Genetic diversity and parentage of Tunisian wild and cultivated grapevines (Vitis vinifera L.) as revealed by single nucleotide polymorphism (SNP) markers

Based on 261 single nucleotide polymorphism (SNP) markers, we analyzed 57 grapevine genotypes, consisting of 29 wild grapevines (Vitis vinifera subsp. sylvestris) prospected from the northwest part of Tunisia and 28 cultivated accessions (V. vinifera subsp. vinifera) maintained in the repository of the Arid Land Institute of Medenine (Tunisia). Pair-wise multilocus comparison with the ICVV SNP database allowed the identification of 13 cultivated genotypes, including ten synonymous groups with known Mediterranean or international varieties, three cases of color sports, and two misnomers. Genotypic analysis showed a high level of genetic diversity for both wild and cultivated groups. Multivariate and structure analyses clearly differentiated wild from cultivated grapevines and showed high average posterior probabilities of assignment to their group of origin. The clustering results largely supported the perceived classification and reflect that most of the present Tunisian cultivated varieties do not derive directly from the local wild populations but could correspond to materials introduced from different locations during historical times. Parentage analysis allowed the determination of the genetic origin of four Tunisian cultivars, “Garai”, “Jerbi” (from Kerkennah), “Mahdoui”, and “Reine de Vignes faux”, and showed that “Heptakilo” and “Planta Fina”, two old and widely distributed varieties in the Mediterranean basin, had an important role in the origin of Tunisian grapevines. The present study demonstrates the efficacy of SNP makers for germplasm characterization and genetic studies in grapevine.     

Molecular characterization and evolutionary pattern of the 9- cis -epoxycarotenoid dioxygenase NCED1 gene in grapevine

This study provides the first analysis of the level and patterns of nucleotide polymorphism of the NCED1 gene in grapevine (Vitis vinifera L.). A total of 123 sequences of the gene were analyzed to give a sample of 50 wild accessions and 73 cultivars. A high single nucleotide polymorphism and haplotype diversity was revealed in the cultivars studied, especially Tunisian germplasms which present an important and diverse reservoir of genetic diversity for grape breeding and conservation. The haplotype distribution highlights two origins of the cultivars studied: one may be related to primary grapevine gene pool domestication while the second seems to be more recent. Thus, besides domestication, gene introgression has also played a role in shaping the current varietal landscape of grape cultivars. Higher nucleotide and haplotype polymorphism was recorded for cultivars. This was accompanied by a higher recombination rate in cultivated grapevines for this gene, a recent selective sweep in wild samples and a balancing selection in cultivars. The conservation of genetic diversity of the endangered wild germplasm is important to ensure that the wild population can be used in future breeding programs of the domesticated cultivars. The high number of alleles discovered can be used as a valuable source for association studies between allele frequencies and phenotypic variations in this gene. In addition to natural selection, molecular evidence shows that genetic variation in this locus appears to be shaped by a combination of mutation and recombination events.     

Insights into the Vitis complex in the Danube floodplain (Austria)

European grapevine populations quickly disappeared from most of their range, massively killed by the spread of North American grapevine pests and diseases. Nowadays taxonomic pollution represents a new threat. A large Vitis complex involves escaped cultivars, rootstocks, and wild grapevines. The study aimed to provide insight into the Vitis complex in the Danube region through field and genetic analyses. Among the five other major rivers in Europe which still host wild grapevine populations, the Danube floodplain is the only one benefiting from an extensive protected forest area (93 km²) and an relatively active dynamic flood pulse. The Donau‐Auen National Park also regroups the largest wild grapevine population in Europe. Ninety‐two percent of the individuals collected in the park were true wild grapevines, and 8% were hybrids and introgressed individuals of rootstocks, wild grapevines, and cultivars. These three groups are interfertile acting either as pollen donor or receiver. Hybrids were established within and outside the dykes, mostly in anthropized forest edges. The best‐developed individuals imply rootstock genes. They establish in the most erosive parts of the floodplain. 42% of the true wild grapevines lived at the edges of forest/meadow, 33.3% at the edges forest/channels, and 23.9% in forest gaps. DBH (Diameter Breast Height) varied significantly with the occurrence of flooding. Clones were found in both true wild and hybrids/introgressed grapevines. The process of cloning seemed to be prevented in places where flooding dynamics is reduced. The current global distribution of true wild grapevines shows a strong tendency toward clustering, in sites where forestry practices were the most extensive. However, the reduced flooding activity is a danger for long‐term sustainability of the natural wild grapevine population.     

