Abstract
Information on the origin of elite table-grape cultivars is essential for breeding programs. A parentage study on table-grape accessions based on analysis of 25 nuclear and five chloroplast microsatellite loci was carried out. The pedigree of 50 cultivars was analyzed, including morphology studies when necessary. The chlorotypes clarify the direction of the crosses in the majority of cases, while likelihood ratios indicate high reliability of the proposed crosses. Nineteen new pedigrees are proposed for varieties from highly diverse geographical origins, such as Circé, Imperial Roja, Misket Vratchanskii, Mistress Hall, and Pizzutello Nero. Mistakes were also detected in previously described pedigrees for 13 cultivars, including Delizia di Vaprio and Madeleine Angevine. Alternative parents were identified for these 13 varieties. For another 18 varieties the previously proposed pedigree was confirmed. The cultivars Muscat of Alexandria, Afus Ali, Muscat Hamburg, and Chasselas were the most frequent parents of these 50 varieties.
Grapevine (Vitis vinifera L.) is an ancient crop with a long history of human selection. Although winegrapes are economically more important, world production of grapes intended for fresh consumption grew by 26% between the end of the 1990s and the beginning of the 2000s (OIV 2005). Table-grape breeding is also becoming more significant, as indicated by the rising number of applications for plant variety rights at the European Community Plant Variety Office (http://www.cpvo.eu.int/main/). Historically, table-grape breeding made significant advances during the 19th century and the beginning of the 20th century because of the work of breeders such as Bruni and Prosperi in Italy, Vibert and Moreau-Robert in France, Foster and Thomson in the United Kingdom, and Mathiasz in Hungary. In Italy, Luigi and Alberto Pirovano produced more than 500 new table-grape cultivars (Galet 2000), some of which, like Italia, are still in use. These breeders often used a limited number of common parents such as Chasselas, Afus Ali, and Muscat of Alexandria. The latter, for instance, is involved in 191 crosses, according to the Vitis International Variety Catalogue (VIVC; www.vivc.bafz.de). Table-grape breeding today is of global interest, and cultivars used in current breeding programs are often progeny from past breeding programs, thus taking advantage of previously selected favorable traits.
Information on the origin of table-grape cultivars is of great value for breeding programs, but often that information is unknown or incorrect. Among widely distributed elite grapevine cultivars there are many synonyms (syn., different names for the same cultivar) and homonyms (different cultivars with the same name) (This et al. 2006). This complication occurs not only in wine cultivars but also in table-grape cultivars (OIV 1987), especially older ones, which makes it difficult to determine the parentage relationships from historical archives. For example, when different homonyms exist for a published parent, it is difficult to know which cultivar is the one cited in the literature. Moreover, some synonyms or homonyms have existed at certain times and places but then disappeared from use, leaving their modern equivalent, if any, unknown.
In grape-breeding programs, there is certainty about the female parent, from which the seeds are collected, but the putative pollen donor cultivar is frequently mistaken, due to, for example, pollen contamination during the pollination process. That is why in some crosses the male parent is unknown or does not correspond to the described progenitor (Ibáñez et al. 2009). Today, molecular markers can help detect mistakes by identifying the putative male parent.
Sequence-tagged microsatellite sites (STMS or microsatellites) are excellent markers for grapevine characterization (Bowers et al. 1996, Thomas and Scott 1993). In general, all plants belonging to the same cultivar (i.e., from a monoclonal origin) show identical genotypes at all microsatellite loci. Co-dominant Mendelian inheritance of microsatellites has been previously used to discover the genetic origin of some important wine cultivars (Bowers and Meredith 1997, Bowers et al. 1999, Sefc et al. 1998). Moreover, chloroplast microsatellites can uncover the direction of the cross, as chloroplasts are maternally inherited in grapevine (Arroyo-García et al. 2002).
Given the great economic importance of this species and the numerous breeding programs in progress worldwide, it is essential to determine the actual genetic origin of historical cultivars. Previously, we confirmed 35 historical pedigrees and suggested eight new pedigrees, paying special attention to seedless grape cultivars (Ibáñez et al. 2009). This expanded work reveals the genetic origins of 50 table-grape cultivars through genotyping of nuclear and chloroplast microsatellite loci. The existence of synonyms and homonyms makes it necessary to accompany molecular analysis with thorough literature revisions and morphological evaluation to ascertain that the described cultivars are those mentioned in the historical records.
