Wild Grapevine Research Articles

Ecological and sanitary characteristics of the Eurasian wild grapevine (Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi) in Georgia (Caucasian region)

This paper shows the results of the investigation on some ecological aspects and on the sanitary status of the wild Euroasiatic native grapevine (Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi) in Georgia (South Caucasus). This taxon is seriously endangered by human activities such as forest cleaning and setting fires. Moreover, invasive Vitaceae of the North American origin, imported after phylloxera (Daktulosphaira vitifoliae Fitch) when vineyards were being replanted, increase the risk to lose these spontaneous vines. The survey includes collection of data on the population structure, the plant sex ratio, the main botanical supporters of the vines and the associated flora, the presence of invasive vines of the North American origin and the incidence of phytophagous arthropods and pathogens. The phytosanitary study showed that monophagous eriophyid mites and exotic fungal diseases, such as downy (Plasmopara viticola (Berkeley and Curtis) Berlease and de Toni) and powdery mildews (Erysiphe necator –(Schweinitz) Burrill), cause symptoms on all the observed populations. The absence of symptoms caused by phylloxera, root-knot nematodes and root rot is remarkable. However, the level of detected injuries caused by these parasitic organisms does not seem to be a real problem for the survival of the populations.

WILD GRAPEVINE (VITIS VINIFERA L. SUBSP. SYLVESTRIS (GMELIN) HEGI) IN ITALY: DISTRIBUTION AND PRELIMINARY GENETIC ANALYSIS

ISHS I International Symposium on Wild Relatives of Subtropical and Temperate Fruit and Nut Crops WILD GRAPEVINE (VITIS VINIFERA L. SUBSP. SYLVESTRIS (GMELIN) HEGI) IN ITALY: DISTRIBUTION AND PRELIMINARY GENETIC ANALYSIS

Identification of microRNAs from Amur grape (vitis amurensis Rupr.) by deep sequencing and analysis of microRNA variations with bioinformatics

BackgroundMicroRNA (miRNA) is a class of functional non-coding small RNA with 19-25 nucleotides in length while Amur grape (Vitis amurensis Rupr.) is an important wild fruit crop with the strongest cold resistance among the Vitis species, is used as an excellent breeding parent for grapevine, and has elicited growing interest in wine production. To date, there is a relatively large number of grapevine miRNAs (vv-miRNAs) from cultivated grapevine varieties such as Vitis vinifera L. and hybrids of V. vinifera and V. labrusca, but there is no report on miRNAs from Vitis amurensis Rupr, a wild grapevine species.ResultsA small RNA library from Amur grape was constructed and Solexa technology used to perform deep sequencing of the library followed by subsequent bioinformatics analysis to identify new miRNAs. In total, 126 conserved miRNAs belonging to 27 miRNA families were identified, and 34 known but non-conserved miRNAs were also found. Significantly, 72 new potential Amur grape-specific miRNAs were discovered. The sequences of these new potential va-miRNAs were further validated through miR-RACE, and accumulation of 18 new va-miRNAs in seven tissues of grapevines confirmed by real time RT-PCR (qRT-PCR) analysis. The expression levels of va-miRNAs in flowers and berries were found to be basically consistent in identity to those from deep sequenced sRNAs libraries of combined corresponding tissues. We also describe the conservation and variation of va-miRNAs using miR-SNPs and miR-LDs during plant evolution based on comparison of orthologous sequences, and further reveal that the number and sites of miR-SNP in diverse miRNA families exhibit distinct divergence. Finally, 346 target genes for the new miRNAs were predicted and they include a number of Amur grape stress tolerance genes and many genes regulating anthocyanin synthesis and sugar metabolism.ConclusionsDeep sequencing of short RNAs from Amur grape flowers and berries identified 72 new potential miRNAs and 34 known but non-conserved miRNAs, indicating that specific miRNAs exist in Amur grape. These results show that a number of regulatory miRNAs exist in Amur grape and play an important role in Amur grape growth, development, and response to abiotic or biotic stress.

