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Exploring mechanisms of resistance to respiratory inhibitors in field strains of Botrytis cinerea, the causal agent of gray mold.

ABSTRACT: Respiratory inhibitors are among the fungicides most widely used for disease control on crops. Most are strobilurins and carboxamides, inhibiting the cytochrome b of mitochondrial complex III and the succinate dehydrogenase of mitochondrial complex II, respectively. A few years after the approval of these inhibitors for use on grapevines, field isolates of Botrytis cinerea, the causal agent of gray mold, resistant to one or both of these classes of fungicide were recovered in France and Germany. However, little was known about the mechanisms underlying this resistance in field populations of this fungus. Such knowledge could facilitate resistance risk assessment. The aim of this study was to investigate the mechanisms of resistance occurring in B. cinerea populations. Highly specific resistance to strobilurins was correlated with a single mutation of the cytb target gene. Changes in its intronic structure may also have occurred due to an evolutionary process controlling selection for resistance. Specific resistance to carboxamides was identified for six phenotypes, with various patterns of resistance levels and cross-resistance. Several mutations specific to B. cinerea were identified within the sdhB and sdhD genes encoding the iron-sulfur protein and an anchor protein of the succinate dehydrogenase complex. Another as-yet-uncharacterized mechanism of resistance was also recorded. In addition to target site resistance mechanisms, multidrug resistance, linked to the overexpression of membrane transporters, was identified in strains with low to moderate resistance to several respiratory inhibitors. This diversity of resistance mechanisms makes resistance management difficult and must be taken into account when developing strategies for Botrytis control.

SUBMITTER: Leroux P

PROVIDER: S-EPMC2950445 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature

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Exploring mechanisms of resistance to respiratory inhibitors in field strains of Botrytis cinerea, the causal agent of gray mold.

Leroux Pierre P Gredt Michel M Leroch Michaela M Walker Anne-Sophie AS

Applied and environmental microbiology 20100806 19

Respiratory inhibitors are among the fungicides most widely used for disease control on crops. Most are strobilurins and carboxamides, inhibiting the cytochrome b of mitochondrial complex III and the succinate dehydrogenase of mitochondrial complex II, respectively. A few years after the approval of these inhibitors for use on grapevines, field isolates of Botrytis cinerea, the causal agent of gray mold, resistant to one or both of these classes of fungicide were recovered in France and Germany. ...[more]

PMID: 20693447

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Project description:Owing to the remarkable antimicrobial potential of these materials, research into the possible use of nanomaterials as alternatives to fungicides in sustainable agriculture is increasingly progressing. Here, we investigated the potential antifungal properties of chitosan-decorated copper oxide nanocomposite (CH@CuO NPs) to control gray mold diseases of tomato caused by Botrytis cinerea throughout in vitro and in vivo trials. The nanocomposite CH@CuO NPs were chemically prepared, and size and shape were determined using Transmission Electron Microscope (TEM). The chemical functional groups responsible for the interaction of the CH NPs with the CuO NPs were detected using the Fourier Transform Infrared (FTIR) spectrophotometry. The TEM images confirmed that CH NPs have a thin and semitransparent network shape, while CuO NPs were spherically shaped. Furthermore, the nanocomposite CH@CuO NPs ex-habited an irregular shape. The size of CH NPs, CuO NPs and CH@CuO NPs as measured through TEM, were approximately 18.28 ± 2.4 nm, 19.34 ± 2.1 nm, and 32.74 ± 2.3 nm, respectively. The antifungal activity of CH@CuO NPs was tested at three concentrations of 50, 100 and 250 mg/L and the fungicide Teldor 50% SC was applied at recommended dose 1.5 mL/L. In vitro experiments revealed that CH@CuO NPs at different concentrations significantly inhibited the reproductive growth process of B. cinerea by suppressing the development of hyphae, spore germination and formation of sclerotia. Interestingly, a significant control efficacy of CH@CuO NPs against tomato gray mold was observed particularly at concentrations 100 and 250 mg/L on both detached leaves (100%) as well as the whole tomato plants (100%) when compared to the conventional chemical fungicide Teldor 50% SC (97%). In addition, the tested concentration 100 mg/L improved to be sufficient to guarantee a complete reduction in the disease's severity (100%) to tomato fruits from gray mold without any morphological toxicity. In comparison, tomato plants treated with the recommended dose 1.5 mL/L of Teldor 50% SC ensured disease reduction up to 80%. Conclusively, this research enhances the concept of agro-nanotechnology by presenting how a nano materials-based fungicide could be used to protect tomato plants from gray mold under greenhouse conditions and during the postharvest stage.

Project description:Gray mold is one of the most important diseases of grapevine in temperate climates. This plant pathogen affects plant growth and reduces wine quality. The use of yeasts as biocontrol agents to apply in the vineyard have been investigated in recent years as an alternative to agrochemicals. In this work, fermenting musts obtained from overripe grape berries, therefore more susceptible to infection by fungal pathogens such as Botrytis cinerea, were considered for the selection of yeasts carrying antifungal activity. Thirty-six isolates were identified as Starmerella bacillaris, a species recently proven to be of enological interest. Among them 14 different strains were studied and antifungal activity against B. cinerea was demonstrated, for the first time, to be present in S. bacillaris species. The production of volatile organic compounds (VOCs), tested in vitro, was found to be the main responsible of S. bacillaris antifungal effects. All the strains were able to reduce B. cinerea decay on wounded grape berries artificially inoculated with gray mold. The colonization level of wound was very high reaching, after 5 days, a concentration of 10(6) cells per ml of grape juice obtained after berry crushing. At this cell concentration S. bacillaris strains were used to ferment synthetic and natural musts. The sequential yeast inoculation, performed by adding S. cerevisiae 48 h after S. bacillaris, was needed to complete sugar consumption and determined a significant increase in glicerol content and a reduction of ethanol and acetic acid concentrations. The high wound colonization ability, found in this work, together with the propensity to colonize grape berry and the interesting enological traits possessed by the selected S. bacillaris strains allow the use of this yeast as biocontrol agent on vine and grape berries with possible positive effects on must fermentation, although the presence of S. cerevisiae is needed to complete the fermentation process. This work introduces new possibilities in wine yeast selection programs in order to identify innovative wine yeasts that are simultaneously antifungal agents in vineyards and alternative wine starters for grape must fermentation and open new perspective to a more integrated strategy for increasing wine quality.

Dataset Information

Exploring mechanisms of resistance to respiratory inhibitors in field strains of Botrytis cinerea, the causal agent of gray mold.

Publications

Exploring mechanisms of resistance to respiratory inhibitors in field strains of Botrytis cinerea, the causal agent of gray mold.

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