Identifcation and relationship of the autochthonous ‘Romé’ and ‘Rome Tinto’ grapevine cultivars

Ana Jiménez-Cantizano

University of Cadiz, Faculty of Sciences, Dept. Chemical Engineering and Food Technology, Vegetal Production Area, Agrifood Campus of International Excellence (CeiA3), IVAGRO, P.O. Box 40, 11510 Puerto Real, Cadiz, Spain.

Antonio Amores-Arrocha

University of Cadiz, Faculty of Sciences, Dept. Chemical Engineering and Food Technology, Vegetal Production Area, Agrifood Campus of International Excellence (CeiA3), IVAGRO, P.O. Box 40, 11510 Puerto Real, Cadiz, Spain.

Rocío Gutiérrez-Escobar

IFAPA, Centro Rancho de la Merced. Ctra. Cañada de la Loba (CA-3102) PK 3.1, 11471 Jerez de la Frontera, Cadiz, Spain.

Víctor Palacios

University of Cadiz, Faculty of Sciences, Dept. Chemical Engineering and Food Technology, Vegetal Production Area, Agrifood Campus of International Excellence (CeiA3), IVAGRO, P.O. Box 40, 11510 Puerto Real, Cadiz, Spain.



The ‘Romé’ variety is considered an Andalusian (southern region in Spain) autochthonous black grape cultivar. However, several white and black grapevine accessions are known by this name, according to Vitis International Variety Catalogue. The aim of the present work was to clarify the identity of the ‘Romé’ and ‘Rome Tinto’ as black grapevine cultivar. Eight accessions known as ‘Romé’ and two as ‘Rome Tinto’ were analyzed using 30 OIV descriptors and 22 SSR loci. The morphologic and genetic analysis showed that all accessions studied presented the same genotype and phenotype and grouped with South Spanish cultivars. This study helps to clarify the confusion over the identity of ‘Romé’ grapevine cultivar, and provides a solid basis to develop a germplasm collection to protect grapevine diversity and to recover cultivars that may be in danger of extinction.

Additional keywords: Vitis vinifera; SSR; ampelography; synonym.

Abbreviations used: SSR (Sample Sequence Repeats); VIVC (Vitis International Variety Catalogue).

Citation: Jiménez-Cantizano, A.; Amores-Arrocha, A.; Gutiérrez-Escobar, R.; Palacios, V. (2018). Short communication: Identifcation and relationship of the autochthonous ‘Romé’ and ‘Rome Tinto’ grapevine cultivars. Spanish Journal of Agricultural Research, Volume 16, Issue 4, e07SC02.

Received: 05 Mar 2018. Accepted: 14 Nov 2018.

Copyright © 2018 INIA. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC-by 4.0) License.

Funding: Ministerio de Ciencia y Tecnología, Spain (grants RF2004-00014-00-00, VIN00-036-C6-5X and RF2006-00011-00-00).

Competing interests: The authors declare no conflict of interest.

Correspondence should be addressed to Antonio Amores-Arrocha:





Material and methods

Results and discussion




Grapevine (Vitis vinifera L.) is a species that presents a wide genetic diversity and is widely conserved in germplasm banks. The existence of synonyms, homonyms and misnomers are one of the major problems for viticulture worldwide (Veloso et al., 2010) and are obstacles for an international network on the conservation of Vitis germplasm in Europe (Maul, 2008). In order to clarify the misidentification and confusion in grapevine variety designations caused by the morphological characterization subjectivity, many collections have already been characterized using Sample Sequence Repeats (SSR) markers (Lopes et al., 1999; Martín et al., 2003, 2011; Ortiz et al., 2004; De Mattia et al., 2007; Dzhambazova et al., 2009; Ibáñez et al., 2009; Vargas et al., 2009; Cipriani et al., 2010; Laucou et al., 2011; Lacombe et al., 2013; Milla-Tapia et al., 2013; Aliquó et al., 2017). Although, this problematic is collected also in The Vitis International Variety Catalogue (VIVC,, which is currently the source of reference to help group the varieties under a common and consensual identifying label (Lacombe et al., 2011). A clear example of mistakes is registered for ‘Romé’ cultivar. The VIVC database (Vitis International Variety Catalogue [VIVC,]) includes two varieties of Spanish origin called ‘Rome’: a white-berry cultivar (VICV 16035), which only is conserved in two Spanish collections [Finca El Encín (Institute Code VIVC ESP080) and Rancho de la Merced Germplasm Bank (Institute Code VIVC ESP074)], and a black-berry cultivar (VIVC 10181), whose conservation location is unknown. This European database also includes a variety with the prime name ‘Rome Tinto’ (VIVC 40905) that only is conserved in the Rancho de la Merced and its origin is uncertain.

