The Eurasian grapevine (
Wild grapevines can still be found in Eastern and Western Europe (Arnold et al. 1998). The South Caucasus (Azerbaijan, Armenia and Georgia), together with eastern Anatolia, has been considered for a longtime as the birth place for viticulture with the earliest examples of wine-making (This et al. 2006, McGovern 2003, Zohary 1995, Olmo 1995, Levadoux 1956, Negrul 1938). A 1998 census (Arnold et al. 1998) showed that wild grapevine were present in Spain, Italy, Switzerland, Romania, Bulgaria, Hungary, Austria, and in the countries of former Yugoslavia (Figure 1). Apparently, Spain and Italy harbor the highest number of recorded populations and they were proposed to work as shelters for
Currently, vines found in natural habitats are considered to be a mixture of wild forms, naturalized cultivated forms and rootstocks escaped from vineyards as well as hybrids derived from spontaneous hybridizations among those species and forms (Laguna 2003, Lacombe et al. 2003, This et al. 2006). Recently, Arrigo and Arnold (2007) compared ecological features and genetic diversity among populations of naturalized rootstocks and native wild grapevines and did not detect the existence of genetic flux between them. The genetic analysis of wild grapevine populations from France and Spain (Di Vecchi et al 2009; Andres et al 2012) detected the existence of gene flow between cultivated and wild grapevine, estimating up to 3% of pollen migration between the cultivated fields and closely located wild grape. These pollen fluxes may have a significant effect on the evolution of those populations. Currently, wild grapevine is endangered throughout all its distribution range, (Di Vecchi et al. 2009) and conservation efforts are required to maintain the genetic integrity and survival of the remnant populations. Within this context, information on the amount and distribution of wild grapevine genetic diversity is crucial for the development of conservation strategies.
The principal key ideas of this chapter is a better understanding of the exact status of the remaining wild grape populations and their relationships with existing varieties using the molecular markers and genetic analysis approaches that it has been published about some wild grapevine populations around the Mediterranean basin.
2. Chlorotype variation and distribution in
V. viniferassp. sylvestris around de Mediterranean basin
The chlorotype variation is based on specific features of the chloroplast genome as well as its conserved gene order and coding sequences in different species and its general lack of heteroplasmy and recombination. Furthermore, chloroplasts are uniparentally transmitted in most species (usually maternal in angiosperms and paternal in gymnosperms). The low mutations rates observed in the chloroplast genome represent a drawback to their wide application in the study of population history and dynamics within a given species. However, this problem has been overcome by the identification of variable intergenic regions and introns flanked by conserved sequences in many species as well as by the identification of chloroplast microsatellites which consist of mononucleotide repeats. Chloroplast microsatellites have been found in all plant species analyzed and they frequently are highly polymorphic (Provan et al. 2001). One problem associated with chloroplast microsatellites is their high homoplasy due to the recurrent generation of alleles of the same length that creates alleles which being identical by state are not identical by descent. High levels of homoplasy can confound estimates of population differentiation and the recurrent generation of alleles could mimic gene flow (Goldstein and Pollock 1997). The risk is however reduced in intraspecific analysis (Arnold et al. 2002).