Materials and Methods
Plant material was obtained from the Vitis Germplasm Bank (BGV) at the Finca El Encín (IMIDRA, Alcalá de Henares, Spain). A total of 94 grape accessions were involved in the studied pedigrees (Table 1⇓). Most are table-grape accessions, but some wine cultivars were included as possible parents of some table-grape cultivars. Among the 94 accessions, 24 in our collection have a name different from the prime name established in the Vitis International Variety Catalogue (VIVC; www.vivc.bafz.de).
DNA extractions were carried out from young leaves using the DNeasy 96 Plant Kit (Qiagen, Hilden, Germany). Twenty-five nuclear STMS were analyzed through three multiplex PCR assays as described previously (Ibáñez et al. 2009). Five additional chloroplast microsatellites were analyzed in 91 accessions through a PCR multiplex (Ibáñez et al. 2009). Chloroplast haplotypes were named using published nomenclature (Arroyo-García et al. 2006).
Pedigree analysis was accomplished using Identity 1.0 software (Wagner and Sefc 1999). Statistical analysis was performed using likelihood ratios (Hagelberg et al. 1991) where parent/offspring combinations were detected. The likelihood ratio (LR) is the quotient between the probability of the progeny genotype if it had the presumptive parents and the probability of the progeny genotype for alternative possibilities.
Results and Discussion
In a previous study of 25 nuclear microsatellites in 381 accessions, compatible crosses were found for 43 table-grape cultivars (Ibáñez et al. 2009). In the present work, the pedigrees of 50 other table-grape cultivars were determined by using the same 25 microsatellite loci (Supplementary Table 1). Because of the existence of synonyms, homonyms, and possible mistakes within the collection, the morphology of many cultivars involved in these pedigrees was studied. These data were compared with published descriptions (Branas and Truel 1965) to confirm, as far as possible, that the accessions used were the correct ones. Chloroplast haplotypes (Supplementary Table 2) were used to uncover the direction of the crosses.
The pedigrees reported were divided into three groups: (1) 19 cultivars for which no pedigree information was available in literature (Table 2⇓), (2) 13 cultivars whose published pedigree was incorrect and for which a corrected pedigree is proposed (Table 3⇓), and (3) confirmation of 18 pedigrees previously described (Table 4⇓). We analyzed the chlorotypes of all parents and progeny and the likelihood ratios for each pedigree using a reference database of 301 mostly table-grape cultivars (data not shown).
New proposed pedigrees.
The Italian breeder Bruni named his new cultivars with his name followed by a number. Bruni 12 and Bruni 36 are described as progeny of open pollinations of Madeleine Angevine (Branas and Truel 1965). We identified Baresana, an Italian cultivar from the region of Bari, as the male parent of Bruni 12, which is also cultivated in this region (Galet 2000). We also identified another Italian cultivar, Luglienga Bianca (Lignan blanc), as the pollen donor of Bruni 36 based on microsatellite genotypes. Furthermore, Bruni 36 is similar to Lignan blanc in traits such as dense bunches and early ripening.
Bruni 707 is compatible with a self-pollination of Cardinal (LR 7.07 x 1021). They differ at only four microsatellite loci and are morphologically very similar. Alternatively, Bruni 707 could be a cross between Cardinal and Koenigin der Weingaerten if a null allele is accepted at locus VMC1B11 (LR 1.25 x 1017). Cardinal is a progeny of Koenigin der Weingaerten (Ibáñez et al. 2009), and thus they share at least one allele per locus, making it difficult to eliminate one of them as a parent, even after analysis of five additional loci (data not shown). Given the higher likelihood ratios and full compatibility, the hypothesis of a self-pollination of Cardinal is more probable.
