Genetic resources, evaluation and screening

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1 Genetic resources, evlution nd screening 5 Reserch on the mening of the enzymtic systems (GPI nd PGM) s prmeters for the definition of vrieties ( Vitis sp.): The Itlin cse of Cbernet frnc A. CALO, A. CosTAcuRTA, R. D1 STEFANO, G. CALO nd G. PALUDETTI lstituto Sperimentle per l Viticoltur, Vile XXVII Aprile, 26, I Coneglino (1V), Itly S u m m r y : Severl studies crried out t Dvis nd Coneglino showed tht isozyme nlysis of the GPI nd PGM enzymtic systems grees with the conventionl definition of the vriety in mpelogrphy. Differences were reponed mong vrieties but not mong biotypes of the sme vrieties. The only exception recorded ws in the popultion of Cbernet frnc in which GPI nd PGM revel two different types (A - the trditionl type encountered in Frnce nd B - type encountered in the Itlin region ofveneto). Further mpelogrphic, mpelometric, phenologicl nd chemicl studies on the polyphenolic nd romtic substnces in fruits hve shown considerble differences between the two types. Such differences demonstrte tht the type B is different vriety nd not clone of Cbernet frnc. Preliminry mpelogrphic nlysis nd the equlity in GPI nd PGM ptterns led to the conclusion tht the type B very probbly is Crmenere. Therefore, the hypothesis of vriety discrimintion bsed on the nlysis ofgpi nd PGM is vlid nd this method is useful to help to chrcterize the vrieties. Key words: vriety of vine, biorype, Itly, mpelogrphy, biometry, nlysis, morphology, lef, berry, enzyme, pyrzine, polyphenol, phenol. Introduction Generlly, mpelogrphy provides the definition of vine vrieties ccording to morphologicl, phenologicl nd chemicl fetures tht my differ in more or less significnt wys. Conventionlly, when the bsic morphologicl fetures of vriety's popultion re the sme, clones re distinguished ccording to the genotypicl vritions of the physiologicl nd/or phenologicl nd/ or chemicl fetures. Studies crried out t Dvis nd Coneglino over 2-yer period ( ), t first on 225 vrieties nd subsequently on 9 vriety popultions nd 63 clones of these (8), showed tht isozyme nlysis of the GPI nd PGM enzymtic systems by strch gel electrophoresis of lef ejurcts grees with the conventionl definition of the v1iety in mpelogrphy. Differences were reported mong vrieties but not mong biotypes of the sme vrieties, probbly becuse the genes of which these enzymes re the min expression, did not cuse ny significnt vritions of the morphologicl, physiologicl nd chemicl fetures, prcticlly nd conventionlly considered in the vriety's definition. So fr, the only exception recorded ws the popultion of Cbernet frnc, widely employed in Itly, in which the GPI nd PGM revel two different types, i. e. A - the trditionl type encountered in Frnce nd B - the type minly encountered in the Itlin region of Veneto. The forementioned types hve significntly different fetures requiring further mpelogrphic, mpelometric, phenologicl nd chemicl studies on the polyphenolic nd romtic substn~es in fruits in order to provide objective differences tht the opertive world is lso suggesting. The Cbernet vrieties seem to dte bck to the Biturie mentioned by PuNrns nd CowMELU., in reltion to the ncient synonyms Viduve nd Vidure~ ccording to PEnT-L'\FITIE (who relied on the uthority ofv1~et, n expert of the l 8th century). Although impossible to prove, it is quite evident tht the vrieties dte hck to very erly ge, fct of which trustworthy evidence is found in Giron de. where Cbernet frnc ws well-known t the times of the Crdinl of RICHELIEC.

