Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity

As a type of multicopper oxidase, laccases play multiple biological roles in entomopathogenic fungi, enhancing their survival, development, and pathogenicity. However, the mechanisms by which laccases operate in these fungi remain under-researched. In this study, we identified two laccase-encoding g...

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Main Authors: Qiaoyun Wu, Yingying Ye, Yiran Liu, Yufan He, Xing Li, Siqi Yang, Tongtong Xu, Xiufang Hu, Guohong Zeng
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Journal of Fungi
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Online Access:https://www.mdpi.com/2309-608X/11/3/176
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author Qiaoyun Wu
Yingying Ye
Yiran Liu
Yufan He
Xing Li
Siqi Yang
Tongtong Xu
Xiufang Hu
Guohong Zeng
author_facet Qiaoyun Wu
Yingying Ye
Yiran Liu
Yufan He
Xing Li
Siqi Yang
Tongtong Xu
Xiufang Hu
Guohong Zeng
author_sort Qiaoyun Wu
collection DOAJ
description As a type of multicopper oxidase, laccases play multiple biological roles in entomopathogenic fungi, enhancing their survival, development, and pathogenicity. However, the mechanisms by which laccases operate in these fungi remain under-researched. In this study, we identified two laccase-encoding genes, <i>Mr-lac3</i> and <i>Mr-lcc2</i>, from <i>Metarhizium robertsii</i>, both of which are highly expressed during conidiation. Knocking out <i>Mr-lac3</i> and <i>Mr-lcc2</i> resulted in a significant increase in the conidial yields of <i>M. robertsii</i>. Furthermore, the relative expression levels of upstream regulators associated with the conidiation pathway were markedly up-regulated in <i>ΔMr-lac3</i> and <i>ΔMr-lcc2</i> compared to the wild-type strain during conidiation, indicating that <i>Mr-lac3</i> and <i>Mr-lcc2</i> negatively regulate conidia formation. qRT-PCR analyses revealed that <i>Mr-lac3</i> and <i>Mr-lcc2</i> are regulated by the pigment synthesis gene cluster, including <i>Mr-Pks1</i>, <i>Mr-EthD</i>, and <i>Mlac1</i>, and they also provide feedback regulation to jointly control pigment synthesis. Additionally, <i>ΔMr-lac3</i> and <i>ΔMr-lcc2</i> significantly reduced the trehalose content in conidia and increased the sensitivity to cell wall-perturbing agents, such as Congo red and guaiacol, which led to a marked decrease in tolerance to abiotic stresses. In conclusion, the laccases Mr-lac3 and Mr-lcc2 negatively regulate conidia formation while positively regulating conidial maturation, thereby enhancing tolerance to abiotic stresses and pathogenicity.
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spelling doaj-art-bc06dc19ddd54dc8ab056b330ef11fe92025-08-20T02:42:32ZengMDPI AGJournal of Fungi2309-608X2025-02-0111317610.3390/jof11030176Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and PathogenicityQiaoyun Wu0Yingying Ye1Yiran Liu2Yufan He3Xing Li4Siqi Yang5Tongtong Xu6Xiufang Hu7Guohong Zeng8Zhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaZhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaAs a type of multicopper oxidase, laccases play multiple biological roles in entomopathogenic fungi, enhancing their survival, development, and pathogenicity. However, the mechanisms by which laccases operate in these fungi remain under-researched. In this study, we identified two laccase-encoding genes, <i>Mr-lac3</i> and <i>Mr-lcc2</i>, from <i>Metarhizium robertsii</i>, both of which are highly expressed during conidiation. Knocking out <i>Mr-lac3</i> and <i>Mr-lcc2</i> resulted in a significant increase in the conidial yields of <i>M. robertsii</i>. Furthermore, the relative expression levels of upstream regulators associated with the conidiation pathway were markedly up-regulated in <i>ΔMr-lac3</i> and <i>ΔMr-lcc2</i> compared to the wild-type strain during conidiation, indicating that <i>Mr-lac3</i> and <i>Mr-lcc2</i> negatively regulate conidia formation. qRT-PCR analyses revealed that <i>Mr-lac3</i> and <i>Mr-lcc2</i> are regulated by the pigment synthesis gene cluster, including <i>Mr-Pks1</i>, <i>Mr-EthD</i>, and <i>Mlac1</i>, and they also provide feedback regulation to jointly control pigment synthesis. Additionally, <i>ΔMr-lac3</i> and <i>ΔMr-lcc2</i> significantly reduced the trehalose content in conidia and increased the sensitivity to cell wall-perturbing agents, such as Congo red and guaiacol, which led to a marked decrease in tolerance to abiotic stresses. In conclusion, the laccases Mr-lac3 and Mr-lcc2 negatively regulate conidia formation while positively regulating conidial maturation, thereby enhancing tolerance to abiotic stresses and pathogenicity.https://www.mdpi.com/2309-608X/11/3/176<i>Metarhizium robertsii</i>laccasesMr-lac3Mr-lcc2conidiationconidial maturation
spellingShingle Qiaoyun Wu
Yingying Ye
Yiran Liu
Yufan He
Xing Li
Siqi Yang
Tongtong Xu
Xiufang Hu
Guohong Zeng
Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
Journal of Fungi
<i>Metarhizium robertsii</i>
laccases
Mr-lac3
Mr-lcc2
conidiation
conidial maturation
title Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
title_full Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
title_fullStr Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
title_full_unstemmed Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
title_short Mr-lac3 and Mr-lcc2 in <i>Metarhizium robertsii</i> Regulate Conidiation and Maturation, Enhancing Tolerance to Abiotic Stresses and Pathogenicity
title_sort mr lac3 and mr lcc2 in i metarhizium robertsii i regulate conidiation and maturation enhancing tolerance to abiotic stresses and pathogenicity
topic <i>Metarhizium robertsii</i>
laccases
Mr-lac3
Mr-lcc2
conidiation
conidial maturation
url https://www.mdpi.com/2309-608X/11/3/176
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