The origin of the grapevine was investigated with archaeobotanical, cultural and historical data. A primary domestication centre was located in the Near East region but there is no agreement on the existence or role of secondary domestication centres. In this work, PCR-based microsatellite analysis has been applied to study the origin of some Italian cultivated grapevines from in situ direct domestication of the wild autoctonous grapevine. Three different Italian locations in Grosseto, Cosenza and Nuoro were identified for this study, and domesticated grapevine as well as wild local accessions growing in these location, were analysed by SSR markers. Cluster analysis performed on Cosenza and Grosseto samples showed a high value of genetic distance between domesticated and wild accessions. On the contrary two cultivars (Bovale Murru and Bovale Muristellu) recovered in Nuoro (in the Sardinia island) were very close to some wild varieties. This suggests that the latter two cultivars may have originated from wild grapevines and consequently that in this location a secondary grapevine domestication event occurred. Six Lambrusco varieties were also included in this analysis as ancient putative ancestors of the cultivated grapevines. The molecular analysis excluded this hypothesis and suggest Lambrusco as an independent Vitis taxon.

In viticulture, biotype identification problems have traditionally been solved using ampelography, ampelometry and chemical traits analysis. However, these tools have resulted in several false attributions, in particular when used at the clonal level. The availability of relatively cheap, reliable and reproducible tools to identify genetic differences at the clonal level would greatly facilitate the work of clonal patenting. In this work, 24 accessions of ‘Traminer’ cultivars were characterized using molecular markers. Three different approaches were applied: simple sequence repeats (SSR), amplified fragment length polymorphism (AFLP) and methyl‐sensitive amplified length polymorphism (MSAP). Results showed that SSRs were not a powerful tool for clonal distinction. In contrast, the AFLP technique was able to distinguish 16 out of the 24 cultivars, even though the average similarity was high (97.1%). The MSAP technique was used to evaluate qualitative differences in the degree of DNA methylation among clones. Results suggest that morphological differences among clones are probably due to the synergetic effect of genetic and epigenetic modifications, and that clonal identification could be greatly improved using molecular tools such as AFLP and MSAP.

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

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The origin of the grapevine, Vitis vinifera L. (ssp.) sativa has been investigated with archaeobotanicalarchaeological, cultural and historical data indicating a unique domestication centre located in the Caucasian and Middel-East regions about 6-7000 years ago, but, events leading to the domestication of this species are still an open issue. In this work, eight universal chloroplast microsatellites are used to assess genetic relationships among varieties selected as representatives of four distinct geographical groups from Middle-East to Western European regions. Results show that two out of the eight analysed chloroplast loci are polymorphic within the 142 individuals. Allele variants of the cpSSR loci combine in a total of six different haplotypes. The analysis of haplotypes distribution and haplotype diversity (HD) suggest that only three out of the six haplotypes are represented in the Caucasian and Middle-East samples, with 90% of individuals sharing the same haplotype. Moreover, the presence of all six haplotypes in the European accessions, with a high level of haplotype diversity, suggests varietal influx in these areas. Concerning the Western European varieties, especially in Spanish accessions, half of the individuals share haplotype VI which is completely absent in the Caucasian and Middle-East cultivars. This result opens the discussion about the existence of a unique and common domestication centre, located in the Caucasian and MiddleEast area, for all the European cultivars. This work suggests the usefulness of chloroplast genome markers to provide information on haplotype distributions that could help to identify further geographical areas for grapevine varietal evolution.Abbreviations: cpSSRs -chloroplast simple sequence repeats

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.

The vintage effect overcomes the terroir effect: a three year survey on the wine yeast biodiversity in Franciacorta and Oltrepò Pavese, two northern Italian vine-growing areas analyses, the year of isolation (vintage) proved to be a factor that significantly affected the biodiversity of the yeast species, whereas the geographical site (terroir) was not. Seventy-five per cent of S. cerevisiae isolates gathered in a unique cluster at a similarity level of 82 %, while the remaining 25 % were separated into minor groups without any evident relationship between d-PCR profile and territory, year or source of isolation. However, in six cases a similar strain appeared at the harvesting time both in Franciacorta and Oltrepò Pavese areas, whereas surprisingly no strain was reisolated in the same vineyard or cellar for consecutive years. INTRODUCTIONIn winemaking, yeasts are essential for the transformation of grape sugars into ethanol and carbon dioxide through alcoholic fermentation; nonetheless, due to their specific enzymic activities and cell autolysis, they can also generate typical sensorial characteristics in wine, like secondary flavours and smoothness (Romano et al., 2003a). Although selected Saccharomyces strains are usually added by oenologists as starter cultures to control the fermentative process, several micro-organisms enter the must from the vineyard environment, winery facilities and cellar equipment, and these can affect the quality of the end product. Nowadays, for a certain style of wines, the use of the 'so called' autochthonous yeasts is considered essential in providing for the valorization and preservation of the environmental microbial biodiversity (Pretorius, 2000). In 3These authors contributed equally to this paper.Abbreviations: ADY, active dry yeast; CE, capillary electrophoresis; ITS, internal transcribed spacer; LSD, least significant difference; PCA, principal component analysis; UPGMA, unweighted pair group method with arithmetic means.

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.