Comparison of grape seedlings population against downy mildew by using different provocation methods.

The common grapevine, Vitis vinifera subsp. sativa L. (2n=6x=38) is one of the major fruit crops worldwide in terms of economic value and cultivated area. In the panorama of Italian viticulture and oenology, Campania region is characterized by a rich biodiversity, which includes a distinctive number of very ancient varieties (more than one hundred years old). This biodiversity needs to be preserved to protect producers and consumers from frauds, and on the counterpart is an essential genetic resource for breeding. The recent sequencing of grape genome represents an essential step forward for structural and functional genomics studies. In this context, one major objective of this research was to investigate the molecular basis of Campania grape germplasm. Microsatellite and retrotransposon-based markers were used in a collection of 62 grapevines. Homonymies and synonymies were found, pointing out that molecular evaluations can provide further insights into genetic structure and differentiation of Vitis germplasm accumulated during centuries of cultivation and selection. Indeed, Procidana and Coglionara are grapes cultivated and sampled on the island of Ischia and we classified them as synonymous. In our effort, we were able to identify 19 grape-specific alleles, which represent useful tools for many purposes such as traceability, typicity preservation and varietal identification. Within the collection analyzed, Aglianico del Taburno is a model for astringency and the strong resveratrol content confers high positive nutriceutical properties to its grapes and wines. For this reason, a transcriptomic analysis of key genes of the phenyl-propanoid pathway was carried out in different tissues of the berry during fruit maturation. This part of the study was correlated with the spectrophotometric assay of total phenols, flavans, anthocyanins and tannins detected in skin and berry. Anthocyanin analysis revealed the highest amount of these molecules in ripening berry skin and the lowest in seed. Correlating the transcriptomic and chemical data, the key-genes able to control production of poly-phenolic compounds in berry tissues were determined. Poly-phenolic compounds are key molecules of grapevine defense and in particular of PTI (PAMP-Triggered Immunity). Indeed, they are produced as defense compounds after pathogen recognition. Nowadays, much has been written about defense mechanisms and chitin perception in Arabidopsis, but little is known in grapevine. Considering powdery mildew as the most destructive disease of grapevines world-wide, an intense structural and functional study of PTI-involved grape genes was conducted. Grapevines are highly resistant to many powdery mildews of other species, called non-adapted (e.g. E. cichoracearum). Using Arabidopsis as model species, we identified and functionally studied the grape genes involved in the perception of chitin in the epidermal cells apoplast, as LysM-RLK, and in the intracellular signal transduction. Our findings suggest that three candidates, VvLYK1, VvLYK2 and VvLYK3, can play a different role in chitin signaling in V. vinifera. We demonstrated that a total resistance to powdery mildew was re-established in the A. thaliana cerk1 mutant only by VvLYK1, but not by VvLYK2 and VvLYK3. Further experiments also suggested an inactive VvLYK2 gene and an active but unknown function of VvLYK3. Regarding the signal transduction pathways, we found that VvMAPK3 and VvMAPK6 are not involved in the biotic response, since they both did not show expression differences in powdery mildew infection and chitin treatment time courses. The transcriptomic data generated in this study allowed us to identify VvWRKY24 as the positive regulatory factor of grape defense to powdery mildew, but excluded VvWRKY16. Indeed, VvWRKY24 had the strongest up-regulation in response to powdery mildew infection and chitin treatment. To better understand the possible interactions between two or more genes and regulatory mechanisms, an over-expression and silencing transformation was conducted. The transformations gave rise to few positive embryos after ten months from co-cultivation with Agrobacterium. This result is strictly correlated with the known recalcitrance of grapevine to transformation. As far as we know, this represents the first complete structural and functional genomics study in grapevine aimed to the identification of the major genes involved in chitin perception and signal transduction.