Ibáñez et al. (2009) characterized a ‘Rome’ conserved in the Finca El Encín using 20 SSR loci. Results showed identical genotype with ‘Muscat of Alexandria’. They proposed that it could be an unknown synonym for this cultivar. Similar results were obtained by García de Luján et al. (1990) with 128 OIV ampelographic descriptors by ‘Rome’ conserved in the Rancho de la Merced collection. However, the genotype of ‘Rome Tinto’ is now unpublished.

The main objective of this work was to identify different ‘Romé’ and ‘Rome Tinto’ grapevine accessions that are conserved in the Rancho de la Merced Germplasm Bank with difference origin. Their genetic and morphologic characterization could help to detect synonyms, homonyms and false attributions. This research is necessary to provide a solid basis to develop a germplasm collection to protect grapevine diversity and to recover cultivars that may be in danger of extinction.

Material and methodsTop

A total of eight accessions, six ‘Romé’ and two ‘Rome Tinto’, were analyzed using 22 SSR loci. Six of the ‘Romé’ accessions were collected during different prospecting trips carried out from 1999-2001 in vineyards situated at the localities of Cómpeta, Torrox and Ronda (province of Málaga, Andalusia community, South Spain). All accessions analyzed are conserved in the Rancho de la Merced Germplasm Bank. Furthermore, four reference cultivars (‘Cabernet Sauvignon’, ‘Chardonnay’, ‘Muscat a Petits Grains Blancs’ and ‘Pinot Noir’) were also included to test for genetic profiles obtained with the different databases published. In order to identify the geographical origin of this ‘Romé’ accessions, the genetic analyses were complemented with other cultivars sampled at the Rancho de la Merced Germplasm Bank that originally came from different geographical areas and included Vitis vinifera varieties (‘Airén’, ‘Flame Seedless’, ‘Garrido Fino’, ‘Graciano’, ‘Mantúo de Pilas’, ‘Michele Palieri’, ‘Muscat of Alexandria’, ‘Muscat Hamburg’, ‘Ohanes’, ‘Palomino Fino’, ‘Pedro Ximenes’, ‘Syrah’, ‘Tempranillo’, ‘Vijiriega Común’, ‘Viura’ and ‘Zalema’ ‘Cabernet Sauvignon’, ‘Chardonnay’, ‘Muscat a Petits Grains Blancs’ and ‘Pinot Noir’) and interspecific hybrids (‘RM2’ and ‘Jacquez’), that were used as an outgroup in the genetic relationship analysis.

DNA extraction from plant material was performed using young leaves collected in spring. Genomic DNAs were isolated from 100 mg of frozen leaf tissue using the DNeasy Plant Mini Kit (Qiagen, Hilden, Germany). A total 22 SSR loci were analysed in order to verify the varietal identity. A first set of 20 microsatellite loci located in the 19 linkage groups of grapevine genome [VMC1B11 (Zyprian & Topfer, 2005), VMC4F3-1 (Di Gaspero et al., 2000); VVMD5, VVMD7, VVMD21, VVMD24, VVMD25, VVMD27, VVMD28, VVMD32 (Bowers et al., 1996, 1999); VVS2 (Thomas & Scott, 1993); VVIB01, VVIH54, VVIN16, VVIN73, VVIP31, VVIP60, VVIQ52, VVIV37, VVIV67 (Merdinoglu et al., 2005)] were analysed using two multiplex PCRs as described in a previous study (Vargas et al., 2007). Another set of 2 microsatellite loci [VrZAG62 and VrZAG79 (Sefc et al., 1999)] were analysed to complete the list of loci authorized by the International Organisation of Vine and Wine (OIV, 2009). These last two loci were used under the conditions detailed in a previous work (Jiménez-Cantizano et al., 2006).