As in other angiosperms, grapevine chloroplasts are maternally inherited (Arroyo-García et al. 2002) and therefore transmitted through seeds and cuttings. The chloroplast genome of grape is 160,928 bp in length and its gene content and gene order are identical to many other unarranged angiosperm chloroplast genomes (Jansen et al. 2006). Genetic diversity at the grape chloroplast has so far only been analyzed at the level of chloroplast microsatellite loci. Polymorphisms were searched by Arroyo-García et al. (2006) with 54 chloroplast microsatellite markers corresponding to 34 different loci in sample sets of four Vitis species (
Very small and isolated populations of
3. Multiple origins for cultivated grapevine
The chlorotype distributions observed among
4. Nuclear diversity in cultivated and wild grapevine
The characterization of the genetic diversity and its distribution throughout the species range is important for our understanding about the adaptation and survival of wild species to ensure that genetic resources are available for use in research and breeding programs (This et al., 2006). Microsatellite markers, being abundant, multi-allelic and polymorphic, provide a means of detecting genetic polymorphism. Due to their co-dominant structure this marker system enables studies on population genetic analysis, assessment of genetic structures and differentiation in germplasm collections and natural populations. The cultivated grapevine (
Cipriani et al., (2010) have analyzed a collection of 1005 grapevine accessions; they were genotyped at 34 microsatellite loci with the aim of analyzing genetic diversity and exploring parentages. This study constitutes the largest analysis of genetic diversity in cultivated grape and confirms previous analyses suggesting that grape is a very diverse species (Martinez et al. 2006; Ibanez et al. 2009). The genetic diversity on average is quite high for
Several studies have described successfully used microsatellite markers to genotype
The total genetic diversity values found in wild grape individuals from Anatolia region are higher than of wild type accessions from other regions such as those described for the Mediterranean basin (Andrés et al., 2012; Di Vecchi et al., 2009; Lopes et al., 2009; Zinelabidine et al. 2010). In general, these values are similar for outcrossing vegetative propagated perennial species (Bejaj et al. 2007). The observed heterozygosity (Ho) is not significantly different (P≤0.01) than expected heterozygosity (He) in the wild group, indicating a random mating population. However, reduction in observed heterozygosity has been observed in wild grapevine populations analyzed in Spain, Portugal, France or Italy (Andrés et al 2012; Lopes et al. 2009; Di Vecchi et al. 2006; Grassi et al. 2003), most likely due to the reduction of these populations by human action. The comparison of the genetic diversity values with the authoctonous grape cultivars from Anatolia region indicated that diversity is greater in the wild grapes than in the cultivated ones. Similar results have been found in other studies (Lopes et al., 2009; Riani
In conclusion, the present study suggests that there is no immediate reason for concern about any demographic bottlenecks facing the wild grape populations of the Anatolian region, and the presence of high number of rare alleles in populations investigated here is clear evidence for this finding. At the same time, the wild population from the western and central Europe pointed out that they are suffering inbreeding depression due to the low level of genetic diversity. For the future, in situ conservation of wild grapevine populations around the Mediterranean basin should be advanced by a dynamic approach to keep the level and composition of genetic diversity as high as possible for safeguarding these precious genetic resources for crop improvement.
5. Genetic relationship: Cultivated versus wild compartment of grape
The picture arising today is of a low but clear genetic differentiation of cultivars and wild grape based either on chloroplast markers (Arroyo-Garcia et al. 2006; Grassi et al. 2006), nuclear microsatellites (Snoussi et al. 2004; Grassi et al. 2003; Lopes et al., 2009; Ergul et al., 2011; Andres et al., 2012) or both (Grassi et al. 2003; Sefc et al. 2003). The wild individuals also cluster according to their populations (Grassi et al. 2008). The positive Fis values observed in the wild grapevine accessions suggest a high level of genetic relationship among the individuals of the same wild populations. In fact, the detection of potential parent-progeny relationships within wild populations supports that possibility (Andres et al., 2012). At the same time, the detection of gene flow between both compartments (Di Vecchi et al. 2009; Andres et al., 2012) could have in the future strong consequences. Therefore, the histories of both compartments are also different and as a consequence linkage disequilibrium is more important in cultivated grape (Barnaud et al. 2006) than in wild individuals (Barnaud et al. 2010).
Until now a systematic genetic and morphological characterization of the individual accessions had been done with some wild grapevine population in order to confirm whether they could correspond to bona fide ssp. sylvestris individuals, naturalized grapevine cultivars, rootstocks, or spontaneous hybrids derived from wild and cultivated forms as previously described (Di Vecchi et al. 2009; Zecca et al., 2011; Andres et al., 2012). The results of the genotypic and phenotypic analyses of wild grapevine accessions from Spain allowed classifying approximately 19% of the samples as naturalized cultivated forms (Andres
Different studies suggest genetic exchange between cultivated and wild grapevines (Cunha et al., 2009; Di Vecchi et al., 2009; Grassi et al., 2003). The genetic relationship between cultivated varieties and wild grapevine populations from Spain suggests a genetic contribution of Southern wild populations in the autochthonous grapevine cultivars varieties (Andres et al., 2012). Therefore, it seems that in opposition to the established dominant theory on the origin of the domestication of grapevine, many of the varieties of the Iberian Peninsula and from other European countries could have local origins.