Demographic and ecogeographic factors limit wild grapevine spread at the southern edge of its distribution range

The spatial distribution of plants is constrained by demographic and ecogeographic factors that determine the range and abundance of the species. Wild grapevine (Vitis vinifera ssp. sylvestris) is distributed from Switzerland in the north to Israel in the south. However, little is known about the ecogeographic constraints of this species and its genetic and phenotypic characteristics, especially at the southern edge of its distribution range in the Levant region. In this study, we explore the population structure of southern Levantine wild grapevines and the correlation between demographic and ecogeographic characteristics. Based on our genetic analysis, the wild grapevine populations in this region can be divided into two major subgroups in accordance with a multivariate spatial and ecogeographical clustering model. The identified subpopulations also differ in morphological traits, mainly leaf hairiness which may imply adaptation to environmental stress. The findings suggest that the Upper Jordan River population was spread to the Sea of Galilee area and that a third smaller subpopulation at the south of the Golan Heights may represent a distinguished gene pool or a recent establishment of a new population. A spatial distribution model indicated that distance to water sources, Normalized difference vegetation index, and precipitation are the main environmental factors constraining V. v. sylvestris distribution at its southern distribution range. These factors in addition to limited gene flow between populations prevent further spread of wild grapevines southwards to semi‐arid regions.     

AFLP analysis of genetic relationships among aromatic grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>)

Genotypic diversity has been detected among aromatic grapevines (Vitis vinifera) by molecular markers (AFLPs). The 22 primer-pairs generated a total of 1,331 bands of which 564 (40%) were polymorphic over all the genotypes. The bootstrap analysis pointed out that a large number of polymorphic bands (200–400) has to be used for a better estimation of the genetic distances among genotypes; 383 polymorphic AFLP bands were used for the cluster and the principal coordinate analyses because they did not present missing data across all the genotypes. The cluster analysis (UPGMA), based on polymorphic AFLP markers, revealed no relationship between the Moscato and Malvasia grapevines. The Malvasias, unlike the Moscatos distinguished by their distinct muscat aroma, have to be considered a more complex group because it includes muscat and non-muscat grapevines. The principal coordinate analysis (PCO) confirmed the pattern of the cluster analysis only for those varieties which presented a low coefficient of dissimilarity, while for the other varieties there was no correspondence between the two analyses. The pattern of aggregation among aromatic grapevines in the cluster and principal coordinate analyses does not support any classification that might include an aromatic grapevine group in V. vinifera. Even though some synonyms and homonyms are present among aromatic grapevines (V. vinifera), genetic diversity exists among genotypes in AFLP markers.Communicated by H.F. Linskens     

Genetic Isolation and Diffusion of Wild Grapevine Italian and Spanish Populations as Estimated by Nuclear and Chloroplast SSR Analysis

Abstract: Genetic structure of six Italian and five Spanish populations of wild grapevine (Vitis vinifera ssp. silvestris) was investigated using nuclear and chloroplast SSR analysis. Results show that the Italian populations are characterised by high genomic diversity within populations, with a peak of heterozygosity ( Ho = 0.7637) for a population collected in southern Italy. However, the low haplotype richness and the high level of genetic distance detected among the Spanish populations, combined with their low gene flow, shows that these populations suffered from a genetic erosion. Genetic relationship between Italian and Spanish populations was investigated and results showed genetic differentiation between the two populations. Using nuclear and chloroplast SSR markers, the ratio between pollen and seed diffusion was estimated. High pollen flow, as compared with seed flow, suggests that seed diffusion is made difficult, probably due to low germinability and survival of seedlings in the wild. Seed germination and young plant survival must be considered a priority target for in situ conservation programmes. By providing information on population history, genetic structure and gene flow, and by identifying areas harbouring high levels of wild grape variability, this study provides the basis for the preservation of biodiversity of the wild grapevine.     

