Curse or blessing: Growth- and laccase-modulating properties of polyphenols and their oxidized derivatives on Botrytis cinerea.

Infection of grapevines with the grey mold pathogen Botrytis cinerea results in severe problems for winemakers worldwide. Browning of wine is caused by the laccase-mediated oxidation of polyphenols. In the last decades, Botrytis management has become increasingly difficult due to the rising number of resistances and the genetic variety of Botrytis strains. During the search for sustainable fungicides, polyphenols showed great potential to inhibit fungal growth. The present study revealed two important aspects regarding the effects of grape-specific polyphenols and their polymerized oxidation products on Botrytis wild strains. On the one hand, laccase-mediated oxidized polyphenols, which resemble the products found in infected grapes, showed the same potential for inhibition of growth and laccase activity, but differed from their native forms. On the other hand, the impact of phenolic compounds on mycelial growth is not correlated to the effect on laccase activity. Instead, mycelial growth and relative specific laccase activity appear to be modulated independently. All phenolic compounds showed not only inhibitory but also inductive effects on fungal growth and/or laccase activity, an observation which is reported for the first time. The simultaneous inhibition of growth and laccase activity demonstrated may serve as a basis for the development of a natural botryticide. Yet, the results showed considerable differences between genetically distinguishable strains, impeding the use of a specific phenolic compound against the genetic variety of wild strains. The present findings might have important implications for future understanding of Botrytis cinerea infections and sustainable Botrytis management including the role of polyphenols.

Culturable yeast community associated with grape must and honey bees sampled from apiaries located in the vineyards.

AIM: In this study, we investigated culturable yeast community, present in grape must sampled from vineyards with apiaries on the borders, and in honey bees collected in these apiaries. METHODS AND RESULTS: To this aim, yeasts isolated from spontaneously fermented grapes randomly collected in two vineyards (P1 and P2) with apiaries on the borders (A1 and A2) were compared to those isolated from spontaneously fermented grapes collected from a vineyard without apiary (P4). At the same time, yeast community was analyzed on bees collected in each apiary placed in the vineyards, in comparison to yeasts isolated from an apiary (A3) located far from the vineyards. The analysis was performed for two consecutive years (2021 and 2022). The isolated yeasts were identified by restriction analysis of amplified ITS region, followed by sequencing of ITS fragment.Our research showed that the presence of apiaries seems to increase yeast counts of grape must, in particular of Saccharomyces cerevisiae; furthermore, the permanence of apiaries in the vineyards allowed the recovering of these yeasts also from bees. CONCLUSIONS: Our findings seem to corroborate the role of bees as vectors and reservoirs of oenologically relevant yeasts, such as a source of non-conventional yeasts with potential biotechnological applications.

Influence of different stress factors during the elaboration of grape must's pieddecuve on the dynamics of yeast populations during alcoholic fermentation.

The pieddecuve (PdC) technique involves using a portion of grape must to undergo spontaneous fermentation, which is then used to inoculate a larger volume of must. This allows for promoting autochthonous yeasts present in the must, which can respect the typicality of the resulting wine. However, the real impact of this practice on the yeast population has not been properly evaluated. In this study, we examined the effects of sulphur dioxide (SO2), temperature, ethanol supplementation, and time on the dynamics and selection of yeasts during spontaneous fermentation to be used as PdC. The experimentation was conducted in a synthetic medium and sterile must using a multi-species yeast consortium and in un-inoculated natural grape must. Saccharomyces cerevisiae dominated both the PdC and fermentations inoculated with commercial wine yeast, displaying similar population growth regardless of the tested conditions. However, using 40 mg/L of SO2 and 1% (v/v) ethanol during spontaneous fermentation of Muscat of Alexandria must allowed the non-Saccharomyces to be dominant during the first stages, regardless of the temperature tested. These findings suggest that it is possible to apply the studied parameters to modulate the yeast population during spontaneous fermentation while confirming the effectiveness of the PdC methodology in controlling alcoholic fermentation.

