) nd 0 mg L-1 ABA t % ve rison on 16 Aug 12, postve rison on 31 Aug 12, nd posthrvest on 28 Sept 12. Note: incresed yellowing of leves s ABA ws pplied lter in the seson from ve rison to posthrvest, which hd the most scised leves Vérison Post-vérison Posthrvest Vérison Post-vérison Posthrvest control nd ABA-treted leves. There were no tretment differences on ll dtes except on 11 Oct (Fig. 3). ABA cused erly nd incresed lef scission nd resulted in nerly complete defolition in post-ve rison-treted vines y hrvest time (Fig. 2). Similr to yer 1, posthrvest ABA cused complete scission dys fter ppliction, while control vines hd % defolition (Fig. 2). Yield, Fruit Qulity, nd Vegettive Growth In oth yers, ABA did not ffect yield components or the sic fruit chemicl composition of Pinot gris. During the 2-yer study, the men rnges of yield components nd fruit compositions t hrvest were 2.62 3.19 kg/vine yield, 24 33 clusters/vine, 91 122 g cluster weight, 55 80
J Plnt Growth Regul (16) 35:245 255 249 Fig. 2 Effect of scisic cid (ABA) on lef scission of Pinot gris grpevines grown in Kingsville in 12 nd 13. In 12,, vérison, post-vérison, nd posthrvest tretments corresponded to ABA ppliction on 16 Aug 12, 31 Aug 12, nd 28 Sept 12, respectively. In 13,, vérison, post-vérison, nd posthrvest tretments corresponded to ABA ppliction on 21 Aug 13, 17 Sept 13, nd 11 Oct 13, respectively. Different letters indicte sttisticl significnce t p B 0.05 Ascised leves (%) Ascised leves (%) 0 90 80 70 60 0 0 90 80 70 60 17-Oct-12 22-Oct-12 25-Oct-12 Oserv on dte c Vérison Post-vérison Posthrvest Vérison Post-vérison Posthrvest 0 6-Sep-13 24-Sep-13 21-Oct-13 Oserv on dte Fig. 3 Effect of scisic cid (ABA) on chlorophyll content of Pinot gris grpevines grown in Kingsville in 13. For clrity, only one ABA plot ws presented which ws the verge of vérison (ABA spryed on 21 Aug 13), postvérison (ABA spryed on 17 Sept 13), nd posthrvest (ABA spryed on 11 Oct 13). Different letters indicte sttisticl significnce t p B 0.05 Chlorophyll content (SPAD units) 35 25 15 ABA 6-Sep-13 18-Sep-13 11-Oct-13 25-Oct-13 Oservtion dte erries/cluster, 1.47 1.66 g erry weight, 19.4 23.5 Brix SS, 3.39 3.59 ph, nd 5.9 6.6 g L -1 TA (Tle 2). The pruning weight, n indictor of vegettive growth, lso showed no response to ABA tretment nd rnged etween 0.5 nd 0.65 kg/vine (Tle 2). The Rvz index, n indictor of lnce etween reproductive nd vegettive growth, rnged etween 4.2 nd 6.4 during the 2-yer study nd ws not different mong tretments (Tle 2). Furthermore, the Rvz index mong ll tretments ws within the optimum rnge of 5 suggesting tht vines were lnced (Smrt nd Roinson 1991). Bud Dormncy In oth yers, DBB rnged from 15 to 45 indicting tht vines trnsitioned from non-dormnt to dormnt stte, respectively. Under forcing conditions, the pttern of udurst ws similr in oth yers nd mong tretments.