Studies have been carried out in 27 vine nurseries in Izmir, Manisa, Usak, Denizli and Canakkale provinces. Disease incidence and prevalence have been determined in the sandboxes, callus chambers, greenhouses (stocks growing in pots), and fields of these nurseries. Determined pathogens are given below: a) In the sandboxes : Armillaria mellea the agent of Armillaria root-rot disease on the buried cane cluster. b) In the callus chambers : Rhizoctonia solani and Phythium sp. which are causing shoots drying and shoot-rot, Diplodia natalensis producing shoot blight c) In the greenhouses : Botrytis cinerea , the agent of grey mold disease has been found at different rates along with D.natalensis, R.solani , and Phythium sp. d) At field : own-rooted (rootstocks), grafted-rooted, and softwood cuttings ( Vitis vinifera var. sultana ) have been observed at two stages : 1 ) At spring- early summer season : Macrophomina phaseolina , A.mellea, D.natalensis, R.solani , and Phythium sp. 2 ) At fall season : Uncinula necator (Powdery mildew) Plasmopara viticola (Downy mildew), A.mellea, D.natalensis, R.solani , and Phythium sp. have been obtained. On the other hand Verticillium dahliae the agent of Verticillium wilt disease have been obtained on the rootstocks 41-B and R-99.

Esca of grapevine is a fungal disease with a complex aetiology that is common in almost all regions of the world where grapes are cultivated. Despite much research, no effective control of the disease has been found. We investigated possible activity of fosetyl-aluminium (fosetyl-Al), an active ingredient in many fungicides against downy mildew, to inhibit development of esca in grapevine and the main pathogens linked to the disease, Phaeomoniella chlamydospora (Pch) and Phaeoacremonium aleophilum (Pal). In greenhouse experiments conducted on vines artificially inoculated with Pch or Pal, we found a reduction in necrosis in the woody tissue. In field experiments conducted over several years statistically significant reductions in the annual and cumulative incidence of the disease symptoms, and in cumulative vine mortality, were seen. The effect of fosetyl-Al treatments on leaf gas exchange, levels of resveratrol and ε-viniferin in the wood, and enzymatic activities were also studied. Hypotheses regarding the mechanism of action of fosetyl-Al against esca are outlined, and the possible use of products containing the chemical, applied on foliage to control downy mildew, is discussed as a strategy for control of esca.

Abstract. The effects of green pruning on fruit quality traits were investigated. In industry food science Lab Science and Technology Park Khorasan Razavi province situated in 10 km of Mashhad is done. And its effect in different situation of berry on the punicle , in terms of seed length growth and fruit growth in Seedless grape varieties of Askari grapes in 2013 in commercial Vineyard place on the Kalakub village (20 km north of Mashhad) were investigated. In the spring of 2013 Green pruning different treatments (1-control, 2- traditional green pruning as a removal 4-5 mature leaves from the lower per branch 3- pound arrangement and remove Seedlings grown on the old branches that in addition charge is grown. And , as well as growing remove tributaries the 4- third treatment+ topping branches growing from 20 seedlings) in a randomly complete block design with two replications for Askari grapes variety was used. The design as a Factorial trial with two factors inserted in above analyze based on randomly complete block design. Which none of factors and their interactions on seed length were not significant effects but on number of seeds formed in factor berry of berry situation in punicle were significant effects in 1% level. Green pruning treatments on fruit quality traits was non-significant effects.

Summary The practical application of pyramiding resistance genes by the use of molecular markers was investigated in a F1 progeny derived from the cross of VHR 3082-1-42 x ‘Regent’. VHR 3082-1-42 is a cross between Muscadinia rotundifolia x Vitis vinifera, backcrossed another four times with V. vinifera (PAUQUET et al. 2001). It carries the Run1-gene which causes resistance to powdery mildew and the Rpv1-gene which is related to resistance against downy mildew. Both genes were introduced from Muscadinia rotundifolia (BOUQUET et al. 2000; WIEDEMANN-MERDINOGLU et al. 2006). ‘Regent’ is a new cultivar with quantitative resistance against downy and powdery mildew (EIBACH and TOPFER 2003) released in Germany in 1996 for commercial use. 119 individuals of the F1 progeny were screened with a molecular marker for the Run1-gene (DONALD et al. 2002), with two SSR-markers for the Rpv1-gene (WIEDEMANN-MERDINOGLU et al. 2006) and with several markers from ‘Regent’ that showed good correlation to powdery and downy mildew resistance (ZYPRIAN et al. 2002, SALAKHUTDINOV et al. 2003, AKKURT 2004, FISCHER et al. 2004, AKKURT et al. 2007). Phenotypic evaluation for downy mildew resistance was done by artificial inoculation of leaf discs, and for powdery mildew by natural infection in a greenhouse. Comparison of the phenotypic data with the results of the molecular marker analyses showed a clear correlation between the degree of resistance and the presence of the resistance related alleles. According to the phenotypic data, 20 genotypes of the offspring were free of powdery and downy mildew infections. Based on a marker-assisted evaluation, out of these 20 genotypes a subset of four carried all the resistance related alleles for powdery and downy mildew indicating that resistance genes from both parents were effectively combined.

