Sensitivity of four isolates of &lt;i&gt;Clonostachys rosea&lt;/i&gt; to pesticides used in the strawberry crop in Brazil

ATP-binding cassette (ABC) transporters mediate active efflux of natural and synthetic toxicants and are considered to be important for drug tolerance in microorganisms. In biological control agents (BCA), ABC transporters can play important roles in antagonism by providing protection against toxins derived from the fungal prey and by mediating the secretion of endogenous toxins. In the present study, we generated deletion and complementation strains of the ABC transporter abcG5 in the fungal BCA Clonostachys rosea to study its role in xenobiotic tolerance and antagonism. Gene expression analysis shows induced expression of abcG5 in the presence of the Fusarium mycotoxin zearalenone (ZEA), secreted metabolites of F. graminearum, and different classes of fungicides. Phenotypic analysis of abcG5 deletion and complementation strains showed that the deletion strains were more sensitive towards F. graminearum culture filtrates, ZEA, and iprodione- and mefenoxam-based fungicides, thus suggesting the involvement of abcG5 in cell protection. The ΔabcG5 strains displayed reduced antagonism towards F. graminearum in a plate confrontation assay. Furthermore, the ΔabcG5 strains failed to protect barley seedlings from F. graminearium foot rot disease. These data show that the abcG5 ABC transporter is important for xenobiotic tolerance and biocontrol traits in C. rosea.

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SREBP-mediated gene expression regulation is essential for the intrinsic fungicide tolerance and antagonism in the fungal biocontrol agentClonostachys rosea

Piombo,

Tzelepis,

Ruus

et al. 2024

Preprint

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Sterol regulatory element-binding proteins (SREBPs) are a family of transcription factors known to regulate sterol biosynthesis and homeostasis in fungi. For this reason they have a role in several biological processes, including virulence, fungicide tolerance, hypoxia adaptation, lipid and carbohydrate metabolisms, and iron homeostasis. While the biological function of SREBPs in yeast and filamentous fungal species pathogenic to humans and plants is known, their role in fungal biocontrol agents (BCAs) is still elusive. This study aimed to investigate the biological and regulatory function of SREBPs in the BCAClonostachys rosea, with a focus on their role in fungicide tolerance, hypoxia adaptation and antagonisms. TheC. roseagenome contains two genes (sre1andsre2) coding for SREBPs and one gene each coding for Insulin induced gene (INSIG) and SREBP cleavage-activating protein (SCAP), required for SREBP-mediated ergosterol biosynthesis in fungi. Deletion ofsre1resulted in mutants with pleiotropic effects, including the reduced ability to grow on media supplemented with proline (active ingredient prothioconazole) and cantus (active ingredient boscalid) fungicides, hypoxia mimicking agent CoCl2, cell wall stressor SDS, and increased growth rate on medium supplemented with caffeine, compared withC. roseawild type (WT). In addition, the antagonistic ability against the fungal hostsBotrytis cinereaandRhizoctonia solaniwas affected whensre1was deleted. However, no significant difference betweensre2deletion strains andC. roseaWT was found for any of the tested phenotypes. To investigate the regulatory role of SRE1, the transcriptome ofC. roseaWT and asre1deletion strain was analyzed. The transcriptome analysis identified differentially expressed genes in thesre1deletion strain associated with carbohydrate and lipid metabolism, respiration, iron homeostasis, and xenobiotic tolerance. Moreover, genes coding for polyketide synthases and chitinases with a proven antimicrobial role were downregulated in the mutant, corroborating the reduced antagonism phenotypes. In summary, this work sheds light on the regulation role of transcription factor SRE1 while also exploring its effect on regulating the antagonistic activity and fungicide resistance ofC. rosea, giving us helpful knowledge to design applications of this organism in IPM strategies.

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Sensitivity of four isolates of <i>Clonostachys rosea</i> to pesticides used in the strawberry crop in Brazil

Cited by 3 publications

References 21 publications

Biodegradation of alkylphenols by rhizosphere microorganisms isolated from the roots of Hosta undulata

Biodegradation of alkylphenols by rhizosphere microorganisms isolated from the roots of Hosta undulata

An ATP-Binding Cassette Pleiotropic Drug Transporter Protein Is Required for Xenobiotic Tolerance and Antagonism in the Fungal Biocontrol Agent Clonostachys rosea

SREBP-mediated gene expression regulation is essential for the intrinsic fungicide tolerance and antagonism in the fungal biocontrol agentClonostachys rosea

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