2 6 Section 1 During the diffusion of these vrieties, strted in the 17th century, severl popultions spred out with different synonymities or distinctions, still existing t the present time. The distinctions involved vrieties, i. e. Cbernet frnc, Cbernet Suvignon, Crmenere nd then series of synonyms reclled by the different mpelogrphies (14, 15, 17, 20). It my be worth underlining the fct tht in the 19th century, the mpelogrphicl expert, Count OnART, mistook Cbernet frnc nd Cbernet Suvignon, since the ltter ws still not very well-known in French regions other thn Gironde. In Itly ( 10, 11, 16 ), pprently Cbernet frnc ws imported for the first time by the Count of SAMBUY in Piedmont t the beginning of the 19th century. By the end of the century, the vriety ws cultivted in pproximtely 45 provinces, from Piedmont to Southern Itly. Noteworthy cultivtions were present in Y eneto. where the vriety hd come directly from Frnce through BoRTOLO CLEMEC\"TI, the Count of ScHio, Count CoRI"ALDI nd other vine-growers. The origins of the vines nd thus of the popultions were nturlly diverse, s still cn be noticed in some vineyrds propgted with this mteril. The lstituto Spe1imentle per l Yiticoltur hs been perfonning selections since 1980 on the bove-mentioned popultions tht led to the identifiction nd now to the chrcteriztion of the types exmined in this report. Mterils nd methods Reserch ws conducted in by the lstituto Spe1imentle per l Yiticoltur in two fields situted on the Estern Venetin plin in entirely different pedoclimtic conditions. In ech field of comprison the two types of Cbernet frnc were represented by 24 vines per biotype with 6 repetitions ech. Exmintion involved the following fetures: 1. Biochemicl fetures The strch gel electrophoretic nlysis of lef extrcts ws performed for the enzymtic systems glucose phosphte isomerse (GPI) nd phosphoglucomutse (PGM). The methods nd procedure employed were those previously reported (3, 18). 2. Morph o 1ogic1 fetures Testing of these fetures ws perfonned ccording to the instructions of the interntionl 0.1.V. desc1iptor list. Fetures re described in Tble 1. The following informtions were lso recorded: - phenologicl ges considering the verge time of the phenomenon's ppernce; - potentil nd ctul fertility of the buds long the fruit bering tendrils (7). 3. Ampelometric mesurements These tests nlyze the fetures provided by mesurements nd the reltions tht define the lef, in prticulr: - rtios between veins - depth of petiole sinus - ngles between veins: - leflength/width - petiole/leflength L/L L/L 1 S/L 1 S/L.., ~between Lnd L 1 f3 between L 1 nd L 2 y between L 2 nd L 3

3 Genetic resources, evlution nd screening 7 Mesurements were performed on dult leves selected in summer from the 5th to the 8th node of the min shoots. The following grpe bunch chrcteristics were lso considered: - berry weighting - ben-y number per bunch - bery volume in ml - petiole length in cm - berry dimensions (length nd width) in cm - bunch size (width nd length) in cm The dt were gthered upon ripening from 10 bunches nd 100 grpes. 4. Berry smples were tken weekly from verison to ripening to estblish the following j u ice components: - totl cidity (g/l) - mlic cid tg/l) - ph - trtric cid (g/l) - sugr(% ml) - potssium (g/l) 5. Voltile berry components nd berry skin phenolic components 5.1. Berry pyrzines 1 kg of berries were homogenized nd stem distilled to estblish the mount of pyrzines. The distillte ws extrcted 3 times with 25 ml of CH, Cl,. After solvent distilltion, the extrct ws nlyzed by gs chromtogrphy-mss spectrometry by mens of the S.l.M. progrmme with cquisition of ions , , , respectively, relted to 2-ethyl-3-methoxy pyrzine, 2-isopropyl-3-methoxy pyrzine, 2-isohutyl 3-methoxy pyrzine Voltile juice components 250 ml of juice ws extrcted with C 5 H 12 : CH 2 Cl 2 in 60: 40 rtio for 12 h. After the solvent's evportion by distilltion~ the extrct ws subjected to gs chromtogrphy Phenolic berry skin components Anthocynins: These were extrcted from 10 berries with ethnol+ 0.1 % of concentrted HCI nd with mixture ethnol: H 2 0: concentrted HCl in the rtio 70: 30: 1. Phenolic cids: The ethnol extrct of skins ws employed for the nlysis of the hydroxy cinnmoyl, trtric cids. 6. 0rgno1 e pt i c tests were perfonned on wines produced by the microvinifiction of the two types of C hem et frnc nd tsting the grpes by mens of the sttisticl duo-trio test. 1. Isoenzymtic nlysis Results nd discussion As my be seen in Fig. 1, the two types of Cbernet frnc hve different GPI nd PGM ptterns. 2. Morpholo~icl fetures Of ll the fetures exmined (see Tble 1). the following proved to be repetedly differenr nd stedy in the two types of Cbernet frnc (Tble 2). Type B hs higher nthocynic pigmenttion levels in the shoot tip, more blisters on the upper lef side, lrger mount of hirs between veins on lower lef side nd looser cluster. These fetures will be emphsized in further detil by nlyzing the mpelometric tests. Moreover. prticulr feture is the specil shpe of stmen tht my be spirl-shped in type B, s previously illustrted (7) on this popultion (see Fig. 10).

