Use of Genetic Markers to Assess Pedigrees of Grape Cultivars and Breeding Program Selections

John Bautista, Gerald S. Dangl, 2 Judy Yang, Bruce Reisch,3 and Ed Stover4

Abstract: In a plant breeding program, an accurate understanding of pedigrees provides useful guidance for future hybridizations. However, plant breeder records occasionally contain errors which may mislead future breeding efforts, and there is considerable value in independently testing reported pedigrees. In this project, SSR markers were used to confirm or correct pedigrees for grape varieties from the Cornell breeding program and a few key varieties used as parents. As expected, most (20 of 24) reported pedigrees were confirmed. Recognizing a heritable null allele from Ontario, at VVMD25, was necessary for parent progeny analysis of several varieties. Suffolk Red and Glenora were shown to be progeny of Black Kishmish and not Black Monukka. The advanced selection NY63.0970,07 is a hybrid of Pinot noir, not Gamay, x . (Ravat 51) is not descended from the reported parentage of Scibel 6905 x Pinot noir.

Key words: crop improvement, microsatellite, null allele, SSR, Vitis

Commercial grape acreage is comprised largely of German grapes and long reported to descend from a cross traditional varieties with recognized market acceptance; of Riesling x Silvaner. Genetic analyses have shown that however, newer varieties from grape breeding programs the actual pedigree is Riesling x Madeleine Royale (Dett- continue to increase in importance. These breeding pro- weiler et al. 2000). The useful high yield, earliness, and grams are primarily directed from universities and other acceptable wine quality of M011er-Thurgau has likely re- public institutions such the Agricultural Research Service sulted in erroneous efforts to duplicate the cross of Ries- of the USDA, but there are significant private breeding ling x Silvaner to achieve similar qualities with somewhat efforts as well. different fruit or vine characteristics. Since 1906 Cornell University has named and released The methodology to assess parentage of Müller-Thur- 56 grape cultivars (Reisch 2007). The primary targets for gau used simple sequence repeat (SSR) DNA markers, these varieties have been vineyards in the northeastern also known as microsatellite markers. These repetitive United States. Therefore, they have been selected for im- regions are seldom associated with functional genes, per- proved cold tolerance, oftentimes fungal disease-resistance, mitting them to accumulate mutations at a relatively high and high fruit quality. In all plant breeding programs, there rate and providing useful diversity for distinguishing in- is considerable use of previous releases as parents and/or dividuals. International adoption of a uniform set of six repeated use of parents that have proven successful in cre- SSR markers in grape (This et al. 2004) has provided an ating earlier varieties. For this reason, it is critical to know extensive database of genotypes from around the world, the actual parentage of previously released varieties. permitting comparison to type fingerprints and providing Almost all plant breeders maintain records of con- an initial framework for testing potential parent/progeny trolled crosses, but there are instances in which the pur- relationships. Confirmation of parent/progeny relation- ported parents ultimately are proven not to be the actual ships requires use of additional SSR markers to reduce parents. The most notable example in the grape world is the likelihood of erroneous conclusions. In all germ- that of MUller-Thurgau, one of the most widely planted plasm collections, it is useful to verify that the genotypes maintained are correctly identified. For many genotypes, this can be done by comparing fingerprints to plants in Laboratory assistant, Curator & research leader, USDA, ARS National a "voucher" collection, such as the one maintained in Clonal Germplasm Repository. University of California, Davis, CA 95616; Montpellier, France. Confirming expected parent/progeny Manager, Plant Identification Lab, Foundation Plant Services. University of relationships provides similar voucher confirmation for California, Davis, CA 95616; and Professor, Deparimentof Horticultural Sci- material released from plant breeders. ences, Cornell University, New York State Agricultural Experiment Station, Geneva, New York 14456. In this study, parent/progeny analyses using SSR Corresponding author (email: ed.stover(aars.usda.gov ; fax: 530 752-5974) fingerprints were applied to 25 grape cultivars and se- Acknowledgment: The authors thank Steve Luce for his careful assistance with lections from the Cornell breeding program as well as the collection of leaf tissue samples in New York. other commercially important cultivars from the eastern Manuscript submitted December 2007; revised March 2008 United States. To our knowledge, this is the first system- Copyright c 2008 by the American Society for Enolo gy and Viticulture All atic study assessing proper identification of parents in a rights reserved. sustained grape breeding program. 248 Am. J. Enol. Vitic. 59:3 (2008) Genetic Markers to Assess Pedigrees - 249