Vibert and his successor Moreau-Robert were active table-grape breeders in Angers, France, during the 19th century. According to microsatellite data, three of their selected cultivars—Sulivan blanc, Circé, and Duc de Magenta—arose from a cross between Schiras and Chasselas. Sulivan blanc is similar to Chasselas (Galet 2000), while Circé has female flowers, like Schiras. Duc de Magenta could also come from the same cross, assuming the presence of a null allele at locus VVIP60. Schiras and Chasselas are also the parents of another cultivar, Black Prince, but we found no available information about its breeder. According to the VIVC (www.vivc.bafz.de), there are three homonymous cultivars named Black Prince: one corresponds with Cinsaut, another with Schiava Grossa, and the third is cultivated in South Africa and corresponds with the actual Black Prince. The latter is presumably the accession present in El Encín, which comes from South Africa and is different from Cinsaut and Schiava Grossa.
Gradisca is another cultivar bred in Angers by Moreau-Robert, using Chasselas as the pollen donor, while Bicane (female flower) was the mother. As Chasselas Doré, Gradisca is a white cultivar with golden tone berries at maturity and has a synonym called Moranet Doré (www.vivc.bafz.de).
Though no pedigree information was found in the literature for the cultivar Malingre Precoce, bred by Malingre in France in 1840, its genotype suggests that it could be derived from a cross between Bicane and Pinot. Malingre Precoce is a white cultivar with loose bunches like Bicane and ripens early like Pinot. Another early ripening French cultivar, Roi des Precoces, could also be a progeny of Pinot, with Cinsaut as the female parent.
Muscat flavor has been a desired trait in many crosses, which sometimes supports a pedigree. Muscat à Petits Grains Blancs may be the male parent of cultivars Duchess of Buccleugh and Misket Vratchanskii, whose female progenitors would be Bicane and Coarna Alba (a female Romanian cultivar), respectively. Duchess of Buccleugh was bred by Thomson in 1863 in Scotland. It has Muscat flavor and dense bunches, as does Muscat à Petits Grains Blancs, while it could have inherited late ripening and the liability of producing millerandage from Bicane. Misket Vratchanskii is a white Muscat-flavored Bulgarian cultivar bearing bunches and berries similar to Muscat à Petits Grains Blancs.
The female-flowered cultivar Marocain noir and Muscat of Alexandria could be the parents of Mistress Hall, an English cultivar. Mistress Hall has a slight Muscat flavor, probably inherited from Muscat of Alexandria, and the dense clusters, dark black berries, and late ripening of Marocain noir. The unknown breeder of this cultivar may be Foster, as he obtained other cultivars in England using Marocain noir as the female progenitor.
The cultivar Vivona 378 (probably bred by Antonino Vivona in Italy) could be the offspring of Muscat of Alexandria and Bruni 415. It would have inherited its loose bunches, flat berries, and early ripening from Bruni 415.
There are two Italian accessions both named Pizzutello Nero with unknown origins and different genotypes. One is synonymous with Olivette Noire and the other is an accession present in different collections (www.vivc.bafz.de). The parents of the second Pizzutello Nero could be Cornichon blanc and Prune de Cazouls. The latter cultivar is a black cultivar of possible Eastern origin cultivated in Sicily (Longo, cited by Galet 2000).
Finally, there exist four Spanish cultivars of unknown origin that could be highly related: Imperial Roja, Ciruela Roja, Colgar Roja, and Almería Nera. Their genotypes reveal that they could all share Ohanes as their mother. This female-flowered cultivar originated in Rágol, a village in the province of Almería (Andalusia, Spain). The cultivar was carried to Ohanes, a village 13 km from Rágol (Alonso et al. 2006). The cultivar Ragol, named after Rágol village, is not a direct relative of Ohanes (Ibáñez et al. 2009). The four accessions proposed here as offspring of Ohanes were collected in Almería, and three of them have Ragol as the male parent. Imperial Roja has been described as a synonym for Ahmeur bou Ahmeur (Galet 2000), likely because of morphological similarity, but the genotype of the accession present in El Encín is distinct from Ahmeur bou Ahmeur and from the rest of the cultivars studied. We found that a cross between Ohanes and Ragol could have produced this cultivar. Ragol has been frequently used as a commercial pollinator for Ohanes in Almería (Hidalgo 1999). On the other hand, Ragol itself is also considered a synonym for Ahmeur bou Ahmeur according to the Vitis International Variety Catalogue. Ragol is called Encarnada de Ragol and Uva de Colgar in Almería (Alonso et al. 2006), and Galet (2000) considers those names synonyms of Ahmeur bou Ahmeur. However, the genotype of Ragol present at El Encín does not match Ahmeur bou Ahmeur. We cannot exclude the existence of two homonyms called Ragol, one a synonym for Ahmeur bou Ahmeur and the other from Almería, studied here.