4 8 Section 1 3. Ampelometric tests Tble 3 illustrtes the fetures tht proved to be significntly different mong those exmined. The nlysis of the results concerning the leves shows tht the dimension of the pex lobe detennined by the rtios L/Lnd S 1 /L 1 re mjor in type A thn in type B. With regrd to the bunch, type B, s lredy mentioned, is looser (longer bunch, fewer berries nd less compct) nd it hs lrger berries (size nd volume). 4. Berry juice components The course of sugr development in the berries. of the degrdtion of totl trtric nd mlic cidity. nd of the ph increse does not differ considerbly from one type ro the other (Fig. 2). Type B, however, ripens erlier nd thus it ttins higher sugr in berries erlier nd, t the sme time, hs significnt degrdtion of totl cidity, relted to the decrese of both mlic nd trtric cid, wheres the ph is higher. In the juice, there is lso considerble difference in potssium concentrtions: higher in type A, n spect tht is probbly interesting in enologicl tenns. Tble 1: Morphologicl fetures exmined o.r.v. CODE FEATURE / / / / / / / young shoot: form of tip intensity of nthocynin colortion of tip hir density ot tips shoot: color of internodes color of nodes density of hirs of nodes mture lef: number of lobes blistering of upper side shpe of teeth generl shpe of petiole sinus prticulrities of petiole sinus density of hirs between the veins Clower side) density of hirs on min veins (upper side) " density of hirs on petiole inflorescence: sex of flower number of inflorescence per shoot bunch: density

5 Genetic resources, evlution nd screening 9 Tble 2: Fetures proved to be repetedly different nd stedy IOIV CODE FEATURE EXPRESSION LEVEL TYPE A TYPE B 003 I young shoot: intensity of nthocynin color-1 tion of tip I mture lef: blistering of up-i per side density of hirs between the veins! Clower side) I bunch density 6,65 5,42 Tble 3: Ampelometric mesurements proved to be significntly different mong those exmined FEATURE CABERNET FRANC CABERNET FRANC TYPE A TYPE B L /L Cin cm) 1 0,893 0,842 S1/L1 Cin cm> 0,544 0,497 S2/L2 Cin cm) 0,628 0,591 o( Cin degrees) 48,62 53,5.6 Cin degrees) 51,52 55,27 r (in degrees) 48,32 53,25 berry weight Cin grmmes) 1,54 1,90 berry volume Cin ml> 1,39 1,67 berry length Cin cm> 1,43 1,52 berry width Cin cm> 1,33 1,45 verge number of grpes 99,44 85,32 bunch length Cin cm) 12,26 13,04

