Grapevine Leafroll-Associated Virus 3 in Single and Mixed Infections Triggers Changes in the Oxidative Balance of Four Grapevine Varieties.

With the aim to characterize changes caused by grapevine leafroll-associated virus 3 (GLRaV-3) singly or in coinfection with other viruses and to potentially determine genotype-specific or common markers of viral infection, thirty-six parameters, including nutrient status, oxidative stress parameters, and primary metabolism as well as symptoms incidence were investigated in 'Cabernet Franc,' 'Merlot,' 'Pinot Noir,' and 'Tribidrag' grapevine varieties. Host responses were characterized by changes in cellular redox state rather than disturbances in nutrient status and primary metabolic processes. Superoxide dismutase, hydrogen peroxide, and proteins were drastically affected regardless of the type of isolate, the host, and the duration of the infection, so they present cellular markers of viral infection. No clear biological pattern could be ascertained for each of the GLRaV-3 genotypes. There is a need to provide a greater understanding of virus epidemiology in viticulture due to the increasing natural disasters and climate change to provide for global food production security. Finding grape varieties that will be able to cope with those changes can aid in this task. Among the studied grapevine varieties, autochthonous 'Tribidrag' seems to be more tolerant to symptoms development despite numerous physiological changes caused by viruses.

Manganese soil and foliar fertilization of olive plantlets: the effect on leaf mineral and phenolic content and root mycorrhizal colonization.

BACKGROUND: The present study aimed to examine the effect of foliar (Mn_fol) and soil Zeolite-Mn (Mn_ZA) application on leaf mineral, total phenolic and oleuropein content, and mycorrhizae colonization of self-rooted cv. Leccino plantlets grown on calcareous soil. RESULTS: The dissolution of zeolite was 97% when citric acid was applied at 0.05 mM dm-3 , suggesting that organic acids excreted by roots can dissolve modified zeolite (Mn_ZA), making Mn available for plant uptake. The leaf Mn concentration was the highest for Mn_fol treatment at 90 days after transplanting (DAT) (172 mg kg-1 ) and 150 DAT (70 mg kg-1 ) compared to other treatments. Mn_ZA soil application increased leaf Mn concentration at 150 DAT compared to control and NPK treatments. The oleuropein leaf content was highest for Mn_fol compared to other treatments at 90 DAT and lowest at 150 DAT. Arbuscular mycorrhizal colonization was higher for Mn_fol treatment at 150 DAT compared to all other treatments. CONCLUSION: Changes in the arbuscular colonization percentage and oleuropein content may be connected to stress conditions provoked by a high leaf Mn concentration in the Mn_fol treatment at 90 DAT. Mn_ZA application increased leaf Mn concentration at 150 DAT compared to control and NPK treatments. It can be assumed that the dominant mechanism in Mn uptake from modified zeolite is Mn_ZA dissolution through root exudates. © 2018 Society of Chemical Industry.

Distribution and diversity of arbuscular mycorrhizal fungi in grapevines from production vineyards along the eastern Adriatic coast.

The colonisation and diversity of arbuscular mycorrhizal fungi (AMF) on roots of grapevines were investigated in production vineyards located along a 500-km-long stretch of karst along the coast of the Adriatic Sea. AMF communities on roots of grapevines were analysed using temporal temperature gel electrophoresis and sequencing of the 18S and internal transcribed spacer segments of the rDNA operon. The AMF colonisation of these grapevines roots was consistent along the whole of this east Adriatic karst region, at 64 to 82% of fine roots. The comparison of the AMF communities on the roots of these grapevines showed that the fungal community associated with grapevine roots seems to be relatively stable, with inter-vineyard variability comparable to intra-vineyard variability. Some of the changes in the fungal communities were attributed to environmental factors (plant-available P) and location of the vineyard, although the latter could also have been influenced by an unmeasured environmental factor. A total of 27 taxa of fungi were identified, including taxa from Glomus group B, based on the sequencing of 18S rDNA. Sequencing of the internal transcribed spacer rDNA yielded 30 different fungal taxa, which comprised eight different Glomeromycota taxa, including Glomus sinuosum and Glomus indicum. To our knowledge, this is the first report of grapevine colonisation by G. indicum.

Arbuscular Mycorrhizal Fungi Induce Changes of Photosynthesis-Related Parameters in Virus Infected Grapevine.