Introduction: Many reports have shown that the grape (Vitis vinifera) seed extract induces antioxidant, hypotensive hypolipidemic, and vasodilatory effects. We have recently shown the relaxatory effect of grape leaf extract on rat's aorta, ileum, uterus and frog's heart rate and contractility. The objective of this study was to investigate the spasmolytic effect of Vitis Vinifera Leaf Hydroalcoholic extract (VLHE) on rat's isolated vas deferens. Materials & Methods: Vas deferens of male adult rat (Sprague Dawley) was removed, placed in an organ bath containing Tyrode’s solution (30oC, pH 7.4) and the bath bubbled with air. Then its contractions under 1 g initial tension, were recorded isometrically. The vas deferens was contracted either by KCl (80 mM) or adrenaline (2 μg/ml). Results: The results demonstrated that the VLHE at 1, 2, 4 and 8 mg/ml reduced the vas deferens contractions evoked by KCl (80 mM) significantly (P<0.0001) and dose-dependently. VLHE (1, 2 and 4 mg/ml) also reduced the adrenaline-induced vas deferens contractions (P<0.0001). In calcium free Tyrod’s solution, KCl-induced contractions were occurred only after adding normal calcium (1.77 mM) to the organ bath solution. VLHE (3mg/ml) reduced the calcium-induced contraction by KCl too. The VLHE-induced vas deferens relaxation was unaffected by the β- adrenoceptor antagonist (propranolol, 3 μM for 5 min). Nitric oxide synthase inhibitor (L-NAME, 300 μM for 10 min) was unable to modify either KCl-induced contraction or the vas deferens relaxation induced by the extract. Conclusion: These results suggested that the spasmolytic effect of Vitis vinifera leaf hydroalcoholic extract on rat's vas deferens occurred through the blockage of voltage dependent calcium channels (VDCC). Also, neither NO nor β-adrenoceptors were involved in this inhibitory effect of the extract.

The researches carried out during the PhD were focused on the effects of different cultural techniques on the phenolic compound of Vitis vinifera L., under the ongoing climate change scenario that has been causing negative effects on grape composition and sensory attributes of wines. In the first trial was evaluated the effectiveness of the delayed winter spur pruning after budburst to slow down sugar accumulation in cv. Merlot, with the aim to reduce the decoupling between technological and phenolic maturity. This technique, if applied in a particular period, resulted able to slow down sugar accumulation without negative effects on berry flavonoids and with acceptable loss of yield. Since the topic of the phenolic maturity became important in the evaluation of the effects that global warming and cultural techniques have on black berry varieties, in the second experiment were investigated the changes that occurs on berry flavonoids and in the properties of skin cell wall material during the last part of ripening in cv. Merlot. Total and extractable anthocyanins increased until harvest, as well as the affinity of cell wall material for the commercial seed tannin. This latter aspect might be correlated to the decrease of astringency during ripening. In the third experiment was investigated the role that sunlight incidence on clusters of cv. Grechetto gentile plays on grape composition and on the phenolic tastes of the resulting wines. Increased light incidence determined higher concentration of flavonols in grape and wine and that was correlated to the greater perception of astringency and bitterness of the wines. The set of results of the present PhD enhanced the knowledge on the behavior of grape flavonoids under the current global warming scenario and proposed innovative approaches in the study of the phenolic maturity.

