Mitteilungen Klosterneuburg 56 (2006): 147-156

Identification and characterization of Swiss grapevine cultivars using microsatellite markers

ANDREA FREI, NAOMI A. PORRET, JUÈ RG E. FREY and JUÈ RG GAFNER

Agroscope ACWChangins-Wa Èdenswil Forschungsanstalt fuÈ r Obst-, Wein- und Gartenbau CH-8820 WaÈdenswil, Postfach 185 E-mail: [email protected]

Microsatellite analysis with up to 12 markers was performed on 463 grapevine samples mostly collected from Swiss cultivar collections, viticulturists, winegrowers and private collectors, using an optimized and fast method based on quick DNA extraction, six-plex PCR and automated fragment analysis. Several already published genetic profiles were confirmed, misnamed plants were identified, and additional new profiles are presented in this study. Some of the misnamed plants could be named correctly, while cultivar allocation of others needs confirmation. We found that 'Briegler' seems to be a synonym for 'Bondola nera', possibly the German name for a cultivar grown in Sou- thern Switzerland. In the case of 'Erlenbacher Schwarz' we found types with two different profiles. The ones from Eastern Switzerland are identical to 'Hitzkircher', while the type collected in Western Switzerland is different. Fur- ther data collection is necessary to elucidate which type is the true-to-type cultivar. Also an old and rare cultivar na- med 'MoÈrchel' was shown to be identical to '' (syn. 'Schiava Grossa', 'Gross-Vernatsch' or 'Frankenthal Noir'). Several 'Chasselas' and '' types were allocated into synonymous groups with identical profiles. Key words: vinifera, , microsatellites, Swiss cultivars, genetic profile.

Identifizierung und Charakterisierung von Schweizer Rebsorten mittels Mikrosatellitenanalyse. 463 Proben von Weinreben aus Schweizer Sortensammlungen, von Weinbauinstituten, Weinbauern und privaten Sammlern wurden mittels Mikrosatellitenanalyse an bis zu 12 Markern untersucht, wobei eine optimierte und schnelle Methode, die aufrascher DNA-Extraktion basierte, sechsfachePCR und automatisierte Fragmentanalyse angewandt wurden. Ei- nige bereits veroÈffentlichte genetische Profile wurden bestaÈtigt, falsch bezeichnete Sorten wurden identifiziert und zusaÈtzliche neue Profile erstellt. Einige der falsch bezeichneten Sorten konnten richtig benannt werden, waÈhrend ei- nige andere Sortenzuweisungen noch einer BestaÈtigung beduÈ rfen. 'Briegler' scheint ein Synonym fuÈ r 'Bondola nera' zu sein, wahrscheinlich der deutsche Name fuÈ r eine Sorte aus der SuÈ dschweiz. Bei 'Erlenbacher Schwarz' wurden Typen mit zwei unterschiedlichen Profilen gefunden. Die aus der Ostschweiz entsprechen 'Hitzkircher', waÈhrend die Typen aus der Westschweiz unterschiedlich sind. Weitere Daten muÈ ssen erhoben werden, um zu klaÈren, welcher Typ der sortentypische ist. Die alte und seltene Sorte 'MoÈrchel' erwies sich als identisch mit 'Trollinger' (syn. 'Schiava Grossa', 'Gross-Vernatsch' oder 'Frankenthal Noir'). Einige Chasselas- und Muscat-Typen wurden synonymen Gruppen mit identischen Profilen zugeordnet. Key words: , Rebe, Mikrosatelliten, Schweiz, Sorten, genetisches Profil

Identification et caracteÂrisation de varieÂteÂs de vignes suisses au moyen de l'analyse de microsatellites. 463 eÂchantil- lons de vignes en provenance des collections de varieÂteÂs de vignes suisses, d'instituts d'únologie, de viticulteurs et de collectionneurs priveÂs ont eÂte examineÂs au moyen de l'analyse de microsatellites en tenant compte de jusqu'aÁ 12 mar- queurs. AÁ cette fin, on a utilise une meÂthode optimiseÂe et rapide baseÂe sur l'extraction rapide de l'ADN, la PCR sex- tuple et l'analyse de fragments automatiseÂe. Quelques profils geÂneÂtiques deÂjaÁ publieÂs ont eÂte confirmeÂs, des varieÂteÂs mal deÂsigneÂes ont eÂte identifieÂes et de nouveaux profils suppleÂmentaires ont eÂteÂeÂtablis. Il a eÂte possible de deÂsigner correctement quelques varieÂteÂs mal deÂsigneÂes, tandis que d'autres attributions de varieÂteÂs doivent encore eÃtre confir- meÂes. Il semble que 'Briegler' soit synonyme de 'Bondola nera', probablement le nom allemand d'une varieÂte du sud