The genetic analysis of wild grapevine from Spain and cultivars from European countries showed the partition in wild and cultivated forms from that region. The STRUCTURE analysis identifies two genetic groups (clusters C1 and C2) which included all the wild accessions from Spain and correspond to Northern and Southern populations and two other (C3 and C4) including the majority of the analyzed cultivars (Figure 4). The existence of two genetic groups within the wild accessions suggests some level of isolation among those genetic lineages. One possible scenario to generate such structure is that it derives from the isolation created by the last Pleistocene glaciations. As reviewed by Gomez and Lunt (2006), the fragmented nature of the Iberian Peninsula habitat favored the occurrence of multiple glacial refugees isolated from each other. Phylogeographic studies of different European species such as olive trees have shown the existence of strong genetic differentiation within the Iberian Peninsula (Belaj et al. 2007). Alternatively, these two genetic groups could represent different colonization events of the Iberian Peninsula by the species
Two different genetic clusters could also be detected within the analyzed cultivars although showing very low genetic differentiation (Fst=0.0048). This low genetic differentiation would result from the high level of gene flow between grapevine cultivars. Myles et al., (2011) have proposed that the genetic structure of the vinifera cultivars represents a large complex pedigree resulting from a number of spontaneous and inter-generation crosses between cultivars that have been vegetatively propagated for centuries. Still within this complex pedigree structure, it could be possible to distinguish different groups of more strongly related cultivars that would vary depending on the set of cultivars analyzed. In this case, an analysis of cluster 3 and 4 identified mainly Iberian cultivars as having higher ancestry in genetic cluster 3 and central European cultivars and Northern Iberian cultivars as having higher ancestry in genetic cluster 4.
Interestingly, the analyses of the ancestry values showed by analyzed cultivars identify some of them with a high ancestry value of cluster 1 and cluster 2. These grapevine cultivars correspond to the Spanish cultivars; Allarén, Benedicto, Listan Negro, Malvasia de Lanzarote and Malvasia Blanca and the European cultivars Cabernet Franc, Petit Verdot, Pinot Meunier and Sangiovese. These cultivars have been described as more closely related to wild accessions (This et al., 2006) or are considered autochthonous cultivars. Therefore, these results support the existence of introgression from Western wild forms of
In conclusion, molecular marker analysis have shown clear divergence between wild and cultivated grapes and low level of introgression (Grassi et al. 2003, Ergul et al. 2011, Andres et al. 2012), but they are still connected through gene-flow (Regner et al. 2000, Lopes et al. 2009). Some studies (Grassi et al. 2003, Arroyo-García et al. 2006, Lopes et al. 2009; Andres et al., 2012) have reported the possibility of multiple domestication events in different geographic locations in the origin of cultivated grape. The several geographic sources of wild and cultivated grapes, supports at least two separate domestication events that gave raise to cultivated grape; one derived from the wild grape from Transcaucasia, and another from the wild grape of southern European and North African origin. Probably, with wider representation of wild grape, one may be able to demonstrate the multiple domestication events supporting diffused center of domestication of cultivated grape.
6. Wild grapes as phytogenetic resource
Genetic erosion was perceived as global scale problem in the middle of the twentieth century. It was found out that the introduction of new grapevine cultivars had rapidly displaced the varieties traditionally cultivated resulting in great uniformity of cultivated crops. Therefore, the genetic diversity of those species became alarmingly scarce. This situation led to the implementation of measures for the conservation of plant genetic resources. In the vine, as in other crops, genetic erosion or loss of variability is occurring. That is, it is reducing dangerously agrobiodiversity, the genetic base on which natural selection acts, increasing dramatically the vulnerability of different cultivars to new environmental changes or the appearance of new pests and diseases (Ocete et al., 2007). It should be noted that the wild forms contain diversity for ongoing feedback to relatives (This et al., 2006). These plant genetic resources are generally not a material that is exploitable in a direct way, but it can be used in plant breeding, because wild populations still conserves an overall important genetic diversity (Grassi et al., 2003). This rich genetic pool can be used to avoid the loss of biodiversity affecting the current viticulture. Indeed, the number of allowed cultivars has been reduced to the detriment of several traditional minority varieties. Some international cultivars, like Cabernet Sauvignon, Merlot, Shyrah, Chardonnay, Sauvignon Blanc and so on are being planted in vineyards of all over the world. At the same time, only few numbers of clones from each cultivar are available (Ocete et al., 2004). These facts contribute to a great extent to speed up the problem of genetic erosion in modern viticulture and mainly lead to increase a risk of rapid propagation of new devastating pests and diseases. Some interesting characteristics of wild plants can be transferred throughout the breeding to cultivars suitable of wine making, table grapes and also rootstocks.
