Search Results (31)

Dikili Tash is a Neolithic settlement that lies next to the ruins of the ancient city of Philippi on the north-eastern part of Greece. A recent archaeological excavation has unearthed charred grapevine pips and pressings together with two-handed clay cups, jugs, and jars that date to 4300 BC. The majority of the pips were found to be Vitis vinifera ssp. sylvestris. Natural populations of this species have been localized in the valley surrounding Dikili Tash and also on Mt Pangaion and Mt Lekani, which flank the valley. Fifty-one samples from these modern populations have been analyzed using microsatellites on twenty microsatellite loci, and a dendrogram has been constructed showing the genetic closeness of the samples analyzed. Cuttings from all the vines analyzed are currently rooted and grown in the Hellenic Agricultural Organization—DIMITRA (ELGO-DIMITRA) greenhouse facilities in Lykovryssi (Athens) with the aim to, eventually, be transplanted in the grapevine, thus establishing the first V. sylvestris ex situ conservation site in Greece. Full article

Background/Objectives: The genus Vitis comprises approximately 70 species with high genetic diversity, among which Vitis vinifera is the most economically significant. Despite numerous studies on the genetic characterizations of V. vinifera, selecting optimal chloroplast DNA barcoding regions for intraspecific differentiation remains unresolved. Most studies have focused on nuclear markers (SSRs, SNPs) or widely used chloroplast loci (e.g., matk, rbcl), which have shown limited resolution at the subspecies level. In this study, the complete chloroplast genomes of 34 V. vinifera accessions from different varieties and hybrids (vinifera, sylvestris, caucasica, and labrusca) were analyzed to identify the key genomic regions for DNA barcoding. Methods: Using bioinformatics tools, we assessed the genome structure, nucleotide variability, microsatellites, codon usage bias, and phylogenetic relationships among the investigated varieties. Results: The chloroplast genomes displayed a quadripartite structure, with lengths ranging from 160,906 to 160,929 bp and a guanine–cytosine (GC) content of ~37.4%. Phylogenetic analysis revealed an unusual position for VV-5 vini and VVVL-3 lab, suggesting potential taxonomic misclassification or hybridization effects. A single locus showed low discrimination power, but the concatenation of five loci (ccsA-trnN-GUU, rpl16, rpl2-rps19, rpoC2, and trnM-CAU) exhibited significantly improved resolution (44.11% K2P), surpassing traditional markers. Conclusions: This study addresses the gap in the literature regarding the use of concatenated chloroplast loci for subspecies research; the results validate these markers across a broader range of Vitis accessions and integrate nuclear and mitochondrial data to achieve a more comprehensive understanding of the evolutionary history and genetic diversity of V. vinifera. Full article

Seed morphological description requires quantitative methods for further comparison. Here, traditional measurements, curvature analysis, and the J-index (percentage of similarity to a geometric model) were applied to the average contours (Acs) of 271 Vitis cultivars from the Spanish collection at IMIDRA (Madrid, Spain), including 9 different Vitis species and several sylvestris seeds (i.e., those derived from plants that once grew in the wild). Acs are graphical representations of the shape in seed populations, which can be obtained either from image analysis programs or computationally opening the way to quantitative analysis. A geometric model is a geometrically defined, closed curve, used as a reference for shape quantification. Based on existing differences between the Hebén cultivar (collected in 2020 and 2024; Hebén model, for morphotype 1) and the European varieties Chenin and Gewurztraminer (Chenin model, for morphotype 2), we created two models. The comparisons were based on a J-index, resulting in four groups: Group 1 contained all seeds with values lower than 90 for both models and included all Vitis species other than V. vinifera and most sylvestris seeds; Groups 2 and 3 contained seeds with J-index values higher than 94 for the Hebén and Chenin models, respectively. Group 4 consisted of seeds not included in the other groups. Based on J-index values, differences in curvature and solidity, and PCA analysis with Fourier coefficients, this work defines two new morphotypes associated with the Hebén (Group 2) and Chenin (Group 3) models, related to Iberian and Western European varieties, respectively. Full article

