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We aimed to test the combination of paraffin oil (PFO) with regular fungicide treatment to assess its efficacy against grape powdery mildew (GPM) in a small spraying experiment on two Vitis vinifera L. cultivars (Chardonnay and Kékfrankos) with different susceptibility to Erysiphe necator. The visual symptoms of GPM on leaves and clusters were examined at three phenological states. The harvest yield was characterized by two methods, data were analyzed with one-way ANOVA and Tukey post-hoc test. Regular fungicide treatment (CT) and its combinations with PFO showed better results in both varieties to repress GPM in 2015 relative to sole PFO treatments. Mean values of combined treatments were often lower than CT but did not differ significantly from each other. The same was observed in 2016, despite the higher pressure of GPM, and missed the third survey. No significant differences were detected between treatments in yield. In contrast, the mean cluster weight of CT and combined treatments resulted in (insignificantly) higher values in each variety and year. In summary, the sole PFO showed some disease control capability as reported earlier, but this effect was greatly affected by the given vintage. Combining PFO with CT resulted in increased protection against GPM relative to the solely applied fungicides. However, this effect was not significant in all cases. It also depended on the vintage and cultivar characteristics. The beneficial impact of paraffin oil as an additive to CT may be due to the induction of plant stress responses and/or its ability to support the adherence and absorption of the combined agents.

Grapevine trunk diseases (GTDs) are a major threat to the wine industry, causing yield loss and dieback of grapevines. While the increasing damage caused by GTDs in recent decades have spurred several studies on grapevine-associated pathogenic fungi, key questions about the emergence and severity of GTDs remain unanswered, including possible differences in plant pathogenic fungal communities in asymptomatic and symptomatic grapevines. We generated fungal DNA metabarcoding data from soil, bark, and perennial wood samples from asymptomatic and symptomatic grapevines sampled in three terroirs. We observed larger compositional differences in plant pathogenic fungi among different plants parts within grapevine plants than among individual grapevines. This is driven by the dominance of GTD-associated fungi in perennial wood and non-GTD pathogens in soil, as well as by the lack of significant differences among asymptomatic and Esca symptomatic grapevines. These results suggest that fungi generally associated with Esca disease belong to the core grapevine microbiome and likely are commensal endophytes and/or latent saprotrophs, some of which can act as opportunistic pathogens on stressed plants. In addition, we found significant compositional differences among sampling sites, particularly in soil, which suggest a certain influence of local edaphic and mesoclimatic factors on plant pathogenic fungal communities. Furthermore, the observed differences among terroirs in plant pathogenic fungal communities in grapevine woody parts indicate that environmental factors likely are important for the development of Esca disease and further studies are needed to investigate the abiotic conditions on fungal compositional dynamics in Esca-affected plants.

Recently, more and more new fungal pathogens have been described as causal agents of grapevine trunk diseases (GTDs), which lead to increasingly significant economic losses in viticulture worldwide. The genus Neofabraea consists of species mainly known as important plant pathogens causing perennial canker and bull’s eye rot, a common postharvest disease of apple (Malus domestica) and pear (Pyrus communis) fruits. Neofabraea kienholzii also causes lesions on pome fruits and canker on woody tissues, but its pathogenicity has not been demonstrated on grapevine yet. In 2015, two strains, identified as N. kienholzii based on ITS sequence data, were isolated from vines showing symptoms of GTDs in Hungary. For an unambiguous taxonomic placement of the isolates, four loci (ITS, LSU, TUB2 and RPB2) were amplified and sequenced. The phylogenetic analysis confirmed that the two isolates represent N. kienholzii. Pathogenicity tests performed on potted grapevines, shoots, and canes confirmed the virulence of these fungi. Their growth and sporulation on different media were also investigated. To the best of our knowledge, this is the first proof of N. kienholzii might cause symptoms on Vitis and might have a role in GTDs.