Materials and Methods for specific samples. The loci analyzed were VVMD5, VVMD6, VVMD7, VVMD2I, VVMD25, VVMD27, Young leaves from near the shoot-tip of rapidly grow- VVMD28, VVMD3I, VVMD32, VVMD34, VVMD36 ing grapevines were collected and rapidly dried between (Bowers et. al. 1996, 1999), VrZAG62. VrZAG79, blotting paper in sealed, labeled envelopes, which were VrZAG93 (Sefc et al. 1999), and VVS2 (Thomas and in plastic bags with approximately 20 grams of place d Scott 1993). The first set of eight markers included the Drierite (W.A. Hammond Drierite Company, Xenia, OH). six internationally adopted reference markers (This et al. The 50 cultivars and breeder selections tested were col- 2004). Forward primers were labeled with one of three lected from various locations; vines of a given cultivar fluorescent dyes. Fragment amplifications were verified were often collected from multiple locations (Table I). Additional data needed to complete family relationships on 2% agarose gels. Samples were prepared for capillary electrophoresis by were taken from previously generated unpublished data. Total DNA was extracted from approximately 20 mg of diluting 1.0 1iL of amplified product and 0.4 1.tL of the internal size standard 400HD ROX (ABI) in 12 l.LL for- dried leaf tissue using a DNeasy Plant Mini Kit (Qia(,en. Valencia, CA) following the manufacturers protocol. maniide. Typically, products from three loci labeled with PCR amplifications were performed on a GcneAmp different fluorescent dyes were multiplexed in PCR and Thermal Cycler (model 9700; Applied Biosystems, Foster thus also in electrophoresis. Amplified fragments were City, CA) in 20-tiL volume reactions following typical separated by electrophoresis on an AB1 Prism 3100 Genet- protocols (Dangl et al. 2005). Each sample was analyzed ic Analyzer (Applied Biosystenis) using 22 cm capillary at eight SSR loci; an additional seven loci were used with 3100 POP-4 as the matrix (see Dangl et al. 2005). Results and Discussion The varieties selected for this study did Table 1 List of grape cultivars and selections used in this study, including source location. not include all 56 releases from the Cornell Accession Source Accession Source, grape breeding program, as many are sel- dom grown or have parents that are no lon- Alden COR Marquis COR, FPS ger available. Instead, this project focused Athens NCGR-D, NCGR-G COR, FPS on recent releases, more popular releases, COR Niabell NCGR-D Bath current advanced selections. and those COR, FPS Black Kishmish NCGRDh used repeatedly as a parent (Table 2). In Black Monukka NCGR-D NY62.0136.01 COR a few instances, a reported parent was no Bronx Seedless NCGR-G NY63.0970.07 COR, FPS longer available, and the SSR data for the Bertille Seyve 4825 NCGR-G NY65.0467.08 COR reported grandparents were used instead. Technical aspects of DNA extraction Canadice COR NY70.0834.06 COR, FPS and allele scoring. DNA extraction was Canner NCGR-D NY76.0844.24 COR, FPS successful for all 69 accessions. For all NY84.0101.04 COR, FPS COR, FPS accessions, all markers provided data used Chancellor COR NY88.051 5.01 COR in the final profiles (Table 3). Final parent/ COR, FPS Ontario COR progeny analysis occasionally required the COR, FPS Ravat 34 COR assumption of the presence of a "heritable Couderc 299-35 NCGR-G Ribier NCGRDb null allele" in which a genotype lacked the sequence to bind one or both primers for Delaware NCGR-G Schuyler NCGR-G an SSR marker. In the codoniinant SSR COR Seibel 6905 NCGR-G Einset Seedless marker system, an individual with a single Seyval NCGR-G Emerald Seedless FPS null allele at a given locus will always ap- Fredonia NCGR-G Steuben COR pear to be homozygous at that locus. Fur- Glenora NCGR-G Suffolk Red COR, NCGRDb thermore, an individual that inherits a null Goff NCGR-G Seyve Villard 18-307 NCGR-G allele will always appear homozygous for an allele present in the other parent at the Himrod COR Thompson Seedless NCGR-D given locus. The assumption of a null allele Himrod 4X COR COR versus hornozygosity for that marker may NCGR-G Valvin Muscat COR, FPS Iona also be supported when the marker peak CORb, NCGR-D Interlaken Seedless NCGR-D Vignoles area reflects a single allele rather than a Joannes-Seyve 23-416 COR Zinfandel NCGR-D larger area typically resulting when two al- Abbreviations of source locations: COR, Cornell University grape breeding program, leles contribute to the PCR product. This Geneva, NY; NCGR-G, USDA—ARS, Plant Genetic Resources Unit, Geneva, NY; type of result combined with matches at all NCGR-D, National Clonal Germplasm Repository Davis, CA; FPS, Foundation Plant other markers allows the confident assess- Services, University of California, Davis. Multiple accessions from this collection were analyzed. ment of null alleles.