The negative effects of viruses and the positive effects of arbuscular mycorrhizal fungi (AMF) on grapevine performance are well reported, in contrast to the knowledge about their interactive effects in perennial plants, e.g., in grapevine. To elucidate the physiological consequences of grapevine-AMF-virus interactions, two different AMF inoculum (Rhizophagus irregularis and 'Mix AMF') were used on grapevine infected with grapevine rupestris stem pitting virus, grapevine leafroll associated virus 3 and/or grapevine pinot gris virus. Net photosynthesis rate (AN), leaf transpiration (E), intercellular CO2 concentration (Ci) and conductance to H2O (gs) were measured at three time points during one growing season. Furthermore, quantum efficiency in light (ΦPSII) and electron transport rate (ETR) were surveyed in leaves of different maturity, old (basal), mature (middle) and young (apical) leaf. Lastly, pigment concentration and growth parameters were analysed. Virus induced changes in grapevine were minimal in this early infection stage. However, the AMF induced changes of grapevine facing biotic stress were most evident in higher net photosynthesis rate, conductance to H2O, chlorophyll a concentration, total carotenoid concentration and dry matter content. The AMF presence in the grapevine roots seem to prevail over virus infection, with Rhizophagus irregularis inducing greater photosynthesis changes in solitary form rather than mixture. This study shows that AMF can be beneficial for grapevine facing viral infection, in the context of functional physiology.

Phosphorus Use Efficiency of Leafy Brassica sp. Grown in Three Contrasting Soils: Growth, Enzyme Activity and Phosphorus Fractionation.

Plant adaptations to low soil phosphorus (P) availability have been intensively studied in Brassica sp. in an attempt to identify the mechanisms involved in P uptake and utilization. The present pot experiment was conducted to evaluate the relationships between plant shoot and root growth, P uptake and use efficiency parameters, and P fractions and enzyme activity, in two species grown in three soil types. The aim of this study was to determine whether adaptation mechanisms are soil-dependent. Two kale species were grown in soils typical for coastal Croatia (terra rossa, rendzina, and fluvisol) with low P availability. Plants grown in fluvisol had the highest shoot biomass and accumulated most P, whereas plants developed the longest roots in terra rossa. Phosphatase activity differed among soils. P use efficiency differed among soils and species. Genotype IJK 17 showed better adaptation to low P availability, which was related to better uptake efficiency. In general, soils differed in inorganic and organic P fractions in rhizosphere soil, but no difference between genotypes was found. The activities of alkaline phosphatase and phosphodiesterase were negatively correlated with most organic P fractions, suggesting their function in the mineralization of soil organic P. Kale species activate different mechanisms of P uptake and utilization when grown in contrasting soil types, suggesting that specific responses to the soil type were more important than the genotypic difference.

High diversity of indigenous populations of dalmatian sage (Salvia officinalis L.) in essential-oil composition.

Essential oils of 25 indigenous populations of Dalmatian sage (Salvia officinalis L.) that represent nearly half of native distribution area of the species were analyzed. Plantlets collected from wild populations were grown in the same field under the same environmental conditions and then sampled for essential-oil analysis. The yield of essential oil ranged from 1.93 to 3.70% with average of 2.83%. Among the 62 compounds detected, eight (cis-thujone, camphor, trans-thujone, 1,8-cineole, ß-pinene, camphene, borneol, and bornyl acetate) formed 78.13-87.33% of essential oils of individual populations. Strong positive correlations were observed between camphor and ß-pinene, ß-pinene and borneol, as well as between borneol and bornyl acetate. The strongest negative correlation was detected between camphor and trans-thujone. Principal component analysis (PCA) on the basis of eight main compounds showed that first main component separated populations with high thujone content, from those rich in camphor, while the second component separated populations rich in cis-thujone from those rich in trans-thujone. Cluster analysis (CA) led to the identification of three chemotypes of S. officinalis populations: cis-thujone; trans-tujone, and camphor/ß-pinene/borneol/bornyl acetate. We propose that differences in essential oils of 25 populations are mostly genetically controlled, since potential environmental factors were controlled in this study.

Lipid Profiles and Cardiovascular Risk in Patients with Oral Lichen Planus.

