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1.
PLoS One ; 16(10): e0258043, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34653213

RESUMO

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.


Assuntos
Ascomicetos , Lacase/metabolismo , Doenças das Plantas/microbiologia , Vitis/microbiologia , Ascomicetos/isolamento & purificação , Ascomicetos/metabolismo , Ascomicetos/patogenicidade , Virulência
2.
BMC Plant Biol ; 21(1): 499, 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34717550

RESUMO

BACKGROUND: Papain-like cysteine proteases (PLCPs), a large group of cysteine proteases, are structurally related to papain. The members belonging to PLCPs family contribute to plant immunity, senescence, and defense responses in plants. The PLCP gene family has been identified in Arabidopsis, rice, soybean, and cotton. However, no systematic analysis of PLCP genes has been undertaken in grapevine. Since Plasmopara viticola as a destructive pathogen could affect immunity of grapes in the field, we considered that the members belonged to PLCPs family could play a crucial role in defensive mechanisms or programmed cell death. We aimed to evaluate the role of PLCPs in 2 different varieties of grapevines and compared the changes of their expressions with the transcriptional data in response to P. viticola. RESULTS: In this study, 23 grapevine PLCP (VvPLCP) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Numerous cis-acting elements related to plant development, hormone, and stress responses were identified in the promoter of the VvPLCP genes. Phylogenetic analysis grouped the VvPLCP genes into nine subgroups. The transcription of VvPLCP in different inoculation time points and varieties indicated that VvPLCP may have vital functions in grapevine defense against Plasmopara viticola. According to transcriptome data and qPCR analysis, we observed the increasing expression levels of VvRD21-1 at 72 h after inoculation in resistant variety, inferring that it was related to grape downy mildew resistance. Meanwhile, 3 genes including VvXBCP1, VvSAG12-1, and VvALP1 showed higher expression at 24 h after pathogen inoculation in the susceptible variety and might be related to the downy mildew phenotype. We nominated these four genes to function during hypersensitive response (HR) process, inferring that these genes could be associated with downy mildew resistance in grapes. CONCLUSIONS: Our results provide the reference for functional studies of PLCP gene family, and highlight its functions in grapevine defense against P. viticola. The results help us to better understand the complexity of the PLCP gene family in plant immunity and provide valuable information for future functional characterization of specific genes in grapevine.


Assuntos
Resistência à Doença/genética , Resistência à Doença/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Vitis/genética , Vitis/microbiologia , Produtos Agrícolas/genética , Produtos Agrícolas/microbiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas
3.
Molecules ; 26(15)2021 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-34361722

RESUMO

Off-flavors produced by undesirable microbial spoilage are a major concern in wineries, as they affect wine quality. This situation is worse in warm areas affected by global warming because of the resulting higher pHs in wines. Natural biotechnologies can aid in effectively controlling these processes, while reducing the use of chemical preservatives such as SO2. Bioacidification reduces the development of spoilage yeasts and bacteria, but also increases the amount of molecular SO2, which allows for lower total levels. The use of non-Saccharomyces yeasts, such as Lachancea thermotolerans, results in effective acidification through the production of lactic acid from sugars. Furthermore, high lactic acid contents (>4 g/L) inhibit lactic acid bacteria and have some effect on Brettanomyces. Additionally, the use of yeasts with hydroxycinnamate decarboxylase (HCDC) activity can be useful to promote the fermentative formation of stable vinylphenolic pyranoanthocyanins, reducing the amount of ethylphenol precursors. This biotechnology increases the amount of stable pigments and simultaneously prevents the formation of high contents of ethylphenols, even when the wine is contaminated by Brettanomyces.


Assuntos
Brettanomyces/metabolismo , Aromatizantes/metabolismo , Tecnologia de Alimentos/métodos , Odorantes/análise , Saccharomycetales/metabolismo , Vinho/análise , Antocianinas/metabolismo , Carboxiliases/metabolismo , Fermentação , Proteínas Fúngicas/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Dióxido de Enxofre/farmacologia , Vitis/metabolismo , Vitis/microbiologia , Vinho/microbiologia
4.
Molecules ; 26(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34443337