Characterization and analysis of dynamic changes of microbial community associated with grape decay during storage.

The rot caused by pathogens during the storage of table grapes is an important factor that affects the development of the grape industry and food safety, and it cannot be ignored. The development of innovative methods for pathogen control should be based on a comprehensive understanding of the overall microbial community changes that occur during grape storage. The study aims to investigate the relationship between the native microbiota (including beneficial, pathogenic and spoilage microorganisms) on grape surfaces and the development of disease during grape storage. In this study, the bacteria and fungi present on grape surfaces were analyzed during storage under room temperature conditions using high-throughput sequencing. During the storage of grapes at room temperature, observable diseases and a noticeable decrease in quality were observed at 8 days. Microbial community analysis showed that 4996 bacterial amplicon sequence variants (ASVs) and 488 fungal ASVs were determined. The bacterial richness exhibited an initial increase followed by a subsequent decrease. However, the diversity exhibited a distinct pattern of gradual decrease. The fungal richness and community diversity both exhibit a gradual decrease during the storage of grapes. Fungal ß-diversity analysis showed that despite the absence of rot and the healthy state of grapes on the first and fourth days, the fungal ß-diversity exhibited a significant difference. The analysis of changes in genera abundances suggested that Candidatus Profftella and Aspergillus exhibited dominance in the rotting grape at 16 days, which are the main pathogens that caused disease in the present study. The co-occurrence networks among the microbial showed that the Candidatus proftella genera has a positive correlation with Aspergillus niger, indicating that they work together to cause disease and promote growth in grapes. Predicting the function of bacterial communities found that the microorganisms associated with lipid metabolism at 4 days play an important role in the process of postharvest decay of grapes.

Development of a qPCR method for classification of botrytized grape berries originated from Tokaj wine region.

One of the best-known Hungarian products on world wine market is Aszú, which belongs to the family of Tokaj wine specialties and is made from aszú berries. An important condition for the formation of aszú berries is the noble rot of technologically mature grapes, which is caused by Botrytis cinerea. At the same time botrytized sweet wines are produced not only in Hungary, but in many locations of wine-producing areas of Europe as well as in certain wine growing regions of other continents. The determination of botrytization is mostly based on sensory evaluations, which is a highly subjective procedure and largely depends on the training and experience of the evaluator. Currently, the classification of aszú berries (class I and class II) is based only on visual inspection and determination of sugar content. Based on these facts the primary goal of our work was to develop a qPCR assay capable for objective rating and classification of aszú berries. The developed qPCR is highly specific and sensitive as can clearly distinguish between B. cinerea and other filamentous fungi and yeast species occur on grapes. Moreover, it is suitable for categorizing berries colonized by B. cinerea to varying degrees. Thus, the developed qPCR method can be a useful technique for classification of the grape berries into four quality groups: healthy, semi-shrivelled, Aszú Class II and Aszú Class I.

Multiple deletions of candidate effector genes lead to the breakdown of partial grapevine resistance to downy mildew.

Grapevine downy mildew, caused by the oomycete Plasmopara viticola (P. viticola, Berk. & M. A. Curtis; Berl. & De Toni), is a global threat to Eurasian wine grapes Vitis vinifera. Although resistant grapevine varieties are becoming more accessible, P. viticola populations are rapidly evolving to overcome these resistances. We aimed to uncover avirulence genes related to Rpv3.1-mediated grapevine resistance. We sequenced the genomes and characterized the development of 136 P. viticola strains on resistant and sensitive grapevine cultivars. A genome-wide association study was conducted to identify genomic variations associated with resistant-breaking phenotypes. We identified a genomic region associated with the breakdown of Rpv3.1 grapevine resistance (avrRpv3.1 locus). A diploid-aware reassembly of the P. viticola INRA-Pv221 genome revealed structural variations in this locus, including a 30 kbp deletion. Virulent P. viticola strains displayed multiple deletions on both haplotypes at the avrRpv3.1 locus. These deletions involve two paralog genes coding for proteins with 800-900 amino acids and signal peptides. These proteins exhibited a structure featuring LWY-fold structural modules, common among oomycete effectors. When transiently expressed, these proteins induced cell death in grapevines carrying Rpv3.1 resistance, confirming their avirulence nature. This discovery sheds light on the genetic mechanisms enabling P. viticola to adapt to grapevine resistance, laying a foundation for developing strategies to manage this destructive crop pathogen.