PCR amplifications were carried out in an Applied Biosystems 9700 thermocycler. Amplified products were separated by capillary electrophoresis using an automated sequencer (ABI Prism 3130, Appl. Biosyst.). Fluorescently labelled fragments were detected and sized using GeneMapper v. 3.7 software (Appl. Biosyst.) and fragment lengths were determined with the help of internal size standards (GeneScan-500 LIZTM, Appl. Biosyst.). SSR profile comparisons were carried out using the Microsatellite toolkit v. 9.0 software package (Park, 2001). Genetic distances between grapevine genotypes were calculated as [-ln (proportion shared alleles)] using Microsat (Minch et al., 1997). The obtained data were used for the construction of a dendrogram using the program EXE from the PHYLIP software package (Felsenstein, 1989) and Treeview (Page, 1996).

Ampelographic analyses were carried out during three years (2012-2015) and using the descriptor list for grapevine cultivars and Vitis species from the OIV (2009) using 5 plants per accession.

Results and discussionTop

All accessions ‘Romé’ and ‘Rome Tinto’ characterized showed identical genotype at 22 SSR loci (Table 1). The genotype obtained was not included in published databases (Ibáñez et al., 2009; Vargas et al., 2009; De Andrés et al., 2012; Lacombe et al., 2013; Jiménez-Cantizano, 2014; and VIVC (, thus, this new genotype corresponds to a new grape cultivar. According to the results obtained in this study, the ‘Rome Tinto’ accession registered in the VIVC database with the variety number “40905”, should be considered a synonym of ‘Rome’ “10181”, which could be the true-to-type of the variety prime name. This variety is conserved only in the Rancho de la Merced Germplasm Bank. On the other hand, morphological characterization showed the same phenotypes in all ‘Romé’ and ‘Rome Tinto’ accessions (Table 2). This variety could be ‘Romé’ (black-berry cultivar) described in Andalusia by several authors in the 16th (Herrera, 1513) and 19th century (Clemente-Rubio, 1807; Abela & de Andino, 1885). In this way, ‘Romé’ (white-berry) characterized by García de Luján et al. (1990) and Ibáñez et al. (2009) identified as ‘Muscat of Alexandria’ could be a misnamed.

Table 1. Genetic profle of ‘Rome’ accessions, reference varieties and other cultivars analyzed at 22 SSR loci. Allele sizes are given in base pairs.

Table 2. Mean values for the OIV (2009) ampelografc and morphologic descriptors observed during three years (2012- 2015).

The resulting dendrogram using the UPGMA method (Fig. 1) defines the genomic relationships among the analyzed cultivars and shows the existence of six groups, denoted A-F. The formation of these groups may be related to the use of the cultivars and regions of origin. According to the results, ‘Romé’ cultivar is grouped (Group A) with white cultivars (‘Zalema’, ‘Pedro Ximenes’, ‘Viura’, ‘Palomino Fino’ and ‘Garrido Fino’) that are of Spanish origin and used in winemaking. Within this group A ‘Romé’ has a greater affinity with ‘Zalema’, ‘Pedro Ximenez’ and ‘Viura’ (Fig. 1). These three varieties share first-degree relationships with ‘Hebén’ (Lacombe et al., 2013; Zinelabidine et al., 2015). In addition, ‘Romé’ presented the same genotype as the ‘Viura’ cultivar for seven SSR loci (Table 1): VVIB01, VVMD7, VVMD28, VVIH54, VVIN16, VVIN73 and VVIP60. These results could indicate that ‘Romé’ variety have ‘Hebén’ as a female parent. 'Hebén' is a variety, already described in the 16th century (Herrera, 1513) as a white variety of grapevine and it was grown in the Andalusian region (García de los Salmones, 1914). The white female ‘Hebén’ proved to be a key genitor in the Iberian Peninsula (Lacombe et al., 2013; Zinelabidine et al., 2015). This variety seems to originate from North Africa (Galet, 2000), which would be consistent with the relationships between Spanish and North African grape gene pools (El Oualkadi et al., 2011). In grapevine numerous changes have occurred as a result of human selection, including the emergence of hermaphroditism and greatly increased variation in berry color (This et al., 2007). Previous research works showed that approximately 800 bp insertion is present in a few red-skinned varieties that are somatic mutants of white-skinned varieties (Kobayashi et al., 2004). In addition, according to Lijavetzky et al. (2006) VvmybA1 gene could be a major determinant of berry colour variation in grapes. The results obtained in a study of the examined allelic variation in VvmybA1 in over 200 accessions of cultivated grapevine including several well characterized fruit color mutants, indicated that the white fruited-allele of VvmybA1 most likely arose a limited number of times and that variation in this gene is likely responsible for the majority of the fruit color variation present in modern grapevine cultivars (This et al., 2007). In this sense, this same effect may have occurred with the ‘Romé’ variety.