RESUMO

Real-time process metrics are standard for the majority of fermentation-based industries but have not been widely adopted by the wine industry. In this study, replicate fermentations were conducted with temperature as the main process parameter and assessed via in-line Oxidation Reduction Potential (ORP) probes and at-line profiling of phenolics compounds by UV-Vis spectroscopy. The California and Oregon vineyards used in this study displayed consistent vinification outcomes over five vintages and are representative of sites producing faster- and slower-fermenting musts. The selected sites have been previously characterized by fermentation kinetics, elemental profile, phenolics, and sensory analysis. ORP probes were integrated into individual fermentors to record how ORP changed throughout the fermentation process. The ORP profiles generally followed expected trends with deviations revealing previously undetectable process differences between sites and replicates. Site-specific differences were also observed in phenolic and anthocyanin extraction. Elemental composition was also analyzed for each vineyard, revealing distinctive profiles that correlated with the fermentation kinetics and may influence the redox status of these wines. The rapid ORP responses observed related to winemaking decisions and yeast activity suggest ORP is a useful process parameter that should be tracked in addition to Brix, temperature, and phenolics extraction for monitoring fermentations.


Assuntos
Fermentação , Espectrofotometria Ultravioleta , Vinho/microbiologia , Oxirredução , Vitis/química , Vitis/microbiologia , Vinho/análise
5.
Molecules ; 26(15)2021 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-34361746

RESUMO

New agricultural strategies aim to reduce the use of pesticides due to their damage to the environment and humans, and the caused resistance to pathogens. Therefore, alternative sources of antifungal compounds from plants are under investigation lately. Extracts from plants have a wide composition of chemical compounds which may complicate the development of pathogen resistance. Botrytis cinerea, causing grey mould, is an important horticultural and ornamental pathogen, responsible for the relevant yield and quality losses. B. cinerea isolated from a different plant host may differ in the sensitivity to antifungal substances from plants. Assessing the importance of research covering a wide range of pathogens for the rapid development of biopesticides, this study aims to determine the sensitivity of the B. cinerea isolate complex (10 strains) to plant extracts, describe morphological changes caused by the extract treatment, and detect differences between the sensitivity of different plant host isolates. The results showed the highest sensitivity of the B. cinerea isolates complex to cinnamon extract, and the lowest to laurel extract. In contrast, laurel extract caused the most changes of morphological attributes in the isolates. Five B. cinerea isolates from plant hosts of raspberry, cabbage, apple, bell pepper, and rose were grouped statistically according to their sensitivity to laurel extract. Meanwhile, the bell pepper isolate separated from the isolate complex based on its sensitivity to clove extract, and the strawberry and apple isolates based on their sensitivity to cinnamon extract.


Assuntos
Antifúngicos/farmacologia , Agentes de Controle Biológico/farmacologia , Botrytis/efeitos dos fármacos , Cinnamomum zeylanicum/química , Hifas/efeitos dos fármacos , Doenças das Plantas/prevenção & controle , Antifúngicos/isolamento & purificação , Agentes de Controle Biológico/isolamento & purificação , Botrytis/crescimento & desenvolvimento , Botrytis/isolamento & purificação , Brassica/microbiologia , Capsicum/microbiologia , Cinnamomum camphora/química , Fragaria/microbiologia , Humanos , Hifas/crescimento & desenvolvimento , Hifas/isolamento & purificação , Malus/microbiologia , Testes de Sensibilidade Microbiana , Doenças das Plantas/microbiologia , Extratos Vegetais/química , Syzygium/química , Vitis/microbiologia
6.
Sci Rep ; 11(1): 16367, 2021 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-34385541

RESUMO

This study is designed to understand the community structure and diversity of fungi in the rhizosphere soil of grape. As the sample for this study, the rhizosphere soil of Crimson seedless grape with different planting years was collected from Shihezi in Xinjiang to carry out high-throughput sequencing, by which the complete sequence of soil fungi DNA was identified, and accordingly, the richness and diversity index of fungi were determined. The results showed that the dominant phyla of fungi in the grape rhizosphere soil with different planting years were Ascomycota and Basidiomycota, and the dominant classes of fungi were Sordariomycetes and Dothideomycetes. Soil organic matter, total potassium, total nitrogen and available phosphorus were the main soil fertility factors affecting the abundance and diversity of soil fungal communities, among which soil organic matter had the most significant influence. In addition, the fungal diversity and richness were highest in the middle layer (20-35 cm) of the grape rhizosphere soil with 12 planting years and lowest in the lower layer (35-50 cm) of the grape rhizosphere soil with 5 planting years. Linear discriminant analysis suggested that there were more biomarkers in the vineyard rhizosphere soil with 10 planting years, which meant there were more fungal communities with significant difference in the soil, especially in the middle layer (20-35). The results of this study can provide data reference and theoretical basis for improving vineyard soil quality, evaluating soil microecological effects and improving ecological environment of vineyard soil.