The formation of volatiles in fruit wine process and its impact on wine quality.

Fruit wine is one of the oldest fermented beverages made from non-grape fruits. Owing to the differences in fruit varieties, growing regions, climates, and harvesting seasons, the nutritional compositions of fruits (sugars, organic acids, etc.) are different. Therefore, the fermentation process and microorganisms involved are varied for a particular fruit selected for wine production, resulting in differences in volatile compound formation, which ultimately determine the quality of fruit wine. This article reviews the effects of various factors involved in fruit wine making, especially the particular modifications differing from the grape winemaking process and the selected strains suitable for the specific fruit wine fermentation, on the formation of volatile compounds, flavor and aroma profiles, and quality characteristics of the wine thus produced. KEY POINTS: • The volatile profile and fruit wine quality are affected by enological parameters. • The composition and content of nutrients in fruit must impact volatile profiles. • Yeast and LAB are the key determining factors of the volatile profiles of fruit wines.

Two Paenibacillus spp. strains promote grapevine wood degradation by the fungus Fomitiporia mediterranea: from degradation experiments to genome analyses.

Ascomycetes, basidiomycetes and deuteromycetes can degrade wood, but less attention has been paid to basidiomycetes involved in Esca, a major Grapevine Trunk Disease. Using a wood sawdust microcosm system, we compared the wood degradation of three grapevine cultivars inoculated with Fomitiporia mediterranea M. Fisch, a basidiomycete responsible for white-rot development and involved in Esca disease. The grapevine cultivar Ugni blanc was more susceptible to wood degradation caused by F. mediterranea than the cultivars Cabernet Sauvignon and Merlot. Solid-state Nuclear Magnetic Resonance (NMR) spectroscopy showed that F. mediterranea preferentially degrades lignin and hemicellulose over cellulose (preferential, successive or sequential white-rot). In addition, co-inoculation of sawdust with two cellulolytic and xylanolytic bacterial strains of Paenibacillus (Nakamura) Ash (Paenibacillus sp. (S231-2) and P. amylolyticus (S293)), enhanced F. mediterranea ability to degrade Ugni blanc. The NMR data further showed that the increase in Ugni blanc sawdust degradation products was greater when bacteria and fungi were inoculated together. We also demonstrated that these two bacterial strains could degrade the wood components of Ugni blanc sawdust. Genome analysis of these bacterial strains revealed numerous genes predicted to be involved in cellulose, hemicellulose, and lignin degradation, as well as several other genes related to bacteria-fungi interactions and endophytism inside the plant. The occurrence of this type of bacteria-fungus interaction could explain, at least in part, why necrosis develops extensively in certain grapevine varieties such as Ugni blanc.

Physical cell-cell contact elicits specific transcriptomic responses in wine yeast species.