Ectopic expression of VpALDH2B4, a novel aldehyde dehydrogenase gene from Chinese wild grapevine (Vitis pseudoreticulata), enhances resistance to mildew pathogens and salt stress in Arabidopsis

Aldehyde dehydrogenases (ALDHs) catalyze the irreversible oxidation of a broad spectrum of reactive aldehydes to their corresponding carboxylic acids. Although the proteins have been studied from various organisms and at different growth stages in plants, their potential roles in pathogen infection have not been examined. Here we isolated and functionally characterized a pathogen-inducible ALDH gene (VpALDH2B4) from Chinese wild grapevine Vitis pseudoreticulata accession Baihe-35-1. When transiently expressed in Arabidopsis leaves, VpALDH2B4 was found to be localized in mitochondria. Escherichia coli expressed GST-VpALDH2B4 exhibited ALDH activity in vitro and was capable of utilizing malondialdehyde (MDA), acetaldehyde and glyceraldehydes as its substrate. Over-expression of VpALDH2B4 in Arabidopsis resulted in hypersensitive response-like cell death, enhanced resistance to downy mildew and powdery mildew presumably via the SA-signaling pathway. The same Arabidopsis transgenic plants also showed enhanced tolerance to salt stress, which is accompanied by less MDA accumulation and upregulation of the stress-responsive superoxide dismutase activity. Taken together, our results suggest that VpALDH2B4 and perhaps its orthologous genes may be involved in responses of plants to stresses imposed by both biotrophic pathogens and high salinity conditions.

Extracts of Phenolic Compounds from Seeds of Three Wild Grapevines—Comparison of Their Antioxidant Activities and the Content of Phenolic Compounds

Phenolic compounds were extracted from three wild grapevine species: Vitis californica, V. riparia and V. amurensis seeds using 80% methanol or 80% acetone. The total content of phenolic compounds was determined utilizing the Folin-Ciocalteu’s phenol reagent while the content of tannins was assayed with the vanillin and BSA precipitation methods. Additionally, the DPPH free radical scavenging activity and the reduction power of the extracts were measured. The RP-HPLC method was applied to identify the phenolic compounds in the extracts, such as phenolic acids and catechins. The seeds contained large amounts of tannins, catechins and gallic acid and observable quantities of p-coumaric acid. The total content of phenolic compounds and tannins was similar in the extracts from V. californica and V. riparia seeds. However, the total content of total phenolic compounds and tannins in the extracts from V. californica and V. riperia seeds were about two-fold higher than that in the extracts from V. amurensis seeds. Extracts from seeds of the American species (V. californica and V. riparia) contained similarly high concentrations of tannins, whereas extracts from seeds of V. amurensis had approximately half that amount of these compounds. The content of catechin and epicatechin was similar in all extracts. The highest DPPH• anti-radical scavenging activity was observed in the acetonic and methanolic extracts of V. californica and V. riparia seeds— while the acetonic extract from the V. californica seeds was the strongest reducing agent.

Genetic diversity of wild grapevine populations in Spain and their genetic relationships with cultivated grapevines

The wild grapevine, Vitis vinifera L. ssp.sylvestris (Gmelin) Hegi, considered as the ancestor of the cultivated grapevine, is native from Eurasia. In Spain, natural populations of V.vinifera ssp.sylvestris can still be found along river banks. In this work, we have performed a wide search of wild grapevine populations in Spain and characterized the amount and distribution of their genetic diversity using 25 nuclear SSR loci. We have also analysed the possible coexistence in the natural habitat of wild grapevines with naturalized grapevine cultivars and rootstocks. In this way, phenotypic and genetic analyses identified 19% of the collected samples as derived from cultivated genotypes, being either naturalized cultivars or hybrid genotypes derived from spontaneous crosses between wild and cultivated grapevines. The genetic diversity of wild grapevine populations was similar than that observed in the cultivated group. The molecular analysis showed that cultivated germplasm and wild germplasm are genetically divergent with low level of introgression. Using a model-based approach implemented in the software structure, we identified four genetic groups, with two of them fundamentally represented among cultivated genotypes and two among wild accessions. The analyses of genetic relationships between wild and cultivated grapevines could suggest a genetic contribution of wild accessions from Spain to current Western cultivars.