BACKGROUND: It has been reported that dyslipidemia prevalence and cardiovascular disease risk were increased in subjects with oral lichen planus diagnose. On the other hand, so far, there is no available data on the topic of cardiovascular risk (CVR) in subjects with oral lichen planus (OLP). The main aim of this study, due to lack of any other study covering this topic, was to investigate lipid profile and assess CVR in patients with OLP. MATERIALS AND METHODS: To create a routine lipid profile, we collected triglyceride serum levels, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol from 63 OLP patients and 63 healthy people representing control subjects. For every patient their individual cardiovascular risk was measured. RESULTS: In comparison with the tested control subjects, patients with OLP had all parameters of the lipid profile elevated, with no differences of statistical importance. Furthermore, the experimental (OLP) and control groups shared similar mean values of the lipid profile parameters. CONCLUSIONS: The association of OLP with cardiovascular risk was not established and further studies with more subjects involved are required to validate this connection.

Prevalence of Virus Infections and GLRaV-3 Genetic Diversity in Selected Clones of Croatian Indigenous Grapevine Cultivar Plavac Mali.

The cultivar Plavac Mali (Vitis vinifera L.), the most important indigenous red grapevine cultivar in Croatia, was tested for the presence of 16 grapevine viruses. Thirty-five samples from the collection vineyard were tested for the presence of grapevine leafroll-associated viruses-1, -2, and -3 (GLRaV-1, GLRaV-2 and GLRaV-3, respectively), grapevine fanleaf virus (GFLV), arabis mosaic virus (ArMV), grapevine virus-A (GVA), -B (GVB), -G (GVG), -H (GVH), -I (GVI), -J (GVJ), grapevine fleck virus (GFkV), grapevine rupestris stem pitting associated virus (GRSPaV), and grapevine pinot gris virus (GPGV) by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, standard PCR was conducted for grapevine badnavirus 1 (GBV-1) and grapevine red blotch virus (GRBV). Mixed infections were most common and GLRaV-3, the most abundant virus found in 85.71% of the vines tested, was further molecularly characterised. Different genomic variants of the heat shock protein homologue (HSP70h) were separated by cloning, detected by single-strand conformation polymorphism (SSCP) analysis, sequenced, and phylogenetically analysed. The presence of phylogenetic groups I and II was only confirmed. This study demonstrates the high virus infection rate of Plavac Mali vines and the heterogeneity of GLRaV-3 present nowadays in a collection vineyard.

Grapevine Leafroll-Associated Virus 3 Replication in Grapevine Hosts Changes through the Dormancy Stage.

Grapevine leafroll-associated virus 3 (GLRaV-3) is a graft-transmissible virus present in every viticultural region of the world and poses a large threat to grapevine production. Frequent coinfections with other viruses, the large number of grapevine varieties, the complexity of processes involved in plant response to virus infection, and the lack of studies on GLRaV-3 replication limit our knowledge of GLRaV-3 damaging effects and their background. In this study, five different inocula, one containing GLRaV-3 and others containing GLRaV-3 in combination with different grapevine viruses were green grafted to 52 different grapevine plants of four varieties to analyze the influence of the phenological stage and virus composition on GLRaV-3 replication. Relative concentration analysis by quantitative PCR conducted over a 16-month period revealed that other viruses as well as plant stage had a significant effect on GLRaV-3 replication and symptoms expression. The replication was most pronounced in the deep dormancy stage at the beginning of the infection, and the least at the exit of the dormancy stage. This study brings new insight into GLRaV-3 replication and discusses about viral interactions in one of the most economically important perennial plants, the grapevine.

Dataset on endophytic and rhizoplane fungi on the roots of wild grapevine in Croatia and Bosnia and Herzegovina.

The data presented in this article are related to the research article entitled "Root-associated community composition and co-occurrence patterns of fungi in wild grapevine". This dataset documents the diversity of endophytic and rhizoplane fungi found on the roots of 38 wild grapevine plants growing at four locations (Krka, Neretva, Psunj and Paklenica) in Croatia and Bosnia and Herzegovina. We record here 48 identified fungal operational taxonomic units (OTUs) from 3 phyla, 23 families and 30 genera. The material in this Data in Brief paper comprised the data on the identification of OTUs with their corresponding ecological niche, abundance distribution of each fungal OTU recorded in each of the host and location, restrictedness and with fit to the Sloan neutral community model.

Fruit quality and volatile compound composition of processing tomato as affected by fertilisation practices and arbuscular mycorrhizal fungi application.