Additionally, crosses between Ohanes and Ragol gave rise to Ciruela Roja and Colgar Roja. Both crosses have the late ripening characteristic of Ohanes. Colgar Roja is listed as an American cultivar in the VIVC, but given its parents, it should be considered of Spanish origin. Furthermore, one synonym for Colgar Roja is Colgar Rioja, named after Rioja village, which is close to the villages of Ohanes and Rágol.
These data point to a natural origin for these three cultivars: spontaneous pollinations in the Almería growing region. Among the millions of natural crosses between Ohanes and Ragol needed to produce grapes in the female Ohanes plants, seedlings growing from unharvested fruits were probably produced. Three of them were selected as new cultivars which have survived until the present time.
Almería Nera is the fourth Ohanes progeny found here. It has been considered a synonym of Alphonse Lavallée (Galet 2000). Nevertheless, Alphonse Lavallée is dark black, while Almería Nera is red, and the berry size and shape are different as are their microsatellite genotypes. Almería Nera was probably confused with Almería Negra (which is the true synonym for Alphonse Lavallée) because of the similarity in their names. According to the Vitis International Variety Catalogue, Almería Nera is Imperial Napoleón, a synonym for Don Mariano. However, Don Mariano is genetically different from Almería Nera. Microsatellite analysis shows that Almería Nera could derive from a cross between Ohanes and Prunesta (syn. Prunesta Nera). Moreover, Almería Nera has late ripening like Ohanes and is red, like Prunesta, from which it could have inherited its second name.
Possible mistakes in literature.
Parentage analyses using microsatellite genotypes also allowed detection of several possible errors in the literature. We used the most complete and extended information sources (Branas and Truel 1965, Galet 2000) unless otherwise stated. Compatible progenitors for the mistaken pedigrees are suggested and their chlorotypes and likelihood ratios are provided (Table 3⇑).
According to the literature, the parents of Foster’s White Seedling are Black Morocco (syn. Marocain noir) and Sweet Water (syn. Chasselas). These two cultivars are also specified as the parents of Lady Downe’s Seedling, also bred by Foster in England in 1835. However, Chasselas is noncompatible as it does not share at least one allele at 10 loci with Foster’s White Seedling and at 12 loci with Lady Downe’s Seedling. Our results show that Marocain noir and Schiava Grossa (syn. Frankenthal) could be the actual parents of Foster’s White Seedling. This confusion between Chasselas and Frankenthal could be due to the existence of a sport of Frankenthal known as Frankenthal blanc, synonym Buckland Sweet Water. This homonym, Sweet Water, could be the source of the mistake. Moreover, Frankenthal (Schiava Grossa) has an herbaceous flavor, as does Foster’s White Seedling. On the other hand, Marocain noir and Muscat of Alexandria could be the real parents of Lady Downe’s Seedling. The accession named Lady Downe’s Seedling in our collection has a slight Muscat flavor in ripe berries, as Perold indicated (Branas and Truel 1965), supporting that one of its parents could be Muscat of Alexandria.
Madeleine Angevine was bred in 1857 and is a presumed progeny of a cross between Madeleine Royale and Malingre Precoce. Although Madeleine Royale is compatible as progenitor, Malingre Precoce is noncompatible at 11 loci, with Circé the most likely other parent. Both Madeleine Angevine and Circé were released from the same breeding program in Angers, France. Madeleine Angevine could have inherited female flowers from Circé.
Several mistakes have also been detected in the described pedigrees of cultivars bred by Pirovano, an Italian who consistently used some cultivars as progenitors, including Delizia di Vaprio, Angelo, and Muscat of Alexandria. Delizia di Vaprio is an Italian cultivar obtained in 1908 from a supposed cross between Muscat of Alexandria and Foster’s White Seedling. Parentage analysis indicates that Sicilien (syn. Panse Precoce) could be its female parent instead of Foster’s White Seedling. Foster’s White Seedling was called Panse Precoce in Italy (Galet 2000), so the existence in Italy of two homonymous Panse Precoce could be responsible for the confusion.