Wild grapevine: silvestris,hybrids or cultivars that escaped from vineyards? Molecular evidence in Sardinia

Vitis vinifera ssp. silvestris, the spontaneous subspecies of V. vinifera L., is believed to be the ancestor of present grapevine cultivars. In this work, polymorphism at 13 SSR loci was investigated to answer the following key question: are wild plants (i) true silvestris, (ii) hybrids between wild and cultivated plants or (iii) or ‘escapes’ from vineyards? In particular, the objective of the present study was to identify truly wild individuals and to search for possible hybridization events. The study was performed in Sardinia, the second largest island in the Mediterranean Sea, which is characterized by a large and well‐described number of both grape cultivars and wild populations. This region was ideal for the study because of its spatial isolation and, consequently, limited contamination from outside material. The results of this study show that domesticated and wild grapevine germplasms are genetically divergent and thus are real silvestris. Pure lineages (both domesticated and wild) show very high average posterior probabilities of assignment to their own clusters, with a low level of introgression.     

Historical isolation and Quaternary range expansion of divergent lineages in wild grapevine

Vitis vinifera subsp. silvestris, the wild subspecies of Vitis vinifera L., is a unique and valuable genetic resource for cultivated grapevines. At present, this wild form is rare and is spread from the Southern Atlantic coast of Europe to the Western Himalayas. In the present study, six nuclear microsatellite DNAs were analysed in 301 wild grape samples, using factorial correspondence analysis and the Bayesian model. The aim was to detect the distribution of genetic variability and admixture proportions and thus to identify the main routes of recolonization after the quaternary glaciation, as well as signals of secondary contacts in Europe. The results obtained show that some Caucasus and Italian areas played key roles as refugial areas. Moreover, the data suggest the existence of two main migration routes through central Europe during the post‐glacial or the inter‐glacial periods. The first started from Southern Italy and moved northward, whereas the second originated from refugia possibly located in eastern areas and migrated westwards. This scenario is most likely associated with the population expansion that followed the European Quaternary glaciations. In accordance with this assumption, central Europe is proposed as an admixture confluence of migration routes radiating from separate grapevine refugia. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 611–619.     

Short-term response of wild grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L<Emphasis Type="Italic">. ssp. sylvestris</Emphasis>) to NaCl salinity exposure: changes of some physiological and molecular characteristics

The physiological and molecular response to salt stress was studied in two wild grapevine (Vitis vinifera L. ssp. sylvestris or Vitis sylvestris) accessions “Khédhayria” and “Houamdia”, previously identified as salt-tolerant and salt-sensitive pair wise. Plants from both accessions were subjected to a progressive salt stress by the use of a nutritional solution containing up to 150 mM NaCl for 2 weeks. Salt stress adversely affected growth and water potential since the first day of exposure to 150 mM NaCl. However, chlorophyll fluorescence parameters were unchanged until 14 days of salt exposure. At that time point the predawn water potential (Ψ_PD), the non-photochemical quenching of fluorescence (NPQ) and the coefficient of photochemical quenching (q_p) were significantly less altered in the tolerant accession. At the molecular level semi-quantitative RT-PCR assays revealed a differential expression of (Vs α-gal/SIP and Vs DHN) genes within these contrasting accessions after exposure to 24 h and 14 days of salt. Comparably, the Vs RD22 gene had increased slightly after only 14 days of treatment in both accessions. These results were the first pieces of information reported on the early and late regulation of salt response genes in wild grapevines. Furthermore, genotype-dependent parameters such as NPQ, q_p, mRNA levels of Vs α-gal/SIP and Vs DHN could be used to screen salt-tolerant wild grapevine genotypes.     