This study aimed to investigate the endophytic microbial populations associated with wild and domesticated grapevines using both culture-based and culture-independent methods. Through culture-based methods, 148 endophytes were identified. The dominant fungal species included Aureobasidium pullulans, Alternaria alternata, and Cladosporium allicinum, while predominant bacterial species were Ralstonia pikettii, Nocardia niigatensis, and Sphingomonas echinoides. Culture-independent methods employed metagenomic techniques to explore microbial biodiversity, focusing on targeted amplification of bacterial 16S rRNA as well as fungal ITS and 26S rRNA gene regions. The main bacterial species identified included Halomonas sp., Sphingomonas sp. and Massilia sp., whereas the fungal population was dominated by Cladosporium sp., Malassezia sp. and Mucor sp. The findings revealed that vineyard management practices did not lead to statistically significant variations in microbial communities. The consistent presence of these genera across all samples suggests that they are stable components of the grapevine endophytic microbiota, remaining relatively unaffected by external environmental factors. Full article

Repeated expeditions across various regions of Georgia in the early 2000s led to the identification of 434 wild grapevine individuals (Vitis vinifera L. subsp. sylvestris (C.C. Gmel.) Hegi) across 127 different sites, with 45% of these sites containing only a single vine and only 7% more than 9 vines. A total of 70 accessions were propagated in a germplasm collection, 41 of them were descripted from the ampelographic point of view and 32 from the phenological one. The geographical and ecological analysis confirmed that wild grapevines primarily grow in humid environments with warm and fully humid climates, often near rivers. They favor deep, fertile, and evolved soils, mainly alluvial and cinnamonic types (80%), with a marginal presence on strongly eroded soils. Their main natural vegetations are forests and open woodlands, with some individuals in the Southeast found in steppes. The altitudinal range spans from 0 to 1200 m, with 80% of vines distributed between 400 and 900 m. The phenological analysis revealed significant differences among the accessions but no difference among populations, with only a slight variation in bud-break timing, indicating a high level of synchronicity overall. Flowering timing proved to be the most uniform stage, suggesting minimal environmental pressure on genetic adaptation. The mature leaf morphology exhibited significant polymorphism, though leaves were generally three- or five-lobed, weak-wrinkling, and -blistering, with a low density of hairs. Bunch and berry morphology were more uniform. Bunches were consistently very small, cylindrical, and never dense or winged. Berries were also very small, mostly globular, always blue-black in color, and non-aromatic. A striking feature was the frequency of red flesh coloration, which ranged from weak to strong, with uncolored flesh being rare. The Georgian population of wild grapevines was found to be fragmented, often consisting of scattered single individuals or small groups. Therefore, we believe it is urgent for Georgia to implement specific protection measures to preserve this vital genetic resource. Full article

By exploring seed samples from vineyards and natural habitats across the Eurasian range of Vitis vinifera, our analysis revealed substantial morphological variation within populations. Through the analysis of domestication index values, probabilities, and entropy, we assessed seed diversity. Samples with high domestication probability values—predominantly from vineyards—exhibited low heterogeneity and entropy, with similar patterns observed in natural habitats, suggesting the presence of feral vines. In parallel, seeds with low domestication index values, found mainly in natural habitats, also displayed low entropy and are likely associated with Vitis sylvestris or other wild Vitaceae species. Intermediate domestication values pointed to hybrid swarms, highlighting the crucial role of hybridization in the development of modern grapevine cultivars. The study identified mixed populations across the Iberian Peninsula, Italy, and the South Caucasus, which act as significant gene reservoirs. A domestication gradient is evident, with higher domestication rates in the South Caucasus compared to Western Europe and East Asia. The results demonstrate the significance of these mixed populations as repositories of genetic diversity, underscoring their conservation value, particularly considering the negative impact of habitat alterations, especially in riparian forests due to major public works. Full article