Grapevine varieties showing partial resistance to downy mildew, caused by Plasmopara viticola, are a promising alternative to fungicides for disease control. Resistant varieties are obtained through breeding programs aimed at incorporating Rpv loci controlling the quantitative resistance into genotypes characterized by valuable agronomic and wine quality traits by mean of crossing. Traditional phenotyping methods used in these breeding programs are mostly based on the assessment of the resistance level after artificial inoculation of leaf discs in bioassays, by using the visual score proposed in the 2nd Edition of the International Organization of Vine and Wine (OIV) Descriptor List for Grape Varieties and Vitis species (2009). In this work, the OIV score was compared with an alternative approach, not used for the grapevine-downy mildew pathosystem so far, based on the measurement of components of resistance (RCs); 15 grapevine resistant varieties were used in comparison with the susceptible variety ‘Merlot’. OIV scores were significantly correlated with P. viticola infection frequency (IFR), the latent period for the downy mildew (DM) lesions to appear (LP50), and the number of sporangia produced per lesion (SPOR), so that when the OIV score increased (i.e., the resistance level increases), IFR and SPOR decreased, while LP50 increased. The relationship was linear for LP50, monomolecular for IFR and hyperbolic for SPOR. No significant correlation was found between OIV score and DM lesion size, sporangia produced per unit area of lesion, length of infectious period, and infection efficiency of the sporangia produced on DM lesions. The correlation between OIV score and area under the disease progress curve (AUDPC) calculated by using the RCs and a simulation model was significant and fit an inverse exponential function. Based on the results of this study, the measurement of the RCs to P. viticola in grapevine varieties by means of monocyclic, leaf disc bioassays, as well as their incorporation into a model able to simulate their effect on the polycyclic development of DM epidemics in vineyards, represents an improved method for phenotyping resistance level.

Turkey is native land of grapes (Vitis spp.) which is the most consumed fruit in the world. Turkey has favorable climatic conditions for the viticulture. The aim of this study was to determine potential cultivation areas of cold durable Amur grape (Vitis amurensis Rupr.) in the districts and villages of Erciş (Van, Turkey) by employing the Geographic Information Systems GIS) that provides climate, soil and topography factors. GIS offers the opportunity to generate and to organize spatial data for the land planning and to analyze more than one layers. In this research, as parameters the sum of effective temperature, the number of days without frost, the lowest winter temperatures, slope, aspect, soil depth, soil drainage and the land use capability were evaluated in terms of viticulture. These parameters were scored according to the level of significance and analyzed by modeling. Possible cultivation areas of Amur Grape (Vitis amurensis Rupr.) were determined and suitability maps were created. It is also aimed to expand vineyard areas and to help the establishment of vineyard in suitable regions which is a costly and laborious efforts, thus preventing waste of time and cost. Considering the criteria evaluated by employing GIS, the agricultural areas in the region have been analyzed and mapped for suitability for the cultivation of Amur grape. Based on the evaluated parameters, possible growing areas of Amur grape have been determined that are far above the areas of traditional viticulture. It has been determined that at three suitability levels Amur Grape cultivation can be performed in a total land of 83.297 hectares in Erciş. Agricultural production depends on multiple factors; therefore planning and programming (product modeling) are not easy. With the contribution of this and similar studies, Agricultural Support and Planning will be carried out more efficiently.

The Amur grape (Vitis amurensis Rupr.) is a wild grape species rich in bioactive compounds, including polyphenols, resveratrol, and iron, making it a potential functional ingredient for food fortification. Despite its proven antioxidant and nutritional benefits, its application in meat product fortification remains unexplored. The present study aims to evaluate the potential use of the Amur grape to fortify mass consumption food products. A systematic review of relevant literature was conducted using the PRISMA algorithm, analyzing sources from Scopus, Web of Science, and Science Direct. Findings indicate that Amur grape extracts exhibit significant antioxidant activity (367.24±0.53 mg ascorbic acid equivalents/g), which can extend the shelf life of meat products and enhance their nutritional profile. Hence, they exhibit anti-anemic and antioxidant effects when added to food products. The polyphenols of Amur grapes, primarily resveratrol, enhance the antioxidant effect of ascorbic acid and exhibit anticarcinogenic, cardioprotective, anti-stressor, and other effects. The fortification of food products with compositions from Amur grape also improves their organoleptic characteristics and extends shelf life due to high antioxidant activity. The conclusion indicates that there are relatively few developments in food product technologies with Amur grapes, and mass consumption food products, including meat products, fortified with Amur grapes are practically nonexistent on the market. The findings contribute to the development of natural, clean-label antioxidant solutions, aligning with consumer trends toward healthier, functional foods. Keywords: Amur grape, Vitis amurensis, functional food ingredient, resveratrol, meat products, antioxidant properties, natural food additives, food preservation.