Pirovano 159 is described as the progeny of a cross between Moscato d’Adda and Olivette Noire. However, the genotype of Pirovano 159 is noncompatible at 5 loci with the former and at 12 with the latter. Our analysis showed that Delizia di Vaprio and Angelo Pirovano (both bred by Pirovano) could be the true parents of this cross. Pirovano 315 is described as a progeny of Delizia di Vaprio and Madeleine Royale, but the genotype of the latter is not consistent with this pedigree. Madeleine Angevine (a descent of Madeleine Royale, see above) is the most likely female progenitor instead. Pirovano 531 is a similar case, where the true parent is likely a progeny of the described parent. The presumed ancestors of Pirovano 531 are Kharistvala Kolkhuri (syn. Gros Colman) and Olivette Noire. However, Olivette Noire is noncompatible at four loci, while Barbableu, a progeny of Olivette Noire and Muscat Hamburg (Ibáñez et al. 2009), could be the actual male progenitor.
Princeps is also a Pirovano release. Its described parents are Angelo Pirovano and Sultana Moscata, but they are noncompatible at seven different loci. Muscat Hamburg and Kharistvala Kolkhuri could be the real progenitors of Princeps.
Antoine Besson bred Olivette Barthelet in 1864 in France. It is described as a cross between Olivette Blanche and Cornichon blanc. However, the accession analyzed in this work arises from crossing Olivette Blanche with Beba. Cornichon blanc is noncompatible at 11 microsatellites, while Beba and Olivette Barthelet share a special trait: their berry pedicels separate easily from the cluster. The mistake could be due to a confusion between Beba and Cornichon blanc, since Beba is an ancient cultivar mainly cultivated in Andalusia and Cornichon blanc was described by a 13th-century Arabian author in Málaga (Andalusia) (Rojas Clemente y Rubio 1807).
Mathiasz bred the cultivar Mathiasz Janosne in 1902 in Hungary, reportedly from a cross between Chasselas rouge and Muscat Ottonel. Muscat Ottonel is noncompatible with Mathiasz Janosne at three loci and at 13 loci when Chasselas rouge is considered as the second parent. Muscat of Alexandria could be the female progenitor. In this case, the male parent was correct, while the mistake occurred in the mother, probably because of confusion between two Muscat cultivars at the breeding center: Muscat Ottonel and Muscat of Alexandria. Mathiasz also bred Thalloczy Lajos from a reported cross between Royal Vineyard and Muscat of Alexandria (Galet 2000, OIV 1987). The prime name for Royal Vineyard is Koenigin der Weingaerten and it is noncompatible with Thalloczy Lajos. Nevertheless, there is a less common homonym for Royal Vineyard whose prime name is Sicilien (syn. Panse Precoce), which is compatible as a parent of this progeny. In this instance, the problem stems from the use of a little-extended synonym for the parent Sicilien. To avoid confusion with Koenigin der Weingaerten, this pedigree has been included as a possible mistake.
Bruni 41 was described as a progeny of Mathiasz Janosne and Panse Precoce. However, Mathiasz Janosne is noncompatible with this pedigree at eight microsatellites. Szauter Gusztav, also bred by Mathiasz, could be the actual male progenitor. Lastly, Prosperi 285 is described as the offspring of Moscato di Terracina and Bicane, but results are noncompatible with Bicane at 12 loci. Muscat of Alexandria could be the male parent.
Confirmation of previously described pedigrees.
We confirmed 18 previously described pedigrees (Branas and Truel 1965, Cosmo 1975, Galet 2000, OIV 1987, Wagner and Truel 1988). The direction of the crosses, frequently unknown, was established through chlorotype analysis (Table 4⇑). The presence in these pedigrees of certain cultivars with revised pedigrees discussed in the previous section, such as Delizia di Vaprio and Mathiasz Janosne, supports findings described above.
The crosses that produced Danam and Bruni 90 are compatible only if a null allele is present at the locus VVIV37. The presence of this possible mutation in two pedigrees described in the literature supports the existence of null alleles at this locus.
Reliability of described crosses.