Vitis vinifera L.: Wild or cultivated? Study of the grape pips found at Petra, Jordan; 150 B.C. – A.D. 40

The identification of carbonised grape pips (Vitis vinifera ssp.) is problematic, and the morphological features generally used to distinguish the wild subspeciesV. vinifera ssp.sylvestris from the cultivated subspeciesV. vinifera ssp.vinifera are not satisfactory. Different biometric studies were carried out on Nabataean and Roman seeds found at Petra, Jordan, dated to 150 B.C. – A.D. 400, and the results were compared to known phytogeographical and climatic data. Depending on the identification method selected, the seeds were attributed either to wild grapevines (based on the ratio of breadth over length, and on discriminant analyses of size variables such as pip length, stalk length, and chalaza position), or to an archaic variety of vine with seeds morphologically close to those of wild grapevines (ratio of stalk length over total pip length). The methods used here were applied to European grape pips; they should be tested on Near Eastern material. Archaeological data did not clarify the situation, and the importance of viticulture, which was prohibited during the Nabataean period, remains difficult to evaluate at Petra.     

Genetic diversity for drought resistance in wild emmer wheat and its ecogeographical associations

Wild emmer wheat (Triticum turgidum spp. dicoccoides (Körn.) Thell.), the tetraploid progenitor of cultivated wheat, is a potential source for various agronomical traits, including drought resistance. The objectives of this study were to characterize (1) the genetic diversity for drought resistance in wild emmer wheat, and (2) the relationship between drought responses of the wild emmer germplasm and the ecogeographical parameters of its collection sites. A total of 110 wild emmer accessions consisting of 25 populations and three control durum wheat cultivars were examined under two irrigation regimes, well-watered (’wet’) and water-limited (’dry’). Wide genetic diversity was found both between and within the wild emmer populations in most variables under each treatment. A considerable number of the wild emmer accessions exhibited an advantage in productivity (spike and total dry matter) over their cultivated counterparts. Most wild emmer wheat accessions exhibited a greater carbon isotope ratio (δ¹³C, indicating higher water-use efficiency) under the dry treatment and higher plasticity of δ¹³C relative to the cultivated controls, which may have contributed to the drought adaptations in the former. The most outstanding drought-tolerance capacity (in term of productivity under the dry treatment and susceptibility indices) was detected in wild emmer populations originated from hot dry locations. The results suggest that wild emmer has the potential to improve drought resistance in cultivated wheat.     

From the cradle of grapevine domestication: molecular overview and description of Georgian grapevine (Vitis vinifera L.) germplasm

Historical information and archaeological and palaeobotanical findings point Georgia, in the South Caucasus, as a cradle for grapevine (Vitis vinifera L.) domestication from its wild form (V. vinifera silvestris Beck.) and subsequent selection and development of varieties with characters suitable for human consumption. The hypothesis of Georgia being a center of domestication, combined with its distance from western countries and the importance of its viticulture and wine production, make Georgian grape germplasm particularly interesting to be investigated under the genetic point of view. Twenty nuclear microsatellite loci were used to genotype 112 Georgian grapevine accessions (V. vinifera sativa Beck.) from germplasm collections and 18 from spontaneous growing plants (V. vinifera silvestris Beck.) found in wild conditions and to compare them to a large international cultivar collection in France. Data analysis shows that Georgian grapevine germplasm has maintained distinctive traits despite arrival of international, foreign varieties and still conserve characteristics of local breeding linked to traditional wine production regions of the country. Results have identified alleles, overall loci, well represented in the Georgian germplasm (cultivated and wild) and absent or poorly represented in other countries, highlighting uniqueness and originality of traits of this viticulture. Moreover, the search for relationships between Georgian and foreign viticulture has evidenced few interesting cases linking the Georgian varieties with Western European ones and with neighboring Caucasian countries, helping to identify the real place of origin in some doubtful cases. In addition, populations or sparse individuals of wild grapevine still preserved in the Georgian natural environments present smaller genetic distances with local cultivars than in other European regions. Principal component analysis (PCA) has also identified special overlapping of the wild compartment with some cultivated varieties. This work provides a highly significant new contribution to applied aspects of Georgian grapevine genetic resources management and use. Uniqueness of the Georgian cultivated grapevine gene pool together with its close relatedness with the wild compartment makes this country a good candidate to address questions regarding domestication and grapevine genetic resource conservation.     