6 10 Section 1 5. Voltile grpe components nd berry skin phenolic components 5.1. Berry pyrzines The two types (A nd B) differ in the 124 ion chromtogrm profile nd prtly the 151 ion profile, relted to 2-isobutyl-3-methoxy pyrzine, the content of which seems to be considerbly higher in type B (Fig. 3). The presence of gret number of peks leds to the conclusion tht the preprtion system genertes dultertions Voltile berry juice components T~e profiles of the smples re prcticlly similr (Fig. 4). There re C 6 components originted by the enzymtic ttck of linoleic nd linolenic cids, besides other unidentified components nd considerble mount ofbenzylic lcohol Phenolic berry skin components Anthocynins: The nlysis of the chromtogrm HPLC of the nthocynins (Fig. 5 nd 6) reveled tht the two types of Cbernet frnc differ in the percentge of peonidin monoglucoside (higher in type B), cettes (higher in type B), nd p-cumrtes (higher in type A). The nthoeynins of both vrieties re trisubstituted. The mount of nthocynins in the skins of type B is pproximtely double the mount of type A (Tble 4). Tble 4: Anthocynins % ANTHOCYANINS!CABERNET FRANC TYPE A CABERNET FRANCI TYPE B IDelphinidin monoslucoside 6,57 ICynidin monoglucoside 0,67 IPetunidin monoslucoside 6,96 IPeonidin monoglucoside 3,66 IMlvidin monoslucoside 41,45 IDelphinidin monoslucoside cette 1,50 ICynidin monoslucoside cette 0, 11 IPeonidin monoglucoside + Mlvidin lmonoglucoside cette 12,99 IDelphinidin monoslucoside p.cum- 1,48 Irte ICynidin monoslucoside p.cumrte 0,06 IPetunidin monoslucoside p.cumrtel 1,83 IPeonidin monoslucoside + Mlvidin lmonoslucoside p.cumrte 21, 16 6,02 1,37 5,96 10,84 38,77 1,63 0,31 20,58 0,87 0,07 0,51 11,28

7 Genetic resources, evlution nd screening 11 Tble 5: Differences between type B of Cbernet frnc nd type A Morphologicl differences 1. Mjor nthocynic pigmenttion of shoot tip 2. Mjor blistering of dult lef upper side 3. Mjor presence of hirs between veins of dult lef lower side 4. Spirl shped flower stmen 5. Minor bunch compctness Ampelometric 1. Lower rtio L /L 1 Lower rtio s 1L 1 1 Lower rtio S2/L2 2. Mjor ngle o(., <between Lnd L 1 > F-> <between Ll nd L2> r <between L2 nd L3> 3. Looser berry bunch: * longer bunch * lower number of berries 4. Mjor berry dimension: * size * weight * volume Biochemicl nd chemicl 1. Different isoenzymes for enzyme systems GP! nd PGM 2. Grpe pyrzines: higher content of 2-isobutyl-3methoxy pyrzine 3. Skin nthocynins: higher content of peonidin monoglucoside 4. Phenolic cids~ higher content Orgnoleptio higher content of cettes lower content of p-cumrte double quntity of totl nthocynins Cler identifiction of the two types nd more herbceous tste in type B.

8 12 Section 1 Phenolic cids: The chromtogrm t 320 nm shows tht type B hs higher incidence of these components. The sme test perfonned t 280 nm (Fig. 7 nd 8) shows tht type B contins lrge mount ofn unidentified component with chrcteristic spectrum (Fig. 9), of which there re no trces in type A or in wine. 6. Orgnoleptic tests Continuous orgnoleptic reserch on wines hs llowed the preprtion of merit list in which Cbernet frnc type B ppers to be more pprecited nd with specific individulity. Furthem10re, grpe tsting performed by mens of the duo-trio test indicted tht the two types my be distinguished quite clerly (80 %). with mjor mrk of the herbceous tste of type B. GJ?I B l u... c GPI-1 3.>... ""' f.c 2 GPI llc I u PGM - A 13 u c.-f B 6 PGIH PGM-2.> r-4 ""' ~ 0 llc 0 A 13 Fig. 1: GPI nd PGM ptterns in Cbernet frnc type A (A) nd type B (B).