Genetic resources in
Together with the genetic variation characterized in the population screened in European countries could be interesting to generate a collection of genotypes that can still represent part of the existent natural genetic variation of the species. This collection could be phenotype in different environments and these genetic tools could be the basis for further studies to establish the relationship between phenotypic variation and nucleotide diversity in grapevine. Understanding grapevine natural genetic variation will help the improvement and breeding of grapevine cultivars.
7. Phenotypic characterization of wild grapevine populations
The analysis of large sets of genetic resources at the morphological level has not been intensive. One of the reasons might be the complexity of the methods available so far or the fact that phenotyping grape is expensive, time consuming and requires a lot of space. Most of the work in the past years has been devoted to the development of methods for many traits from composition of berries to disease resistance and abiotic stresses tolerance but development of rapid methods and non-destructives ones should still be a priority in order to speed up the analysis of genetic resources.
7.1. Enological characterization of wild grapevine populations from Spain
The anthocyanin composition of female grape accessions, mostly Spanish, preserved at El Encin Germoplasm Bank (Madrid, Spain) was analysed during several years. After the extraction from grape skins, total anthocyans were determined by spectrophotometry, and the anthocyanin fingerprint of grapes by HPLC, considering the relative amount of 15 anthocyanins (Revilla et al., 2010). Some typical chromatograms are shown in Figure 5.
The anthocyanin fingerprint of grapes revealed the presence of three types of accessions (Revilla et al., 2010; Revilla
Nevertheless, the intensity of acylation is quite variable in this group of accessions, and in about 30% of them the proportion of acylated derivatives is <15%, revealing that the expression of genes involved in the acylation of anthocyanins is quite variable among the accessions.
Results obtained by two-factor ANOVA (accession and year) of the 15 variables used to describe the anthocyanin fingerprint of grapes, using a group of 21 accessions sampled during three consecutive years, suggest that variations in the anthocyanin profile among wild grape accessions were more important than differences among years for a given accession (Revilla et al., 2010). Weather conditions affect to some extent the relative proportion of primitive anthocyanins (DpGl and CyGl) and of some acylated derivatives. Similar results were obtained previously in studies with cultivated varieties (Ryan and Revilla 2003, Revilla
In conclusion, the maintenance of genetic variability and the phenotypic characterization within wild grape populations has become a priority primarily due to the concurrent risks of increased human impact on flood-plain areas and the spread of new pests. Fragmentation of species habitat will reduce both the number and size of the population, and decrease the genetic variation within populations. So the existence of different genetic pools within this population is remarkable and the conservation of this germplasm becoming more interesting. This population, as the rest situated in Spain, has not a specific preservation statute. It is necessary to take into account that Spain is the country with the largest area of vineyards all over the world, and it is affected by a heavy process of genetic erosion (Ocete et al. 2007). In consequence, there is an urgent need to bring this material that could be propagated to nurseries for use in the restoration of riparian forests and undertake breeding programs of cultivars and rootstocks. Particularly, the low incidence of pests and diseases is remarkable, the high acidity of the wines and their high intensity of color total, interesting characteristics can be transferred by crossing with cultivars from Mediterranean areas. On the other hand, the immersion tolerance, absence of rot root and symptoms caused by nematodes could be interesting for obtaining new rootstocks, hybriding with traditional rootstocks, especially when many vineyards have fertirrigation or are planted on clayey soils under a rainy climate, as it was indicated by Ocete et al (2010). These phenotypic data will be used to incorporate the wild populations found to the European Vitis Data Base, according to the postulates of the COST Action FA-1003 of Viticulture (EU).