CHLOROPLAST DNA POLYMORPHISM SUGGESTS NIGERIAN CENTER OF DOMESTICATION FOR THE COWPEA,VIGNA UNGUICULATA (LEGUMINOSAE)

Twenty-one independent chloroplast DNA polymorphisms were identified in Vigna unguiculata defining 19 different chloroplast DNA molecules (plastome types). Two plastome types, differing by a single character, were found among 32 accessions of cultivated cowpea (Vigna unguiculata ssp. unguiculata). Eighteen different plastome types were found among 26 accessions of wild cowpea (V. unguiculata ssp. dekindtiana). The very low level of chloroplast DNA diversity found in cultivated accessions relative to wild cowpea suggests that 1) the domesticated form was derived from a narrow selection of the wild germplasm and 2) chloroplast gene flow between wild and cultivated types has been very limited. Cladistic analysis of the cpDNA data generated a robust tree completely lacking homoplasy. Three wild accessions from Nigeria possessed a plastome type indistinguishable from one present in cultivated accessions, suggesting that Nigeria represents one center of domestication of the cowpea. The other plastome type within the cultivated germplasm was not found among wild accessions.     

Genetic relationships in Lens species and parentage determination of their interspecific hybrids using RAPD markers

Broadening of the genetic base and systematic exploitation of heterosis in cultivated lentils requires reliable information on genetic diversity in the germplasm. The ability of random amplified polymorphic DNA (RAPD) to distinguish among different taxa of Lens was evaluated for several geographically dispersed accessions/cultivars of four diploid Lens species. This study was carried out to assess whether RAPD data can provide additional evidence about the origin of the cultivated lentil and to measure genetic variability in lentil germplasm. Three cultivars of Lens culinaris ssp. culinaris, including one microsperma, and two macrosperma types, and four wild species (L. culinaris ssp. orientalis, L. odemensis and L. nigricans) were evaluated for genetic variability using a set of 1 11-mer and 14 random 10-mer primers. One hundred and fifty-eight reproducible and scorable DNA bands were observed from these primers. Genetic distances between each of the accessions were calculated from simple matching coefficients. Split decomposition analysis of the RAPD data allowed construction of an unrooted tree. This study revealed that (1) the level of intraspecific genetic variation in cultivated lentils is narrower than that in some wild species. (2) L. culinaris ssp. orientalis is the most likely candidate as a progenitor of the cultivated species, (3) L. nigricans accession W6 3222 (unknown) and L. c. ssp. orientalis W6 3244 (Turkey) can be reclassified as species of L. odemensis and (4) transmission of genetic material in Lens interspecific hybrids is genotypically specific, as identified by the RAPD markers in our study.     

Genetic diversity for grain nutrients in wild emmer wheat: potential for wheat improvement

Background and Aims

Micronutrient malnutrition, particularly zinc and iron deficiency, afflicts over three billion people worldwide due to low dietary intake. In the current study, wild emmer wheat (Triticum turgidum ssp. dicoccoides), the progenitor of domesticated wheat, was tested for (1) genetic diversity in grain nutrient concentrations, (2) associations among grain nutrients and their relationships with plant productivity, and (3) the association of grain nutrients with the eco-geographical origin of wild emmer accessions.