9 Genetic resources, evlution nd screening 13 TOTAL ACIDITY,.. -' ~ ""... :n :.;...,. u &I,_ Cl 4g e DATE ptl ""'--~ ~~~----~~-~ ~~ DATE 6 7 Q b SUGAR... i b Dnl Fig. 2: Berry components of Cbernet frnc type A (A) nd type B (B). (Continued overlef.)

10 14 Section ~ TARTARIC ACID 4~~~~~~~~~~~~~ DATE L... - ' ~ ~ z HJ 5 e HALIC ACID = L DATE POTASSIUH,.. -' ~... E :I... Ill., "' lls m--~- / / '..----,.,.,,,,.. /-_..._.. --= ~ L DATE Fig. 2 (continued).

11 Genetic resources, evlution nd screening 15 I o n I mu f r o m DAT A: C FIS IM. D 4. 0E4 4.0[4 3. 0E4 3.0E4 2.0[4 l 2'. 0E L:W k~l J ~... ~ CV u c: tt1 I o n I 5 I. (30 mu. f r o m DAT A: C ri 5 IH. D -0 c: ~4. 0E4 4. 0[4 II 3.0[4 3.0[4 II.' u c: (tj 2. (1[4 2.0[4 I C10C11J 100(1( TI rne ( rr1 i n. ) Ion mu. from DATA:CTFSIM.D 1.2[4 1.2[ CABERNET FRANC type A 1000( 9080 BG ;~~~ ~ J.bw~~L. J-~ ~ :.~~: ~I. 2 E4 i~ IOOOO Bfl80 6('.J(l~J Ion 151.0D.mu. from DATA:crrsrn.o I. 2E4 I 00Dl'. -B OCIC1 Fig. 3: Ion chromtogrm of the CH,Cl, extrct of the homogenizted grpes.~= Ions 124 nd 151 re the ones rel<ited to 2-isobutyl-3-methoxy pyrzine.

12 16 Section 1 C8brnt Fr8nc type B 7.P' TRilH5-2-E5EHALE TRAH5-3-E5EHOLO ESAHllLO 17.tll r-9~,~ ]'j.9f _r l"i 'J - f6 ~ju. 4.,,, i:2 CAPRllHIC ACID TRAHS-2-ESEHDIC ACID 4'Ufi Fig. 4: Chromtogrm of grpe juice voltile components.

13 Genetic resources, evlution nd screening DELPHIHIDIH PIOHOGUJCOSIDE 8.14 DELPHIHIDIH "OHOGWCOSIDE ACETATE PETUHIDIH "OHOGLUCOSIDE ACETATE ACETATE Fig. 5: HPLC chromtogrm of the nthocynins of the skins of Cbernet frnc type A. A= 520 nm.

14 18 Section ::;: 1 5 DELPHIHIDIH "OHOGUJCOSIDE 10- l 0, :3 2 CVAHIDIH "OHOGLUCOSIDE PEOHIDIH "OHOGLUCOSIDE ~========================~==-- 15.:?-7 c MALIJIDIH "OHOGWCOSIDE 1 :i 3 DELPHIHIDIH "OllOGWCOSIDE ACETATE I " I :1: C _ CYAH!D!N """""LOCOSlll< OCETOTt PETUHIDIH MOHOGLUCOSIDE ACETATE PEOHIDIH "OtlOGUJCOSIDE ACETATE "ALIJIDIH MOtlOGLUCOSIDE ACETATE s=:: "" I ; DELPHIHIDIH "OHOGWCOSIDE p - CU"ARATE CVAHIDIH ltohogldcoside p - CUMARATE PETUHIDIH MOHOGWCOSIDE p - CU"ARATE PEOHIDIH "OHOGLUCOSIDE + "ALIJIDIH "OHOGUJCOSIDE p - CUMARATE l ' Fig. 6: HPLC chromtogrm of the nthocynins of the skins of Cbernet frnc type B..A = 520 nm.

