Overexpression of
            <i>ShCYP51B</i>
            and
            <i>ShatrD</i>
            in Sclerotinia homoeocarpa Isolates Exhibiting Practical Field Resistance to a Demethylation Inhibitor Fungicide

Hulvey, Jon; Popko, James T.; Sang, Hyunkyu; Berg, Andrew; Jung, Geunhwa

doi:10.1128/aem.00417-12

Cited by 81 publications

(80 citation statements)

References 42 publications

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“…CYP51 is involved in the biosynthesis of fungal sterols which are required for membrane stability [101]. It has been speculated that overexpression of CYP51 or its paralogues in S. homoeocarpa and Monilinia fructicola is one of the mechanisms that decrease sensitivity to DMI [91, 102]. SS1G_04805 is the paralogue of S. homoeocarpa CYP51 and was up-regulated at 1 to 12 hpi in the current study.…”

Section: Resultsmentioning

confidence: 50%

“…The S. sclerotiorum orthologue of BcAtrA (SS1G_06715) was slightly induced at 1 hpi. The ABC transporter AtrD has been implicated in resistance to demethylation inhibitor fungicides in B. cinerea [90] and S. homeocarpa [91], but the S. sclerotiorum orthologue (SS1G_02407) was not up-regulated during B. napus infection in the current study. The S. sclerotiorum orthologue of BMR1 (SS1G_04483), which is involved in resistance to polyoxin and iprobenfos toxicants in B. cinerea [92], was up-regulated at 48 hpi in the current study.…”

Section: Resultsmentioning

confidence: 54%

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Changes in the Sclerotinia sclerotiorum transcriptome during infection of Brassica napus

et al. 2017

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Background Sclerotinia sclerotiorum causes stem rot in Brassica napus, which leads to lodging and severe yield losses. Although recent studies have explored significant progress in the characterization of individual S. sclerotiorum pathogenicity factors, a gap exists in profiling gene expression throughout the course of S. sclerotiorum infection on a host plant. In this study, RNA-Seq analysis was performed with focus on the events occurring through the early (1 h) to the middle (48 h) stages of infection.ResultsTranscript analysis revealed the temporal pattern and amplitude of the deployment of genes associated with aspects of pathogenicity or virulence during the course of S. sclerotiorum infection on Brassica napus. These genes were categorized into eight functional groups: hydrolytic enzymes, secondary metabolites, detoxification, signaling, development, secreted effectors, oxalic acid and reactive oxygen species production. The induction patterns of nearly all of these genes agreed with their predicted functions. Principal component analysis delineated gene expression patterns that signified transitions between pathogenic phases, namely host penetration, ramification and necrotic stages, and provided evidence for the occurrence of a brief biotrophic phase soon after host penetration.ConclusionsThe current observations support the notion that S. sclerotiorum deploys an array of factors and complex strategies to facilitate host colonization and mitigate host defenses. This investigation provides a broad overview of the sequential expression of virulence/pathogenicity-associated genes during infection of B. napus by S. sclerotiorum and provides information for further characterization of genes involved in the S. sclerotiorum-host plant interactions.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3642-5) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 50%

Section: Resultsmentioning

confidence: 54%

Changes in the Sclerotinia sclerotiorum transcriptome during infection of Brassica napus

et al. 2017

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“…Propiconazole sensitivity previously determined by Hulvey et al . (): I, insensitive; PFR, practical field resistance; S, sensitive.…”

Section: Resultsmentioning

confidence: 97%

“…Isolates are named according to Hulvey et al . (). Location abbreviations: HF, Hartford Golf Club; HR, Hickory Ridge Country Club; JT, Joseph Troll Turf Research Center at University of Massachusetts, Amherst, MA, USA; SM, Shuttle Meadow Country Club; WB, Wintonbury Hills Golf Club.…”

Section: Resultsmentioning

confidence: 99%

A pleiotropic drug resistance transporter is involved in reduced sensitivity to multiple fungicide classes in Sclerotinia homoeocarpa (F.T. Bennett)