Methods

A total of 154 genotypes, including wild emmer accessions from across the Near Eastern Fertile Crescent and diverse wheat cultivars, were characterized in this 2-year field study for grain protein, micronutrient (zinc, iron, copper and manganese) and macronutrient (calcium, magnesium, potassium, phosphorus and sulphur) concentrations.

Key Results

Wide genetic diversity was found among the wild emmer accessions for all grain nutrients. The concentrations of grain zinc, iron and protein in wild accessions were about two-fold greater than in the domesticated genotypes. Concentrations of these compounds were positively correlated with one another, with no clear association with plant productivity, suggesting that all three nutrients can be improved concurrently with no yield penalty. A subset of 12 populations revealed significant genetic variation between and within populations for all minerals. Association between soil characteristics at the site of collection and grain nutrient concentrations showed negative associations between soil clay content and grain protein and between soil-extractable zinc and grain zinc, the latter suggesting that the greatest potential for grain nutrient minerals lies in populations from micronutrient-deficient soils.

Conclusions

Wild emmer wheat germplasm offers unique opportunities to exploit favourable alleles for grain nutrient properties that were excluded from the domesticated wheat gene pool.     

Application of AFLPs to the characterization of grapevine Vitis vinifera L. genetic resources. A case study with accessions from Rioja (Spain)

 AFLPs were used to characterize 67 different grapevine accessions from a collection of D.O.Ca. Rioja in Spain. A correct selection of primers and selective nucleotides allowed us to maximize the number of amplified fragments analyzed per reaction yielding an average of 100 per reaction, 49% of which were polymorphic. Based on the presence or absence of amplified fragments for each genotype resulting from a reaction with two primer combinations, we have established the genetic similarity between the different accessions in the collection. These results allowed us to resolve different genotypes maintained under the same name (homonyms) and to identify the same genotype under different names (synonyms) thus permitting the elimination of redundant germplasm. Furthermore, by providing information on more than 50 polymorphic loci per reaction, a few reactions were sufficient to identify distinct AFLP patterns characteristic of specific clones, with different agronomic and organoleptic features, belonging to the same cultivar. The possibility for clonal identification, shown here for grapevines, can have important implications in the protection and management of clonal selections. Received: 1 February 1998/Accepted: 25 February 1998     

Effects of competition with Bromus tectorum on early establishment of Poa secunda accessions: can seed source impact restoration success?

When landscapes are heavily impacted by biological invasion, local populations of native plant species may no longer be adapted to altered environmental conditions. In these cases, it is useful to investigate alternative sources of germplasm, such as cultivated varieties, for planting at restoration sites. This study compared cultivated and wild (local) varieties of the native perennial bunchgrass, Poa secunda J. Presl, grown with and without the exotic, invasive Bromus tectorum L. in a greenhouse setting. While P. secunda cultivars emerged and grew more rapidly than wild seed sources, this advantage declined in the presence of B. tectorum and cultivated germplasm did not outperform wild accessions in the presence of an invasive species. Given the novel genetic background of cultivars and their potential to alter patterns of dominance in native plant communities, we recommend the use of local or regional wild seed sources when possible to conserve regional patterns of genetic diversity and adaptation. Use of multiple seed sources may increase the potential for capturing vigorous genotypes in the restoration seed mix. In cases where sites are heavily impacted by exotic, invasive species, other control measures will be necessary to improve establishment of native species in grassland restoration programs.     

Genetic diversity of taro, Colocasia esculenta (L.) Schott, in Southeast Asia and the Pacific