Figure 1. Dendrogram representing relationships among the different studied cultivars, based on molecular data using UPGMA as grouping method. A-F, formed groups; see text for comments.

Autochthonous cultivars are a genetic resource which could play an important role in the future, to study their warm climatic conditions adaptation capacity and their oenological potential into new wines. According to Fraga et al. (2016), adapting to future climates may comprise a selection of more resilient varieties to warming and drying. In this regard, studies carried out with Portuguese varieties (Lopes et al., 2008; Fraga et al., 2016) shows that genetic, morphological and physiological differences between each variety allows a better adaptation to different climates resulting in of singular wines production. According to the EU and Spanish normative only registered plant material is possible to be used in new plantations. As a conclusion, ‘Romé’ cultivar could be considered as an Andalusian autochthonous black grape cultivar and its correct identification could facilitate ‘Romé’ inclusion in the Official Register of Spanish grapevine varieties.


The authors thank José Antonio Pérez Ortiz and Mª José Serrano Albarrán for technical assistance with ampelographic description.


Abela EJ, de Andino S, 1885. El libro del viticultor. Impresor de la Real Casa, Madrid, Spain.

Aliquó G, Torres R, Lacombe T, Boursiquot JM, Laucou V, Gualpa J, Fanzone M, Sari S, Pérez-Peña J, Prieto JA, 2017. Identity and parentage of some South American grapevine cultivars present in Argentina. Aust J Grape Wine Res 23: 452-460.

Bowers JE, Dangl GS, Vignani R, Meredith CP, 1996. Isolation and characterization of new polymorphic simple sequence repeat loci in grape (Vitis vinifera L.). Genome 39: 628-633.

Bowers JE, Dangl GS, Meredith CP, 1999. Development and characterization of additional microsatellite markers for grape. Am J Enol Viticult 50: 243-246.

Cipriani G, Spadotto A, Jurman I, Di Gaspero G, Crespan M, Meneghetti S, Frare E, Vignani R, Cresti M, Morgante M, Pezzotti M, Pe E, Policriti A, Testolin R, 2010. The SSR-based molecular profile of 1005 grapevine (Vitis vinifera L.) accessions uncovers new synonymy and parentages, and reveals a large admixture amongst varieties of different geographic origin. Theor Appl Genet 121: 1569-1585.

Clemente Rubio S de R, 1807. Ensayo sobre las variedades de vid que vegetan en Andalucía. Imp. Villalpando, Madrid.

De Andrés MT, Benito A, Pérez-Rivera G, Ocete R, López MA, Gaforio L, Muñoz G, Cabello F, Martínez-Zapater JM, Arroyo-García R, 2012. Genetic diversity of wild grapevine populations in Spain and their genetic relationships with cultivated grapevines. Mol Ecol 21 (4): 800-816.

De Mattia F, Imazio S, Grassi F, Lovicu G, Tardaguila J, Failla O, Maitt CH, Scienza A, Labra M, 2007. Genetic characterization of Sardinia grapevine cultivars by SSR markers analysis. J Int Sci Vigne Vin 41 (4): 175-184.

Di Gaspero G, Peterlunger E, Testolin R, Edwards KJ, Cipriani G, 2000. Conservation of microsatellite loci within the genus Vitis. Theor Appl Genet 101: 301-308.