Short-term response of wild grapevines (Vitis vinifera L. ssp. sylvestris) 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 “Khedhayria” 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 (ΨPD), the non-photochemical quenching of fluorescence (NPQ) and the coefficient of photochemical quenching (qp) 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, qp, mRNA levels of Vs α-gal/SIP and Vs DHN could be used to screen salt-tolerant wild grapevine genotypes.

Study of genetic relationships between wild and domesticated grapevine in the north of Morocco

In Morocco, knowledge about the wild grapevines has been absent. Until now, no accurate inventory was available and the characterization of this wild grapevine group was lacking. In the present work, prospecting conducted in the North of Morocco (Rif) permitted us to inventory 18 sites with wild grapevines (10 to 50 km away from each other). A total of 168 individuals have been found. This material was analyzed with the 20 nuclear, 3 chloroplast SSR markers and 2 genes involved in the anthocyanin metabolic pathway. We compared the diversity of this material with 48 individuals from cultivated grapevines prospected in the same region, 50 cultivated grapevines from Tunisia and 30 from Algeria along with 128 wild grapevines from France. The diversity observed in the Moroccan wild group was slightly higher compared to other groups. Studies of the genetic structure were then carried out. Within the wild grapevines, the French and Moroccan samples were well differentiated. The wild group (of French or Moroccan origin) was also differentiated from the cultivated, thus revealing the wild status of this indigenous material. The analyses of the sequences showed the same SNP in the different group but the haplotypes reconstruction revealed the presence of specific haplotypes in the Moroccan material when compared with the cultivars group used in this study. This work, confirms the existence and the interest in the conservation of the wild grapevine in Morocco. We suppose that these wild vines contain interesting genes which facilitate the adaptation of grape to its environment. Key words: Vitis vinifera, gene flow, genetic diversity, Morocco, plant conservation genetics.

Use of chloroplast microsatellite markers as a tool to elucidate polymorphism, classification and origin of Tunisian grapevines

Chloroplast microsatellite markers were used in this study to genotype 43 grapevines accessions grown in Tunisia. Size variation was observed for the three cpSSR loci, both in the sample of cultivars and in wild accessions. The seven alleles observed in the sample of cultivars for the three loci are present in wild accessions except that their distribution is different. Levels of genetic diversity obtained for the Tunisian grapevines either in wild or cultivated gene pools are high and comparable with values obtained with other studied samples of Vitis vinifera. The distribution of haplotypes within the two samples is differential. Indeed, the chlorotype A is most abundant in the wild sample, whereas the chlorotype C is majority in the sample of cultivars. Haplotypes frequencies for cultivated grapevine distinguish haplotypes B and C as the most frequent (28% and 44% respectively) and haplotypes A and D as the least frequent (16% and 12% respectively). For wild grapevines, the seven alleles combined in three haplotypes, A, C and D. The haplotype A is the most frequent (44%) in the analyzed sample of wild accessions while haplotypes C and D show a frequency of 28%. Chlorotype distribution in Tunisian cultivars is comparable with that of cultivars in the Eastern Region representing the primary centre of domestication of the species. These results agree with the higher relevance of table grape cultivars in Tunisian viticulture and support an oriental origin of a large part of autochthons cultivars. Our results agree with other studies based in nuclear and chloroplast microsatellite markers and suggest independent domestication events for V. vinifera L. species.