Assuntos
DNA Fúngico/genética , Fungos/genética , Micobioma/genética , Vitis/microbiologia , Biodiversidade , Nitrogênio/metabolismo , Fósforo/metabolismo , Rizosfera , Solo , Microbiologia do Solo
7.
Food Microbiol ; 100: 103859, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34416959

RESUMO

Beta-glucosidase is an important enzyme for the hydrolysis of grape glycosides in the course of winemaking. Yeasts are the main producers of ß-glucosidase in winemaking, therefore play an important role in determining wine aroma and flavour. This article discusses common methods for ß-glucosidase evaluation, the ß-glucosidase activity of different Saccharomyces and non- Saccharomyces yeasts and the influences of winemaking conditions, such as glucose and ethanol concentration, low pH environment, fermentation temperature and SO2 level, on their activity. This review further highlights the roles of ß-glucosidase in promoting the release of free volatile compounds especially terpenes and the modification of wine phenolic composition during the winemaking process. Furthermore, this review proposes future research direction in this area and guides wine professionals in yeast selection to improve wine quality.


Assuntos
Proteínas Fúngicas/metabolismo , Compostos Orgânicos Voláteis/metabolismo , Vinho/análise , Leveduras/enzimologia , beta-Glucosidase/metabolismo , Proteínas Fúngicas/genética , Odorantes/análise , Fenóis/química , Fenóis/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Vitis/química , Vitis/microbiologia , Compostos Orgânicos Voláteis/química , Vinho/microbiologia , Leveduras/genética , Leveduras/metabolismo , beta-Glucosidase/genética
8.
Food Microbiol ; 100: 103865, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34416965

RESUMO

The purpose of this study was to evaluate the inhibitory effect of allyl-isothiocyanate (AITC) and benzyl-isothiocyanate (BITC) on fungal growth and Ochratoxin A (OTA) production by Aspergillus ochraceus, A. carbonarius and A. niger. Here, we found that spore germination and fungal growth of the three fungi were significantly inhibited when the concentration of AITC and BITC was higher than 1.25 µg/mL. The inhibitory effect of AITC or BITC on A. carbonaceus and A. ochraceus was significantly stronger than that of A. niger. Scanning electron microscopy showed that the mycelia of all three fungi were changed by AITC and BITC. Compared with A. ochraceus and A. carbonarius, the damage to A. niger was lower. For OTA production, AITC and BITC could significantly down-regulated the expression of all five OTA biosynthesis genes in A. niger and A. carbonarius. In A. ochraceus, although several OTA biosynthesis genes were up-regulated, the key PKS gene was down-regulated by AITC and BITC. Twenty-five µg/mL of AITC or BITC could reduce the infection of the three fungi on grapes with inhibition rates of 28%-36% during 14 days and prolong the shelf life of grapes. In maize, the OTA production of the three fungi was significantly reduced by 25 µg/mL of AITC and BITC with the inhibition rates 68.04%-93.49% and 65.87%-75.45%, respectively. These results suggest that AITC and BITC can be used as natural fungicides to prevent A. niger, A. carbonarius and A. ochraceus from infecting grapes and maize and control OTA contamination.


Assuntos
Fungos/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Isotiocianatos/farmacologia , Ocratoxinas/biossíntese , Vitis/microbiologia , Zea mays/microbiologia , Contaminação de Alimentos/análise , Fungos/crescimento & desenvolvimento , Fungos/metabolismo
9.
Plant Sci ; 310: 110989, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34315603