Part of the work described in this chapter was funded by the grant number RTA2008-00032-C02-01 and the grant number RTA2011-00029-C02-01. We are grateful to Rafael Ocete and Osvaldo Failla for their collaboration in the COST Action FA1003 and for sharing their expertise in wild grapevine.
Andrés M. T. Benito A. Perez-Rivera G. Ocete R. MA Lopez Gaforio. L. Muñoz G. Cabello F. Martínez-Zapater J. M. Arroyo-García R. 2012Genetic diversity of wild grapevine populations in Spain and their genetic relationship with cultivated grapevines. Molecular Ecology 21; 800 816
Aradhya MK, Dang GS, Prins BH, Boursiquot JM, Walker MA, Meredith CP, Simon CJ 2003 Genetic structure and differentiation in cultivated grape Vitis vinifera L.Genetic Resources, 81 179 192
( Arnold C. Gillet F. Gobat J. M. 1998) Situation de la vigne sauvage Vitis vinifera ssp silvestris en Europe. Vitis, 41 159 170.
Arnold C. Rossetto M. Mcnally J. Henry R. J. 2002The application of SSRs characterized for grape (V. vinifera) to conservation studies in Vitaceae. American Journal of Botany 89 22 28
Arroyo-García R. Lefort F. de Andrés M. T. et al. 2002Chloroplast microsatellites polymorphisms in Vitis species. Genome, 45 1142 1149
Arroyo-García R. Ruiz-García L. Bolling L. Ocete R. MA Lopez et. al 2006Multiple origins of cultivated grapevine (Vitis vinifera L ssp sativa) based on chloroplats DNA polymorphims. Molecular Ecology, 15 3707 3714
Arrigo N. Arnold C. 2007 Naturalised Vitis Rootstocks in Europe and Consequences to Native wild GrapevinePLos One, 2, (6) 521e.
Barnaud A. Lacombe T. Doligez A. 2006 Linkage disequilibrium in cultivated grapevine, Vitis vinifera LTheoretical Applied Genetics 112 708 716
Barnaud A. Laucou V. This P. Lacombe T. Doligez A. 2010Linkage disequilibrium in wild European grapevine, Vitis vinifera L. ssp. silvestris. Heredity 104 431 437
Belaj A. Muñoz-Diez C. Baldoni L. Procedí A. Barranco D. Satovic Z. 2007 Genetic Diversity and Populations Structure of Wild Olives from the North-western Mediterranean Assessed by SSR Markers. Annals of Botany, 100 449 458
Bryan G. J. Mc Nicoll J. Ramsay G. Meyers R. C. De Jong W. S. 1999Polymorphic simple sequence repeat markers in chloroplastgenomes of Solanaceous plants. Theoretical and Applied Genetics, 99 859 867
Chung SM, Staub JE 2003 The development and evaluation of consensus chloroplast primer pairs that possess highly variable sequence regions in a diverse array of plant taxa.Theoretical and Applied Genetics, 107 757 767
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. 2010The 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. Theoretical Applied Genetics 121 1569 1585
Cunha J. Balerias-Couto M. Cunha J. P. Banza J. Soveral A. Carneiro L. C. Eiras-Dias J. E. 2007 Characterization of Portuguese populations of Vitis vinifera L ssp sylvestris (Gmelin) Hegi.
Dangl GS, Mendum ML, Prins BH, Walker MA, Meredith CP, Simon CJ 2001 Simple sequence repeat analysis of a clonally propagated species: a tool for managing a grape germplasm collection
Di Vecchi M. Lucou V. Bruno G. Lamcombe T. Gerber S. Bourse T. Boselli M. This P. 2009 Low level of Pollen-mediated gene flow from cultivated to wild grapevine: Consequences for the evolution of the endangered subspecies Vitis vinifera L. ssp silvestris.
De Andres M. T. Cabezas J. A. Cerveza M. T. Borrego J. Martinez-Zapater J. M. Jouve N. 2007 Molecular characterization of grapevine rootstocks maintained in germplasm collectionsAmerican Journal of Enology and Viticulture 58 75 86
Ergul A. Perez-Rivera G. Soybelezoglu G. Kazan K. Arroyo-Garcia R. 201 2011 Genetic diversity in Anatolian wild grapes (Vitis vinifera subsp sylvestris) estimated by SSR markers.Plant Genetic Resources 9 3 375 383
Esselink G. D. Smulders M. J. M. Vosman B. 2003Identification of cutrose (Rosa hybrida) and rootstock varieties using robust sequence tagged microsatellite site markers. Theoretical Applied Genetics 106 277 286
Galet P. 2000 Dictionnaire encyclopédique des cépages.Hachette, Paris, France.