The flowering and fruit set of grapevines are determined by many morphological, physiological, and environmental factors. Although the elemental composition of pollen grains plays a crucial role in the fruit set, there is still a considerable gap in our knowledge. To date, no study has been conducted on the elemental composition of Vitis vinifera subsp. sylvestris (hereafter V. sylvestris) pollen grains. The aim of this work was to investigate the elemental composition of pollen grains of V. sylvestris using scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). The pollen grains of ten V. sylvestris individuals (eight male and two female) and one hermaphrodite cultivar ‘Plavac mali crni’ were analyzed. SEM-EDX analysis revealed the presence of eight elements (carbon—C, oxygen—O, magnesium—Mg, phosphorus—P, potassium—K, calcium—Ca, molybdenum—Mo, and aluminum—Al) in the pollen grains. Interestingly, aluminum was detected exclusively in the pollen of the cultivated grape cultivar ‘Plavac mali crni’, while it was not present in the genotypes of V. sylvestris. No significant differences between genotypes were found for oxygen and phosphorus, while significant differences were found for other elements. Pollen dimorphism was not associated with differences in element composition, although principal component analysis separated the genotypes into two distinct groups, with two female individuals (Pak10 and Pak12) and one male (Im19) tending to form separate clusters. This study is the first report on the elemental composition of pollen grains of V. sylvestris genotypes and provides valuable insights for further studies on pollen functionality. Full article

The earliest archaeological evidence of wine came from ceramic vessels of the Transcaucasian ‘Shulaveri-Shomutepe’ or ‘Aratashen-Shulaveri-Shomutepe culture’ (SSC/AShSh: c. 6000–5200 BC). Western European ‘Bell Beaker culture’ (BB: c. 2500–2000 BC) is characterized by bell-shaped pottery vessels but has so far not been found with residues consistent with wine. Knowing that wild grapes populated both habitats, the absence of wine during the Bell Beaker period remains to be explained. The main goal of this work was to investigate whether the shape of the vessels could influence the performance of spontaneous fermentation, specifically regarding the production of volatile acidity. Crushed grapes or juices from various grape cultivars were fermented in two types of vessels: (i) borosilicate glass beakers (4–5 L) to imitate bell beakers and (ii) Erlenmeyer flasks (5 L) to imitate SSC/AShSh vessels. Fermentations occurred spontaneously, and the wines were analyzed for their conventional physical–chemical parameters (e.g., ethanol content, total acidity, volatile acidity, pH), chromatic characteristics (e.g., wine color intensity, wine hue), and volatile composition by gas-chromatography-flame ionization detection (GC-FID). At the end of fermentation, the yeast species were identified by molecular methods. In addition, wine yields and phenolic composition (e.g., total phenols, anthocyanins, total pigments) were determined for wild grapes in comparison with six red varieties Vitis vinifera L. subsp. sativa (Vinhão, Marufo, Branjo, Melhorio, Castelão and Tempranillo Tinto), chosen as a function of their genetic relatedness with the wild counterpart. Wines produced from V. sylvestris grapes showed higher total acidity and color intensity when compared to the cultivated varieties. Saccharomyces cerevisiae dominated at the end of all spontaneous fermentations in all types of vessels and conditions. Wines fermented in Erlenmeyers showed ethanol concentrations as high as 14.30% (v/v), while the highest ethanol level was 12.30% (v/v) in beakers. Volatile acidity increased to a maximum of 4.33 g/L (acetic acid) in Erlenmeyers and 8.89 g/L in beakers. Therefore, the shape of the vessels influenced the performance of fermentation, probably due to the different exposures to air, leading to vinegary ferments more frequently in open mouths than in conical-shaped flasks. These results provide a hypothesis based on fermentation performance for the absence of wine produced in the Iberian Peninsula until the arrival of Phoenician settlers. Full article

Studying the evolution of seed morphology and, in turn, the evolution of cultivars across time and space is of fundamental importance to agriculture and archaeology. The identification of ancient and modern grapevine (Vitis vinifera L.) cultivars is essential for understanding the historical evolution of grape cultivation. Grape seed morphology provides valuable information to explore the evolution of grape cultivars over time and space. The main aim of our study was to build a comprehensive regional database of grape seed morphological traits from modern and archaeological wine cultivars and wild grape species. We aimed to identify which seeds of modern grape cultivars exhibited morphological similarities to archaeological cultivars. This study focused on fifteen distinct modern types of seeds and two archaeological samples from the Byzantine-to-Early Medieval period. We acquired digital images of seeds using a flatbed scanner. For each sample, 100 seeds were randomly selected, and morphometric data on each seed were gathered using ImageJ. Differences among the seed cultivars were investigated using linear discriminant analysis. Archaeological seeds were found to be more similar to cultivated V. vinifera cultivars rather than V. sylvestris populations. Among the cultivated cultivars, Sangiovese and Tosta antica resulted to be cultivars most similar cultivars to the archaeological ones. The morphometric analysis of grape seeds proved to be a valuable resource for investigating the evolution of vine cultivars throughout history. Combining image analysis techniques with genetic data will open new perspectives for studying the origins of and variations in grape cultivars, contributing to the conservation and enhancement of viticultural heritage. Full article