147 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

de la Suisse. Des types avec deux profils diffeÂrents ont eÂte trouveÂs pour 'Erlenbacher Schwarz'. Ceux provenant de l'est de la Suisse correspondent aÁ 'Hitzkircher', tandis que les types provenant de l'ouest de la Suisse sont diffeÂrents. Il est neÂcessaire de saisir encore plus de donneÂes afin de deÂterminer quel type est celui typique pour la varieÂteÂ. La va- rieÂte ancienne et rare 'MoÈrchel' s'est reÂveÂleÂe eÃtre identique au 'Trollinger' (syn. 'Schiava Grossa', 'Gross-Vernatsch' ou 'Frankenthal Noir'). Quelques types de Chasselas et de Muscat ont eÂte attribueÂs aux groupes synonymes preÂsen- tant des profils identiques. Mots cleÂs : Vitis vinifera, vigne, microsatellites, Suisse, varieÂteÂs, profil geÂneÂtique

Despite the relatively small size of the country, many but related cultivars (VOUILLAMOZ et al., 2004). Other different grapevine cultivars are grown in Switzerland, mentioned synonyms are, e.g. 'Briegler' - 'Hitzkircher' as a result of historically strong influences of surroun- (as formerly named in cultivar collection in Au, Swit- ding regions. Samples of these grapevine cultivars were zerland), 'Hitzkircher' - 'Grosser blauer MoÈ rsch' (AE- collected in order to identify and genetically characte- BERHARD, 2005), 'Schwarzer Erlenbacher' - 'GaÈnsfuÈ sser' rize them, and to arrange the results in a database. The - 'Argant' (AMBROSI et al., 1994). It is of great import- basis for the construction of any database is the reliabi- ance that cultivars are clearly characterized and possible lity of the data included. This means that the analyzed synonyms and naming errors are found. Genetic me- plants must be true-to-type representatives of a cultivar. thods such as microsatellite analysis enable clarification That might sound obvious, but misnamed grapevine if two plants are identical or related. plants are found even in cultivar collections. Misnamed In the present study, special emphasis was placed on old cultivars have a negative impact on research, grapevine Swiss cultivars focusing on those cultivated in the ¹hi- breeding and the organization of grapevine collections storical vineyardª at the wine production museum in (THIS and DETTWEILER, 2003). Errors in nomenclature Au, Switzerland. These vines were originally planted are then propagated worldwide through material ex- by W. EGGENBERGER of the University of Applied Sci- change. It is known that about 5% of the types in the ences of WaÈdenswil in 1978. Several old varieties can world grapevine collections are misnamed and hence be found there, such as 'Elbling weiss', 'Elbling rot', not true-to-type (DETTWEILER, 1992). Therefore, identi- 'MoÈ rchel', 'Completer', 'Erlenbacher Schwarz', 'Brieg- fication and true-to-type assignment of cultivars in a ler', 'Thuner blau', 'RaÈuschling rot', and 'RaÈuschling database is a prerequisite for any parental analysis or weiss'. Many of these cultivars are very rare. For exam- other studies. ple, it is supposed to be the only planting of 'MoÈ rchel' When analyzing typical and historical Swiss grapevine in Switzerland (AEBERHARD, 2005). cultivars, we faced the above mentioned difficulties. The central geographical situation and the intensive tra- Many cultivars were introduced to different areas over ding connections of Switzerland over hundreds of years a long period of time and were given either similar or led to a melting pot of different and rare grape varieties totally different names. Also, due to limited travelling from all over Europe. In order to safeguard the genetic possibilities in ancient times, ampelographers were diversity in grapevine, Pro Specie Rara and SKEK forced to rely on descriptions, and so errors were pas- (Schweizerische Kommission zur Erhaltung von Kul- sed on from one report to another. There is not much turpflanzen) are running several programs. One mea- information in ampelographic books about historical sure is a so-called positive-list of all (historical) grapevi- Swiss cultivars. Several diploma theses were conducted nes in Switzerland that are worth safeguarding. These at our institute on the subject (BALMER, 2000; GREUTER cultivars will be planted in collections in different areas and WUNDERLIN, 1998). Recently, a book about the hi- of Switzerland. An inventory of all existing cultivar col- story of old grapevine cultivars has been published lections is under way. that includes many Swiss grapevine cultivars (AEBER- Especially for rare cultivars it is therefore very impor- HARD, 2005). Several names of historical Swiss cultivars tant to try to find many different, but originally inde- are mentioned as synonyms in literature, but errors re- pendent types of the same cultivar and so to find the garding synonyms can be found. For instance, 'Lafnet- true-to-type cultivar and their corresponding genetic scha' and 'Completer' often are mentioned as or su- profiles. spected to be synonyms (AMBROSI et al., 1994; EGGEN- The present study describes the current situation of the BERGER and LENNERT, 1996; HILLEBRAND et al., 1998), efforts to name, characterize and identify Swiss culti- but this was proven incorrect; they are two different vars. The results of this study contribute to the charac-

148 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

terization of true-to-type grape cultivars. They will Tab. 1: Collection sites (mostly in Switzerland) and number of enable the clear identification of unknown, rare or mis- analyzed grape cultivar accessions named plants by comparing their genetic profile to the database.