Grapevine trunk diseases (GTDs) are a major threat to the wine industry, causing yield loss and dieback of grapevines. While the increasing damage caused by GTDs in recent decades have spurred several studies on grapevine-associated pathogenic fungi, key questions about the emergence and severity of GTDs remain unanswered, including possible differences in plant pathogenic fungal communities in asymptomatic and symptomatic grapevines. We generated fungal DNA metabarcoding data from soil, bark, and perennial wood samples from asymptomatic and symptomatic grapevines sampled in three

Fungal diseases in agronomically important plants such as grapevines result in significantly reduced production, pecuniary losses, and increased use of environmentally damaging chemicals. Beside the well-known diseases, there is an increased interest in wood-colonizing fungal pathogens that infect the woody tissues of grapevines. In 2015, a traditional isolation method was performed on grapevine trunks showing symptoms of trunk diseases in Hungary. One isolate (T15142) was identified as Kalmusia longispora (formerly Dendrothyrium longisporum) according to morphological and phylogenetic analyses. To evaluate the pathogenicity of this fungus on grapevines, artificial infections were carried out under greenhouse and field conditions, including the CBS 824.84 and ex-type CBS 582.83 strains. All isolates could be re-isolated from inoculated plants; however, varying virulence was observed among them in terms of the vascular necrosis caused. The incidence and severity of this symptom seemed to be congruent with the laccase-producing capabilities of the isolates. This is the first report on the ability of Kalmusia longispora to cause symptoms on grapevines, and on its possible dependence on laccase secretion.

Background Mineral oils have been widely used in the pest control of several crops. However, their mode of action is poorly understood, especially in the case of their antifungal properties. The possible direct fungicidal activity and the stress-inducing capability of paraffin oil on grapevine were examined using Vitis vinifera L. cv 'Kekfrankos' cuttings and the fungus Erysiphe necator the causal agent of powdery mildew. Results Our experiments demonstrated that paraffin oil does not have fungicide activity on E. necator, but induces significant stress-related changes in grapevine physiology. Elevated H2 O2 production and the accumulation of the phytohormone salicylic acid were measured. Secondary thickening of the cell wall by lignin deposition and the accumulation of phenolic compounds were also observed. Some enzyme activities related to the detoxification of reactive oxygen species, disease response, and the synthesis of lignin were changed in accordance with the determined changes in cell wall composition and H2 O2 levels. Conclusion The results suggest that paraffin oil induces stress responses on grapevine leaves through oxidative burst, which response is systemized by salicylic acid. The accumulation of lignin and water-soluble phenolics may be directly responsible for the paraffin oil-induced resistance of grapevine against powdery mildew. This article is protected by copyright. All rights reserved.

Grapevine (Vitis vinifera) is a reservoir of fungal endophytes that may affect its growth, health status and grape production. Although there is growing interest in comparing fungal communities of mainly red grape varieties across various factors using only high-throughput sequencing, the small-scale mycobiome variations in geographically close vineyards need further examination. We aimed to characterize the fungal microbiome of the above-ground tissues of V. vinifera cv. Furmint in different plant parts, seasons and sites using culture-dependent and culture-independent methods, and in planta fluorescent microscopic visualization techniques. Samples were collected from four sites of the Tokaj wine region in Mád and two reference sites in Eger, Hungary, across different seasons for 2 years. Fungal endophytes of young and mature leaves, flowers and grape bunches were collected at different phenological stages. Based on each technique, Aureobasidium pullulans, Cladosporium spp. and the complex species Alternaria alternata dominated the community at every site, season and plant organ. We found no significant difference among communities in distinct neighbouring vineyards, nor when compared with the distant reference sites. We can conclude that the different shoot parts of the Furmint grapevines harbour a common core group of fungal community in these regions.