15 Genetic resources, evlution nd screening 19 ~~~~===~---2.2: lludentified conponent CAFFEIL TARTARIC ACID p - CUMRIL TARTARIC GUICDSIDE ACID p - cunaril TARTARIC ACID CAFFEIC ACID FERULIL TARTARIC ACID z ~ P 400 Fig. 7: HPLC Chromtogrm of the phenolic cids of the skins of Cbernet frnc cype A..l = 280 nm.

16 20 Section CAFFEIL TARTARIC ACID p - CUflARIL TARTARIC GLIJCOSIDE ACID l '? 6 l p - CUPIARIL TARTARIC ACID CAFFElC ACID FERULIL TARTARlC ACID Fig. 8: HPLC chromtogrm of the phenolic cids of the skins of Cbernet frnc rype B..l 280 run.

17 Genetic resources, evlution nd screening 21 Conclusions A series of studies crried out on different vrieties nd on clones of the sme vrieties proved tht isozyme nlysis of the GPI nd PGM enzymtic systems could be used s discriminting fctor when identifying vrieties since different ptterns my occur between vrieties but not between the clones of the vriety. However, the popultion of Cbernet frnc employed in Itly exhibited two different types (A nd B) for GPI nd PG M. In this report the forementioned types were crefully nlysed for severl morphologicl, mpelomenic nd chemicl fetures. Considerble differences were recorded between type B (minly ~iffused in Veneto) nd type A (the trditionl rype populr in Frnce). Such differences, illustrted in Tble 5, led to the conclusion tht it is different vriety nd not clone of Cbernet frnc. Type B is very probbly Crmenere, whose popultion spred in Veneto since lst century nd hs been frequently confused with Cbernet frnc. This hypothesis is confirmed by preliminr mpelogrphic nlysis s well s the equlity in GPI nd PGM ptterns nd it will be moreover exminted with further checks. Fig. 9: Spectrum of the unidentified component presem in the skins of Cbernet frnc type B.

18 22 Section 1 Fig. 10: Shpe of the stmen in the two popultions of Cbernet frnc. Left: stmen of type A; right: spirl-shped stmen of type B. Therefore. the hypothesis of vriety disc1imintion bsed on the nlysis ofgpi nd PGM is bsiclly vlid nd thus the method is presently useful to help to chrcterize the vrieties nd in future will be suitble for the definition. References ALBACH, G.; KEPNER, R.: WEBB, A.; 1959: Comprison of nthocynic pigments of red Vinifer grpes. Amer. J. Eno!. Viticult. 10, 164. ALLEWELDT, G.; DETT\\1:ILER, E.; 1986: Ampelogrphic studies to chrcterize grpevine vrieties. Atti IV Simp. Intern. Genetic dell Vite, ARt.:LSEKAR, S.; PARFITT, D.E.: 1986: Isozyme nlysis procedures for stonefruits, lmond, grpe, wlnut, pistchio nd fig. HortScience 21, ASTEGIA'.'O, V.; C!OLFI, G.; 1974: Indgine sui costituenti ntocinici dei vini rossi piemontesi. Riv. Viticolt. Enol. 27, BACHMA::-Jl'\, 0.; 1989: lsoenzymes s tool for grpe cultivr identifiction. Riv. Viticolt. Enol. 42, BouRSIQl;oT, J. M.; VJG:S:AC, L.; BOULET, J. C.; 1989: Ricerche sull'utilizzzione dell'mpelometri. Riv. Viticolt. Eno!. 42, CALO, A.; COSTAct;RTA, A.; CERSOSIMO, A.; CA'\CELLIER, S.; Ml'CIG'\AT, D.; 1988: L selezione clonle dei vitigni bordolesi: Cbernet Frnc N, Cbernet Suvignon N, Merlot N. Riv. Viticolt. Eno!. 41, ; -- - ; PALUDETTI, G.; CALO, G.; ARliLSEKAR, S.; PARFITT, D. E.; 1989: The use ofisozyme mrkers to chrcterize grpe cultivrs. Riv. Viticolt. Eno!. 42, ; Lit.:l'\I, C.S.; 1963: Indgini sull fertilit de! Cbernet frnc. Ani Accd. Itl. Vite e Vino, Vol. XV. CosMo, I.; 1940: Rilievi mpelogrfici comprtivi su vriet di Vitis rinifer. I vitigni bordolesi. Itli Agricol 7, ; POLSIXELLI, M.; 1960: Principli vitigni d vino coltivti in Itli: Cbernet frnc. Vol. I. Ministero Agricoltur e Foreste, Rom. DI ROVASEXDA, G.; 1877: Sggio di un Ampelogrfi Universle. Tori.no. DI STEFAXO, R.: CoRr::-.:o, L.: 1984: Terpeni ed ntocini di uve rosse romtiche. Riv. Viticolt. Enol. 37, GALET, P.; 1962: Cepges et Vignobles de Frnce. Imprimerie du Pysn du Midi, Montpellier.