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Each of the 50 cultivars and selections in this study grees were confirmed. New York cultivars and advanced (Table I) had unique profiles at the 8 or 15 markers an- selections were shown to have a pedigree consistent with alyzed with the exception of Himrod and Himrod 4X, the reported parentage based on the SSR markers used, which appeared identical at all 15 markers. These pro- with just three notable exceptions, Suffolk Red, Glenora, files combined with previously generated profiles result and NY63.0970.07 (groups Q, V, and W, Table 3). For in 54 unique profiles which are arranged in 24 sets of groups N and 0, one reported parent was unavailable, relationships in Table 3. Three accessions (NY65.0467.08, and the associated grandparents were assessed using the NY88.0515.01, and Niabell) reproducibly amplified more same marker system, with the resulting data supporting than two alleles at one or more markers. That is unex- the reported pedigrees. pected for NY65.0467.08, but not unexpected for the lat- Null alleles have been previously reported in SSR ter two genotypes, which are both tetraptoids. marker analysis (Dangl et al. 2005, Vouillamoz et at. Interpretation of parent/progeny relationships. Con- 2003). The SSR fingerprints for groups Q, R, and S dis- firmation of a parent/progeny relationship may require played data consistent with a heritable null allele from numerous SSRs when elucidating previously unknown Ontario at VVMD25. For Canadice (group T) results are relationships. However, the probability is very small that consistent with Bath x Himrod as parents if both Bath a variety that a plant breeder may erroneously report as a and Canadice have a null allele at VVMD5 and VVMD6. parent also has SSR markers supporting a parent/progeny Examination of these data showed that the null allele at relationship, when tested using informative loci such as VVMD5 has been passed from Fredonia (group V) to the international marker set (This et al. 2004). Therefore, Bath (group T) and on to Canadice (group T). The simi- allele matches in this study are reported as pedigree con- larity of leaf, cluster, berry color/shape, and flavor of firmations. As expected, most (20 of 24) reported pedi- Canadice to Delaware has raised suspicions that Delaware may be a parent, but that was inconsis- tent with the SSR marker data. Table 2 Reported pedigrees of cultivars from the New York State Agricultural For Alden (group S) the original pedi- Experiment Station, Geneva (Reisch 2007), addressed in this study.______gree specified Ontario x Gros Guillaume, Name released Year Parentage Berry color and it appeared from this analysis that Alden 1952 Ontario x Gros Guillaume blue Gros Guillaume may be a synonym for Athens 1938 Hubbard x Portland blue Ribier, which confirms the suggestion (Slate et al. 1962) that Gros Guillaume Bath 1952 Fredonia x NY10805 (Chasselas Rose blue Violet x Mills) may have been Ribier, based on corn- Bronx Seedless 1937 (Goff x Iona) NY8536 x Sultanina red parisons with published descriptions. Canadice 1977 Bath x Himrod red Accounting for these factors, reported Cayuga White 1972 Seyval x Schuyler white pedigrees are confirmed in this group. Advanced seedless table-grape selec- Chardonel 1991 Seyval x Chardonnay white tion NY88.0515.01 and its two reported Corot noir 2006 Seyve Villard 18-307 x Steuben blue parents (group U) are all tetraploids. Einset Seedless 1985 Fredonia x Canner (Hunisa x Sultanina) red As expected, the spontaneously doubled Fredonia 1927 Champion x Lucile blue tetraploid Himrod 4x displays only two Glenora 1977 Ontario x Russian Seedless blue alleles for each marker. The hybrid tet- Goff 1906 Goff #19 Open Pollinated red raploid parent Niabell (Campbell 4x x Himrod 1952 Ontario x Sultanina white 4x; Olmo and Koyarna 1962) has Interlaken Seedless 1947 Ontario x Sultanina white three alleles at two of the eight markers

Marquis 1996 Athens x Emerald Seedless white tested. Inheritance in NY88.0515.01 is

Melody 1985 Seyval x GW5 (Pinot blanc x Ontario) white consistent with the reported parentage, with three alleles at three markers and Noiret 2006 NY65.0467.08 x Steuben blue four at one marker: this is consistent with NV63.0970.07 Gamay x Chancellor blue random assortment of two chromosomes NY70.0834.06 Vignoles x Cayuga White white from each linkage group to each gamete. NY76.0844.24 Traminette x Ravat 34 white The presence of a third allele at the single NY84.0101.04 NY62.0136.01 x Ravat 34 white marker VVMD28 for NY65.0467.08 may NY88.0515.01 Niabell x Himrod 4x blue be the result of a somatic mutation giving Schuyler 1947 Zinfandel x Ontario blue rise to a chimera (Riaz et al. 2002). Steuben 1947 Wayne x Sheridan blue There were four instances where the Suffolk Red 1972 Fredonia x Russian Seedless 136 red reported parentage was confused or mis- leading, three related to Cornell culti- Traminette 1996 J.S. 23-416 x Gewurztraminer white vars and selections. Suffolk Red was Fe- Valvin Muscat 2006 Couderc 299-35 x Muscat Ottonel white portedly a hybrid of Fredonia x Russian