Understanding the origins and evolution of modern grapevine varieties in the Iberian Peninsula and western Europe necessitates an examination of the proportions of Vitis vinifera cultivars, their relationships with wild grapevine populations, and the utilization of seedless cultivars in al-Andalus. Employing morphometric studies, domestication indices, multivariate analysis, and Bayesian hypothesis testing, this study investigates several distinct seed types identified in materials from Roman and medieval deposits. These seeds exhibit a spectrum from highly domesticated to purely wild. Our findings reveal the predominance of Proles Occidentalis Negrul, and the presence of feral-like grapevines associated with Proles Euphratica. Additionally, we observe the continuous presence of wild grapevines related to Vitis sylvestris CC Gmelin throughout the studied period. Seeds exhibiting intermediate characteristics are documented, alongside the identification of “stenosperms”, suggesting anomalies in seed formation. Notably, the presence of Vitis vinifera raisins “stenospermocarpics” of the sultana type is suggested, potentially elucidating the absence of table grapes and raisins of the Proles Orientalis Negrul in the archaeological record, despite frequent mentions by medieval agronomy writers from al-Andalus. Full article

The discovery of well-preserved fossil Vitis L. seeds from the Gelasian stage in Italy has provided a unique opportunity to investigate the systematics of fossilized Vitis species. Through seed image analyses and elliptical Fourier transforms of fossil Vitis seeds from the sites Buronzo−Gifflenga and Castelletto Cervo II, we pointed out a strong relationship to the group of extant Eurasian Vitis species. However, classification analyses highlighted challenges in accurately assigning the fossil grape seeds to specific modern species. Morphological comparisons with modern Vitis species revealed striking similarities between the fossil seeds and V. vinifera subsp. sylvestris, as well as several other wild species from Asia. This close morphological resemblance suggests the existence of a population of V. vinifera sensu lato in Northen Italy during the Gelasian. These findings contributed to our understanding of the evolution and the complex interplay between ancient and modern Vitis species. Full article

Vitis vinifera L. subsp. sylvestris (sylvestris) is the only native wild grapevine in Eurasia (Europe and western Asia) and is the existing ancestor of the grapevine varieties (for wine and table grape production) belonging to the subsp. sativa. In Slovenia, the prevailing opinion has been that there are no Slovenian sylvestris habitats. This study describes sylvestris in Slovenia for the first time and aims to present an overview of the locations of the wild grapevine in the country. In this project, a sample set of 89 accessions were examined using 24 SSR and 2 SSR markers plus APT3 markers to determine flower sex. The accessions were found in forests on the left bank of the Sava River in Slovenia, on the border between alluvial soils and limestone and dolomite soils, five different sites, some of which are described for the first time. The proportion of female to male accessions differed between sites. At two sites, female plants dominated; at others, the ratio was balanced. The plants’ genetic diversity and structure were compared with autochthonous and unique varieties of subsp. sativa from old vineyards in Slovenia and with rootstocks escaped from nature from abandoned vineyards. Sylvestris was clearly distinguishable from vinifera and the rootstocks. Based on genetic analyses, it was confirmed that Slovenian sylvestris is closest to the Balkan and German sylvestris groups. Meanwhile, a safety duplication of the wild grapevine accessions has been established at the University Centre of Viticulture and Enology Meranovo, Faculty of Agriculture and Life Sciences at the University of Maribor. Full article