Dzhambazova T, Tsvetkov I, Atanassov I, Rusanos K, Martínez Zapater JM, Atanassov A, Hvarleva T, 2009. Genetic diversity in native Bulgarian grapevine germplasm (Vitis vinifera L.) based on nuclear and chloroplast microsatellite polymorphisms. Vitis 48: 115-121.

El Oualkadi A, Ater M, Messaoudi Z, El Heit K, Laucou V, Boursiquot JM, Lacombe T, This P, 2011. Genetic diversity of Moroccan grape accessions conserved ex situ compared to Maghreb and Europe gene pools. Tree Genet Genome 7 (6): 1287-1298.

Felsenstein J, 1989. Phylogeny inference package. Cladistics 5:164-166.

Fraga H, Santos JA, Malheiro AC, Oliveira AA, Moutinho-Pereira J, Jones GV, 2016. Climatic suitability of Portuguese grapevine varieties and climate change adaptation. Int J Climatol 36 (1): 1-12.

Galet P, 2000. Dictionnaire encylcopédique des cépages. Hachette, Paris.

García de Luján A, Puertas B, Lara M, 1990. Variedades de vid en Andalucía. Junta de Andalucía, Sevilla.

García de los Salmones N, 1914. Memoria General de las Sesiones del Congreso y Ponencias Presentadas. Imprenta Provincial, Pamplona.

Herrera A, 1513. Agricultura General, edición facsimil (1981). Servicio de Publicaciones del Ministerio de Agricultura y Pesca, Madrid.

Ibáñez J, Vargas MA, Palancar M, Borrego J, De Andrés MT, 2009. Genetic relationships among table-grape varieties. Am J Enol Viticult 60(1): 35-47.

Jiménez-Cantizano A, Martínez-Zapater JM, García de Luján A, Arroyo-García R, 2006. Caracterización molecular de accesiones de vid del banco de germoplasma del Rancho de la Merced. 29th World Congress of Vine and Wine, 25-30 Jun, Logroño, Spain.

Jiménez-Cantizano A, 2014. Caracterización molecular del banco de germoplasma de vid del Rancho de la Merced. Doctoral thesis. Universidad de Cádiz, Cádiz, Spain.

Kobayashi S, Goto-Yamamoto N, Hirochika H, 2004. Retrotransposon-induced mutations in grape skin color. Science 304: 982.

Lacombe T, Audeguin I, Boselli M, Bucchetti B, Cabello F, Chatelet P, Crespan M, D'onofrio C, Eiras-Dias J, Ercisli S, et al., 2011. Grapevine European Catalogue: Towards a Comprehensive List. Vitis 50 (2): 65-68.

Lacombe T, Boursiquot JM, Laucou V, Di Vecchi-Staraz M, Péros JP, This P, 2013. Large-scale parentage analysis in an extended set of grapevine cultivars (Vitis vinifera L.). Theor Appl Genet 126: 401-414.

Laucou V, Lacombe T, Dechesne F, Siret R, Bruno JP, Dessup M, Dessup J, Ortigosa P, Parra P, Roux C, Santoni S, Varès D, Péros JP, Boursiquot JM, This P, 2011. High throughput analysis of grape diversity as a tool for germplasm collection management. Theor Appl Genet 122 (6): 1233-1245.

Lijavetzky D, Ruiz-García L, Cabezas, J A, De Andrés M T, Bravo G, Ibáñez A, Carreño J, Cabello F, Ibáñez J, Martínez-Zapater JM, 2006. Molecular genetics of berry colour variation in table grape. Mol Genet Genomics 276 (5): 427-435.

Lopes M, Sefc K, Eiras-Dias J, Steinkellner H, Da Câmara Machado M, Da Câmara Machado A, 1999. The use of microsatellites for germplasm management in a Portuguese grapevine collection. Theor Appl Genet 99: 733-739.

Lopes J, Eiras-Dias JE, Abreu F, Climaco P, Cunha JP, Silvestre J, 2008. Thermal requirements, duration and precocity of phenological stages of grapevine cultivars of the Portuguese collection. Ciência Téc Vitivinic 23 (1): 61-71.