2 J Plnt Growth Regul (16) 35:245 255 Tle 2 Effect of scisic cid (ABA) on yield components, fruit composition, nd vine size in Pinot gris grpevines grown in Kingsville, Ohio Tretment Yield/vine (kg) Clusters/ vine Cluster wt (g) Berries/cluster Berry wt (g) Pruning wt/ vine (kg) Rvz index SS ( Brix) ph Titrtle cidity (g L -1 ) Hrvest: 19 Sept 12 (no 3.11 31 0 63 1.60 0.53 5.9 23.4 3.53 6.5 ABA) Vérison 2.70 91 55 1.66 0.65 4.2 23.5 3.54 6.5 Postvérison 3.13 33 95 61 1.55 0.62 5.1 23.2 3.59 6.5 Posthrvest 2.91 31 95 58 1.64 0.58 5.0 23.5 3. 6.6 Significnce d NS NS NS NS NS NS NS NS NS NS Hrvest: 24 Sept 13 (no 2.94 26 113 72 1.58 0.59 5.0 19.8 3.39 6.5 ABA) Vérison 3.17 26 122 80 1.53 0.64 5.0 19.6 3. 6.4 Postvérison 3.19 27 118 75 1.57 0. 6.4 19.7 3.51 5.9 Posthrvest 2.62 24 9 74 1.47 0. 5.2 19.4 3.43 6.3 Significnce NS NS NS NS NS NS NS NS NS NS SS Solule solid, NS Not significnt Vérison, post-vérison, posthrvest: ABA spryed t % vérison, nd dys fter % vérison, respectively Rvz index ws clculted y dividing totl yield per vine y the pruning weight recorded during the winter following ech seson Vlues of DBB of single-node cuttings strted low in lte summer nd then incresed stedily towrd the utumn seson reching mximum vlues in mid-(yer 12) to lte-(yer 13) Octoer, indicting tht Pinot gris entered the deepest dormncy stte (Fig. 4). Vlues of DBB typiclly decresed rpidly fter peking in Octoer (17 Oct in 12 nd Oct in 13) nd reched lowest vlues on 21 Mrch 13 (Fig. 4, ). Even though the dormncy pttern ws the sme mong ll tretments, ABA-treted vines hd delyed udurst compred to control, nd the mgnitude of the dely vried with tretment nd seson. In the first study yer, post-vérison ppliction showed the most significnt dely of udurst under forcing conditions, nd verged 6 dys from mid- Octoer to erly Jnury (Fig. 4). In the second study yer, vérison ppliction cused the most udurst dely verging 7 dys from erly Septemer to lte Novemer. The dely of udurst is indictive tht ABA-treted vines entered deeper dormncy thn the untreted vines. In oth yers, the lter ABA ppliction during posthrvest did not ffect ud dormncy (dt not shown). Freezing Tolernce During the 12 13 dormnt seson, the first fll frost occurred on 12 Oct 12, nd the lowest temperture (-16.3 C) occurred on 18 Fe 13 (Fig. 5). Throughout the dormnt seson, the minimum ir tempertures were higher thn the lowest LT indicting tht the criticl tempertures to cuse tissue dmge were not reched. Exogenous ABA incresed ud FT of Pinot gris throughout the dormnt seson prticulrly during the cold cclimtion nd decclimtion stges (Fig. 6). Vérison nd post-vérison pplictions were the most effective nd incresed FT y n verge of 2.3 C during the utumn cclimtion (lte August mid-decemer) nd 2 C during the lte winter decclimtion (lte Ferury lte Mrch) (Fig. 6). During the 13 14 seson, the first fll frost occurred on 29 Nov 13 (Fig. 5). During the period etween Jnury nd Mrch, minimum tempertures elow - C occurred nine times nd rnged from -21.0 to -25.9 C. These tempertures were colder thn midwinter LT, indicting tht the criticl tempertures to cuse tissue dmge were reched. Therefore, freezing tests were only conducted from 6 Sept. to 25 Nov 13 nd were interrupted fter the lethl freezing events. Bud smples were collected fter these su-freezing events to determine tissue dmge. In the second study yer, ABA pplictions t vérison nd post-vérison were gin the most effective nd incresed the FT of Pinot gris y n verge of 2.2 C during the utumn cclimtion (erly Septemer to lte Novemer). Following the successive temperture events elow - C, primry, secondry, nd tertiry uds sustined 0 % dmge. However, phloem cne