Fermenting grape juice provides a habitat for a well-mapped and evolutionarily relevant microbial ecosystem consisting of many natural or inoculated strains of yeasts and bacteria. The molecular nature of many of the ecological interactions within this ecosystem remains poorly understood, with the partial exception of interactions of a metabolic nature such as competition for nutrients and production of toxic metabolites/peptides. Data suggest that physical contact between species plays a significant role in the phenotypic outcome of interspecies interactions. However, the molecular nature of the mechanisms regulating these phenotypes remains unknown. Here, we present a transcriptomic analysis of physical versus metabolic contact between two wine relevant yeast species, Saccharomyces cerevisiae and Lachancea thermotolerans. The data show that these species respond to the physical presence of the other species. In S. cerevisiae, physical contact results in the upregulation of genes involved in maintaining cell wall integrity, cell wall structural components, and genes involved in the production of H2S. In L. thermotolerans, HSP stress response genes were the most significantly upregulated gene family. Both yeasts downregulated genes belonging to the FLO family, some of which play prominent roles in cellular adhesion. qPCR analysis indicates that the expression of some of these genes is regulated in a species-specific manner, suggesting that yeasts adjust gene expression to specific biotic challenges or interspecies interactions. These findings provide fundamental insights into yeast interactions and evolutionary adaptations of these species to the wine ecosystem.IMPORTANCEWithin the wine ecosystem, yeasts are the most relevant contributors to alcoholic fermentation and wine organoleptic characteristics. While some studies have described yeast-yeast interactions during alcoholic fermentation, such interactions remain ill-defined, and little is understood regarding the molecular mechanisms behind many of the phenotypes observed when two or more species are co-cultured. In particular, no study has investigated transcriptional regulation in response to physical interspecies cell-cell contact, as opposed to the generally better understood/characterized metabolic interactions. These data are of direct relevance to our understanding of microbial ecological interactions in general while also creating opportunities to improve ecosystem-based biotechnological applications such as wine fermentation. Furthermore, the presence of competitor species has rarely been considered an evolutionary biotic selection pressure. In this context, the data reveal novel gene functions. This, and further such analysis, is likely to significantly enlarge the genome annotation space.

ADP-ribosylation factor 1 (ARF1) protein interacts with elicitor PvNLP7 from Plasmopara viticola to mediate PvNLP7-triggered immunity.

Revealing the effector-host molecular interactions is crucial for understanding the host immunity against Plasmopara viticola and devising innovative disease management strategies. As a pathogenic oomycete causing grapevine downy mildew, Plasmopara viticola employs various effectors to manipulate the defense systems of host plants. One of these P. viticola derived effectors is necrosis- and ethylene-inducing peptide 1 (Nep1) -like protein (PvNLP7), which has been known to elicit cell death and immune responses in plants. However, the underlying molecular mechanisms remain obscure, prompting the focus of this study. Through yeast two-hybrid screening, we have identified the Vitis rotundifolia ADP-ribosylation factor (VrARF1) as a host interactor of PvNLP7. This interaction is corroborated through bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assays. Heterologous expression of VrARF1 in Nicotiana benthamiana verifies its accumulation in both the cytoplasm and nucleus, and induction of cell death. Moreover, the VrARF1 gene is strongly induced during early P. viticola infection and upon PvNLP7 transient expression. Overexpression of the VrARF1 gene in grapevine and N. benthamiana enhances resistance to P. viticola and Phytophthora capsici, respectively, via induction of defense related genes PR1 and PR2. Conversely, virus-induced gene silencing (VIGS) of NbARF1 in N. benthamiana, homologous to VrARF1, markedly attenuates PvNLP7-triggered cell death and reduces the expression of four PTI marker genes (PTI5, Acre31, WRKY7 and Cyp71D20) and two defense related genes (PR1 and PR2), rendering plants transiently transformed with PvNLP7 more susceptible to oomycete P. capsici. These findings highlight the role of ARF1 in mediating PvNLP7-induced immunity and indicate its potential as a target for engineering disease-resistant transgenic plants against oomycete pathogens.

De Novo Synthesis of Resveratrol from Sucrose by Metabolically Engineered Yarrowia lipolytica.