The reliability of the 50 crosses studied was quantified through the calculation of likelihood ratios. The values ranged from 1016 to 1022 for the new and amended pedigrees (Table 2⇑, Table 3⇑), while ratios for the confirmed pedigrees were lower (1013 to 1021, Table 4⇑), supporting the new proposed pedigrees. When considering the possibility of close relatives involved in the crosses (rel(2)x(1) and rel(1)x(2) in Tables 2, 3, 4⇑⇑⇑), likelihood ratios are much lower, but still above 103 for most crosses. Again, there are no substantial differences between the values obtained for the new and for the previously described pedigrees. Low likelihood ratios are expected as a consequence of the high endogamy existing within table-grape cultivars, but our values are in the same order as published pedigrees for wine cultivars such as Chardonnay, with 32 microsatellite loci (Bowers et al. 1999). In general, the values are similar or slightly higher than those obtained by other authors (Bowers et al. 1999, di Vecchi Staraz et al. 2007, Piljac et al. 2002, Sefc et al. 1998, Vouillamoz et al. 2003), although lower than that obtained for the pedigree of Lafnetscha, an alpine grape cultivar, using 31 microsatellites (Vouillamoz et al. 2004).
Only four chloroplast haplotypes (A, B, C, and D) were found among the studied cultivars. Despite this (expected) low diversity, the chlorotypes allowed discrimination of the female parent in 42 of the 49 outcrosses. In two instances unsolved by chlorotypes, maternity could be resolved because one parent was a female (Madeleine Angevine). We found no prospective pedigree with full compatibility for the nuclear microsatellites and no compatibility for the chlorotype of both parents, which also supports the reliability of the suggested crosses.
Historical table-grape breeding.
The 50 cultivars for which compatible parents were found in this work as well as those previously published (Ibáñez et al. 2009) were not chosen or selected for any particular reason and could be then considered as representative results of breeding efforts during the 19th and 20th centuries. Hence, some general remarks can be made regarding historical table-grape breeding by pooling data for the 90 pedigrees examined in these two studies. While 54 cultivars were used as parents, only 10 of them were used six or more times: Sultanina (15), Muscat of Alexandria (14), Muscat Hamburg (12), Afus Ali (10), Bicane (9), Delizia di Vaprio (8), Chasselas (8), Koenigin der Weingaerten (7), Alphonse Lavallée (6), and Angelo Pirovano (6). Together, these 10 cultivars appear 95 times in the 90 crosses studied. There is a balance between the use of cultivars of natural origin (such as Sultanina, Afus Ali, and Chasselas) and bred cultivars (such as Muscat Hamburg, Alphonse Lavallée, and Angelo Pirovano). Sultanina is involved in most seedless table-grape crosses, while the two main Muscat table-grape cultivars (Muscat of Alexandria and Muscat Hamburg) appeared in 26 of the 90 different crosses, indicating the importance of Muscat flavor in table-grape breeding. Female cultivars have also been relevant: Bicane, Madeleine Angevine, and Ohanes appeared in 18 crosses, most of which produced seeded cultivars.
Conclusions
This work contributes to the definition and clarification of the parentage of 50 table-grape cultivars and provides additional information on the existence of possible synonyms and homonyms for cultivars involved in the crosses. Nineteen new pedigrees with high likelihood were suggested. Thirteen mistakes in the literature were detected and putative parents identified. Finally, 18 pedigrees previously described were confirmed by our results. In most instances, the direction of the cross was determined using chloroplast microsatellites that allow definition of chlorotypes. Results confirm that several cultivars like Muscat of Alexandria, Muscat Hamburg, and Afus Ali have contributed very significantly to the present table-grape gene pool.
Footnotes
↵2 present address, Instituto de Ciencias de la Vid y del Vino (ICVV), Complejo Científico Tecnológico, C/Madre de Dios 5, 26006 Logroño, Spain.
Acknowledgments: This study was made possible with the funding from the RF99-009 project (INIA, Agriculture Ministry of Spain) and of the GrapeGen project (joint venture between Genome Canada and Genoma España). A.M. Vargas was funded by a predoctoral fellowship from IMIDRA.
Supplementary data is freely available with the online version of this article at www.ajevonline.org.
- Received March 2009.
- Revision received May 2009.
- Accepted June 2009.
- Published online December 2009
- Copyright © 2009 by the American Society for Enology and Viticulture