J Plnt Growth Regul (16) 35:245 255 251 Fig. 4 Effect of scisic cid (ABA) on ud dormncy (dys to % udurst) on Pinot gris grown in Kingsville during 12 13 nd 13 14 sesons. For clrity, only control nd ABA tretments with significnt effect re presented. In 12, ABA (postvérison) ws spryed on 31 Aug 12. In 13, ABA (vérison) ws spryed on 21 Aug 13. Different letters indicte sttisticl significnce t p B 0.05 Dys to % udrust 45 35 25 15 ABA (post-vérison) 5-Oct-12 17-Oct-12 14-Nov-12 12-Dec-12 9-Jn-13 28-Fe-13 21-Mr-13 Collec on dte Dys to % udrust 45 35 25 15 ABA (vérison) 6-Sep-13 4-Oct-13 -Oct-13 21-Nov-13 Collec on dte injury ws highest in control ( %) nd lowest in ABAtreted (0 %) vines (Tle 3). Field Budurst On 6 My, udurst in ABA-treted ws lower (verge 34 %) thn tht in control vines (56 %), indicting dely of udurst. However, the udurst dely ws not oserved on My 8 nd susequent dtes (Tle 4). Discussion Influence of ABA on Yield, Fruit Qulity, Dormncy, Freezing Tolernce, nd Budurst ABA did not impct negtively yield components, fruit composition, or vegettive growth of Pinot gris. These findings re consistent with previous reports on oth tle grpes (Lurie nd others 09; Peppi nd others 07) nd wine grpes including Cernet frnc (Zhng nd Dmi 12), Chmourcin (Zhng nd Dmi 12), nd Chrdonny (Dmi nd others 15). The lef senescence pttern in Pinot gris ws similr to tht reported in Cernet frnc (Zhng nd others 11; Zhng nd Dmi 12) nd Chmourcin (Zhng nd Dmi 12). These studies reported reduction of chlorophyll concentrtion in Octoer, which ws the sme time s with Pinot gris. The decrese of chlorophyll content is indictive of the slowdown of photosynthesis nd initition of lef senescence (Evns 1996). The ppliction of exogenous ABA cused erly lef senescence nd scission in Pinot gris. ABA s effects on lef color chnge nd scission incresed when pplied t lter phenologicl stges corresponding to pplictions on older leves (134 148 DAB). These findings support previous studies tht demonstrted tht only older leves (DAB C 1) were responsive to ABA, y ccelerting lef scission in treted versus untreted greenhousegrown Vitis Cernet frnc grpevines (Zhng nd others 11). Interestingly, ABA seems to e most effective in reducing chlorophyll content thus hstening senescence t the sme time of the yer nd corresponding to DAB C 1 regrdless of vriety used. It hs een reported tht lef senescence is linked with ABA ccumultion (Dong nd others 08), y upregulting senescence genes (Prk nd others 1998). The dely of udurst in ABA-treted vines suggests tht ABA induced deeper dormncy thn in untreted vines. These results concur with previous studies with pple (Guk
252 J Plnt Growth Regul (16) 35:245 255 Mx Air Temp ( C) Min Air Temp ( C) BB (6-My-13) Temperture ( C) 0 - FF (12-Oct-12) - - Dte Mx Air Temp ( C) Min Air Temp ( C) Temperture ( C) 0 - FF (29-Nov-13) - - Fig. 5 Dily minimum nd mximum tempertures nd dtes of fll frost (FF) nd udurst (BB) recorded t the Pinot gris reserch vineyrd in Kingsville in study yer 1 (12 13), nd study yer 2 (13 14) Dte nd Fuchigmi 01) nd grpe (Zhng nd others 11; Zhng nd Dmi 12, ). Furthermore, the ppliction timing (vérison nd post-vérison) of ABA effectiveness ws similr to tht reported with two other grpe cultivrs, Cernet frnc (Zhng nd Dmi 12) nd Chmourcin (Zhng nd Dmi 12). It is known tht ABA is positive regultor of dormncy induction (Kucer nd others 05) nd tht its concentrtion increses during dormncy nd is highest t the deepest dormncy stte (Or nd others 