Materials and Methods

Plant material A total of 463 grapevine leaf samples were collected from several sites, mostly in Switzerland (Table 1). Cul- tivar collections as Rheinhof/FruÈ msen, Sortengarten Au and the collection of Agroscope ACWChangins in Pully contain types from different parts of Switzerland. Of the 463 cultivars that were finally analyzed, the lar- gest part came from collections in Au and Agroscope ACWWa Èdenswil (38.7%). In total, 373 types have been sampled from cultivar collections, 33 from a pri- vate collection in Berne, 44 from nurseries and vi- neyards, and ten from private gardens (Table 1). One type is an unknown wild grape found in an ancient vi- neyard. Types from vineyards and individual gardens came from seven different cantons from all over Swit- zerland. Twenty-eight types were from Germany or Austria, 22 of them from a 'Gutedel'/'Chasselas' collec- when trying to identify the true-to-type profile of a tion in Badenweiler, Germany. The aim was to find out cultivar. if 'Chasselas' was identical with some of the Muscat All cultivars were analyzed at six marker sites interna- cultivars, e.g. 'Muskat Gutedel'. We also compared the tionally proposed for grapevine cultivar characteriza- results to other Muscat profiles. tion: VVS2 (THOMAS and SCOTT, 1993), VVMD5, Cultivars were named according to the French and VVMD7 (BOWERS et al., 1996), VVMD27 (BOWERS et German names, generally known synonyms are: al., 1999), VrZAG62 and VrZAG79 (SEFC et al., 1999). 'Chasselas' = syn. 'Gutedel'; 'Pinot noir' = syn. 'Blau- It is generally accepted that these six markers are suffi- burgunder'; 'Gouais blanc' = syn. 'GwaÈss' = syn. cient to clearly distinguish between different cultivars 'Heunisch weiss'; 'Savagnin blanc' = syn. 'PaõÈen' = (REGNER et al., 2001). These markers have also been syn. 'Heida' = syn. 'Traminer weiss'. 'Cornalin' was chosen to establish a standard set of microsatellite refe- named 'Cornalin du Valais' according to VOUILLAMOZ rence alleles in the EU Project GenRes CT96 No81 et al. (2003) to avoid confusion with 'Cornalin d'Ao- (THIS et al., 2004). Some of the cultivars were analyzed ste' which is a synonym of 'Humagne rouge' (VOUIL- at six additional markers (VVS4 (THOMAS and SCOTT, LAMOZ et al., 2003). 1993), VVMD6, VVMD8 (BOWERS et al., 1996), VrZAG21, VrZAG 67 and VrZAG112 (SEFC et al., DNA extraction, PCR and sample analysis 1999)). Generally, methods were as described in FREI et al. PCR reaction was performed with the Qiagen Multi- (2004). Extraction of DNA had started using a common plex PCR kit, using a final PCR volume of 10 ml(5ml method (THOMAS and SCOTT, 1993), with some modifica- of 2x Qiagen multiplex PCR mastermix, 1 mlof5xQ- tions. For some of the samples, a quick DNA extrac- Solution, 1 ml of 10x primer mix containing 2 mMof TM tion method was applied, using the Extract-N-Amp each F and R primer (six-plex), 1 ml of pure H2O and Plant PCR kit (Sigma). Thirty-two types were analyzed 2 ml of template DNA, diluted 1:10 with pure H2O). using both DNA extraction methods, and the results PCR was done on a Techne Genius thermocycler (Wi- were always identical (data not shown). Several types tec AG, Littau, Switzerland) under the following cy- were analyzed for most cultivars, which is important cling conditions: 15 min at 95 8C (for initial denatura-