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0) CO Co CO CO 0) C--N- U) N-- (0(0(0 L0 U) (0(0(0(0 = =Co 0) = = CO = M U) U) = 0 U) c-icc C,) CS) C C CFC>)C>) C C C>) CO Co CO CO Co Lo ((0(0(0 1> (0(0(0 U) U) (0 (0Co 0) Co (0 (0 (0(00 0)05)0) 0)0000 C>) C) C) ) N (0 U) (0 U) (0 U) It 054 CO CO CO C) C) C>) (0 (0 0 CD 0)0)>) 0) 0) 0) (.0 (0 (0 (0 U) (0 (0 0 - - - 0 - - c-i.0000 CONc-iC C-i (0(0 0) U)>0 0) C) U)U) Z > - 0) 0) 0 0 0 --00) (C)U)(0C-i U)U) C) > 0) C) 00 0)0)0-i 00-V- 00.- CO C) CO C) C) 00 CO U) C) .0) C 0 C) CO CO Co 0 0 C000 02 (0(0(0 (0(0(0 (0(0(0 (0(0 CO 00 cc U) 00(0 (0 0 0-iç000j (0St(0(000 (0 (0(0 > cCic-iç 0)CO,) COO doo 0 o Ô 0 ( C) C)> > (0(0(0 (0(0(0 (0 (0 (0 (0 dod o > 0)000>) C) C) C) 0-i C) 0-i C>) (0(0 (0 CU 0-i (0 (5) C,) > CO C,) CO 2 co (U CO CI) ,- (F) (0(0(0 N-- (0 gCOCo ( Co 0) 0) N- .9 Co 0) N- C CO c\1 (0 (0 Co CO (0 0 4COC C,) 0)COCO 0) 0) 0)-,- 0)0) (0 c-i c (0 Co co C0(0 (0 CO Co C>) r 0)000>) 0) (0 C) C) C>) C-i CO 0 C>) C>) C>) C>) 0 COCOCO2 0)0)C>)C>) E CO CO C) -C 0) 0 (I 0)0)0) 0) (0(0 0) 0) 0) 0) 1) 0 (0 (0_ (0 (0(0(0 (0 (0(0(0- 0) CO CO 0) 0)0) CO CC>) (0U)U) (0 (0(0(C) c.cc-ic>)j cj > CO 0)0) N-N-CO co CO Co 0) It (0 C) (0 (0 (0 (0 (0 0)0) - - 0) C > C) 054 CJ 0) C>) C) (0 (0 (0(0 (0(0(0 C>) CO C) CO C) CO C) 0 a> C-) C N. 0). 0) 0) U) U) (00)0) 0) 0) 0) N- (0 0) 0 Lo U) (0 (0 "-U) U). 0) N- N- N- 0) 0) 0) >0(0>) U) U) (0 IT a; (0 CO (0 (0 U) U) U) ,-N- C U) U) 0) (0 (0 It (0 U) U) (0 (0 U) (C) C (0 CO U) c c c c.)C cc005Jc-i U 04 C) - - 0) CO cOi.COç-i 0 N 0)0) - (0 U) 0) 0) N- N- N- N- N-N-U) c ço (0(0(0 U)U) U) 0) N- N- 0) CO U) U) (C) (0 (0 (0 (0 Co 0) 0) o N- 0) N- 0 > C>) c\J C>) 000-ic>) (0(0(0 (0 Co CoCO 0) (0(0(0 (0 U) (0 (0 0)0-iC>) 0)0)0-i C,> (54 0>) C>) U CO O-iC>) (0(0(0 0)04(5) COCO C>) 04 0-iC-iCO )1) (0 0) 0) 0 I CD N- (.0 U-) ) 0) U) U) C-) Co U) U) CoCo Co C) (0 C 00)0) 0) MU) (0 :o 0) Co Co Co C) 0) 0) 0 00)0) 0 0 0 0 (0 CO Co 0) (C) U) U)0) - - 0) 00 0 0) CO 0 0 0 0 0 0 Co IN CO - -0)00 0 0 0 cc - 00(0 0) 0) 0) 0) 0) 0) c-ij C>JZ 0 -ccc>)c 0)0)05)0) N c) c) (0 0) COCo - 0) Co 0) 0) 0) Co U) 0 COCO -.27 U, a) (0 (0 (0 (0 0) 0) C" U) CO 0) 0) 0) Cood C cc 0) (0 0) 0) cc cc 0 (0 > 0) 0 (0 -) 0 (0) (0 cc 0(0 cc 2 — (0 --C)-, 2co - C>) - C-i-,- -.22 C>) .- ,- 0 ca C 0 C a- CO 0) 0) 0) co C) 0) 0) CO o)(r)(oCo CO N- (C) 0) 0) (.0 Co Co (0(0(0 -COCos 0) 0) CO U) - .