00). Even though ABA concentrtion ws not mesured in this study, it is likely tht the ppliction of folir ABA led to n increse in endogenous ABA, which induced mximum dormncy. Moreover, ABA-treted vines reched the sme level of dormncy (DBB *35) 26 dys erlier thn untreted vines, suggesting tht ABA dvnced dormncy induction in Pinot gris (Fig. 4). This response is desirle ecuse it would llow n erly cold cclimtion nd the cquisition of mximum FT. Our findings tht exogenous ABA incresed the ud FT of Pinot gris support the results reported in severl woody
J Plnt Growth Regul (16) 35:245 255 253 plnts including grpes (Bertrnd nd others 1997; Dmi nd others 15; Guk nd Fuchigmi 01; Zhng nd Dmi 12, ). In Pinot gris, vérison nd post-vérison LT ( C) LT ( C) -.0-13.0-16.0-19.0-22.0-25.0 0.0-5.0 -.0-15.0 -.0-25.0 c 31-Aug-12 6-Sep-13 c 5-Oct-12 17-Oct-12 4-Oct-13 14-Nov-12 12-Dec-12 Collec on dte Collec on dte Vérison Post-vérison Posthrvest -Oct-13 9-Jn-13 28-Fe-13 25-Nov-13 21-Mr-13 Vérison Post-vérison Post-hrvest Fig. 6 Effect of ABA on freezing tolernce (LT ) in Pinot gris grpevines grown in Kingsville during 12 13 nd 13 14 sesons. In 12,, vérison, post-vérison, nd posthrvest tretments corresponded to ABA ppliction on 16 Aug 12, 31 Aug 12, nd 28 Sept 12, respectively. In 13,, vérison, post-vérison, nd posthrvest tretments corresponded to ABA ppliction on 21 Aug 13, 17 Sept 13, nd 11 Oct 13, respectively. Different letters indicte sttisticl significnce t p B 0.05 ABA tretments hd the lowest LT s nd gined n verge of 2 C in FT during the utumn cold cclimtion in oth yers nd during decclimtion in the first yer. These findings re similr to those reported in other grpe vrieties. ABA pplied t vérison nd post-vérison incresed ud FT y up to 4.2 C in Cernet frnc etween lte Octoer nd lte Ferury (Zhng nd Dmi 12) nd 3.5 C in Chmourcin etween erly Novemer nd lte Ferury (Zhng nd Dmi 12). A multi-loction study with Chrdonny confirmed the effectiveness of ABA, nd its ppliction timing with FT increses etween 1 nd 3.3 C (Dmi nd others 15). The freezing events in 14 provided n opportunity to evlute the effectiveness of ABA on field-tested Pinot gris vines. Unfortuntely, the dmge ws totl mong ll types of uds nd ll tretments. The resons for the ineffectiveness of ABA cn e explined s follows. The lowest FT of Pinot gris or LT ws -21.3 C prior to the occurrence of -25.3 C, which ws 4 C, lower thn the ud tissue criticl or lethl temperture. Our l freezing tests consistently showed tht ABA incresed FT y n verge of 2 C, compred to the control during tht time of the yer. Therefore, it is possile tht ABA provided 2 C of cold protection in treted uds, ut n dditionl Tle 4 Effect of scisic cid (ABA) on udurst (%) of Pinot gris grpevines t different oservtion dtes in 13 Tretment 6 My 8 My My 13 My 16 My (no ABA) 56 88 91 93 94 Vérison 43 87 92 94 96 Post-vérison c 80 86 89 91 Posthrvest 28c 81 89 90 92 Significnce * NS NS NS NS Vérison, post-vérison, posthrvest: ABA spryed t % vérison, nd dys fter % vérison, respectively NS, *, **, nd *** not significnt, significnt t p B 0.05, 0.01, nd 0.001, respectively Tle 3 Bud nd cne phloem dmge of Pinot gris fter sufreezing events (-25.3 C) on 7 Jn 14 Tretment Bud dmge (%) Cne phloem dmge (%) Primry ud Secondry ud Tertiry ud (no ABA) 0 0 0 Vérison 99 99 97 5 Post-vérison 0 0 0 Posthrvest 0 0 99 0c Significnce NS NS NS * Vérison, post-vérison, posthrvest: ABA spryed t % vérison, nd dys fter % vérison, respectively NS, *, **, nd *** not significnt, significnt t p B 0.05, 0.01, nd 0.001, respectively Smples were collected on 13 Jn 14