149 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

tion), followed by 40 cycles of 40 sec at 94 8C, 90 sec at compared to THIS et al. (2004); but they were 0 to 2 bp 57 8C and 90 sec at 72 8C; then 30 min at 60 8C and fi- longer compared to GRANDO and FRISINGHELLI (1998). nal hold at 10 8C(FREY et al., 2004). A particular situation was observed for the markers A mastermix containing 15 ml formamide and 0.5 mlof VVMD6 and VVMD8. Six bp had to be added to allele fluorescent GeneScanTM-500 ROXTM standard dye lengths found in this study in order to be able to com- (Applied Biosystems) per sample was mixed and 15 ml pare them to allele lengths mentioned in VOUILLAMOZ were pipetted in each well of a 96 well plate. Then, 1 et al. (2003 and 2004). At the same marker sites, the ml of the 1:10 diluted PCR product was added. After lengths had to be minimized by 2 bp when compared centrifugation, heat denaturation for 2 min at 96 8C to GRANDO and FRISINGHELLI (1998). and rapid cooling in the freezer followed. The plates The allele lengths of cultivars from this study were were then transferred to a 3100 Genetic Analyzer (Ap- shown to be identical in most marker sites compared plied Biosystems) and run on 50 cm capillaries with a to literature profiles, with a difference of one allele at high resolution polymer (POP-6; Applied Biosystems). one marker each in 'Merlot', 'Muscat blanc a petits The data produced by the Genetic Analyzer were im- grains', 'Silvaner', 'Himbertscha', and 'Plantscher' (Ta- ported into the software GeneMapperTM version 3.0 ble 2). The first three showed a difference at (Applied Biosystems) and automatically sized and ana- VVMD27, a 2 and 1 bp shorter allele, respectively (in lyzed. Bins were created based on the first results and this study, after the parallel shift), while 'Himbertscha' autobinning was performed with the rest of the data. and 'Plantscher' were heterozygous in this study at The values were rounded to the nearest base and then VVS4 or VVMD8, respectively. These differences are exported into the spreadsheet program Excel (Micro- probably due to the use of different methods, i.e. capil- soft). The program Identity version 1.0 (WAGNER and lary analysis vs. polyacrylamide gel readings. For 'Hu- SEFC, 1999) was used to compare and find identical pro- magne Rouge', we confirmed a heterozygous allele at files among the results. marker VVMD 8, as mentioned by VOUILLAMOZ et al. (2003), while its Italian synonym 'Cornalin d'Aoste' is Results and Discussion homozygous at this site. These results show that a pro- file or a database from literature can be adapted to High genetic variability was found among the analyzed other results or be used for comparison with unknown grapevine cultivars, which is in agreement with earlier profiles, after a parallel shift of known numbers of studies for grapevine cultivars from Central Europe, base pairs, for the purpose of comparing, characterizing e.g. by SEFC et al. (1999). From the 463 types analyzed, or identifying unknown grapevine cultivars. 195 different profiles could be identified. Most of the profiles could be assigned to a cultivar, while in a few Misnamed plants cases plants remain unidentified. Profiles of several, The assigned names of 58 analyzed grapevine types mostly well known, cultivars could be confirmed from were unknown or wrong (Table 3). Compared to the literature (Table 2). Nine profiles were identical to 463 analyzed types, this corresponds to a proportion true-to-type profiles of the European Vitis database of 12.5%. If we only take into account the 23 misnamed (THIS et al., 2004) and 15 profiles were found to be iden- cultivars from cultivar collections (not counting the ob- tical compared to several publications (GRANDO and viously propagated material) and compare it to the total FRISINGHELLI, 1998; SEFC et al., 1997; SEFC et al., 2000; of 373 types from cultivar collections, the percentage THIS et al., 2004; VOUILLAMOZ et al., 2003; VOUILLAMOZ drops to 6.2% of falsely named plants. This number is et al., 2004). The profiles were compared at six to nine close to the presumed number of about 5% of misna- marker sites, depending on the markers analyzed in med plants in cultivar collections (Dettweiler, 1992). A each study. Table 2 provides additional allele lengths at total of 44 individual misnamed plants as well as 12 cul- more marker sites. tivars of unknown or uncertain status could be cor- To enable direct comparison of the allele lengths be- rectly identified by comparing their profile to the data- tween this and previous studies, a parallel shift of base base. Seven cultivars still remain unidentified (Table 3). pairs (bp) was necessary. Allele lengths in this study We found that even in cultivar collections, the situation were generally 0 to 3 bp shorter compared to VOUILLA- is not always consistent. Of three plants of supposedly MOZ et al. (2003 and 2004), 0 to 2 bp shorter compared the same cultivar according to the name tag, all three to SEFC et al. (1997 and 2000), and 0 to 3 bp shorter can be wrong as in the case of 'Humagne Blanc' which

150 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

Tab. 2: Grape cultivar profiles analyzed, com- pared and confirmed with literature data, including allele lengths at additional mar- ker sites. Numbers in bold are different to literature but have been confirmed se- veral times in this study. In literature, the allele length is +2bp (a), or +1 bp (b), re- spectively, compared to the results in this table. Numbers in italics (c) are homozy- gous in literature, while heterozygous in this study. The markers that were compa- red are underlined. turned out to be 'Lafnetscha ', or two out of three can be wrong ('Silvaner') and the third one is correct, as in the case of 'GwaÈss' (Table 3). So it is important to ana- lyze repetitions of the same cultivar, espe- cially if they seem to be ampelographically different, and if they will be propagated or used as examples in cultivar collections.