- (U (0(0(0 Co (0 (0 Co Co Co C-i U) U) U) Co Co C 0) CO Co (0 C) U) U) U) C) U) U) 0) Cl) -—-(0 U) > C CO (OCOCO CoCOCO COO>) CO (000 CO Co Co Co CO CI) CO CoU).-,-- > Co C) CO F IT C-i (5) CoCo Co0 -1 = 0) C,) U) (0 CO CO (0 0 0) - 0) U .0 co (/) 1) 0) = U) (F) C., Co Co = E CO CO U) N- N-CoCo0 Co Co Co C- (0 >0 (0 N-- N- N- (0 C. -CoCo 0)0 N- COCO C 0 U U) U) N- N-- N- N- U) N-- (0 04 cli C\1 N- N- (0 0) (0 N- P, U) U) U) U) - 4) a. CO C) C) C) ) a. > (0(0(0 (0(0(0 (0(0(0. N- -U CO C-i CO 0)0005) >0(0(0(0(0 U)(0 -T (0 U (0(0(0(00 (0(0(0 - 00(004. 0)0)0) 00CJC>) 0) (0 U) U) (0 0 (0(0>0 o C) (5) C-i C,) CO 0-i C,) >< CO C>) C>) CO N- C) C) C) (0 U, I- (4 0 C., (0(0(0 (0 0 0 (0 C >0 (0(0 (0> (0 (0 CO C,) 0 0 0 (0 (0 - C>) -COO>) C-i- -r 0)0)C>) (0(0a (0(0(0(1 0 5-C - ,- - co (0 co CO .2 -J•çc>i C-i (5) C-i C) CO C) c-i.cj (N00C C): çJj c (0(0(0 0) (0 (0 e c c > 0 0 0 U) C>) (0(0 >- C) (0(0(O 04(00) 0) 0 0 > CD C) . (0 C) (0 C) (00-i 0) (Si (Sic-i ON - C) CO (0 - 0 0 - 0 - 0 0-0) (0 C) C) C-i CO C>) C>) C) CO —Q = .0 0)0)04 N. (I 0) (0(00) >00)(3). 0) N- N- N-- 0)0) (00).. (0 (1) U) U) - 2? 0 F, 0) (0 (0 (0 U) (0 cc cc cc cc c cc cc co cc 0) 0) (.0 - (0(0(0 >0(0(0- -Coçj 0) CO a) cc a) (0 C CO 0) 0) 0) C 0) 0) 0) C Lo L0 Lo ---(0 -0) 0)0) à N- (0 (1) (0 (4) Lo ) ( cc> cc cc cc (0 N-- cc U) U) U) U) ,- 0) cc cc cc cc cc cc cc a) co co N- 9 cc - C - - -co C cc co cc co co co 0 = - C 0 0 C.) (-I 0 0 Co CO C) C-) 0 (0>,") - •- N- (0 CO Co N- Co (I >0>) (0 (0 (0 (0 (0 Coo) N- Co CO Co 0) N- N- 0 >0(0(0 (0(0-U•(0 a (0 Co 0) (0 (0 (0 (0(0(0 (0(0(0(0 (0 (0 U) c c c-i4 o a; a; Lo CoCO 0)(O U) N- 0)0) c j c c c-i.> c 0)J ),)C c4 i CO > CO It CO(0 CO U) N- U) U) 0) 0) 0) - U) (0 CO CO L0Co(0 CO Co Co CO Co U) U) 0) Co (0 0) 0) 0)000>) (Sic>) 054 Co (0 (0 Co U) CO Co C-i CO C-i 0)0)0-i 0) CO CO C) C) Co Co (0 U) Co CO CO 00(0(5,) 0) COC) 0) 0)0)0-i 0) CO CO C,) C C C C 0 C C C (0(0(0 (0(0(0 (0 (5) co 0 0 •0 0 Co (0 (0 0) a) (0 0-i 0>) cc cc - (0 0) U) 0) C) CO (00>) CO (0 Co C Co CO Co Co (0 (0 (0 a)cc(00 U, U) (I) CO Co CoCo COCO = Co cc c q IN c-.• CO 0)0-iC>) c-i15 00(0 CO Co = Co CoCo(0 Co Co CO ) IN IN C) C CO (0000>) (0 (0 (0 >)75 N c)l5 c\1 co coc N ç5 IN I ) c c 0)0000 000-i c>) C Co C5)01 C-i C) CO Co 0 C) COCoC>J 0 >o (0 (0 0 C) (0 C\) CO 05) 0>) (5) C) C,) C, COCOC,J COO C>) CO C) 0 Co CO Co Co 0 C-i C) (0 C) 0 C-i 04 CO C) 0 (5) C) CO Co Co 0 CO C) 0 C) C) CO 0 C-i C) 00 CO 0 C) C>) CO 0