Resveratrol, a phenylpropanoid compound, exhibits diverse pharmacological properties, making it a valuable candidate for health and disease management. However, the demand for resveratrol exceeds the capacity of plant extraction methods, necessitating alternative production strategies. Microbial synthesis offers several advantages over plant-based approaches and presents a promising alternative. Yarrowia lipolytica stands out among microbial hosts due to its safe nature, abundant acetyl-CoA and malonyl-CoA availability, and robust pentose phosphate pathway. This study aimed to engineer Y. lipolytica for resveratrol production. The resveratrol biosynthetic pathway was integrated into Y. lipolytica by adding genes encoding tyrosine ammonia lyase from Rhodotorula glutinis, 4-coumarate CoA ligase from Nicotiana tabacum, and stilbene synthase from Vitis vinifera. This resulted in the production of 14.3 mg/L resveratrol. A combination of endogenous and exogenous malonyl-CoA biosynthetic modules was introduced to enhance malonyl-CoA availability. This included genes encoding acetyl-CoA carboxylase 2 from Arabidopsis thaliana, malonyl-CoA synthase, and a malonate transporter protein from Bradyrhizobium diazoefficiens. These strategies increased resveratrol production to 51.8 mg/L. The further optimization of fermentation conditions and the utilization of sucrose as an effective carbon source in YP media enhanced the resveratrol concentration to 141 mg/L in flask fermentation. By combining these strategies, we achieved a titer of 400 mg/L resveratrol in a controlled fed-batch bioreactor. These findings demonstrate the efficacy of Y. lipolytica as a platform for the de novo production of resveratrol and highlight the importance of metabolic engineering, enhancing malonyl-CoA availability, and media optimization for improved resveratrol production.

Eicosapentaenoic acid: New insights into an oomycete-driven elicitor to enhance grapevine immunity.

The widespread use of pesticides in agriculture remains a matter of major concern, prompting a critical need for alternative and sustainable practices. To address this, the use of lipid-derived molecules as elicitors to induce defence responses in grapevine plants was accessed. A Plasmopara viticola fatty acid (FA), eicosapentaenoic acid (EPA) naturally present in oomycetes, but absent in plants, was applied by foliar spraying to the leaves of the susceptible grapevine cultivar (Vitis vinifera cv. Trincadeira), while a host lipid derived phytohormone, jasmonic acid (JA) was used as a molecule known to trigger host defence. Their potential as defence triggers was assessed by analysing the expression of a set of genes related to grapevine defence and evaluating the FA modulation upon elicitation. JA prompted grapevine immunity, altering lipid metabolism and up-regulating the expression of several defence genes. EPA also induced a myriad of responses to the levels typically observed in tolerant plants. Its application activated the transcription of defence gene's regulators, pathogen-related genes and genes involved in phytoalexins biosynthesis. Moreover, EPA application resulted in the alteration of the leaf FA profile, likely by impacting biosynthetic, unsaturation and turnover processes. Although both molecules were able to trigger grapevine defence mechanisms, EPA induced a more robust and prolonged response. This finding establishes EPA as a promising elicitor for an effectively managing grapevine downy mildew diseases.

Temporal and spatial dynamics within the fungal microbiome of grape fermentation.

Over 6 years, we conducted an extensive survey of spontaneous grape fermentations, examining 3105 fungal microbiomes across 14 distinct grape-growing regions. Our investigation into the biodiversity of these fermentations revealed that a small number of highly abundant genera form the core of the initial grape juice microbiome. Consistent with previous studies, we found that the region of origin had the most significant impact on microbial diversity patterns. We also discovered that certain taxa were consistently associated with specific geographical locations and grape varieties, although these taxa represented only a minor portion of the overall diversity in our dataset. Through unsupervised clustering and dimensionality reduction analysis, we identified three unique community types, each exhibiting variations in the abundance of key genera. When we projected these genera onto global branches, it suggested that microbiomes transition between these three broad community types. We further investigated the microbial community composition throughout the fermentation process. Our observations indicated that the initial microbial community composition could predict the diversity during the early stages of fermentation. Notably, Hanseniaspora uvarum emerged as the primary non-Saccharomyces species within this large collection of samples.

Advancements and challenges for brewing aroma-enhancement fruit wines: Microbial metabolizing and brewing techniques.