This study aimed to investigate, for the first time, the chemical composition and antioxidant activity of fluid extracts obtained from three Romanian cultivars of haskap berries (Lonicera caerulea L.) var. Loni, bitter cherries (Prunus avium var. sylvestris Ser.) var. Silva, and pomace from red grapes (Vitis vinifera L.) var. Mamaia, and their capacity to modulate in vitro steatosis, in view of developing novel anti-obesity products. Total phenolic, flavonoid, anthocyanin, and ascorbic acid content of fluid extracts was spectrophotometrically assessed and their free radical scavenging capacity was evaluated using Trolox Equivalent Antioxidant Capacity (TEAC) and free 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition assays. The Pearson coefficients showed a moderate correlation between the antioxidant activity of fluid extracts and their phenolic content, but a strong correlation between anthocyanin and ascorbic acid content. HPLC analysis identified and quantified the main phenolic compounds of chlorogenic and syringic acid, catechin, and glycosylated kaempferol, apigenin, and quercetin, in variable proportions. An in vitro experimental model of steatosis was developed in HepG2 hepatocytes treated with a mixture of free fatty acids. Cell culture analyses showed that cytocompatible concentrations of fluid extracts could significantly reduce the lipid accumulation and inhibit the reactive oxygen species, malondialdehyde, and nitric oxide secretion in stressed hepatocytes. In conclusion, these results put an emphasis on the chemical compounds’ high antioxidant and liver protection capacity of unstudied fluid extracts obtained from Romanian cultivars of bitter cherries var. Silva and pomace of red grapes var. Mamaia, similar to the fluid extract of haskap berries var. Loni, in particular, the positive modulation of fat deposition next to oxidative stress and the lipid peroxidation process triggered by fatty acids in HepG2 hepatocytes. Consequently, this study indicated that these fluid extracts could be further exploited as hepatoprotective agents in liver steatosis, which provides a basis for the further development of novel extract mixtures with synergistic activity as anti-obesity products. Full article

The biochemical response and gene expression in different grapevine cultivars to water deficit are still not well understood. In this study, we investigated the performance of four traditional Croatian Vitis vinifera L. cultivars (‘Plavac mali crni’, ‘Istrian Malvasia’, ‘Graševina’, and ‘Tribidrag’), and one wild (Vitis vinifera subsp. sylvestris) genotype exposed to water deficit (WD) for nine days under semi-controlled conditions in the greenhouse. Sampling for biochemical and gene expression analyses was performed at days six and nine from the beginning of WD treatment. The WD affected the accumulation of metabolites with a significant increase in abscisic acid (ABA), salicylic acid (SA), and proline in the leaves of the stressed genotypes when the WD continued for nine days. Lipid peroxidation (MDA) was not significantly different from that of the control plants after six days of WD, whereas it was significantly lower (297.40 nmol/g dw) in the stressed plants after nine days. The cultivar ‘Istrian Malvasia’ responded rapidly to the WD and showed the highest and earliest increase in ABA levels (1.16 ng mg⁻¹ dw, i.e., 3.4-fold increase compared to control). ‘Graševina’ differed significantly from the other genotypes in SA content at both time points analyzed (six and nine days, 47.26 and 49.63 ng mg⁻¹ dw, respectively). Proline level increased significantly under WD (up to 5-fold at day nine), and proline variation was not genotype driven. The expression of aquaporin genes (TIP2;1 and PIP2;1) was down-regulated in all genotypes, coinciding with the accumulation of ABA. The gene NCED1 (9-cis-epoxycarotenoid dioxygenase) related to ABA was up-regulated in all genotypes under stress conditions and served as a reliable marker of drought stress. This work suggests that the stress response in metabolite synthesis and accumulation is complex, treatment- and genotype-dependent. Full article

The wild grapevine, Vitis vinifera subsp. sylvestris, grows naturally throughout the northern hemisphere, including the Mediterranean region. Wild grapevines have also been observed sporadically across the southern Levant and are considered a non-native feral plant. Nevertheless, no formal characterization has been conducted for wild grapevines in this region; thus, its taxonomical assignment remains elusive. Previously, we have shown that the wild grapevine populations growing in northern Israel are genetically separated from the feral domesticated forms. This work aimed to comprehensively describe the morphological, anatomical, and ecological traits of wild grapevines naturally thriving in two distinct habitats in Israel. The dioicous nature of the wild grapevine, the flower and pollen morphology, and the characteristic Sylvestris fruit and seed morphology, in addition to the occurrence of the natural germination of seeds in close vicinity of the mother plant, have all led to the conclusion that these plants belong to Vitis vinifera subsp. sylvestris and should be included in the Flora Palaestina. These findings, combined with the recently published genetic evidence for these populations, significantly advance our understanding of the species’ ecology and the importance of its preservation. Full article