CO 4) 0) Lo U) I) Co C) 0) U-, C) co 0) (0- CO_ (4) (0(0 -D (0 (0 OD 0) C) 0 0 00) >0) (I) U 0) C.) 0 >, > 0) >1) (0 N- 0) >, U) (0 Co E (0 C (1) 0) , C (0(0 (0 00) C c 0 CO 0)0(0 C =00) .2? 0 0 -( U) CU a) - >a1 CL 0 o N- > C,) CU 0 C C a C N->. CO a; 2E 2>- co 0 _ (0 (0(0 0(0 >- 0 C .020 C C 2? (U CO —z >z0 ZZC/) 0 • , H(002 DIN- I Am. J. Enol. Vitic. 59:3 (2008)

Genetic Markers to Assess Pedigrees - 253

C., C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) Seedless 136. Russian Seedless C, C) - C) C) C) C) C) C) - C) C) CD C) C) C) 0 --C)-- C) is not considered a prime name N C) C) C) C)C))N-- N- N- C) C) C)C)C)C) C) C) C) C) C) C) C) C) C) (C C) (CO (CO C) C) C) C) C) C)Co > - C) (Vitis International Variety Cata- (0 logue, 2007; http://wwwvivc.bafz. C) (C (C o C) C) Co Co CoC) (C(C(C(C C.) C)C)C) N- C) C) C) N- N- C) C) C) C) C) C) (C Co Co C) C) C) C) de/index.php) but is considered a CJC)ç•C)j C) C) IN (C (C (C (C (C 66 66 (C (C (C Co (C (C Co Co (CQ > (oC)C) (C C) C) C) (C(C(CC) C) C) (C (C C)(CC) synonym for both Black Monukka > C) CJCJ C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) and Black Kishmish, In our evalu- U, = = C) C) Co Co C) N- C) Co C) Co Co C) N- co o) CC) (C C) C) (C (C(CC)C) (C C) (C (C C) C) C) ation, Black Kishmish and Black C C C) IC) C) C) C) C) C)C)C)C) C) C) C) 5 C) C) Co C) Co Co Co C) CoCoC) Monukka are unique cultivars, > (CC)(C (C (C C) (C (C(C(CC) (C (C (C (C (C C) (C > C) C) CS-I C)CIC)CJ C) C) C) C) C)C)C)C) C) C) C) with distinct SSR fingerprints