Aroma, a principal determinant of consumer preference for fruit wines, has recently garnered much attention. Fruit wines brewing was concomitant with complex biochemical reactions, in which a variety of compounds jointly contribute to the aroma quality. To date, the mechanisms underlying the synthesis of aroma compounds and biological regulation methods in fruit wines have remained ambiguous, hindering the further improvement of fruit wines sensory profiles. This review provides a detailed account of the synthesis and regulatory mechanisms of typical aroma compounds and their contributions to the characteristics of wines. Additionally, Comprehensive involves between microflora and the formation of aroma compounds have been emphasized. The microflora-mediated aroma compounds evolution can be controlled by key fermentation techniques to protect and enhance. Meanwhile, the genes impacting key aroma compounds can be identified, which provide references for the rapid screening of aroma-enhanced strains as well as target formation of aroma by modifying relative genes.

Antifungal activity of Serratia plymuthica against the phytopathogenic fungus Alternariatenuissima.

The antifungal activity of Serratia plymuthica CCGG2742, a bacterial strain isolated from grapes berries skin, against a phytopathogenic fungus isolated from blueberries was evaluated in vitro and in vivo. In order to characterize the wild fungal isolate, phylogenetic analysis using concatenated DNA sequences from the RPB2 and TEF1 genes and of the ITS region was performed, allowing the identification of the fungal isolate that was called Alternaria tenuissima CC17. Hyphae morphology, mycelium ultrastructure, conidia and reproductive structures were in agreement with the phylogenetic analysis. The antifungal activity of the S. plymuthica strain was dependent on the composition of the culture medium. The greatest inhibition of mycelial growth of A. tenuissima CC17 by S. plymuthica CCGG2742 was observed on YTS medium, which lacks of an easily assimilable carbon source. Fungal growth medium supplemented with 50 % of bacterial supernatant decreased the conidia germination of A. tenuissima CC17 up to 32 %. Preventive applications of S. plymuthica CCGG2742 to blueberries and tomato leaves at conidia:bacteria ratio of 1:100, protected in 77.8 ± 4.6 % and 98.2 ± 0.6 % to blueberries and tomato leaves from infection caused by A. tenuissima CC17, respectively. To the best of our knowledge, this is the first report on the antifungal activity of S. plymuthica against A. tenuissima, which could be used as a biological control agent of plant diseases caused by this fungal species. In addition, the results of this work could be a starting point to attribute the real importance of A. tenuissima as a pathogen of blueberries in Chile, which until now had been considered almost exclusively to A. alternata. Likewise, this research could be relevant to start developing highly effective strategies based on S. plymuthica CCGG2742 for the control of this important phytopathogenic fungus.

Membrane protein Bcsdr2 mediates biofilm integrity, hyphal growth and virulence of Botrytis cinerea.

Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSV II were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea. KEY POINTS: • The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins. • Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea. • Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.

Is wine perception influenced by sustainability information? Insights from a consumer experiment with fungus resistant grape and organic wines.

Literature has highlighted that the organic attribute has heuristic value for many consumers, representing an overarching signifier of positive characteristics. Nowadays a plethora of alternative systemic approaches side organic production in the aim to improve the overall sustainability of the agrifood sector. Current study, based on blind and informed tasting, measured sustainability information influence on respondents' (n = 162) perceptions of organic and fungus-resistant grape (FRG) white wines. Findings of the within-subject non hypothetical experiment revealed that information has a stronger, positive impact on participants' perception of organic wine (increasing 13 % monetary preferences) compared to FRG wine (+9%). Additionally, attitudinal characteristics driving consumers' preferences towards FRG wine diverge from organic core motivations.

Potential to take root in viticulture? An evaluation of mycorrhizal inoculants on the growth and nutrient uptake of young wine grapes planted in live field soil.