RESUMO

Resveratrol (3,5,4'-trihydroxy-stilbene) is a phytoalexin that can prevent plants from pathogen attacks. Piceid is the glycosylation product of resveratrol and the main storage form of stilbenes in grapevines. Here, we reported the function of a ß-glycoside hydrolase gene, VqBGH40a, from the Chinese wild grapevine Vitis quinquangularis accession Danfeng-2 in the regulation of plant resistance to powdery mildew (Uncinula necator). VqBGH40a belonging to ß-glycoside hydrolase family 1 encoded 506 amino acids and was located on the cytomembrane. Its optimal induction condition was 28 or 30℃, for 4 h, with 0.1 mM IPTG in a prokaryotic expression system. Enzyme activity detection showed that purified VqBGH40a could hydrolyze trans-piceid to form trans-resveratrol in vitro. VqBGH40a was transiently overexpressed in Danfeng-2 leaves and then artificially inoculated with powdery mildew showed that VqBGH40a protein could hydrolyze trans-piceid in vivo. Additionally, a comparative family analysis between VqBGH40a and 38 VviBGHs was performed. Overall, these results demonstrate that VqBGH40a can hydrolyze trans-piceid, enhance trans-resveratrol content, and participate in the defense mechanism of grapevine against powdery mildew.


Assuntos
Resveratrol/farmacologia , Vitis/química , Resistência à Doença , Erysiphe/patogenicidade , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Vitis/microbiologia
10.
Fitoterapia ; 153: 105000, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34303765

RESUMO

Six new polyketides, alternaritins A-D [(±)-1-4] and isoxanalteric acid I (8), and 25 known Alternaria toxins were isolated from the culture of an endophytic fungi Alternaria sp. MG1. 3 is a rare fungal metabolite. 6 is a new natural product, and 5, 7, and 9 are known previously but their absolute configurations have not been determined. Three enantiomers [(±)-1, (±)-7, and (±)-15] were separated via chiral HPLC resolution. The structures of those polyketides (1-9) were elucidated by spectrometric analysis using MS and NMR. The absolute configurations were established using X-ray diffraction analysis and statistical comparative analysis of the experimental ECD and OR data, in conjunction with quantum mechanical calculations. All of the compounds were evaluated for their bioactivities. Known compound 27 exerted the most potent cytotoxic activities against HT-1080 and NCI-H1299 cell lines. The new compounds, 2 and 3, showed moderate inhibition on COX-2, while a pair of isomers, 8 and 9, exhibited medium activity on COX-2 and uropathogenic Escherichia coli.


Assuntos
Alternaria/química , Anti-Inflamatórios/farmacologia , Antineoplásicos/farmacologia , Policetídeos/farmacologia , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Anti-Inflamatórios/isolamento & purificação , Antineoplásicos/isolamento & purificação , Produtos Biológicos/isolamento & purificação , Produtos Biológicos/farmacologia , Linhagem Celular Tumoral , Inibidores de Ciclo-Oxigenase 2/isolamento & purificação , Inibidores de Ciclo-Oxigenase 2/farmacologia , Endófitos/química , Humanos , Estrutura Molecular , Policetídeos/isolamento & purificação , Estereoisomerismo , Escherichia coli Uropatogênica/efeitos dos fármacos , Vitis/microbiologia
11.
Molecules ; 26(14)2021 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-34299533

RESUMO

Grapevine is susceptible to fungal diseases generally controlled by numerous chemical fungicides. Elicitors of plant defence are a way of reducing the use of these chemicals, but still provide inconsistent efficiency. Easy-to-analyse markers of grapevine responses to elicitors are needed to determine the best conditions for their efficiency and position them in protection strategies. We previously reported that the elicitor sulphated laminarin induced the emission of volatile organic compounds (VOCs) by grapevine leaves. The present study was conducted to characterise and compare VOC emissions in response to other elicitors. Bastid® was first used to test the conditions of VOC collection and analysis. Using SBSE-GC-MS, we detected several VOCs, including the sesquiterpene α-farnesene, in a time-dependent manner. This was correlated with the induction of farnesene synthase gene expression, in parallel with stilbene synthesis (another defence response), and associated to resistance against downy mildew. The other elicitors (Redeli®, Romeo®, Bion®, chitosan, and an oligogalacturonide) induced VOC emission, but with qualitative and quantitative differences. VOC emission thus constitutes a response of grapevine to elicitors of various chemical structures. Therefore, VOC analysis is relevant for studying the impact of environmental factors on grapevine defence responses and optimising the performance of elicitors in vineyards.


