The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>

The necrotrophic pathogen <i>Sclerotinia sclerotiorum</i> is widely distributed and infects a broad range of hosts, making it one of the most economically damaging plant pathogens. This study demonstrated that pyrimethanil, an anilinopyrimidine fungicide, exhibited potent activity agains...

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Main Authors: Yanfen Wang, Zeyuan Chen, Tiancheng Liu, Xupeng Gao, Yanchao Shi, Honghui Wu, Runqiang Liu, Yunchao Kan, Hao Yu, Feng Zhou
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Journal of Fungi
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Online Access:https://www.mdpi.com/2309-608X/11/5/344
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author Yanfen Wang
Zeyuan Chen
Tiancheng Liu
Xupeng Gao
Yanchao Shi
Honghui Wu
Runqiang Liu
Yunchao Kan
Hao Yu
Feng Zhou
author_facet Yanfen Wang
Zeyuan Chen
Tiancheng Liu
Xupeng Gao
Yanchao Shi
Honghui Wu
Runqiang Liu
Yunchao Kan
Hao Yu
Feng Zhou
author_sort Yanfen Wang
collection DOAJ
description The necrotrophic pathogen <i>Sclerotinia sclerotiorum</i> is widely distributed and infects a broad range of hosts, making it one of the most economically damaging plant pathogens. This study demonstrated that pyrimethanil, an anilinopyrimidine fungicide, exhibited potent activity against <i>S. sclerotiorum</i>, with EC<sub>50</sub> values ranging from 0.411 to 0.610 μg/mL. Four highly pyrimethanil-resistant mutants were obtained through chemical taming, with EC<sub>50</sub> values of 7.247 to 24.718 μg/mL. These mutants exhibited significantly reduced mycelial growth, sclerotia production, and pathogenicity compared to their wild-type parental isolates, indicating that pyrimethanil resistance suffered a fitness penalty in <i>S. sclerotiorum</i>. Notably, three mutants (DDJH-Pyri-R1, DDJH-Pyri-R3, and DDJH-Pyri-R4), completely lose the capacity to infect detached tomato leaves. Point mutations that cause amino acid changes in the predicted sequence of cystathione-γ synthase (CGS) and cystathione-β lyase (CBL), encoded by <i>SsCGS1</i> and <i>SsCGS2</i>, were identified in three mutants. However, one mutant (DDJH-Pyri-R2) showed no mutations in these genes, suggesting an alternative resistance mechanism. Molecular docking revealed that mutations in SsCGS1-R3, SsCGS1-R4, and SsCGS2-R1 reduced the binding affinity between pyrimethanil and SsCGSs. No cross-resistance was observed between pyrimethanil and other commonly used fungicides, including carbendazim, fludioxonil, prochloraz, tebuconazole, pyraclostrobin, boscalid, fluazinam, and cyprodinil. These findings provide valuable insights for designing resistance inhibitors and suggest that pyrimethanil has significant potential for controlling soybean sclerotinia stem rot (SSR) caused by <i>S. sclerotiorum</i>.
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spelling doaj-art-7e01f6a153c04ff081abe7238d8b61b72025-08-20T02:33:50ZengMDPI AGJournal of Fungi2309-608X2025-04-0111534410.3390/jof11050344The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>Yanfen Wang0Zeyuan Chen1Tiancheng Liu2Xupeng Gao3Yanchao Shi4Honghui Wu5Runqiang Liu6Yunchao Kan7Hao Yu8Feng Zhou9School of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaSchool of Plant Protection and Environment, Henan Institute of Science and Technology, Xinxiang 453003, ChinaThe necrotrophic pathogen <i>Sclerotinia sclerotiorum</i> is widely distributed and infects a broad range of hosts, making it one of the most economically damaging plant pathogens. This study demonstrated that pyrimethanil, an anilinopyrimidine fungicide, exhibited potent activity against <i>S. sclerotiorum</i>, with EC<sub>50</sub> values ranging from 0.411 to 0.610 μg/mL. Four highly pyrimethanil-resistant mutants were obtained through chemical taming, with EC<sub>50</sub> values of 7.247 to 24.718 μg/mL. These mutants exhibited significantly reduced mycelial growth, sclerotia production, and pathogenicity compared to their wild-type parental isolates, indicating that pyrimethanil resistance suffered a fitness penalty in <i>S. sclerotiorum</i>. Notably, three mutants (DDJH-Pyri-R1, DDJH-Pyri-R3, and DDJH-Pyri-R4), completely lose the capacity to infect detached tomato leaves. Point mutations that cause amino acid changes in the predicted sequence of cystathione-γ synthase (CGS) and cystathione-β lyase (CBL), encoded by <i>SsCGS1</i> and <i>SsCGS2</i>, were identified in three mutants. However, one mutant (DDJH-Pyri-R2) showed no mutations in these genes, suggesting an alternative resistance mechanism. Molecular docking revealed that mutations in SsCGS1-R3, SsCGS1-R4, and SsCGS2-R1 reduced the binding affinity between pyrimethanil and SsCGSs. No cross-resistance was observed between pyrimethanil and other commonly used fungicides, including carbendazim, fludioxonil, prochloraz, tebuconazole, pyraclostrobin, boscalid, fluazinam, and cyprodinil. These findings provide valuable insights for designing resistance inhibitors and suggest that pyrimethanil has significant potential for controlling soybean sclerotinia stem rot (SSR) caused by <i>S. sclerotiorum</i>.https://www.mdpi.com/2309-608X/11/5/344<i>Sclerotinia sclerotiorum</i>pyrimethanilresistance mechanismcystathione-γ synthase
spellingShingle Yanfen Wang
Zeyuan Chen
Tiancheng Liu
Xupeng Gao
Yanchao Shi
Honghui Wu
Runqiang Liu
Yunchao Kan
Hao Yu
Feng Zhou
The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
Journal of Fungi
<i>Sclerotinia sclerotiorum</i>
pyrimethanil
resistance mechanism
cystathione-γ synthase
title The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
title_full The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
title_fullStr The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
title_full_unstemmed The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
title_short The Resistance Mechanisms of Anilinopyrimidine Fungicide Pyrimethanil in <i>Sclerotinia sclerotiorum</i>
title_sort resistance mechanisms of anilinopyrimidine fungicide pyrimethanil in i sclerotinia sclerotiorum i
topic <i>Sclerotinia sclerotiorum</i>
pyrimethanil
resistance mechanism
cystathione-γ synthase
url https://www.mdpi.com/2309-608X/11/5/344
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