The aim of the present study was to examine the efficiency of paraffin oil against powdery mildew in Eger wine region. The experiment has been carried out in 2013 and 2014 with Chardonnay and Kékfrankos grape varieties, which have different resistance against powdery mildew. The effectiveness of the oil was examined on leaves and clusters (frequency and intensity). This oil was effective against Erysiphe necator infection on field trials in Chile and Brazil. The spread of downy mildew (Plasmopara viticola) was also inhibited by this material in some experiments conducted in Spain and France. The differences between oil treatments represented the sensitivity of the grape varieties in accordance with the applied dosages. The oil was effective against powdery mildew with different extent as a result of the so called ”vintage effect”. In 2013, the treatment of the highest dosage (D3) didn’t differed significantly in frequency and intensity of infection from the regular treatment (clusters of Kékfrankos, leaves of Chardonnay). In 2014, the oil was not so effective against powdery mildew compared to 2013. No remarkable differences were detected between the treatments due to the strong pressure of powdery mildew. Furthermore, no any effect of the lowest dosage (D1) was detected in the case of the sensitive clusters of Chardonnay and leaves of Kékfrankos in both experimental years. In summary, the oil treatment has an effect against powdery mildew, however this efficiency largely depends on the vintage characteristics and the pressure of powdery mildew. Further investigations are neccessary, for example field trials with combinations of other sprays. The oil can be useable as fungicide with proper care in eco-friendly integrated and bio (ecological) viticulture.

HmbB, a predominantly mitochondrial high-mobility group box (HMGB) protein, of Aspergillus nidulans affects diverse biological activities, such as sterigmatocystin production, the maintenance of mitochondrial DNA copy number, germination of asexual and sexual spores, and protection against oxidative stress agents. We hypothesized that the latter correlates with an unbalanced intracellular redox state, in which case, a not yet fully characterized physiological function could be attributed to this mitochondrial HMGB protein. Here, we studied the intracellular redox environment and oxidative stress tolerance in hmbB+ and hmbBΔ strains under normal and oxidative stress conditions by measuring glutathione redox couple, intracellular reactive oxygen species (ROS) content and ROS-protecting enzyme activities. Our results revealed that the intracellular redox environment is different in hmbBΔ conidia and mycelia from that of hmbB+, and shed light on the seemingly contradictory difference in the tolerance of hmbBΔ mycelia to diamide and menadione oxidative stressors.

Seven HMG-box proteins of Aspergillus nidulans have been identified in the genomic databases. Three of these have the characteristics of non-specific DNA-binding proteins. One of these, AN1267 (HmbB), comprises one canonical HMG-box in its C-terminus and upstream of the canonical box two structurally related boxes, to be called Shadow-HMG-boxes. This protein defines, together with the Podospora anserina mtHMG1, a clade of proteins present in the Pezizomycotina, with orthologues in some of the Taphrinomycotina. HmbB localizes primarily to the mitochondria but occasionally in nuclei. The deletion of the cognate gene results in a number of pleiotropic effects, including those on hyphal morphology, sensitivity to oxidative stress, absence of sterigmatocystin production and changes in the profile of conidial metabolites. The most striking phenotype of deletion strains is a dramatic decrease in conidial and ascospore viability. We show that this is most likely due to the protein being essential to maintain mitochondrial DNA in spores.

Application of fungicides have advantages and also some direct or indirect disadvantages, such as imbalance and/or fungicide resistance in microbe population. To avoid these problems the development of alternative, eco-friendly methods like mostly spraying with oils are in the focus nowadays. The investigations of the effects of fungicides on microbiota in some cultivations can give a more complex view to this topic and developmental possibilities. In the present study, our aim was testing of the effects of paraffine oil (as alternative fungicide) on microbial properties (CFU and rate of filamentous fungi and yeasts) of Chardonnay and Kékfrankos leaves and berries. Our results from 2014 showed that the application of paraffine oil as sole spray agent can decrease the presence of saprophytic filamentous fungi on the berries of Chardonnay (susceptible for fungal infections). In the case of Kékfrankos berries opposite properties were observed, which may be the result of the absorption of oil by the thick wax layer of this variety. The oil treatment did not affect the yeast population of Chardonnay and Kékfrankos berries contrary to negative effect of the regular pesticide treatment. The selective fungicide effect of paraffine oil against filamentous fungal population caused the accumulation of yeast cells in the mycobiota of grape berries. The careful use of this yeasts in spontaneous fermentation can improve the aroma profile of wines. The year of 2015 did no prefer the growth of fungi, therefore no interesting properties were detected in the mycobiota of grape varieties. The occurence of the harmful saprophytic filamentous fungi predicted to be increased in mild climate agricultures as the result of the climate change. In summary, the paraffine oils are seem to be promising tools for the eco-friendly control of harmful fungi of grapes.