Assuntos
Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Vitis/microbiologia , Compostos Orgânicos Voláteis/análise , Fungicidas Industriais/análise , Cromatografia Gasosa-Espectrometria de Massas , Folhas de Planta/química , Sesquiterpenos/análise , Vitis/química
12.
Sci Rep ; 11(1): 13924, 2021 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-34230575

RESUMO

Powdery mildews are notorious fungal plant pathogens but only limited information exists on their genomes. Here we present the mitochondrial genome of the grape powdery mildew fungus Erysiphe necator and a high-quality mitochondrial gene annotation generated through cloning and Sanger sequencing of full-length cDNA clones. The E. necator mitochondrial genome consists of a circular DNA sequence of 188,577 bp that harbors a core set of 14 protein-coding genes that are typically present in fungal mitochondrial genomes, along with genes encoding the small and large ribosomal subunits, a ribosomal protein S3, and 25 mitochondrial-encoded transfer RNAs (mt-tRNAs). Interestingly, it also exhibits a distinct gene organization with atypical bicistronic-like expression of the nad4L/nad5 and atp6/nad3 gene pairs, and contains a large number of 70 introns, making it one of the richest in introns mitochondrial genomes among fungi. Sixty-four intronic ORFs were also found, most of which encoded homing endonucleases of the LAGLIDADG or GIY-YIG families. Further comparative analysis of five E. necator isolates revealed 203 polymorphic sites, but only five were located within exons of the core mitochondrial genes. These results provide insights into the organization of mitochondrial genomes of powdery mildews and represent valuable resources for population genetic and evolutionary studies.


Assuntos
Ascomicetos/fisiologia , Erysiphe/fisiologia , Genoma Mitocondrial , Íntrons/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Vitis/genética , Vitis/microbiologia , DNA Complementar/genética , DNA Mitocondrial/genética , Erysiphe/isolamento & purificação , Genes Mitocondriais , Genes de Plantas , Anotação de Sequência Molecular , Fases de Leitura Aberta/genética , Filogenia , Polimorfismo Genético
13.
Sci Rep ; 11(1): 14098, 2021 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-34239007

RESUMO

Grapevine trunk diseases (GTDs) are well-known and significant fungal diseases of Vitis vinifera with a worldwide distribution. During August to November 2016 in a survey to characterize fungi associated with grapevine trunk diseases in Kermanshah Province (west of Iran) vineyards, 286 fungal isolates were obtained. Based on morphology and DNA sequences data eight species were identified, of which Biscogniauxia rosacearum, Neoscytalidium hyalinum and Phaeoacremonium minimum were the most aggressive fungal pathogenic species characterized in this research. N. hyalinum was the most prevalent species. N. hyalinum and Ph. minimum have previously been reported from Vitis vinifera. Thus far, there are two records of Biscogniauxia mediterranea and Biscogniauxia capnodes on grapevine in the world with no data on pathology aspects. To our knowledge, it is the first time B. rosacearum is reported from grapevine across the globe. Pathogenicity test with three strains of B. rosacearum on 2-year-old potted grapevines confirmed the pathogenicity of B. rosacearum on grapevine. The proximity of vineyards to the oak trees in Zagros forests as one of the plant hosts of Biscogniauxia spp. further highlights the need for extensive studies on B. rosacearum as a new fungal pathogen.


Assuntos
Ascomicetos/patogenicidade , Doenças das Plantas/microbiologia , Vitis/microbiologia , Ascomicetos/genética , Ascomicetos/isolamento & purificação , DNA Intergênico/genética , Geografia , Irã (Geográfico) , Filogenia
14.
BMC Plant Biol ; 21(1): 265, 2021 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-34103007