Profiles of Swiss grapevine cultivars To identify the true-to-type profile of Swiss cultivars, it is important to analyze types from different origins. This reduces the chances to include a misnamed cultivar in the database. An interesting example where we successfully confirmed profiles is the case of 'Completer' and 'Lafnetscha', two ancient Swiss cultivars believed to be syno- nyms. As shown by VOUILLAMOZ et al. (2004), these varieties belong to two diffe- rent cultivars. We analyzed 16 plants na- med 'Completer' from five collection sites and 11 plants named 'Lafnetscha' from four collection sites. We found the correct profiles for all 'Completer' and for six 'Laf- netscha', while three 'Lafnetscha' turned out to be 'Completer', one was 'Gouais blanc'/'Heunisch weiss' and one was 'Heida' (Table 3). Interestingly, the 'Heida' came from the same site as two correctly named 'Lafnetscha'. Table 4 shows the genetic profiles of 27 cul- tivars, analyzed at 6 or 12 marker sites, re- spectively. True-to-typeness has been esti- mated according to the number of types analyzed, compared to literature, or of con- firmed origin (e.g., cultivars that have been bred by Agroscope ACWChangins). If

151 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

Tab. 3: Falsely named grape cultivar accessions from different analyzing more types. Two known synonym collection sites that were identified with the grapevine data- groups ('Heida' / 'Savagnin blanc' / 'PayÂen' / base 'Weisser Traminer' and 'Heunisch weiss' / 'GwaÈss' / 'Gouais blanc') were confirmed with types from four and five collection sites, respec- tively. For five cultivars bred by Agroscope ACWChangins ('Charmont', 'Doral', 'Dioli- noir', 'Gamaret' and 'Garanoir'), a genetic profile was established. These profiles can be added to the identification files of the cultivars. A unique profile was determined for several historical Swiss cultivars. Finding the true-to-type profile for rare cultivars sometimes turned out to be difficult. Historical cultivars are often only grown at one or a few si- tes or originate all from the same site. Both types of 'Eyholzer rot' from the cultivar collections in FruÈ msen and Au were misnamed and turned out to be 'Completer' and 'Doral', respectively. Only the two types from a vineyard in Valais and the cultivar collection of Agroscope ACW Changins, Pully, showed the same profile which was unique and therefore was assigned the true- to-type profile. '', from two different collection sites (collected from a vineyard and a nursery, respectively), is a cultivar that had been introduced by Agroscope ACWChangins from Italy, showed two false profiles. One turned out to be 'Diolinoir', while the other was a 'Cabernet Franc'. We were not able to clearly assign the genetic profile of all cultivars. Of six types of 'AÈ ugstler weiss' from three type sites (Au, Agroscope ACWWa Èdenswil and FruÈ msen), none could be assigned a typical profile. One type of 'AÈ ugstler weiss' was identical to 'Scheurebe', and five were identical to 'Madeleine Angevine'. This suggests that 'AÈ ugstler weiss' is not a unique cul- tivar, but might be a synonym of 'Madeleine An- gevine'. Another possibility is that the types of 'AÈ ugstler weiss' found so far in Switzerland ori- ginated from one plant that was confused with a 'Madeleine Angevine' in the past. In the case of 'GaÈnsfuÈ sser', we had two types with two diffe- rent, unique profiles. According to the expe- rience of a versed ampelographer, the 'GaÈnsfuÈs- ser' in the cultivar collection Au is the true-to- type one. It was also identical with 'Argant'. only one or a few types could be analyzed, the true-to- The ¹falseª 'GaÈnsfuÈ sser' turned out to be identical to typeness is supposed, but will have to be confirmed by 'Douce Noire' (J. VOUILLAMOZ, pers. comm.).

152 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

Tab. 4: Profiles of grape culti- vars found in Switzerland, analyzed at 6 or 12 micro- satellite mar- ker sites, re- spectively.

New syno- nyms Several new sy- nonyms were found among the analyzed culti- vars. All culti- vars named 'Hitzkircher' (four types) sho- wed the same profiles as 'Er- lenbacher Schwarz' collec- ted in Eastern Switzerland (four types). La- ter, two 'Erlen- bacher Schwarz' with a different profile were col- lected in Western Switzerland. Four of six mar- kers were identi- cal, and one of them hetero- and homozy- gous, respecti- vely. There are two possible ex- planations for this: 1) 'Hitzkir- cher' and 'Erlen- bacher Schwarz' are synonyms for the same cul- tivar, and the type from We- stern Switzer-