Martín JP, Borrego J, Cabello F, Ortiz JM, 2003. Characterization of Spanish grapevine cultivar diversity using sequence-tagged microsatellite site markers. Genome 46: 10-18.

Martín JP, Arranz C, Castro ID, Yuste J, Rubio JA, Pinto-Carnide O, Ortiz JM, 2011. Prospection and identification of grapevine varieties cultivated in north Portugal and northwest Spain. Vitis 50: 29-33.

Maul E, 2008. Synonymy, homonymy and misnaming are obstacles for an international network on the conservation of Vitis germplasm in Europe. In: Report of a Working Group on Vitis. First Meeting, 12-14 Jun 2003, Palic, Serbia and Montenegro. Bioversity International, 109-115, Rome, Italy.

Merdinoglu D, Butterlin G, Bevilacqua L, Chiquet V, Adam-Blondon AF, Decroocq S, 2005. Development and characterization of a large set of microsatellite markers in grapevine (Vitis vinifera L.) suitable for multiplex PCR. Mol Breed 15: 349-366.

Milla-Tapia A, Gómez S, Moncada X, León P, Ibacache A, Rosas M, Carrasco B, Hinrichsen P, Zurita-Silva A, 2013. Naturalised grapevines collected from arid regions in Nothern Chile exhibit a high level of genetic diversity. Aust J Grape Wine Res 19: 299-310.

Minch E, Ruíz-Linares A, Goldstein D, Feldman M, Kidd JR, Cavalli-Sforza LL, 1997. Microsat 1.5: a computer program for calculating various statistics on microsatellite data. Washington State Univ., Pullman, WA, USA.

OIV, 2009. OIV descriptor list for grape varieties and Vitis species (2nd edition), Dedon, Paris.

Ortiz JM, Martín JP, Borrego J, Chávez J, Rodríguez I, Muñoz G, Cabello F, 2004. Molecular and morphological characterization of a Vitis gene bank for the establishment of a base collection. Genet Resour Crop Evol 51: 403-409.

Page RDM, 1996. TreeView: An application to display phylogenetic trees on personal computers. Comput Appl Biosci 12 (4): 357-358.

Park SDE, 2001. Trypanotolerance in West African cattle and the population genetic effects of selection. Doctoral thesis. University of Dublin, Dublin, Ireland.

Sefc KM, Regner F, Tureschek E, Glössl J, Steinkellner H, 1999. Identification of microsatellite sequences in Vitis riparia and their application for genotyping of different Vitis species. Genome 42: 367-373.

This P, Lacombe T, Cadle-Davidson M, Owens CL, 2007. Wine grape (Vitis vinifera L.) color associates with allelic variation in the domestication gene VvmybA1. Theor Appl Genet 114 (4): 723-730.

Thomas MR, Scott NS, 1993. Microsatellite repeats in grapevine reveal DNA polymorphisms when analyzed as sequence-tagged sites (STSs). Theor Appl Genet 86: 985-990.

Vargas AM, Velez MD, De Andrés MT, Laucou V, Lacombe T, Boursiquot JM, Borrego J, Ibáñez J, 2007. Corinto bianco: A seedless mutant of Pedro Ximenes. Am J Enol Viticult 58: 540-543.

Vargas AM, De Andrés MT, Borrego J, Ibáñez J, 2009. Pedigrees of fifty tables-grape cultivars. Am J Enol Viticult 60 (4): 525-531.

Veloso MM, Almandanim MC, Baleiras-Couto M, Pereira HS, Carneiro LC, Fevereiro P, Eiras-Dias J, 2010. Microsatellite database of grapevine (Vitis vinifera L.) cultivars used for wine production in Portugal. Cienc Tec Vitivinic 25: 53-61.

Zinelabidine LH, Cunha J, Eiras-Dias JE, Cabello F, Martínez-Zapater JM, Ibáñez J, 2015. Pedigree analysis of the Spanish grapevine cultivar 'Hebén'. Vitis 54: 81-86.

Zyprian E, Topfer R, 2005. Development of microsatellite-derived markers for grapevine genotyping and genetic mapping. NCBI, GeneBank.