RESUMO

BACKGROUND: Long non-coding RNAs (lncRNAs) are regulatory transcripts of length > 200 nt. Owing to the rapidly progressing RNA-sequencing technologies, lncRNAs are emerging as considerable nodes in the plant antifungal defense networks. Therefore, we investigated their role in Vitis vinifera (grapevine) in response to obligate biotrophic fungal phytopathogens, Erysiphe necator (powdery mildew, PM) and Plasmopara viticola (downy mildew, DM), which impose huge agro-economic burden on grape-growers worldwide. RESULTS: Using computational approach based on RNA-seq data, 71 PM- and 83 DM-responsive V. vinifera lncRNAs were identified and comprehensively examined for their putative functional roles in plant defense response. V. vinifera protein coding sequences (CDS) were also profiled based on expression levels, and 1037 PM-responsive and 670 DM-responsive CDS were identified. Next, co-expression analysis-based functional annotation revealed their association with gene ontology (GO) terms for 'response to stress', 'response to biotic stimulus', 'immune system process', etc. Further investigation based on analysis of domains, enzyme classification, pathways enrichment, transcription factors (TFs), interactions with microRNAs (miRNAs), and real-time quantitative PCR of lncRNAs and co-expressing CDS pairs suggested their involvement in modulation of basal and specific defense responses such as: Ca2+-dependent signaling, cell wall reinforcement, reactive oxygen species metabolism, pathogenesis related proteins accumulation, phytohormonal signal transduction, and secondary metabolism. CONCLUSIONS: Overall, the identified lncRNAs provide insights into the underlying intricacy of grapevine transcriptional reprogramming/post-transcriptional regulation to delay or seize the living cell-dependent pathogen growth. Therefore, in addition to defense-responsive genes such as TFs, the identified lncRNAs can be further examined and leveraged to candidates for biotechnological improvement/breeding to enhance fungal stress resistance in this susceptible fruit crop of economic and nutritional importance.


Assuntos
Resistência à Doença/genética , Resistência à Doença/imunologia , Erysiphe/patogenicidade , Peronospora/patogenicidade , Doenças das Plantas/genética , Imunidade Vegetal/genética , RNA Longo não Codificante , Vitis/genética , Produtos Agrícolas/genética , Produtos Agrícolas/imunologia , Produtos Agrícolas/microbiologia , Erysiphe/imunologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Estudo de Associação Genômica Ampla , Peronospora/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Vitis/imunologia , Vitis/microbiologia
15.
Int J Mol Sci ; 22(11)2021 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-34067363

RESUMO

Grapevine downy mildew, caused by the biotrophic oomycete Plasmopara viticola, is one of the most severe and devastating diseases in viticulture. Unravelling the grapevine defence mechanisms is crucial to develop sustainable disease control measures. Here we provide new insights concerning fatty acid's (FA) desaturation, a fundamental process in lipid remodelling and signalling. Previously, we have provided evidence that lipid signalling is essential in the establishment of the incompatible interaction between grapevine and Plasmopara viticola. In the first hours after pathogen challenge, jasmonic acid (JA) accumulation, activation of its biosynthetic pathway and an accumulation of its precursor, the polyunsaturated α-linolenic acid (C18:3), were observed in the leaves of the tolerant genotype, Regent. This work was aimed at a better comprehension of the desaturation processes occurring after inoculation. We characterised, for the first time in Vitis vinifera, the gene family of the FA desaturases and evaluated their involvement in Regent response to Plasmopara viticola. Upon pathogen challenge, an up-regulation of the expression of plastidial FA desaturases genes was observed, resulting in a higher content of polyunsaturated fatty acids (PUFAs) of chloroplast lipids. This study highlights FA desaturases as key players in membrane remodelling and signalling in grapevine defence towards biotrophic pathogens.


Assuntos
Resistência à Doença/genética , Ácidos Graxos Dessaturases/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Vitis/genética , Vitis/microbiologia , Vias Biossintéticas/genética , Cloroplastos/genética , Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Genótipo , Lipídeos/genética , Oomicetos/patogenicidade , Oxilipinas/metabolismo , Peronospora/patogenicidade , Folhas de Planta/genética , Folhas de Planta/microbiologia
16.
Food Microbiol ; 99: 103824, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34119109

RESUMO

Rapid yeast identification is of particular importance in monitoring wine fermentation and assessing strain application in winemaking. We used MALDI-TOF MS analysis supported by 26 S rRNA gene sequence analysis and Saccharomyces-specific PCR testing to differentiate reference and field strains recovered from organic wine production facilities in Waipara, New Zealand, in which Pinot Noir wine was produced by spontaneous fermentations in the vineyard and in the winery. Strains were isolated from each of four key stages of each ferment to evaluate changes in taxonomic diversity. MALDI-TOF MS analysis was confirmed as an excellent yeast identification method, with even closely related Saccharomyces species readily distinguished. A total of 13 indigenous species belonging to eight genera were identified from Pinot Noir ferments, with taxonomic diversity generally reducing as fermentation progressed. However, differences between the taxa recovered were observed between the vineyard and winery ferments, despite the grapes used being from the same batch. Furthermore, some consistent proteomic differences between strains of S. cerevisiae, Hanseniasporum uvarum, Candida californica, Pichia membranifaciens and Starmerella bacillaris correlated with the different fermentation systems used. The high speed, low cost, taxonomic resolution and ability to characterise subtle changes in phenotype that may result from variations in environmental conditions makes MALDI-TOF analysis an attractive tool for further and wider applications in the wine industry. Such applications may include monitoring wine fermentation to actively support the consistency of high-quality wine products, and potentially for the development of such products too.