Sang

Hulvey

Popko

et al. 2014

Molecular Plant Pathology

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Dollar spot, caused by Sclerotinia homoeocarpa, is a prevalent turfgrass disease, and the fungus exhibits widespread fungicide resistance in North America. In a previous study, an ABC-G transporter, ShatrD, was associated with practical field resistance to demethylation inhibitor (DMI) fungicides. Mining of ABC-G transporters, also known as pleiotropic drug resistance (PDR) transporters, from RNA-Seq data gave an assortment of transcripts, several with high sequence similarity to functionally characterized transporters from Botrytis cinerea, and others with closest blastx hits from Aspergillus and Monilinia. In addition to ShatrD, another PDR transporter showed significant over-expression in replicated RNA-Seq data, and in a collection of field-resistant isolates, as measured by quantitative polymerase chain reaction. These isolates also showed reduced sensitivity to unrelated fungicide classes. Using a yeast complementation system, we sought to test the hypothesis that this PDR transporter effluxes DMI as well as chemically unrelated fungicides. The transporter (ShPDR1) was cloned into the Gal1 expression vector and transformed into a yeast PDR transporter deletion mutant, AD12345678. Complementation assays indicated that ShPDR1 complemented the mutant in the presence of propiconazole (DMI), iprodione (dicarboximide) and boscalid (SDHI, succinate dehydrogenase inhibitor). Our results indicate that the over-expression of ShPDR1 is correlated with practical field resistance to DMI fungicides and reduced sensitivity to dicarboximide and SDHI fungicides. These findings highlight the potential for the eventual development of a multidrug resistance phenotype in this pathogen. In addition, this study presents a pipeline for the discovery and validation of fungicide resistance genes using de novo next-generation sequencing and molecular biology techniques in an unsequenced plant pathogenic fungus.

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“…It has been demonstrated in many fungi, including plant and human pathogens, that certain ABC transporter genes were up-regulated in response to azole treatment, and loss of genes encoding PDR-type ABC transporters resulted in hypersensitivity to azoles [35–37]. According to phylogenetic analysis, the A .…”

Section: Discussionmentioning

confidence: 99%

A Novel Zn2-Cys6 Transcription Factor AtrR Plays a Key Role in an Azole Resistance Mechanism of Aspergillus fumigatus by Co-regulating cyp51A and cdr1B Expressions

et al. 2017

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Successful treatment of aspergillosis caused by Aspergillus fumigatus is threatened by an increasing incidence of drug resistance. This situation is further complicated by the finding that strains resistant to azoles, the major antifungal drugs for aspergillosis, have been widely disseminated across the globe. To elucidate mechanisms underlying azole resistance, we identified a novel transcription factor that is required for normal azole resistance in Aspergillus fungi including A. fumigatus, Aspergillus oryzae, and Aspergillus nidulans. This fungal-specific Zn2-Cys6 type transcription factor AtrR was found to regulate expression of the genes related to ergosterol biosynthesis, including cyp51A that encodes a target protein of azoles. The atrR deletion mutant showed impaired growth under hypoxic conditions and attenuation of virulence in murine infection model for aspergillosis. These results were similar to the phenotypes for a mutant strain lacking SrbA that is also a direct regulator for the cyp51A gene. Notably, AtrR was responsible for the expression of cdr1B that encodes an ABC transporter related to azole resistance, whereas SrbA was not involved in the regulation. Chromatin immunoprecipitation assays indicated that AtrR directly bound both the cyp51A and cdr1B promoters. In the clinically isolated itraconazole resistant strain that harbors a mutant Cyp51A (G54E), deletion of the atrR gene resulted in a hypersensitivity to the azole drugs. Together, our results revealed that AtrR plays a pivotal role in a novel azole resistance mechanism by co-regulating the drug target (Cyp51A) and putative drug efflux pump (Cdr1B).

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Overexpression of ShCYP51B and ShatrD in Sclerotinia homoeocarpa Isolates Exhibiting Practical Field Resistance to a Demethylation Inhibitor Fungicide

Cited by 81 publications

References 42 publications

Changes in the Sclerotinia sclerotiorum transcriptome during infection of Brassica napus

Changes in the Sclerotinia sclerotiorum transcriptome during infection of Brassica napus

A pleiotropic drug resistance transporter is involved in reduced sensitivity to multiple fungicide classes in Sclerotinia homoeocarpa (F.T. Bennett)

A Novel Zn2-Cys6 Transcription Factor AtrR Plays a Key Role in an Azole Resistance Mechanism of Aspergillus fumigatus by Co-regulating cyp51A and cdr1B Expressions

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