CD C) (C (C = = (C (C = = (C (C (CC)C)C) C) C) C) (Table 3), Only Black Kishmish. Co - = = C Co - C C C) C) C C C) C)C)C)• C.N. not Black Monukka, produced S C) C) (C (C C) C) C) C) (C (C C) )C)C)C) C) - > Co Co C) Co Co Co Co Co Co oLll - - > C) C) - C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) SSR fingerprints consistent with parentage of Suffolk Red (group C., C) co co Co C) C) C) C) Co Co a) C) Co Co Co Co C) Co IT (C (C (C(C (C(C (C (C (C (C (C(C(C(C C) C) C) C) C) C) IT It (C V), Foundation Plant Services of cj C) (I C) C) C) C), cli c)cIj S CoCC Codd CoC OQQCo Co Co Co > (C (C (C (C (C (C (C (C (C (C (C (C(C(C(C (C(C (C the University of California, Da- C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) C)C)C) vis once maintained a selection co CM CON-N- C) C) C) C) C- C) C) N- C) C) ,- C)Co Co (C (C C) C) C) N- (C(C (C (C D (C Co (C (C C) C) C) named Russian Seedless with a C) C) C) cJ• cJj ç-4C) C)C)C)C)Cs) J CJC) (C C)C) S C)CON C)C) C)C)--- C) fingerprint matching Black Kish- > C) Co (C C) C) C) Co C) C) C) C) -2 C) Co Co C) (C C) > C) C) C) C) C) C) C) C) C) C) C) C) C) C) C) 2 C) (C mish (Dangl et al. 2001), Russian 0 (-I C) C) C) (0 C) C) C) C) C) (D C) " C) C) C) C) C) C) C) Seedless was also reported to be 0 (C (C (C C) (0 (C (0 (C C) C) (CC_ (C (C - (C 5C4C) C) : c C) C) C) C) C) C) 6 "`d" c7] C) a parent of (ilenora (group Q) but S - C) C) C)C)-Co 1C)C)C)C)C)C)C)Co C C)CoC) > (C (C (C (C (C (C Co (C (C It (C (C (C (C (C (C (C > C)C)C) C) C) C) C) C) C) .5 C) C) C) C) C) C C) C) C) C) C) C) C) that, too, can be corrected by the C) C) evidence gathered here showing 0) C) N- C) C)- C) L-) C) C) C) C) C) N- N- .= C) C) C) C) (C C) C) C) C) C) C) C) C) C) (C N- C) C) (C C) ).0 (C (C (C that Black Kishmish was the likely 2 clj LD E C) C) C) C) C) -.- C) C) C) C) C) C)C), C)C) ( çJ•çJçIC) (C N N- C) C) (C N- N- C) N- C) N- N- N- N- C) CoC)C)C)(J C),-C) parent of Glenora. In both of the (C Co Co 2 (C ICT C) C) Co Co C) (C (C (C 2 (C Co (C (C Co C) IT > C) C) C) = C) C) C) C) C) C) C) C) C) (-I C) Fn C) C) C) C) C) C) C) (C (C original publications (Einset 1973, 2 C S N- (C Co .co Pool et al, 1977), the Russian c-I C c) (C (0 C) Co C) Co C) C) C) Co Co Co C) C) C) C) Co C) C) C) C, Co Co Co c- Co 0 Co Co oLo Co Co Co Co Co C) 5 C) Co C) C) Co C) C) co (C Seedless parent was described as IN C) C) IN C) C) ^4 INC)-(C (C N Co N- N- 0 N- Co C) Co C) C) Co C) C) C)C) C) C) C) C) C) V Co C) C) U C) 0 C) Co 0 C) C) (:> C) (C) (C) C) C) C) C) C) C) C) (C "probably" being Black Monukka. > C),-,- (C ,_ C),- C) C) - - C).-.- .0 ,- - . - V C S We can correct this based on the (C C) C) (C C) C) C) - Co --Co Co - C) C)C)- N-.- ,- C) ._C) C) (C present data. C) CO Co Co C) C) C) L0 C) n (C (I) (C The advanced selection NY CoO) CO2 C)COLC)C) Co C) ) > > C) C) Co C) Co C) C) C) C) C) C) > C) C) (C C0 c_2 Co...... Co ,2)c Co C) (C C) cu = 63.0970.07 was reported in breed- CoCo 0 N-N-- S ing program records to he a hybrid (I ç-•ç.Jj (C C) C., - Co C) Co Co ,- C) C) - CoCo-N- - Co Co Co N- C Co.- Co 0 N- N- C) (C (C C) N-N-C) C) N- N- N- C) N- C) N- of Gamay x Chancellor. Possibly C) C) C) C),ç) C) C) C) C) C) C) C) C) C) C) 0 C) NJ C) S - Co Co .- - > (C(CC) N-C) C) (C C) C) C) C) - (C (C C) C) C) (C the clone of Pinot noir long mis- > C) C) C) C) C) C) C) C) C) C) C) C) C) C) C C) C) C) C) Q_ (CC) C) C) 2 named Gamay Beaujolais in Cali- a C.) CD C) (0 to C) C) C) C) C) (C C) C) co C) C) C) (C C) fornia was the parent (group W). 0 C)C)C)j C) C) C) çJ• ç çJj C)C)C)C) (C C)C)C)C) C S (C(CCo (C (C C) - C)C) cr (CC) C) Vignoles (Ravat 51), a variety of > Co Co CoCoCo 00 Co Co C) - . ,- ,- C) C) - > C) C) C) C) C) C) C) C) C\1 C) C) C) - C) _5 C) C) - 0 C) C) C) importance in the eastern United (C N-- C) States and in grape breeding pro- (I C) C) C) - C) C) (C C) (C C)C)(C(C C) C) C) C) - C) (C C) 0 C) C) C) 2 C) (0 C) C) C) C) C) C) C) C) C) C) 2 C) C) CC - - (C C C) grams, has been used in the Cor- S C) C) C) 2 C)C) )CU( C)ÔiC) C) C) > C) C) C) C) C) > a C) C) C) -(C S C) nell grape breeding program and C -(C C C = C 0 C C) is a parent of NY70,0834,06, Vig- I- N- C)C) u) N- N- C) Co Co C) C) Co.- - (C Co-Co 0 (C(CC) (C (C (C (C (C (C (C Co C) C) C) (C (C C) C) - (C C) (C noles was reported to be a hybrid C;.JC)C)•, C)c,Jj C)C) C)CJC) C) C) C) C) -(C CJC)C) S C)C)C) C) C) C) Co - C) (3) 0 C) C) C) C)à)I - > Co Co (C (C CoCo Co (C (C Co Co CoCo (C C) 2 Co Co Co C) (C CoCo Co of Seibel 6905 x Pinot de Corton > C) C) C) C)C) C) C)( C) C) C) C) C) C) C) C) C) C) C)C) C) C) C) = C) C C (Galet 1979). (There is no grape C Co 0 0 C)C)C) a = = C) (C (0 C) Co (D (C Co S C) C) C) (C S (C U) C C) co - C) variety with a prime name of Pinot a Co Co Co C C Co (C Co Co C) Co co (C 5 Co Co Co Co C Co (C Co S N IN cC)cJ IN IN IN - de Corton; thus we assume that the > C) C) C) C ID C)(C C) (0 (C (0 (C (C (C C (C aD (C C (0(0(0 Co Co C) 0 Co Co C) Co ( Co COCOCOCO C Co C) C) C) C C) C) C) > C) C) C) 0 C)C) 11)C)C)C C) C) (-I C C) C) C) C) 0 C)C)C)C)0 C) C) C) pollen parent designated was the

(C variety Pinot noir from the Corton = (C _(C N-- C - o -- Co C) wine region of Burgundy.) Howev- 2x (C ,C C o Co C 0 C) I 0 N- C C) C C) (C (CCr 5 C) C) = = Co 2 > C Co (C er, the marker data are not consis- C.) W g (Co 0 0 - (C_ Os 0 (C c V -.- 0 0 E0C)5 0 C V tent with either of these varieties C-) (C(C -- (C 2222 (C .E >- - a < C)010 z I LL CC) C) C) 0 CL z a_ >0) being parents of Vignoles (group I— Cs) I- > >< X). Two additional independent