254 J Plnt Growth Regul (16) 35:245 255 4 C of protection ws needed. Previously, we demonstrted tht ABA ws effective to protect Cernet frnc from n ctul freezing event (Zhng nd Dmi 12). The sitution then ws different ecuse the minimum ir tempertures were wrmer (-19 to -23 C in two loctions) in 11 thn in 14 (-25.3 C). Further, the minimum tempertures experienced in 11 were ner the LT of Cernet frnc. This ws not the cse in 14 where the minimum tempertures were much lower thn the LT. Nevertheless, it ws interesting to oserve the lest injury of the vsculr tissue (phloem) in ABA-treted vines. This response could e very significnt where ABA could protect the permnent structures such s the cordons nd trunks of the vine from freezing injury. However, this needs further investigtion. ABA Effectiveness in Reltion to Appliction Timing In this study, the timing of ABA ppliction ws determined using three methods: (1) phenologicl stges, (2) GDD, nd (3) DAB corresponding to the respective phenologicl stges. Even though ABA ws pplied t the sme phenologicl stges (tht is, vérison, post-vérison, nd posthrvest) in oth yers of the study, the corresponding GDDs were different. GDD vried t vérison (943 1138, or.7 % vriility from yer 1 to yer 2), post-vérison (1292 1296, or 0.3 % vriility), nd posthrvest (17 1493, or 14.2 % vriility) (Tle 1). However, when time of ABA ppliction ws expressed s DAB, the vriility ws not s lrge s with GDD. DAB t vérison corresponded to 5 7 (or 1.9 % vriility from yer 1 to yer 2), DAB t post-vérison ws 1 134 (or 11.7 % vriility), nd DAB t posthrvest ws 148 158 (or 6.7 % vriility). Becuse ABA ws most effective etween vérison (yer 2) nd post-vérison (yer 1), its ppliction corresponded to DAB etween 7 nd 134, respectively. The rnge of DAB, in which leves were responsive to exogenous ABA in Pinot gris, ws similr to tht reported in Chrdonny (DAB = 95 137) (Dmi nd others 15), Cernet frnc (DAB = 1 1) (Zhng nd Dmi 12), nd Chmourcin (DAB = 1 1) (Zhng nd Dmi 12). The consistency of the effectiveness of ABA timing sed on DAB suggests tht lef ge influenced the sensitivity to exogenous ABA nd its role on susequent physiologicl chnges including dormncy induction nd cquisition of FT. Becuse the effectiveness of exogenous ABA is optimum t nrrow time period nd DAB showed the lest vriility, it is criticl to define more ccurtely the timing nd the conditions involved during the spry ppliction sed on DAB. Therefore, it is suggested tht lef ge estimte using DAB is more ccurte indictor for timing of ABA ppliction thn the phenologicl stge of the vine. In conclusion, exogenous ABA ppliction t concentrtions of 0 mg L -1 effectively dvnced nd induced deeper dormncy nd improved the ud FT of Pinot gris without dversely ffecting vine size, yield, or fruit qulity. The optimum time to spry ABA on Pinot gris ws etween vérison nd 3 weeks post-vérison corresponding to lef ge etween DAB = 7 nd 134. The findings of this study confirmed tht ABA might provide n dditionl tool to protect Pinot gris from freezing dmge in cold regions. Acknowledgments This reserch ws funded y the Deprtment of Horticulture nd Crop Science, USDA/NIFA Specilty Crops Reserch Inititive Awrd Numer -51181-21599 USDA-NIFA, nd Ohio Grpe Industry Progrm. We thnk Mr. Greg Jones, Ms. Yvonne Woodworth for mintining the reserch vineyrd t AARS. Ms. Dine Kinney, Mr. Thoms Todro, Ms. Ntlie Fry, Ms. Lis Roins, Mr. Steven Prk, nd Ms. Biley Miller deserve specil thnks for their ssistnce throughout the durtion of this project. 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