Downy mildew is a highly destructive disease in grapevine production. A gene encoding pathogenesis-related (PR) thaumatin-like protein was isolated from the downy mildew-resistant grapevine "Zuoshan-1," a clonal selection from wild Vitis amurensis Rupr. The predicted thaumatin-like protein (VaTLP) has 225 amino acids and it is acidic, with a calculated isoelectric point of 4.8. The full length of the VaTLP gene was transformed into somatic embryogenic calli of V. vinifera 'Thompson Seedless' via Agrobacterium tumefaciens. Real-time RT-PCR confirmed that the VaTLP gene was expressed at a high level in the transgenic grapevines. Improved resistance of the transgenic lines against downy mildew was evaluated using leaf disks and whole plants inoculated with Plasmopara viticola, the pathogen causing grapevine downy mildew disease. Bioassay of the pathogen showed that both hyphae growth and asexual reproduction were inhibited significantly among the transgenic plants. Histological analysis also confirmed this disease resistance by demonstrating the inhibition and malformation of hyphae development in leaf tissue of the transgenic plants. These results indicated that the accumulation of VaTLP could enhance resistance to P. viticola in transgenic 'Thompson Seedless' grapevines.

Armenia is an important area of crop origins including grapes. The objectives of this study were the molecular characterization of 38 Armenian grape genotypes and the detection of the genetic relationships between the accessions. In total, 164 alleles were obtained at 18 SSRs loci. The number of alleles per locus ranged from 5 to 15 with a mean number of 9.17 alleles per locus. The expected heterozygosity ranged from 0.514 (VVIN73) to 0.90 (VVS2) and the observed heterozygosity from 0.417 (VVIV37) to 0.947 (VVMD7 and VMC1B11). In our research the highest values of polymorphic information content (PIC) were obtained using markers VVS2, VRZAG62 and VRZAG79, while the least informative ones were VVIN73 and VVIB01. A high level of gene diversity was observed between ancient and new bred cultivars. Higher values were recorded for the new grape varieties, (He = 0.792) and lower values for ancient cultivars (He = 0.739). The mean number of alleles (MNA) for all loci per population ranged from 7.39 in ancient to 8,00 in new crosses. A clear separation was observed for the groups of ancient aboriginal cultivars, Vitis vinifera L. xVitis vinifera L. and Vitis vinifera L. x Vitis amurensis Rupr . crosses, which were bred in Armenia and seedless cultivars. Very close relationships with high similarity were determined for 5 pairs of cultivars. Two cases of possible homonymy were also detected. Our results are the basis for future MAS selection and target breeding.

Corresponding author: N. Salem Fax: + 1530 7525674 E-mail: nmsalem@ucdavis.edu Grapevine (Vitis vinifera L.) is the second major crop after olive in Jordan. It is distributed throughout the country on an area of about 39,000 ha (Anonymous, 2003). The health status of Jordanian viticulture is little known as there are few published reports; those that are available record the occurrence of leaf roll, rugose wood and fan leaf (Boscia et al., 1995; AlTamimi et al., 1998). Several virus-like disease symptoms, including yellowing, leaf distortion, stunted growth and decline have been observed during the last few years. A survey was conducted between March and July 2003 in the most important grapevine growing areas of Jordan. A total of 1,088 leaf samples were collected from 60 commercial vineyards and 10 nurseries. The samples were individually tested for Tomato ringspot virus (ToRSV) by a double-antibody sandwich enzyme-linked immunosorbent assay (DASELISA), using a commercial kit from Bioreba (Bioreba AG, Reinach, Switzerland) following manufacturer’s instructions. About 6.5% of the Occurrence of Tomato ringspot virus on grapevines in Jordan