153 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

land is incorrectly named; or 2) The profile of the type by Spring from Agroscope ACWChangins (pers. from Western Switzerland belongs to the true-to-type comm.). 'Trollinger' is also a synonym of 'Schiava cultivar of 'Erlenbacher Schwarz', while in Eastern Grossa', 'Gross-Vernatsch', and 'Frankenthal noir' Switzerland, an error was made and 'Hitzkircher' (HILLEBRAND et al., 1998). The only original Swiss plants are falsely named 'Erlenbacher Schwarz'. Hitz- planting of 'MoÈ rchel' is the one at Au (AEBERHARD, kirch and Erlenbach are two villages in German spea- 2005). king Switzerland, one at the lake of Baldegg, Luzern, According to AEBERHARD (2005), the 'RaÈuschling rot' is the other at the lake of ZuÈ rich. Little is known about an ampelographic rarity, not included in any ampelo- their and whether these two cultivars could graphy nor in any foreign cultivar collection. In the am- have anything to do with each other historically. The pelographic descriptions, the plants are identical with grape cultivar might simply have been given a different the 'RaÈuschling weiss', except for the pink grapes. As name at two different growing sites. According to AE- expected, the genetic profile of 'RaÈuschling rot' is iden- BERHARD (2005), only very few plantings of 'Hitzkir- tical to the one of 'RaÈuschling weiss'. These plants are cher' still exist. In his own private collection, he grows berry color mutants, identical cultivars except for the a plant originally purchased from H. Zulauf sen., berry color. The same situation applies to 'Elbling rot' Schinznach. This plant is also called 'Grosser blauer and 'Elbling weiss', as well as for 'Heunisch weiss' and MoÈ rsch'. The cultivar 'Hitzkircher' had been described 'Heunisch dreifarbig'. But, in several other cases, culti- by H. ZULAUF sen. as very resistant to vars with the same name and different berry colors tur- (Plasmopara viticola)(AEBERHARD, 2005) and might be ned out to be different cultivars. This applies to 'Bon- of interest as a genetic resource for disease resistance dola bianca', 'Elbling blau' and 'Heunisch blau'. A rare breeding. plant from a vineyard in Tessin called 'Bondola bianca' Another synonym is proposed for 'Briegler' and showed a different profile to 'Bondola nera'. 'Elbling 'Bondola nera'. All ten 'Briegler' and five 'Bondola blau' and 'Heunisch blau' both showed a unique pro- nera' types analyzed showed the same profile. Accor- file, not identical to 'Elbling rot/weiss' or 'Heunisch ding to the identification plate in the historical garden weiss', respectively. This result confirms the known in Au, it was supposed that 'Briegler' and 'Hitzkir- fact that these cultivars are not berry color mutants to cher' were synonyms, which can be clearly denied ba- the 'Elbling' and 'Heunisch'. The profile for 'Heunisch sed on our results. 'Briegler' may be the German blau' established in our study showed the same profile name of 'Bondola nera', a red grape cultivar from as 'Heunisch noir' in the database of VOUILLAMOZ Tessin in the southern part of Switzerland, which (pers. comm.) and also 'Heunisch schwarz' from Ger- was grown widely in ancient times (HILLEBRAND et many (JUNG, pers. comm.). al., 1998). 'Briegler' is neither identical to 'Balsamina Based on these data we propose 'Bondola nera' - nera' nor to 'Ancellotta', as suggested by ampelogra- 'Briegler' to be synonyms. The few plants of 'MoÈr- phic similarities and found in old literature (AEBER- chel' of one type site studied here might be either HARD, 2005). A unique genetic profile was determined misnamed, or 'MoÈ rchel' and 'Trollinger' are syno- for 'Thuner blau', different from the profiles of 'Heu- nyms. The same situation applies to 'Weisser AÈ ugst- nisch Blau', and 'Hitzkircher' as was already mentio- ler' - 'Madeleine Angevine'. In the case of 'Hitzkir- ned by AEBERHARD (2005). 'Thuner blau' is not men- cher' - 'Erlenbacher Schwarz' there might have been tioned in any nor in the global grape- a confusion of cultivars in the Eastern part of Swit- vine cultivar register from Geilweilerhof. The compa- zerland ('Hitzkircher' cultivars falsely named 'Erlen- rison with an international database suggested a syno- bacher'). Further studies are necessary to decide syno- nymy with 'Peloursin' (VOUILLAMOZ, pers. comm.). nymy or true-to-typeness of 'Erlenbacher Schwarz'. Unfortunately, only one type site is known at the hi- 'Hitzkircher' and 'Grosser blauer MoÈ rsch' seem to be storical garden in Au. local names of the cultivar 'Grossblau' (AEBERHARD, A rare cultivar named 'MoÈ rchel' was located by the 2005); other synonyms such as 'Urbanitraube', 'Volo- founder of the cultivar collection Au, W. EGGENBER- vina', 'Kriecher' are proposed by the same author. GER. Comparison of the genetic profile in this study According to these results, the synonym 'GaÈnsfuÈ sser' showed that it has the same profile as true-to-type - 'Erlenbacher' (proposed by AMBROSI et al., 1994) 'Trollinger' (VOUILLAMOZ, pers. comm.). This result must be rejected. The synonym 'GaÈnsfuÈ sser' - 'Ar- was verified based on ampelographical comparison gant' was confirmed.

154 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

Tab. 5: Profiles of Chasselas and the Muscat groups at 6 microsatellite marker sites, comparison to literature profiles and synonymous names.