The effects of different fertilisation treatments with arbuscular mycorrhizal fungi (AMF) inoculation on AMF root colonisation, fruit yield, nutrient and total phenol contents, volatile compound composition, and sensory attributes of tomato (Solanum lycopersicum L.) were investigated. Mineral, organic, and mineral + organic fertiliser application positively affected tomato yield (35%-50%) and phosphorus concentration (24%-29%) compared with controls. AMF application had a significant impact on the total nitrogen (+9%), manganese (+12%), and hydrophilic phenol (+8%) contents in the fruit. Volatile compounds were affected by the interactive effects of fertilisation and AMF application. The response of tomato fruit sensory quality indicators was relatively modest, with only a few sensory characteristics affected to a lesser extent. Although tomato showed susceptibility to field-native AMF, particular combinations of fertilisation and AMF inoculation were more effective at improving the quality parameters of tomatoes under field conditions applied in this study.

Predominance and Diversity of GLRaV-3 in Native Vines of Mediterranean Croatia.

Sixteen grapevine cultivars from Mediterranean Croatia were surveyed for the presence of 10 of the most economically important grapevine viruses. The presence of Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Grapevine leafroll associated virus-1, -2, and -3 (GLRaV-1; GLRaV-2 and GLRaV-3), Grapevine virus A (GVA) and B (GVB), Grapevine fleck virus (GFkV), Grapevine rupestris stem pitting associated virus (GRSPaV), and Grapevine Pinot gris virus (GPGV) were tested by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). All 71 analyzed clones were positive for the presence of one or more viruses. The most abundant one, detected in almost 95% of samples was GLRaV-3. In most of cases it was reported in mixed infections with GVA, GRSPaV, and GPGV. Virus genomes of GLRaV-3 infected vines were further characterized molecularly in order to determine their genetic diversity. Different genomic variants of heat shock 70 protein homologue (HSP70h) were identified by single-strand conformation polymorphism (SSCP) and sequenced. Sequence analysis confirmed their clustering into phylogenetic group I and/or phylogenetic group II. This study emphasizes the wide virus heterogenicity in Mediterranean vines and the predominant presence of GLRaV-3 phylogenetic groups I and II, either individually or in combination.

Genetic Structure and Relationships among Wild and Cultivated Grapevines from Central Europe and Part of the Western Balkan Peninsula.

The genetic diversity and relationship between wild (Vitis vinifera L. subsp. sylvestris (Gmel.) Hegi and cultivated (V. vinifera L. subsp. vinifera) grapevine in the western Balkan region and Central Europe have not been studied together previously, although this area has a rich viticultural past. Here, we studied wild grapevine populations sampled from their natural habitats in several countries of the western Balkan region and Central Europe. Their genetic diversity and structure were compared to cultivars that are traditionally in use in this region. A sample set of 243 accessions was genotyped at 20 nuclear microsatellite loci, including 167 sylvestris and 76 diverse vinifera cultivars. The genetic diversity of the wild grapevines was lower than that of cultivars by all genetic parameters. Both hierarchical and nonhierarchical clustering methods differentiated two main groups, indicating clear separation between wild and cultivated vines but also revealed clear gene flow between the cultivated and wild gene pools through overlaps and admixed ancestry values in the graphs. There was greater affinity to the wild grapes in Central European cultivars than in Balkan cultivars. Fine arrangement of the structure among cultivated grapevines showed differentiation among Central European and Balkan cultivars. These results confirm the divergence of wild grapes from vinifera and highlight the "crossroad" role of the western Balkan peninsula in the broader context of European viticulture.

Marine bacterioplankton production of polysaccharidic and proteinaceous particles under different nutrient regimes.

The influence of inorganic nutrient concentrations on the ability of bacterioplankton to produce and degrade polysaccharidic transparent exopolymer particles (TEPs) and proteinaceous Coomassie-stained particles (CSPs) was investigated in an 11-day experiment. The dynamics of these particles were followed in prefiltered (1 microm) northern Adriatic seawater enclosures enriched either with 1 microM orthophosphate (main limiting nutrient in this area), 10 microM ammonium or both orthophosphate and ammonium. These enclosures were referenced to a nonenriched control. A high potential for bacterial TEP and CSP production was observed (10(4) - 10(5) L(-1) for particles larger than 4 microm). In conditions of high orthophosphate concentration (either orthophosphate enriched or both orthophosphate and ammonium enriched), lower abundances and surface areas of CSPs were obtained, whereas TEP dynamics were more affected by unbalanced enrichments where only orthophosphate or ammonium was added. The impact of unbalanced nutrient ratios on TEPs was indicated by their higher abundance but low capacity for Alcian blue absorption, implying a change in their structure. Inorganic nutrient availability was thus proven to affect the bacterial potential for producing and degrading bacterially derived TEPs and CSPs.