Gao LZ, Zhang CH, Li DY, Pan DJ, Jia JZ, Dong YS 2006Genetic diversity within Oryza rufipogon germplasms preserved in Chinese fi eld gene banks of wild rice as revealed by microsatellite markers. Biodiver Conserv 15 4059 4077
García-Beneytez E. Revilla E. Cabello F. 2002 Anthocyanin pattern of several red grape cultivars and wines made with them.European Food Research and Technology, 215 32 37
Grassi F. Labra M. Imazio S. Spada A. Sgorbati S. Scienza A. Sala F. 2003 Evidence of a secondary grapevine domestication centre detected by SSR analysis.Theoretical and Applied Genetics, 107 1315 1320
Grassi F. Labra M. Imazio S. Ocete Rubio. R. Failla O. Scienza A. Sala F. 2006 Phylogeographical structure and conservation genetics of wild grapevine
Grassi F. De Mattia F. Zecca G. Sala F. Labra M. 2008 Historical isolation and Quaternary range expansion of divergent lineages in wild grapevineBiological Journal of the Linnea Society, 95 611 619
Gomez A. Lunt D. 2006Refugia within refugia: patterns of phylogeographic concordance in the Iberian Peninsula. Edited by S Weiss and N Ferrand. Phylogeography of Southern European Refugia. Springer Dordrecht, 155 188The Netherlands.
Goldstein DB, Linares AR, Cavalli-Sforza LL, Feldman MW 1995An evaluation of genetic distances for use with microsatellitesloci. Genetics, 139 463 471
Goldstein DG, Pollock DD 1997Launching microsatellites: a review of mutation processes and methods of phylogenetic inference. Journal Heredity 88 335 342
Gómez-Alonso S. Férnandez-González M. Mena A. Martínez J. García-Romero E. 2007 Anthocyanin profile of Spanish Vitis vinifera L. red grape varieties in danger of extinction
( Heywood V. Zohary D. 1991) A catalogue of wild relatives of cultivated plants native to Europe. Flora Mediterranea 5 375 415.
Hopf M. 1991In: Die funde der Südostspanishchen Bronzezeit aus der Sammlung Siret (eds Schubart H, Ulreich H), 397 413Philipp von Zabern, Mains.
Ibanez J. MD Velez de Andres. M. T. Borrego J. 2009 Molecular markers for establishing distinctness in vegetatively propagated crops: a case study in grapevineTheoretical Applied Genetics 119 1213 1222
Imazio S. Labra M. Grassi F. Winfield M. Bardini M. Scienza A. 2002 Molecular tools for clone identification: the case of the grapevine cultivar ‘traminer’’. Plant Breed 121 531 535
IUCN( 1997) A Global Overview of Forest Protected Areas on The World Heritage List. Jim Thorsell and Todd Sigaty. (Eds.) IUCN.
Jansen R. K. Kaittanis C. Saski C. Lee S. B. Tomkins J. Alverson A. J. Daniell H. 2006 Phylogenetic analyses of Vitis (Vitaceae) based on complete chloroplast genome sequences: effects of taxon sampling and phylogenetic methods on resolving relationships among rosidsBMC Evol Biol 6: 32.