Interesting alleles 'Chasselas' aus SiebenbuÈ rgen, 'Chasselas noir', 'Chasse- In two of the analyzed cultivars, we found special alle- las MusqueÂ', 'Gutedel Schweiz', 'Weisser Gutedel' les. One was 'Madera', which shows a 176 bp long al- (100jaÈhriger Stock, Meissen), 'Roter Gutedel' (100jaÈhri- lele at VVMD27. This allele was not found in any other ger Stock, Meissen), 'Chasselas rouge', 'Chasselas vio- cultivar profile so far, yet is not included in the standard let', 'Chasselas graurot', 'Chasselas rose royal', 'Chasse- reference alleles proposed by THIS et al. (2004). Accor- las de Fontainebleau'. Another group profile was that ding to the coding proposed in that study, this allele of 'Muscat Ottonel', also found in the Chasselas collec- would be n+2 for VVMD27. The other cultivar with a tion. Interestingly, two of the 'Muskat Gutedel' showed special allele, 'Uccellina nera', showed a rare allele at the same profile as 'Muscat Ottonel' ('Muskat Gutedel the marker VVMD7, with a length of 263 bp. This allele Eg' and 'Muskat Gutedel FR'), while 'Musque Richter' is the largest found in the standard set of alleles for this and 'Muskat Gutedel' from cultivar collection FruÈ msen marker (VVMD7, n+34) and was found in the two showed the same profile as 'Chasselas'. Morphologi- rootstock cultivars 'Kober 5BB' and 'Teleki 5C' (THIS cally, 'Muskat Gutedel' is very similar to 'Muscat Otto- et al., 2004). The rootstock cultivars 'Kober 5BB' and nel' and therefore can easily be confused. So, the two 'Teleki 5C' are crosses between Vitis berlandieri and Vi- 'Muskat Gutedel' must be correctly named 'Muscat Ot- tis riparia. Morphologically it is obvious that 'Madera' tonel'. Within the Muscats, another profile was the one and 'Uccellina nera' do not belong to the Vitis vinifera of 'Muscat blanc aÁ petits grains'. Synonyms in Switzer- species, but are interspecific crosses of unknown origin. land were found: 'Moscato bianco', 'Moscato di Ca- Our results indicate that 'Uccellina nera' from Tessin is nelli', 'Muscat du Valais', 'Muscat roseÂ', 'Muskateller not the same cultivar as the one from Italy, which be- roter' and 'Muskateller violetter'. Another Muscat longs to Vitis vinifera. found in Valais, called 'Muscat du pays' or 'Muscat vert', was found to be a 'Moscato giallo', identical to Chasselas and Muscat group the profile of CRESPAN and MILANI (2001). Five more In the Chasselas and the Muscat group, we found nine cultivars, 'Muscat bleu', 'Muscat preÂcoce de Gordola', different profiles (Table 5). The Muscat profiles were 'Muscat Olivier', 'Muskateller weissgelber' and 'Mus- compared to the profiles in CRESPAN and MILANI kat New York', each had an individual profile not (2001), and three matches with literature were found. found elsewhere in our database. Most of the cultivars named 'Chasselas', 'Gutedel' or a Together with the University of NeuchaÎtel, our re- combined name, belong to the Chasselas profile. These search institute is working on a microsatellite characte- include (local names): 'Roter Gutedel Freiburg', 'Kra- rization of all Swiss cultivars. The complete database cher Gutedel', 'Gutedel Malaga', 'Diamant Gutedel', comprising genetic profiles of Swiss cultivars is availa-