&lt;p style="text-align: justify;"&gt;&lt;strong&gt;Aim:&lt;/strong&gt; To investigate the antifungal efficacy of grape cane extracts enriched in stilbenes against &lt;em&gt;Plasmopara viticola&lt;/em&gt; by &lt;em&gt;in vivo&lt;/em&gt; experiments on grape plants.&lt;/p&gt;&lt;p style="text-align: justify;"&gt;&lt;strong&gt;Methods and results&lt;/strong&gt;: Experiments on grape plants were carried out from 2007 to 2008 at commercial vineyards in « Le Tourne » (southwest region of France) on &lt;em&gt;Vitis vinifera&lt;/em&gt; L. cv Merlot plants and in a greenhouse, where 45-day-old plants (Merlot) were used. Grapevine plants were treated with stilbene extract (STE) or « Bordeaux mixture » (BM) as positive control. STE, prepared from &lt;em&gt;Vitis vinifera&lt;/em&gt; canes, contained 33% of total stilbenes ranging from monomers to tetramers and was used at 5 g L&lt;sup&gt;-1&lt;/sup&gt;. The degree of protection against downy mildew was determined as the percentage of infected leaf surface area and pathogen attack frequency. On young plants in greenhouse, &lt;em&gt;P. viticola&lt;/em&gt; inoculation and three treatments with STE were carried out. After the treatments, the disease reduction was from 59 to 69% for pathogen attack frequency and from 83 to 88% for infected leaf surface for the two modes of infestation (artificial and secondary). STE triggered a similar intense decrease in downy mildew infection as recorded for BM. In the vineyard assay, downy mildew infection was allowed to occur naturally. Field grown plants were treated every 7&lt;sup&gt;th&lt;/sup&gt; day for eight weeks or every 14&lt;sup&gt;th&lt;/sup&gt; day for eight weeks during June and July. We observed that STE was also able to prevent &lt;em&gt;P. viticola&lt;/em&gt; infection in vineyard but to a lesser extent. Treatment with STE every 7&lt;sup&gt;th&lt;/sup&gt; day provided better results in terms of infection frequency and infected leaf area than that every 14&lt;sup&gt;th&lt;/sup&gt; day. STE treatment of plants every 7&lt;sup&gt;th&lt;/sup&gt; day in vineyard reduced the frequency of attack by 16 to 39% and leaf surface infection by 57 to 61%, which was lower than that recorded for BM. STE also displayed &lt;em&gt;in vitro&lt;/em&gt; inhibitory effects on the release of zoospores from sporangia and the germination of zoospores of &lt;em&gt;P. viticola&lt;/em&gt;. Zoospore germination was completely inhibited at a concentration of 100 mg L&lt;sup&gt;-1&lt;/sup&gt; and zoospore release was inhibited at 330 mg L&lt;sup&gt;-1&lt;/sup&gt;.&lt;/p&gt;&lt;p style="text-align: justify;"&gt;&lt;strong&gt;Conclusions&lt;/strong&gt;: Treatment of young grapevine plants in greenhouse with stilbene extract from grapevine canes offered considerable protection against &lt;em&gt;P. viticola&lt;/em&gt; that was identical to that provided by Bordeaux mixture. STE was also able to prevent &lt;em&gt;P. viticola&lt;/em&gt; infection in vineyard but to a lesser extent. Treatment with STE every 7&lt;sup&gt;th&lt;/sup&gt; day in vineyard provided better results than that every 14&lt;sup&gt;th&lt;/sup&gt; day.&lt;/p&gt;&lt;p style="text-align: justify;"&gt;&lt;strong&gt;Significance and impact of the study&lt;/strong&gt;: Stilbenes from grapevine canes are good candidates as natural fungicides against downy mildew. Given the very large quantity of grape canes available each year, this strategy could be scaled up to control this grapevine disease in a sustainable manner.&lt;/p&gt;

Grape (Vitis vinifera L.) cultivation is gaining importance under tropical and sub-tropical conditions of India by adapting advance crop techniques. The problems of detoriated soil, poor and shortage of water conditions affecting the grape production on own rooted vines, which results into use of rootstock in grape cultivation. There are large number of rootstocks available in grape cultivation according to different production constraints. Rootstock not only helps in withstanding vineyard in adverse climatic conditions but also help in improving yield and quality of. The rootstock Dogridge became popular among the grape growers due to its capabilities of overcoming abiotic stresses and better stionic combination. The rootstock suitable for one variety in a given location may not be suitable for the same variety on other location. Hence, choice of rootstock is varied among the different cultivars. In a view of better yield and quality of grapes, there should have better stock-scion compatibility which imparts high vigour into the scion.