AIMS: Incorporating biofertilizers, such as arbuscular mycorrhizal fungal (AM) fungal inoculants, into vineyard management practices may enhance vine growth and reduce environmental impact. Here, we evaluate the effects of commercially available and local AM fungal inoculants on the growth, root colonization, and nutrient uptake of wine grapes (Vitis vinifera) when planted in a field soil substrate. METHODS AND RESULTS: In a greenhouse experiment, young wine grapes were planted in a field soil substrate and inoculated with one of three commercially available mycorrhizal inoculant products, or one of two locally collected whole soil inoculants. After 4 months of growth, inoculated vines showed no differences in plant biomass, colonization of roots by AM fungi, or foliar macronutrient concentrations compared to uninoculated field soil substrate. However, vines grown with local inoculants had greater shoot biomass than vines grown with mycorrhizal inoculant products. CONCLUSIONS: Although effects from inoculations with AM fungi varied by inoculant type and source, inoculations may not improve young vine performance in field soils with a resident microbial community.

Isolation and Enzymatic Characterization of Fungal Strains from Grapevines with Grapevine Trunk Diseases Symptoms in Central Mexico.

Grapevine production is economically indispensable for the global wine industry. Currently, Mexico cultivates grapevines across approximately 28 500 hectares, ranking as the 26th largest producer worldwide. Given its significance, early detection of plant diseases' causal agents is crucial for preventing outbreaks. Consequently, our study aimed to identify fungal strains in grapevines exhibiting trunk disease symptoms and assess their enzymatic capabilities as indicators of their phytopathogenic potential. We collected plant cultivars, including Malbec, Shiraz, and Tempranillo, from Querétaro, Mexico. In the laboratory, we superficially removed the plant bark to prevent external contamination. Subsequently, the sample was superficially disinfected, and sawdust was generated from the symptomatic tissue. Cultivable fungal strains were isolated using aseptic techniques from the recovered sawdust. Colonies were grown on PDA and identified through a combination of microscopy and DNA-sequencing of the ITS and LSU nrDNA regions, coupled with a BLASTn search in the GenBank database. We evaluated the strains' qualitative ability to degrade cellulose, starch, and lignin using specific media and stains. Using culture morphology and DNA-sequencing, 13 species in seven genera were determined: Acremonium, Aspergillus, Cladosporium, Dydimella, Fusarium, Sarocladium, and Quambalaria. Some isolated strains were able to degrade cellulose or lignin, or starch. These results constitute the first report of these species community in the Americas. Using culture-dependent and DNA-sequencing tools allows the detection of fungal strains to continue monitoring for early prevention of the GTD.

Carbon trade-offs in the fruits of fungus-tolerant Muscadinia × Vitis hybrids exposed to water deficit.

Adopting disease-tolerant grapevines is an efficient option to implement a smarter management strategy limiting the environmental impacts linked to pesticide use. However, little is known on their production of fruit metabolites regarding expected future climate fluctuations, such as increased water shortage. Moreover, previous studies about how water deficit impacts grape composition, lack accuracy due to imprecise timing of fruit sampling. In this study, we phenotyped six new fungus-tolerant genotypes exposed to varying water status in field-grown conditions. The accumulation of water, main cations, primary and secondary metabolites were precisely monitored at the arrest of phloem unloading in fruits, which was targeted at the whole cluster level. The goal was to decipher the effects of both genotype and water deficit on the allocation of carbon into soluble sugars, organic acids, amino acids and anthocyanins. The results revealed that the effect of decreased water availability was specific to each berry component. While fruit sugar concentration remained relatively unaffected, the malic/tartaric acid balance varied based on differences among genotypes. Despite showing contrasted strategies on carbon allocation into berry metabolites, all genotypes reduced fruit yield and the amount of compounds of interest per plant under water deficit, with the extent of reduction being genotype-dependent and correlated with the response of berry volume to plant water status. This first set of data provides information to help reasoning the adaptation of these varieties according to the expected risks of drought and the possibilities of mitigating them through irrigation.