Am. J. Enol. Vitic. 59:3 (2008) 254 - Bautista et al. sources of Vignoles were obtained and analyzed, yielding Bowers, i.E., G.S. Dangl, R. Vignani, and C.P Meredith. 1996. Iso- the same SSR profile. At this point, we do not have suf- lation and characterization of new polymorphic simple sequence ficient information to suggest what might be the correct repeat loci in grape (Vilis vinifera L.). (ienonle 39:628-633. parentage of Vignoles. To the best of our knowledge, the Dangl. G.S., M.L. Mendum, B.H. Prins, M.A. Walker, C.P. Mer- Seibel 6905 sample is true-to-type, but there is only one edith, and C.J. Simon. 2001. Simple sequence repeat analysis known source in the United States, and there is a small of a clonally propagated species: A tool for managing a grape likelihood that this sample is not true-to-type. It is also germplasm collection. Genome 44:432-438. possible that what we have as Vignoles in this country is Dangl, G.S., K. Woeste, M.K. Aradhya, A. Koehmstedt, C. Simon, not Ravats actual selection 51. D. Potter, C.A. Leslie, and G. McGranahan. 2005. Character- ization of fourteen microsatellite markers for genetic analysis Athens, but not its offspring Marquis (group A), has a and cultivar identification of walnut. J. Am. Soc. Hortic. Sci. rare allele (290 bp) at VVMD5. This allele is also found 130:348-354. in Fredonia. Athens is reported to be Hubbard Seedless x Dettweiler, E., A. Jung, E. Zyprian, and R. Topfer. 2000. Grape- Portland. Portland is a half sib of Fredonia, both resulting vine cultivar MQIIer-Thurgau and its true to type descent. Vitis from a cross with Champion, so it is reasonable to hypoth- 39:63-65. esize that Champion is the source of this rare allele. Einset, J. 1973. Lakemont, Suffolk Red and Cayuga White, In the present report, we are able to correct the re- new grapes from New York. Fruit Varieties J. 27:12-15. ported parentage of one New York breeding program se- lection and two named cultivars. In the case of Glenora Galet, P. 1979. A Practical Ampelography: Grapevine Identifica- tion. Cornell University Press, Ithaca, NY. and Suffolk Red, a V. vinifera grape that arrived with the label Russian Seedless was used as the male parent. Olmo, H.P., and A. Koyama. 1962. Niabell and Early Niabell, new tetraploid varieties of the Concord type. Bull. 790:10. California The breeders inferred that the actual name of the culti- Agric. Experiment Station, Berkeley. var was Black Monukka, but we are able to confirm that it was actually Black Kishniish. In the case of the New Pool, R.M., J.P. Watson, K.H. Kimball, and J. Einset. 1977. Ca- nadice and Glenora seedless grapes named. N.Y. Food and Life York selection, NY63.0970.07, the correction to the par- Sciences. Bull. 68. New York State Agric. Experiment Station, entage was also likely a matter of use of an inaccurately Geneva. named parent. None of the 24 breeding program releases Reisch, B. 2007. Varieties named at the New York State Agricul- evaluated in this study resulted from contamination with tural Experiment Station, Geneva, New York (http://www.nysaes. foreign pollen during the crossing process. That is sig- cornell.edu/hort/faculty/reisch/nyreleases.htmi). nificant since it is often assumed that adequate cautions Riaz, S., K.E. Garrison, G.S. Dangl, J.M. Boursiquot, and C.P. are taken to exclude foreign pollen, but the possibility Meredith. 2002. Genetic divergence and chimerism within ancient exists for foreign pollen to contaminate crosses despite asexually propagated winegrape cultivars. J. Am. Soc. Hortic. preventive efforts. With this report, we can definitively Sci. 127:508-514. say that the commercially valuable cultivar Vignoles, also Sefc, K.M., F. Regner, E. Turetsheck, J. Glössl, and H. Steinkellner. known as Ravat 51, is not descended from the reported 1999. Identification of microsatellite sequences in Vilis riparia parentage of Seibel 6905 x Pinot noir. and their applicability for genotyping of different Vitis species. Genomc 42:367-373. Conclusions Slate, G.L., J. Watson, and J. Einset. 1962. Grape varieties intro- SSR marker analysis permitted evaluation of reported duced by the New York State Agricultural Experiment Station, pedigrees for 24 grape varieties from the Cornell breed- 1928-1961. Bull. 794. New York State Agric. Experiment Sta- ing program and a few key varieties used as parents. As tion, Geneva. expected, most (20 of 24) reported pedigrees were con- This P.. et at. 2004. Development of a standard set of microsatel- firmed, but several key errors were discovered, provid- lite reference alleles for identification of grape cultivars. Theor. ing valuable information to future grape breeding efforts. AppI. Genet. 109:1448-1458. Recognizing a heritable null allele was necessary for par- Thomas, MR.. and N.S. Scott. 1993. Microsatellite repeats in grape- ent progeny analysis of several varieties. vine reveal DNA polymorphisms when analyzed as sequence- tagged sites (STSs). Theor. AppI. Genet. 86:985-990. Literature Cited Vouillamoz, J., D. Maigre, and C.P. Meredith. 2003. Microsatellite analysis of ancient alpine grape cultivars: Pedigree reconstruc- Bowers, J.E., G.S. Dangl, and C.P. Meredith. 1999. Development tion of Vitis vinfera L. Cornalin du Valais. Theor. AppI. Genet. and characterization of additional microsatellite DNA markers 107:448-454. for grape. Am. J. Enol. Vitic. 50:243-246.

Am. J. EnoI. Vitic. 59:3 (2008)