The genetic diversity of 255 taro (Colocasia esculenta) accessions from Vietnam, Thailand, Malaysia, Indonesia, the Philippines, Papua New Guinea and Vanuatu was studied using AFLPs. Three AFLP primer combinations generated a total of 465 scorable amplification products. The 255 accessions were grouped according to their country of origin, to their ploidy level (diploid or triploid) and to their habitat—cultivated or wild. Gene diversity within these groups and the genetic distance between these groups were computed. Dendrograms were constructed using UPGMA cluster analysis. In each country, the gene diversity within the groups of wild genotypes was the highest compared to the diploid and triploid cultivars groups. The highest gene diversity was observed for the wild group from Thailand (0.19), the lowest for the diploid cultivars group from Thailand (0.007). In Malaysia there was hardly any difference between the gene diversity of the cultivars and wild groups, 0.07 and 0.08, respectively. The genetic distances between the diploid cultivars groups ranges from 0.02 to 0.10, with the distance between the diploid accessions from Thailand and Malaysia being the highest. The genetic distances between the wild groups range from 0.05 to 0.07. First, a dendrogram was constructed with only the diploids cultivars from all countries. The accessions formed clusters largely according to the country from which they originated. Two major groups of clusters were revealed, one group assembling accessions from Asian countries and the other assembling accessions from the Pacific. Surprisingly, the group of diploid cultivars from Thailand clustered among the Pacific countries. Secondly, a dendrogram was constructed with diploid cultivated, triploid cultivated and wild accessions. Again the division of the accessions into an Asian and a Pacific gene pool is obvious. The presence of two gene pools for cultivated diploid taro has major implications for the breeding and conservation of germplasm.     

High throughput analysis of grape genetic diversity as a tool for germplasm collection management

Using 20 SSR markers well scattered across the 19 grape chromosomes, we analyzed 4,370 accessions of the INRA grape repository at Vassal, mostly cultivars of Vitis vinifera subsp. sativa (3,727), but also accessions of V. vinifera subsp. sylvestris (80), interspecific hybrids (364), and rootstocks (199). The analysis revealed 2,836 SSR single profiles: 2,323 sativa cultivars, 72 wild individuals (sylvestris), 306 interspecific hybrids, and 135 rootstocks, corresponding to 2,739 different cultivars in all. A total of 524 alleles were detected, with a mean of 26.20 alleles per locus. For the 2,323 cultivars of V. vinifera, 338 alleles were detected with a mean of 16.9 alleles per locus. The mean genetic diversity (GDI) was 0.797 and the level of heterozygosity was 0.76, with broad variation from 0.20 to 1. Interspecific hybrids and rootstocks were more heterozygous and more diverse (GDI?=?0.839 and 0.865, respectively) than V. vinifera cultivars (GDI?=?0.769), Vitis vinifera subsp. sylvestris being the least divergent with GDI?=?0.708. Principal coordinates analysis distinguished the four groups. Slight clonal polymorphism was detected. The limit between clonal variation and cultivar polymorphism was set at four allelic differences out of 40. SSR markers were useful as a complementary tool to traditional ampelography for cultivar identification. Finally, a set of nine SSR markers was defined that was sufficient to distinguish 99.8% of the analyzed accessions. This set is suitable for routine characterization and will be valuable for germplasm management.     

Phylogeographical structure and conservation genetics of wild grapevine

The distribution of Vitis vinifera subsp. silvestris, the wild grapevine subspecies of Vitis vinifera L., has been dramatically reduced in its major sites of diffusion, at first by the spread, over the last 150 years, of pathogens from North America and, more recently, with fragmentation of habitat and disbranching by humans. In this work, 418 wild grapevine samples, belonging to 78 populations, were collected in their main Mediterranean distribution areas, including the Caucasus area, and the extent of their genetic variability evaluated by analysing plastid microsatellite DNA polymorphism. Results show low haplotype diversity value, with five haplotypes detected within the analysed populations. The highest within-population haplotypic diversity, with the presence of all five detected haplotypes, was found in the Caucasus regions and in the central regions of Italy. The distribution of all detected haplotypes suggests the Caucasian region as the possible center of origin of Vitis vinifera subsp. silvestris. A principal plastid lineage was found to be fixed in several locations, in the Northernmost European countries and in the Southern island of Sardinia. These results draw attention to two different refugium sites in the Mediterranean basin and suggest that conservation priority should be given to grapevine populations still preserved in hotspots of these regions.