VpRFP1, a novel C4C4-type RING finger protein gene from Chinese wild Vitis pseudoreticulata, functions as a transcriptional activator in defence response of grapevine

RING finger proteins comprise a large family and play important roles in regulation of growth and development, hormone signalling, and responses to biotic and abiotic stresses in plants. In this study, the identification and functional characterization of a C4C4-type RING finger protein gene from the Chinese wild grapevine Vitis pseudoreticulata (designated VpRFP1) are reported. VpRFP1 was initially identified as an expressed sequence tag (EST) from a cDNA library constructed from leaves of V. pseudoreticulata inoculated with the grapevine powdery mildew Uncinula necator. Sequence analysis of the deduced VpRFP1 protein based on the full-length cDNA revealed an N-terminal nuclear localization signal (NLS) and a C-terminal C4C4-type RING finger motif with the consensus sequence Cys-X2-Cys-X13-Cys-X1-Cys-X4-Cys-X2-Cys-X10-Cys-X2-Cys. Upon inoculation with U. necator, expression of VpRFP1 was rapidly induced to higher levels in mildew-resistant V. pseudoreticulata plants. In contrast, expression of VpRFP1 was down-regulated in mildew-susceptible V. vinifera plants. Western blotting using an antibody raised against VpRFP1 showed that VpRFP1 was also induced to higher levels in V. pseudoreticulata plants at 12–48 hours post-inoculation (hpi). However, there was only slight increase in VpRFP in V. vinifera plants in the same time frame, even though a more significant increase was observed at 96–144 hpi in these plants. Results from transactivation assays in yeast showed that the RING finger motif of VpRFP1 exhibited some activity of transcriptional activation; however, no activity was seen with the full-length VpRFP1. Overexpression of VpRFP1 in Arabidopsis plants was found to enhance resistance to Arabidopsis powdery mildew Golovinomyces cichoracearum, which seemed to be correlated with increased transcript levels of AtPR1 and AtPR2 in the pathogen-infected tissues. In addition, the Arabidopsis transgenic lines showed enhanced resistance to a virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Taken together, the results suggested that VpRFP1 may be a transcriptional activator of defence-related genes in grapevines.

Environmental, sanitary and ampelographic characterization of wild grapevine in Western Pyrenées (Spain, France)

<p style="text-align: justify;"><strong>Aims</strong>: The aim of this paper was to locate and describe the wild grapevine as a phytogenetic resource in Western Pyrenees, including Navarre, Aragon (Spain) and the department of Pyrénées atlantiques (France).</p><p style="text-align: justify;"><strong>Methods and results</strong>: An ampelographic description of wild grapevine populations was made as well as a study of their oenological potential and an evaluation of their sanitary condition. The ampelographic results confirmed the dioecious nature of all the specimens. The microvinification results of two wine samples showed low alcohol content, high acidity (low pH) and high intensity of colour. The sanitary study revealed that the main parasites of the vines were eriophyids (mites), powdery and downy mildews (fungus), with aerial organs being most susceptible (leaves, bunches). No symptoms of phylloxera, nematodes or root destroying fungi were detected. The paper is complemented by a list of the main accompanying botanical species.</p><p style="text-align: justify;"><strong>Conclusion</strong>: This phytogenetic resource is well represented in the area of study, exhibiting a good sanitary status.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: This paper could provide the basis for future genetic studies addressing the possible links between the wild grapevines and the cultivated varieties of the region. Wild grapevines could also be used in future breeding programs.</p>

Spliceosomal intron size expansion in domesticated grapevine (Vitis vinifera)