Assuntos
Técnicas de Tipagem Micológica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Vinho/microbiologia , Leveduras/isolamento & purificação , Leveduras/metabolismo , Fermentação , Frutas/microbiologia , Nova Zelândia , Vitis/microbiologia , Vinho/análise , Leveduras/química , Leveduras/classificação
17.
World J Microbiol Biotechnol ; 37(7): 112, 2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-34081209

RESUMO

Grapevine cultivars are distributed worldwide, nevertheless the fermentation of its grape berries renders distinct wine products that are highly associated to the local fungal community. Despite the symbiotic association between wine and the fungal metabolism, impacting both the terroir and mycotoxin production, few studies have explored the vineyard ecosystem fungal community using both molecular marker sequencing and mycotoxin production assessment. In this study, we investigated the fungal community of three grapevine cultivars (Vitis vinifera L.) in two tropical vineyards. Illumina MiSeq sequencing was performed on two biocompartments: grape berries (GB) and grapevine soil (GS); yielding a total of 578,495 fungal internal transcribed spacer 1 reads, which were used for taxonomic classification. GB and GS fungal communities were mainly constituted by Ascomycota phylum. GS harbors a significant richer and more diverse fungal community than GB. Among GB samples, Syrah grape berries exclusively shared fungal community included wine-associated yeasts (e.g. Saccharomycopsis vini) that may play key roles in wine terroir. Mycotoxin production assessment revealed the high potential of Aspergillus section Flavi and Penicillium section Citrina isolates to produce aflatoxin B1-B2 and citrinin, respectively. This is the first study to employ next-generation sequencing to investigate vineyard associated fungal community in Brazil. Our findings provide valuable insights on the available tools for fungal ecology assessment applied to food products emphasizing the coexistence between classical and molecular tools.


Assuntos
DNA Espaçador Ribossômico/genética , Fungos/classificação , Micotoxinas/metabolismo , Análise de Sequência de DNA/métodos , Vitis/microbiologia , Brasil , DNA Fúngico/genética , Fazendas , Fungos/genética , Fungos/isolamento & purificação , Fungos/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Microbiologia do Solo , Clima Tropical
18.
Int J Food Microbiol ; 350: 109225, 2021 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-34023678

RESUMO

To address a knowledge gap about the grape berry mycobiome from Washington State vineyards, next-generation sequencing of the internal transcribed spacer region (ITS1) was used to identify native yeast and fungal species on berries of cultivar 'Cabernet Sauvignon' from two vineyards at veraison and harvest in 2015 and 2016. Four hundred fifty-six different yeast amplicon sequence variants (ASV), representing 184 distinct taxa, and 2467 non-yeast fungal ASV (791 distinct taxa) were identified in this study. A set of 50 recurrent yeast taxa, including Phaeococcomyces, Vishniacozyma and Metschnikowia, were found at both locations and sampling years. These yeast species were monitored from the vineyard into laboratory-scale spontaneous fermentations. Taxa assignable to Metschnikowia and Saccharomyces persisted during fermentation, whereas Curvibasidium, which also has possible impact on biocontrol and wine quality, did not. Sulfite generally reduced yeast diversity and richness, but its effect on the abundance of specific yeasts during fermentation was negligible. Among the 106 recurring non-yeast fungal taxa, Alternaria, Cladosporium and Ulocladium were especially abundant in the vineyard. Vineyard location was the primary factor that accounted for the variation among both communities, followed by year and berry developmental stage. The Washington mycobiomes were compared to those from other parts of the world. Sixteen recurrent yeast species appeared to be unique to Washington State vineyards. This subset also contained a higher proportion of species associated with cold and extreme environments, relative to other localities. Certain yeast and non-yeast fungal species known to suppress diseases or modify wine sensory properties were present in Washington vineyards, and likely have consequences to vineyard health and wine quality.