155 Mitteilungen Klosterneuburg 56 (2006): 147-156 Frei et al.

ble on the internet (http://www.unine.ch/nccr/svmd). GREUTER, A. und WUNDERLIN, P. (1998): Historische Rebsorten der Schweiz, eine Dokumentation. Semesterarbeit Hoch- Identification of Swiss grapevine cultivars, especially schule WaÈdenswil, Fachhochschule ZuÈ rich (unpublished) all the autochthonous and typical cultivars, is far from FREI, A., BAUMGARTNER, D., FREY, J.E. and GAFNER, J. 2004: Op- being completed. Of all errors in naming grapevines, timized identification of grapevine cultivars using multi- plex microsatellite markers. Bot. Helvetica 114(2): 169- the most severe are the ones in cultivar collections, 179 since they represent standard plants for ampelographers FREY, J.E., FREY, B., SAUER, C. and KELLERHALS, M. 2004: Effici- as well as for establishing genetic profiles. Continuing ent low-cost DNA extraction and multiplex fluorescent PCR method for marker-assisted selection in breeding. efforts for correct cultivar identification and quality Plant Breed. 123: 554-557 control are therefore imperative. HILLEBRAND, W., LOTT, H. und PFAFF, F. (1998): Taschenbuch der Rebsorten. 12. Aufl. Mainz: Fraund, 1998 Acknowledgements REGNER, F., E ISENHELD, C., KASERER, H. und STADLBAUER, A. 2001: Weitere Sortenanalysen bei Rebe mittels geneti- The authors would like to thank J.L. SPRING (Agroscope scher Marker. Mitt. Klosterneuburg 51: 3-14 ACWChangins) and M. A EBERHARD, Berne, for ampe- SEFC, K.M., STEINKELLNER, H., WAGNER, H.W., GLOÈ SSL, J. and REGNER, F. 1997: Application of microsatellite markers lographic information, Dr. J. VOUILLAMOZ (University to parentage studies in grapevine. Vitis 36: 179-183 of NeuchaÃtel) for the possibility of comparing the data SEFC, K.M., REGNER, F., T URETSCHEK, E., GLOÈ SSL, J. and STEIN- with his large database and helpful comments, Dr. M. KELLNER, H. 1999: Identification of microsatellite sequen- ces in and their applicability for genotyping PFUNDER for critically reading the manuscript, and Dr. of different Vitis species. Genome 42: 367-373 B. DUFFY (both Agroscope ACWWa Èdenswil) for Eng- SEFC, K.M., LOPES, M.S., LEFORT, F., B OTTA, R., ROUBELAKIS-AN- lish corrections. Thanks also to A. LEHMANN, N. IN- GELAKIS, K.A., IBAÂ NÄ EZ, J., PEJIC, I., WAGNER, H.W., GLOÈ SSL, J. and STEINKELLNER, H. 2000: Microsatellite variability in and M. ZAMPICCOLI for technical assistance. AUEN grapevine cultivars from different European regions and evaluation of assignment testing to assess the geographic References origin of cultivars. Theor. Appl. Genet. 100: 498-505 THIS, P. and DETTWEILER, E. 2003: EU-project GENRES ct96 no81: European Vitis database and results regarding the AEBERHARD, M.(2005): Geschichte der alten Traubensorten : use of a common set of microsatellite markers. Acta Hor- Entstehung und Herkunft der Namen unserer Trauben- tic. 603: 59-66 sorten. - Solothurn: Aarcadia-Verl., 2005 THIS, P., J UNG, A., BOCCACCI, P., B ORREGO, J., BOTTA, R., CO- AMBROSI, H., DETTWEILER-MUÈ NCH, E., RUÈ HL, E.H., SCHMID, J. STANTINI, L., CRESPAN, M., DANGL, G.S., EISENHELD, C., and SCHUMANN, F. (1994): Farbatlas Rebsorten. - Stutt- F MONTEIRO, F., G RANDO, S., IBAÂ NÄ EZ, J., LACOMBE, gart: Ulmer, 1994 ERREIRA- T., L AUCOU, V., M AGALHAÄ ES, R., MEREDITH, C.P., MILANI, BALMER, M. (2000): Untersuchung der Verwandtschaft histori- N., PETERLUNGER, E., REGNER, F., Z ULINI, L. and MAUL, scher Rebsorten der Schweiz. Semesterarbeit Hochschule E. 2004: Development of a standard set of microsatellites WaÈdenswil, Fachhochschule ZuÈ rich (unpublished) reference alleles for identification of grape cultivars. BOWERS, J.E., DANGL, G.S., VIGNANI, R. and MEREDITH, C.P. Theor. Appl. Genet. 109: 1448-1458 1996: Isolation and characterization of new polymorphic THOMAS, M.R. and SCOTT, N.S. 1993: Microsatellite repeats in simple sequence repeat loci in grape (Vitis vinifera L.). grapevine reveal DNA polymorphisms when analyzed Genome 39: 628-633 as sequence-tagged sites (STSs). Theor. Appl. Genet. 86: BOWERS, J.E., DANGL, G.S. and MEREDITH, C.P. 1999: Develop- 985-990 ment and characterization of additional microsatellite VOUILLAMOZ, J., MAIGRE, D. and MEREDITH, C.P. 2003: Microsa- DNA markers for grape. Am. J. Enol. Vitic. 50(3): 243- tellite analysis of ancient alpine grape cultivars: pedigree 246 reconstruction of Vitis vinifera L. 'Cornalin du Valais'. CRESPAN, M. and MILANI, N. 2001: The Muscats : A molecular Theor. Appl. Genet. 107: 448-454 analysis of synonyms, homonyms and genetic relations- VOUILLAMOZ, J., MAIGRE, D. and MEREDITH, C.P. 2004: Identity hips within a large family of grapevine cultivars. Vitis and parentage of two alpine grape cultivars from Switzer- 40: 23-30 land (Vitis vinifera L. 'Lafnetscha ' and 'Himbertscha'). DETTWEILER, E. 1992: The grapevine herbarium as an aid to Vitis 43: 81-87 grapevine identification - First results. Vitis 31: 117-120 WAGNER, H.W. and SEFC, K.M. (1999): Identity 1.0. Centre for (Newsletter 4) Applied Genetics, University of Agricultural Sciences. - EGGENBERGER, W. and LENNERT, J. (1996): Lexikon des Schwei- Vienna, 1999 zer Weins. - Aarau: AT Verl., 1996 GRANDO, M.S. and FRISINGHELLI, C. 1998: Grape microsatellite markers : Sizing of DNA alleles and genotype analysis of some grapevine cultivars. Vitis 37: 79-82 Received August 23, 2006

156