Lacombe T. Laucou V. Di Vecchi M. Bordenave L. Bourse T. Siret R. David J. Boursiquot J. M. Bronner A. Merdinoglu D. This P. 2003Inventory and characterization of Vitis viniferassp. silvestris in France. Acta Horticulturae 553 557
Laguna A. 2003 Sobre las formas naturalizadas de Vitis en la Comunidad Valenciana I. Las especies. Flora Montiberica, 23 46 82
Laucou V. Lacombe T. Dechesne F. Siret R. Bruno J. P. Dessup M. Dessup T. Ortigosa P. Parra P. Roux C. Santoni S. Vare`s D. Pe´ros J. P. Boursiquot J. M. This P. 2011High throughput analysis of grape genetic diversity as a tool for germplasm collection management. Theor Appl Gene 122 1233 1245
Levadoux L. 1956 Les populations sauvages et cultivées de Vitis vinifera L. Annales d´Amelloration del Plante, 1 59 118
MS Lopes Mendoça. D. do Rodrigues Santos. J. E. Eiras-Dias J. E. da Camara. Machado A. 2009 New insights on the genetic basis of Portuguese grapevine and on grapevine domestication
Mattia F. Imazio S. Grassi F. Doulati H. Scienza A. Labra M. 2008 Study of genetic relationships between wild and domesticated grapevine distributed from middle east regions to European countries
Mattivi F. Guzzon R. Vrhovsek U. Stefanini M. Velasco R. 2007 Metabolite profiling of grape: flavonols and anthocyanins.
Martinez L. E. Cavagnaro P. F. Masuelli R. W. Zuniga M. 2006SSR-based assessment of geneticdiversity in South American V. vinifera varieties. Plant Science 170 1036 1044
Mc Govern P. E. Glusker D. L. Exener L. J. MM Voigt 1996Neolithic resin wine. Nature, 381(6528), 480-481.
McGovern PE 2003 Ancient wine. The search for the origins of viniculture.Princeton University Press, Princeton, NJ.
Myles S. Boyko A. R. Owens C. Brown P. Grassi F. Aradhya M. K. Prins B. Reynolds A. Chia J. M. Ware D. Bustamante C. D. Buckler E. 2011Genetic structure and domestication history of the grape. Proceedings National Academic of Science USA, 108, (9), 3530-5.
Negrul AM 1938Evolucija kuljturnyx form vinograda. Doklady Akademii nauk SSSR, 8 585 585
Ocete R. MA Lopez Gallardo. A. MA Perez Troncoso. A. Cantos M. Arnold C. Perez F. 2004Las poblaciones anadaluzas de vid silvestre, Vitis vinífera L subspecie sylvestris (Gmelin) Hegi: estudio ecológico, ampelográfico, sanitario, y estrategias de conservación. Ed Consejeria de Medio Ambiente, Junta de Andalucia, Sevilla (Spain).
Ocete R. Cantos M. MA López Gallardo. A. MA Pérez Troncoso. A. Lara M. Failla O. Ferragut F. J. Liñán J. 2007Caracterización y conservación del recurso fitogenético vid silvestre en Andalucía. Ed. Falcor. Sevilla (Spain).
Ocete R. Arroyo-Garcia R. Morales M. L. Cantos M. Gallardo A. MA Perez Gomez. I. MA Lopez (2011. 2011Characterization of Vitis vinifera L. subspecies sylvestris (Gmelin) Hegi in the Ebro river Basin (Spain). Vitis 50 1 11 16
Olmo HP 1995The origin, domestication of the vinifera grape In: PE Mc Govern, SJ Fleming,SH Katz (eds) The Origins and Ancient History of Wine. Gordon and Breach Publishers, Philadelphia, USA, 23 30
Papa R. Gepts P. 2003Asymetry of gene flow and differential geographical structure of molecular diversity in wild and domesticated common bean (Phaseolus vulgaris) from Mesopotamia. Theoretical Applied Genetics, 106 239 250
Pico F. X. Mendez-Vigo B. Martinez-Zapater J. M. Alonso-Blanco C. 2008 Natural genetic variation of Arabidopsis thaliana is geographically structured in the Iberian Peninsula
Pomar F. Novo M. Masa A. 2005 Varietal differences among the anthocyanin profiles of 50 red table grape cultivars studied by high performance liquid chromatographyJournal of Chromatography 1094 34 41
Provan J. Soranzo N. Wilson N. J. Mc Nicol J. W. Forrest G. I. Cottrell J. Powell W. 1998Genepool variation in Caledonian and European Scots pine (Pinus sylvestris L.) revealed by chloroplast simple sequence repeats. Proc Roy Soc London B Biol Sci 265 1697 1705
Provan J. Soranzo N. Wilson N. J. Goldstein D. B. Powell W. 1999A low mutation rate for chloroplasts microsatellites. Genetics, 153 943 947
Provan J. 2000 Novel chloroplast microsatellites reveal cytoplasmic variation in Arabidopsis thaliana
Provan J. Powell W. Hollingsworth P. M. 2001 Chloroplast microsatellites: new tools for studies in plant ecology and evolutionTrends Ecology Evolution 16 142 147
Revilla E. MJ González-Reig Garcinuño. P. García-Beneytez E. 2005 Role of anthocyanins in the differentiation of Tempranillo winesIn Food Flavor and Chemistry: Exploration into the 21st Century. A.M. Spanier et al. (Eds.), 72 81Royal Society of Chemistry, Cambridge.