19 Genetic resources, evlution nd screening 23 MASSE; PULLIAT; 1877: Le Vignoble, Vol. 11. G. Msson Editeur, Pris. MoLOl\, G.; 1906: Ampelogrfi. Vol. I, II. Milno. ODART, A. P.; 1849: Ampelogrphie Universelle ou Trire des Cepges. Libririe Agricole De Dusco, Pris. PALUDETTI, G.; CALO, G.; 1988: Elenroforesi di estrni enzimtici in Vitis sp.: not metodologic. Riv. Viticolt. Eno!. 41, RAPP, A.; HASTRICH, H.; E1'GEL, L.; 1978: Anlyse pr chromtogrphie cpillire des constitunts romtiques des vins et des risins; possibilites d'identifiction des vrietes. Genetique et Ameliortion de l Vigne, Bordeux, VIAL-\, P.; VERMOREL, V.; 1901: Ampelogrphie, Vol. II, Pris. Evlution nd utiliztion of Vitis ripri s source of genes for extreme cold hrdiness P. R. Hn1sTAD nd J. J. LrnY Depnmem ofhoniculturl Science nd Lndscpe Architecture, University of Minnesot, St. Pul, MN, 55108, USA Abstrct: The gol of the University of Minnesot grpe breeding progrm is to develop high qulity wine nd tble culrivrs tht tolerte winter temperrures s low s -35 C nd ripen in reltively cool. short growing seson. ::\onhern clones of Vi1is ripri hve been collected, e\ lured nd used in the breeding progrm. Genotypes of V ripri from the northern portion of the species rnge experience winrer tempertures of -40 "C or lm\ er. In ddition to extreme cold hrdiness, these genotypes lso.exhibit erly ripening, high sugr levels. nd resistnce to powdery mildew nd phylloxer. Deleterious trits of mny V ripri clones include high cidity, smll cluster nd ben-y size. very drkjuice, strong chrcteristic flrnr. nd excessive vegettive growth. Over 100 V. ripri clones hve been selected nd evluted in Minnesot over the pst decde. These clones exhibit substntil vribility for the viriculturl trits mentioned bove. For exmple. sugr levels typiclly rnge from 23 to 28 Brix: tirrrble cidity rnges from under 2 % to over 4 %;juice color vries from very drk purple ro light red: rime oflefsenescence cn vry by 2 to 3 weeks. Controlled environment srudies indicte tht erly cclimtion in these northern clones is phoroperioj-induced. Superior clones crossed with I~ l'in/(er nd inrerspecifrc hybrid cultivrs produced selections in the F 1 nd subsequent genertions tht were virici.llrurlly nd enologiclly cceptble. Some of these selections hve suffered miniml injury fter experiencing midwinter tempertures s low s -35 C, both in the \ ineyrd nd in lbortory freezing studies.