Transparent exopolymeric particles' distribution in the northern Adriatic and their relation to microphytoplankton biomass and composition.

Transparent exopolymer particles (TEP) are generated during transformation processes within the non-living organic matter continuum in the seawater. Seasonal dynamics of TEP along the Po River Delta-Rovinj transect was followed during the 3-year period, using a spectrophotometric method of determination of TEP. Relationships between TEP and phytoplankton biomass (chlorophyll a), abundance and composition, as well as phaeophytin, were analysed. Microphytoplankton spring blooms are confirmed to be an important source of TEP, with some species more related to TEP concentration dynamics, such as Chaetoceros sp. and Skeletonema costatum. Notable spatial west to east gradient of autotrophic biomass and consequently of TEP was observed, as well as gradual decrease with depth. In addition, TEP concentration increments were observed up to 2 months before the mucilage events followed by the decrease of TEP concentration during the event itself. Pattern of TEP dynamics in the years with mucilage events was the most pronounced in 2002, when the event was spatially and temporally the most extended.

Relationships between heterotrophic bacteria and cyanobacteria in the northern Adriatic in relation to the mucilage phenomenon.

High variability of heterotrophic bacterial (HB; 0.1.10(9)-6.10(9) cells L(-1)), nanoflagellates (HNF; 0.02.10(6)-2.4.10(6) cells L(-1)) and cyanobacterial (CB; 10(6)-700.10(6) cells L(-1)) abundances were observed during approximately monthly measurements at six stations along the transect Po Delta-Rovinj from March 1999 to August 2002. Substantially higher values were observed during the stratification period (June-September) in the surface layer of the western stations that were under more direct influence of Po River discharges. Changes of the HB abundance were significantly correlated with temperature in the entire water column and chlorophyll a (Chla) concentration in the surface layer. The nutrients did not look to have directly influenced the HB growth, except orthophosphate in the eastern, more oligotrophic part of transect, where probably HB efficiently competed with phytoplankton for this nutrient. Temperature was also important for CB growth that, however, appeared to occur more intensively in waters with low nutrient concentrations. Probably, in these conditions CB can still develop, while larger autotrophs are strongly nutrient limited. In the upper water column of reduced salinity (35-37), the contributions of CB carbon biomass to the total picoplankton biomass (CB+HB) were mostly larger than 30% (up to 80%) in years with mucilage events (1991, 2000-2002) than in other years. For example, in 1999 only a few values were higher than 30% (up to 50%). In abundant presence of mucilaginous aggregates (e.g. in June 2000 and late June 2002) the chlorophyll a ascribed to CB accounted for much larger portions of total measured Chla. From these results it was concluded that cyanobacteria during mucilage events may play a substantially increased role both as primary producers and prey within the microbial loop of the northern Adriatic.

Phytoplankton and bacterial alkaline phosphatase activity in the northern Adriatic Sea.

The importance of bacterial, phytoplankton and dissolved alkaline phosphatase activity (APA) in the northern Adriatic was investigated during 2006. In surface waters total APA increased from early spring (0.07-0.08 micromol l(-1) h(-1)) to late spring (up to 4.64 micromol l(-1) h(-1)) and remained relatively high during the summer (0.46-0.71 micromol l(-1) h(-1)), due to an increase in specific phytoplankton (up to 30 nmol microg C(-1) h(-1)) and bacterial APA (up to 17.11 nmol microg C(-1) h(-1)). Activity of free enzymes was not important. During late spring and summer both communities exploited dissolved organic phosphorus although, taking into account biomass, phytoplankton activity usually dominated over bacterial activity. In autumn an extra P supply from deeper waters drastically reduced phytoplankton APA, though not bacterial APA, in upper waters. Probably in these months bacteria that were degrading phytoplankton produced organic matter were P limited. In deeper waters APA was low and mainly due to the activity of free enzymes.