BackgroundSpliceosomal introns are important components of eukaryotic genes as their structure, sizes and contents reflect the architecture of gene and genomes. Intron size, determined by both neutral evolution, repetitive elements activities and potential functional constraints, varies significantly in eukaryotes, suggesting unique dynamics and evolution in different lineages of eukaryotic organisms. However, the evolution of intron size, is rarely studied. To investigate intron size dynamics in flowering plants, in particular domesticated grapevines, a survey of intron size and content in wine grape (Vitis vinifera Pinot Noir) genes was conducted by assembling and mapping the transcriptome of V. vinifera genes from ESTs to characterize and analyze spliceosomal introns.ResultsUncommonly large size of spliceosomal intron was observed in V. vinifera genome, otherwise inconsistent with overall genome size dynamics when comparing Arabidopsis, Populus and Vitis. In domesticated grapevine, intron size is generally not related to gene function. The composition of enlarged introns in grapevines indicated extensive transposable element (TE) activity within intronic regions. TEs comprise about 80% of the expanded intron space and in particular, recent LTR retrotransposon insertions are enriched in these intronic regions, suggesting an intron size expansion in the lineage leading to domesticated grapevine, instead of size contractions in Arabidopsis and Populus. Comparative analysis of selected intronic regions in V. vinifera cultivars and wild grapevine species revealed that accelerated TE activity was associated with grapevine domestication, and in some cases with the development of specific cultivars.ConclusionsIn this study, we showed intron size expansion driven by TE activities in domesticated grapevines, likely a result of long-term vegetative propagation and intensive human care, which simultaneously promote TE proliferation and repress TE removal mechanisms such as recombination. The intron size expansion observed in domesticated grapevines provided an example of rapid plant genome evolution in response to artificial selection and propagation, and may shed light on the important genomic changes during domestication. In addition, the transcriptome approach used to gather intron size data significantly improved annotations of the V. vinifera genome.

Genetic diversity in Anatolian wild grapes (Vitis viniferasubsp.sylvestris) estimated by SSR markers

Anatolia (Asia Minor) is considered to be the cradle of viticulture, but wild grape accessions from this region have not been subjected to any genetic analysis. We present the first genetic characterization of wild grapes (Vitis viniferasubsp.sylvestris) from this region. Using 15 nuclear microsatellites, we genotyped 84 wild grape accessions collected from three Anatolian locations. The unweighted pair group method with arithmetic mean analysis revealed four genetic clusters that partially separated Anatolian wild grape populations. In general, accessions from geographically closer locations showed higher genetic similarities than those from more distant locations. In some cases, accessions from one population showed close genetic relationships to accessions from a different population. The genetic diversity between natural populations from both ends of the Mediterranean basin has shown higher genetic diversity in the Anatolian Peninsula than peripheral populations, suggesting that this area could be one centre of diversity of the species. The genetic relationship between wild and cultivated grapevine from Anatolian Peninsula indicated a clear separation between them. However, we cannot discarded a local genetic contribution. Overall, this study reveals useful information for management and potential utilization of Anatolian wild grape germplasm.

Plant economy at a late Neolithic lake dwelling site in Slovenia at the time of the Alpine Iceman

We present the results of a plant macroremain study of the late Neolithic lakeshore settlement Stare gmajne (SG) at Ljubljansko barje, Slovenia, with cultural horizons that ended around 3330 and 3110 cal. b.c., as obtained by dendrochronological and radiocarbon dating of the most frequent construction timbers of Quercus sp. (oak) and Fraxinus sp. (ash). Fourteen systematically taken samples were investigated, using standard methods for studying waterlogged plant remains, which had been developed during lake dwelling research north of the Alps. Most of the remains were preserved in a waterlogged state, and we identified a total of 93 taxa. The most important cultivated plants were Triticum dicoccum (emmer), Hordeum vulgare (six-rowed naked barley), T. monococcum (einkorn), Linum usitatissimum (flax) and Papaver somniferum (opium poppy). The numerous possibly gathered plants also included Trapa natans (water chestnut) and Vitis vinifera ssp. sylvestris (wild grapevine). Chenopodium album (goosefoot) and Brassica rapa (turnip) with seeds/fruits rich in oil and starch were probably gathered as well. Comparisons of the Stare gmajne results with contemporary north Alpine sites (NA) showed, among other things, that Triticum durum/turgidum (tetraploid naked wheat), frequent at NA, was not found at SG. Trapa natans (water chestnut) was rare and Vitis (grapevine) was not found at NA. The observed differences in the wild plant spectra may have ecological causes, for example a warmer climate south of the Alps, but differences in cultivar spectra are more likely for cultural-historical reasons.