The occurence of atmospherical drought causes serious water-supply problems in the most cases of our domestic agricultural plant species. This paper was studied, how can we quantificate the atmospherical drought, with the help of a low input (relative humidityof the air, temperature) index. If this index (LSZI) characterized the atmospherical drought well, it will suitable to estimate the yield amount of agricultural plants.The index elaborated by the authors was tested on county average crop yield of 14 agricultural plant species. Moreover we compared the atmospherical drought index (LSZI) to other aridity parameters, how suitable for estimate the yield amount.Result of experiments show that, the atmospherical drought index (LSZI) can be used well by several agricultural plant species in especially coern and sugar-beet to estimate yield amount. Excellent results were found by comparison to other aridity indexes, this means it is worth using in the aridity researches in the future.

Seven HMG-box proteins of Aspergillus nidulans have been identified in the genomic databases. Three of these have the characteristics of non-specific DNA-binding proteins. One of these, AN1267 (HmbB), comprises one canonical HMG-box in its C-terminus and upstream of the canonical box two structurally related boxes, to be called Shadow-HMG-boxes. This protein defines, together with the Podospora anserina mtHMG1, a clade of proteins present in the Pezizomycotina, with orthologues in some of the Taphrinomycotina. HmbB localizes primarily to the mitochondria but occasionally in nuclei. The deletion of the cognate gene results in a number of pleiotropic effects, including those on hyphal morphology, sensitivity to oxidative stress, absence of sterigmatocystin production and changes in the profile of conidial metabolites. The most striking phenotype of deletion strains is a dramatic decrease in conidial and ascospore viability. We show that this is most likely due to the protein being essential to maintain mitochondrial DNA in spores.

Neosartorya fischeri antifungal protein (NFAP) is a β-defensin-like peptide produced by the N. fischeri NRRL 181 isolate. In this study, we investigated the manifestation of the antimicrobial effect of NFAP via heterologous expression of the nfap gene in an NFAP-sensitive fungus, Aspergillus nidulans. Heterologous expression of the nfap gene was carried out in A. nidulans CS2902 using a pAMA1-based autonomous replicative vector construct. The effect of the produced NFAP on the germination of A. nidulans conidia was investigated by scanning electron microscopy (SEM), and by DAPI and Calcofluor white (CFW) staining. 2',7'-Dichlorodihydrofluorescein diacetate staining and an Annexin V-FITC Apoptosis Detection kit were used to reveal the accumulation of reactive oxygen species (ROS) and the possible apoptotic, necrotic effect. The impact of mono- and divalent cations on the antimicrobial activity of NFAP was also examined. Transformants expressing the nfap gene showed reduced hyphal growth compared with the untransformed strain. This effect was absent in the presence of mono- and divalent cations (50 and 100 mM KCl, Mg(2)SO(4), Na(2)SO(4)). Delayed and abnormal germination was observed in the case of transformants. Conidia developed short branching germination tubes with swollen tips. The great majority of germinating conidia were destroyed after 8 h of cultivation, although a few survived and developed into abnormal hyphae. Damage in the organization of the cell wall, the destruction of chitin filaments and the accumulation of nuclei at the broken hyphal tips were detected by SEM, DAPI and CFW staining. The accumulation of ROS and more frequent apoptotic, necrotic events were also observed in the case of the NFAP-producing A. nidulans strain.