Revilla E. García-Beneytez E. Cabello F. 2009 Anthocyanin fingerprint of clones of Tempranillo grapes and wines made with them
Revilla E. Carrasco D. Benito A. Arroyo-Garcia R. 2010 Anthocyanin composition of several wild grape accessions
Revilla E. Carrasco D. Carrasco V. Benito A. Arroyo-García R. 2011Composición antociánica de la vid silvestre (Vitis vinífera spp. sylvestris). Proceedings of the 34th World Congress of Vine and Wine, Porto, Portugal.
Revilla E. Carrasco D. Carrasco V. Benito A. Arroyo-García R. 2012On the absence of acylated anthocyanins in some wild grapevine accessions. Vitis (in press).
Ranc N. Munos S. Santoni S. Causse M. 2008 A clarified position for Solanum lycopersicum var. cerasiforme in the evolutionary history of tomatoes (Solanaceae)BMC Plant Biol 8:130
Regner F. Stadlbauer A. Eisenheld C. Kaserer H. 2000 Genetic relationships among Pinots and related cultivars
Riahi L. Soghlami N. El -Heir K. Laucou V. Cunff L. L. Boursiquot J. M. Lacombe T. Mliki A. Ghorbel A. This P. 2010Genetic structure and differentiation among grapevines (Vitis vinifera) accessions from Maghred region. Genetic Resources and Crop Evolution, 57 255 272
Ryan J. M. Revilla E. 2003 Anthocyanin composition of Cabernet Sauvignon and Tempranillo grapes at different stages of ripening.
Sefc K. M. Steinkellner H. Lefort F. Botta R. Machado A. D. Borrego J. Maletic E. Glossl J. 2003 Evaluation of the genetic contribution of local wild vines to European grapevine cultivarsAmerican Journal of Enology and Viticulture 54 15 21
Snoussi H. Slimane M. H. Ruiz-Garcia L. Martinez-Zapater J. M. Arroyo-Garcia R. 2004 Genetic relationship among cultivated and wild grapevine accessions from TunisiaGenome, 47 6 1211 19
Smulders M. J. M. Cottrell J. E. Le fever F. van der Shoot J. Arens P. Vosman B. et al. 2008 Structure of the genetic diversity in black poplar (Populus nigra L.) populations across European river systems: consequences for conservation and restorationFor Ecol Manag 255 1388 1399
This P. Lacombe T. Thomas M. R. 2006 Historical origins and genetic diversity of wine grapes
Weising K. Gardner R. C. 1999A set of conserved PCR primers for the analysis of simple sequence repeat polymorphisms in chloroplast genomes of dicotyledoneous angiosperms. Genome, 42 9 19
Wenzel K. Dittrich H. H. Heimfarth M. 1987Anthocyanin composition in berries of different grape varieties. Vitis 26 65 78
Zecca G. De Mattia F. Lovicu Gm. Labra M. Sala F. Grassi F. 2010Wild grapevine:silvestis, hybrids or cultivars that escaped from vineyards? Molecular evidence in Sardinia. Plant Biology, 12 558 562
Zinelabine L. H. Haddioui A. Bravo G. Arroyo-Garcia R. Martinez-Zapater J. M. 2010Genetic origins of cultivated and wild grapevines from Morocco. American Journal of Enology and Viticulture, 61:1.
Zohary D. 1995Domestication of the Grapevine Vitis vinifera L. in theNear East. In: PE Mc Govern, SJ Fleming, SH Katz (eds) The Origins and Ancient History of Wine. Gordon and Breach Sciences Publisher, New York, USA, 23 30