Characterization of Vitis vinifera L. subspecies sylvestris (Gmelin) Hegi in the Ebro river Basin (Spain)

A wild grapevine population of 76 vines of Vitis vinifera L. subspecies sylvestris (Gmelin) Hegi was found along the Iregua river valley (Northeastern Spain), located in the Iberica mountain-range. The characterization of this phytogenetic resource was based on an ampelographic description of the male and female individuals and an evaluation of their sanitary state. This shows that the imported North American downy and powdery mildews are the main pathogens, but no symptoms caused by phylloxera on roots were found. Different degrees of infestation caused by Eryophid mites, Colomerus vitis and Calipitrimerus vitis , were also registered on leaves. On the other hand, the red wines obtained show an alcoholic degree situated between 8.5 and 10.8, pH around 3.5 and a high intensity of colour, between 10.2 and, 11.3. The vinegar had only 3.5° of acetic acid. Its phenolic composition is similar to those reported from red and Sherry vinegars produced from cultivars. The genetic analysis based on 18 samples, using 16 nuclear microsatellites, shows a low genetic diversity (He = 0.45). This might be due to inbreeding caused by mating among siblings in this isolated population. The genetic comparison with the allowed cultivars of this Guarantee of Origin showed two distinct gene pools. So it indicated that there is no genetic contribution of these native vines to current instead of actual local varieties. It is necessary to preserve this phytogenetic resource to be used in breeding programs and to restore its destructed habitats by human different impacts.

Distribution of Wild and Cultivated Grapes in Turkey

Turkey is one of main gene centers in the world for grapes. It is believed that cultivated grapes have their origins in Turkey and the surrounding countries. Vitis vinifera ssp sylvestris is the only wild grape species in this region. That is why Turkey has a very large amount of wild grapevine populations and grape cultivars which offer to grapevine breeders a valuable gene pool. Wild grapevines have significant characters for inducing the resistence to biotic and abiotic stress factors, such as resistance to lime, drought, pests and diseases. Turkey has over 1.600 local grape cultivars, among which the majority of them are conserved at the national grape collection vineyard in Tekirda?. They are mostly used as table grapes, dried grapes or for local consumptions. Wild grapes are distributed all over the country territory, mainly in the river basins and forests. Wild grape collection vineyards were established at some universities in Turkey. These grapevines will be screened for the resistance to biotic and abiotic stress factors.

Anthocyanin Composition of Several Wild Grape Accessions

This study investigated the anthocyanin composition of 21 mostly Spanish wild grapevine accessions preserved at El Encin Germoplasm Bank and selected in consideration of observed ampelographic differences and molecular characterization. Sampling was carried out in 2006, 2007, and 2008. After extraction from grape skins, total anthocyanins was determined by spectrophotometry and the anthocyanin fingerprint of grapes, based on 15 anthocyanin variables, was determined by HPLC. Total anthocyanin concentration was similar to that found in winegrape cultivars. The accessions studied showed considerable variability in their anthocyanin fingerprints and it was possible to distinguish several groups, similar to previous reports on the anthocyanin fingerprint of winegrapes. The anthocyanin composition of wild grapevine accessions was similar to that of cultivated grapes. Nevertheless, the presence of wild accessions with anthocyanin fingerprints uncommon or nonexistent in Spanish cultivated varieties suggests that the genetic variability related to anthocyanins in Spanish wild grapevine populations may be higher than that of cultivated varieties commonly considered of Spanish origin.

The cytoskeleton enhances gene expression in the response to the Harpin elicitor in grapevine.