Assuntos
Ascomicetos/classificação , Basidiomycota/classificação , Frutas/microbiologia , Micobioma/genética , Vitis/microbiologia , Vinho/microbiologia , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Basidiomycota/genética , Basidiomycota/isolamento & purificação , DNA Intergênico/genética , Fazendas , Fermentação , Sequenciamento de Nucleotídeos em Larga Escala , Washington , Fermento Seco , Leveduras/classificação , Leveduras/genética , Leveduras/isolamento & purificação
19.
Commun Biol ; 4(1): 637, 2021 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-34050254

RESUMO

Grapevine fanleaf disease, caused by grapevine fanleaf virus (GFLV), transmitted by the soil-borne nematode Xiphinema index, provokes severe symptoms and economic losses, threatening vineyards worldwide. As no effective solution exists so far to control grapevine fanleaf disease in an environmentally friendly way, we investigated the presence of resistance to GFLV in grapevine genetic resources. We discovered that the Riesling variety displays resistance to GFLV, although it is susceptible to X. index. This resistance is determined by a single recessive factor located on grapevine chromosome 1, which we have named rgflv1. The discovery of rgflv1 paves the way for the first effective and environmentally friendly solution to control grapevine fanleaf disease through the development of new GFLV-resistant grapevine rootstocks, which was hitherto an unthinkable prospect. Moreover, rgflv1 is putatively distinct from the virus susceptibility factors already described in plants.


Assuntos
Resistência à Doença/genética , Nepovirus/patogenicidade , Vitis/genética , Agricultura/métodos , Animais , Genótipo , Nematoides/virologia , Nepovirus/genética , Melhoramento Vegetal/métodos , Doenças das Plantas/genética , Doenças das Plantas/virologia , Vitis/metabolismo , Vitis/microbiologia
20.
mBio ; 12(3)2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33975945

RESUMO

Botrytis cinerea is one of the most important plant-pathogenic fungus. Products based on microorganisms can be used in biocontrol strategies alternative to chemical control, and mycoviruses have been explored as putative biological agents in such approaches. Here, we have explored the mycovirome of B. cinerea isolates from grapevine of Italy and Spain to increase the knowledge about mycoviral diversity and evolution, and to search for new widely distributed mycoviruses that could be active ingredients in biological products to control this hazardous fungus. A total of 248 B. cinerea field isolates were used for our metatranscriptomic study. Ninety-two mycoviruses were identified: 62 new mycoviral species constituting putative novel viral genera and families. Of these mycoviruses, 57 had a positive-sense single-stranded RNA (ssRNA) genome, 19 contained a double-stranded RNA (dsRNA) genome, 15 had a negative-sense ssRNA genome, and 1 contained a single-stranded DNA (ssDNA) genome. In general, ssRNA mycoviruses were widely distributed in all sampled regions, the ssDNA mycovirus was more frequently found in Spain, and dsRNA mycoviruses were scattered in some pools of both countries. Some of the identified mycoviruses belong to clades that have never been found associated with Botrytis species: Botrytis-infecting narnaviruses; alpha-like, umbra-like, and tymo-like ssRNA+ mycoviruses; trisegmented ssRNA- mycovirus; bisegmented and tetrasegmented dsRNA mycoviruses; and finally, an ssDNA mycovirus. Among the results obtained in this massive mycovirus screening, the discovery of novel bisegmented viruses, phylogenetically related to narnaviruses, is remarkable.IMPORTANCE The results obtained here have expanded our knowledge of mycoviral diversity, horizontal transfers, and putative cross-kingdom events. To date, this study presents the most extensive and wide diversity collection of mycoviruses infecting the necrotrophic fungus B. cinerea The collection included all types of mycoviruses, with dsRNA, ssRNA+, ssRNA-, and ssDNA genomes, most of which were discovered here, and some of which were previously reported as infecting B. cinerea or other plant-pathogenic fungi. Some of these mycoviruses are reported for the first time here associated with B. cinerea, as a trisegmented ssRNA- mycovirus and as an ssDNA mycovirus, but even more remarkablly, we also describe here four novel bisegmented viruses (binarnaviruses) not previously described in nature. The present findings significantly contribute to general knowledge in virology and more particularly in the field of mycovirology.


Assuntos
Botrytis/virologia , Micovírus/classificação , Micovírus/genética , Genoma Viral , Filogenia , Viroma , Micovírus/isolamento & purificação , Itália , Doenças das Plantas/microbiologia , RNA Viral/genética , Vitis/microbiologia
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