The cytoskeleton undergoes dramatic reorganization during plant defence. This response is generally interpreted as part of the cellular repolarization establishing physical barriers against the invading pathogen. To gain insight into the functional significance of cytoskeletal responses for defence, two Vitis cell cultures that differ in their microtubular dynamics were used, and the cytoskeletal response to the elicitor Harpin in parallel to alkalinization of the medium as a fast response, and the activation of defence-related genes were followed. In one cell line derived from the grapevine cultivar ‘Pinot Noir’, microtubules contained mostly tyrosinylated α-tubulin, indicating high microtubular turnover, whereas in another cell line derived from the wild grapevine V. rupestris, the α-tubulin was strongly detyrosinated, indicating low microtubular turnover. The cortical microtubules were disrupted and actin filaments were bundled in both cell lines, but the responses were elevated in V. rupestris as compared with V. vinifera cv. ‘Pinot Noir’. The cytoskeletal responsiveness correlated with elicitor-induced alkalinization and the expression of defence genes. Using resveratrol synthase and stilbene synthase as examples, it could be shown that pharmacological manipulation of microtubules could induce gene expression in the absence of elicitor. These findings are discussed with respect to a role for microtubules as positive regulators of defence-induced gene expression.

Evaluation of DNA polymorphism among cultivated and wild grapevine accessions from Azerbaijan

To estimate genetic relationships among 31 cultivated and 34 wild grape accessions originating from regions near the Caspian Sea in the Azerbaijan Republic, RAPD analysis was performed with 27 decamer primers selected from a total of 55 primers. The most discriminating primers were OPC-16, OPF-18 and OPA-17, which showed the highest values of genetic diversity (0.927, 0.914 and 0.909, respectively). The lowest values of diversity pertained to the markers OPA-1 (0.615) and V-20 (0.624). The cluster analysis representing genetic similarity among all selected samples divided the genotypes into nine separate groups at similarity index 0.508. Within the studied Azerbaijan grape populations the highest genetic diversity belonged to the population of cultivated samples originating from the Absheron peninsula, with a diversity index 0.852 and the next ranks were assigned to the wild populations originating from Nabran and Guba regions, with a diversity index 0.824 and 0.793, respectively. The lowest diversity was observed within Davachi individuals, with a diversity index 0.765. The wild population from Azerbaijan was molecularly similar to the cultivated gene pool from this area. This result supported the hypothesis that the southwest of the Caspian Sea is a region where grape was brought into culture. During the analysis a special band was observed which could be used in identifying wild and cultivated grape accessions with high or low resveratrol content. The results of this work clearly indicated that the RAPD analysis can be used to estimate genotypic similarities, genetic diversity and for clustering cultivated and wild grape accessions.

Morphological variation and relationships of Georgian populations of Vitis vinifera L. subsp. sylvestris (C.C. Gmel.) Hegi

Georgia is considered as one of the regions where for the first time wild growing grapevine plants were used and taken into cultivation by men. An ampelographic and ampelometric study of leaf characters and flower landmark morphometrics was carried out in seven populations of wild grapevine located in four river basins of three geographic regions – Western (1 popul.), Eastern (5 popul.) and Southern (1 popul.) Georgia, and then compared using principal component analysis and multivariate discriminant analysis allowing the populations to be distinguished. Lengths of main leaf veins and nectaries in male flowers were the variables, which allowed the best discrimination among populations. A mutation leading to white-fruited grapevine is found in one population. Pubescence varies within populations and even within an individual genome and could be used as a discriminant trait only in combination with shapes and sizes of leaves and other morphological characters. The results revealed significant differences among populations located in different geographic regions of the Southern Caucasus area, but the five investigated populations from Eastern Georgia differ only insignificantly. The high morphological variability of wild grapevine in Georgia might be considered as characterizing the large gene pool that contributed to the domestication process of grapevine.