Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>

ABSTRACT In filamentous fungi, 1,8-dihydroxynaphthalene (DHN) melanin is a major component of the extracellular matrix, endowing fungi with environmental tolerance and some pathogenic species with pathogenicity. However, the subcellular location of the melanin biosynthesis pathway components remains...

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Autores principales:	Xue Chen, Chuanxi Zhu, Yantao Na, Dandan Ren, Chenghua Zhang, Yifan He, Yiwen Wang, Sheng Xiang, Weiheng Ren, Yina Jiang, Ling Xu, Pinkuan Zhu
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	Botrytis cinerea DHN melanin scytalone subcellular trafficking endosome peroxisome Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3168d767944c473e8d6b35cf2dc8e69f
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spelling	oai:doaj.org-article:3168d767944c473e8d6b35cf2dc8e69f2021-11-10T18:37:48ZCompartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>10.1128/mBio.00007-212150-7511https://doaj.org/article/3168d767944c473e8d6b35cf2dc8e69f2021-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00007-21https://doaj.org/toc/2150-7511ABSTRACT In filamentous fungi, 1,8-dihydroxynaphthalene (DHN) melanin is a major component of the extracellular matrix, endowing fungi with environmental tolerance and some pathogenic species with pathogenicity. However, the subcellular location of the melanin biosynthesis pathway components remains obscure. Using the gray mold pathogen Botrytis cinerea, the DHN melanin intermediate scytalone was characterized via phenotypic and chemical analysis of mutants, and the key enzymes participating in melanin synthesis were fused with fluorescent proteins to observe their subcellular localizations. The Δbcscd1 mutant accumulated scytalone in the culture filtrate rather than in mycelium. Excessive scytalone appears to be self-inhibitory to the fungus, leading to repressed sclerotial germination and sporulation in the Δbcscd1 mutant. The BcBRN1/2 enzymes responsible for synthesizing scytalone were localized in endosomes and found to be trafficked to the cell surface, accompanied by the accumulation of BcSCD1 proteins in the cell wall. In contrast, the early-stage melanin synthesis enzymes BcPKS12/13 and BcYGH1 were localized in peroxisomes. Taken together, the results of this study revealed the subcellular distribution of melanin biosynthetic enzymes in B. cinerea, indicating that the encapsulation and externalization of the melanin synthetic enzymes need to be delicately orchestrated to ensure enzymatic efficiency and protect itself from the adverse effect of the toxic intermediate metabolite. IMPORTANCE The devastating gray mold pathogen Botrytis cinerea propagates via melanized conidia and sclerotia. This study reveals that the sclerotial germination of B. cinerea is differentially affected by different enzymes in the melanin synthesis pathway. Using gene knockout mutants and chemical analysis, we found that excessive accumulation of the melanin intermediate scytalone is inhibitory to B. cinerea. Subcellular localization analysis of the melanin synthesis enzymes of B. cinerea suggested two-stage partitioning of the melanogenesis pathway: the intracellular stage involves the steps until the intermediate scytalone was translocated to the cell surface, whereas the extracellular stage comprises all the steps occurring in the wall from scytalone to final melanin formation. These strategies make the fungus avert self-poisoning during melanin production. This study opens avenues for better understanding the mechanisms of secondary metabolite production in filamentous fungi.Xue ChenChuanxi ZhuYantao NaDandan RenChenghua ZhangYifan HeYiwen WangSheng XiangWeiheng RenYina JiangLing XuPinkuan ZhuAmerican Society for MicrobiologyarticleBotrytis cinereaDHN melaninscytalonesubcellular traffickingendosomeperoxisomeMicrobiologyQR1-502ENmBio, Vol 12, Iss 2 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Botrytis cinerea DHN melanin scytalone subcellular trafficking endosome peroxisome Microbiology QR1-502
spellingShingle	Botrytis cinerea DHN melanin scytalone subcellular trafficking endosome peroxisome Microbiology QR1-502 Xue Chen Chuanxi Zhu Yantao Na Dandan Ren Chenghua Zhang Yifan He Yiwen Wang Sheng Xiang Weiheng Ren Yina Jiang Ling Xu Pinkuan Zhu Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
description	ABSTRACT In filamentous fungi, 1,8-dihydroxynaphthalene (DHN) melanin is a major component of the extracellular matrix, endowing fungi with environmental tolerance and some pathogenic species with pathogenicity. However, the subcellular location of the melanin biosynthesis pathway components remains obscure. Using the gray mold pathogen Botrytis cinerea, the DHN melanin intermediate scytalone was characterized via phenotypic and chemical analysis of mutants, and the key enzymes participating in melanin synthesis were fused with fluorescent proteins to observe their subcellular localizations. The Δbcscd1 mutant accumulated scytalone in the culture filtrate rather than in mycelium. Excessive scytalone appears to be self-inhibitory to the fungus, leading to repressed sclerotial germination and sporulation in the Δbcscd1 mutant. The BcBRN1/2 enzymes responsible for synthesizing scytalone were localized in endosomes and found to be trafficked to the cell surface, accompanied by the accumulation of BcSCD1 proteins in the cell wall. In contrast, the early-stage melanin synthesis enzymes BcPKS12/13 and BcYGH1 were localized in peroxisomes. Taken together, the results of this study revealed the subcellular distribution of melanin biosynthetic enzymes in B. cinerea, indicating that the encapsulation and externalization of the melanin synthetic enzymes need to be delicately orchestrated to ensure enzymatic efficiency and protect itself from the adverse effect of the toxic intermediate metabolite. IMPORTANCE The devastating gray mold pathogen Botrytis cinerea propagates via melanized conidia and sclerotia. This study reveals that the sclerotial germination of B. cinerea is differentially affected by different enzymes in the melanin synthesis pathway. Using gene knockout mutants and chemical analysis, we found that excessive accumulation of the melanin intermediate scytalone is inhibitory to B. cinerea. Subcellular localization analysis of the melanin synthesis enzymes of B. cinerea suggested two-stage partitioning of the melanogenesis pathway: the intracellular stage involves the steps until the intermediate scytalone was translocated to the cell surface, whereas the extracellular stage comprises all the steps occurring in the wall from scytalone to final melanin formation. These strategies make the fungus avert self-poisoning during melanin production. This study opens avenues for better understanding the mechanisms of secondary metabolite production in filamentous fungi.
format	article
author	Xue Chen Chuanxi Zhu Yantao Na Dandan Ren Chenghua Zhang Yifan He Yiwen Wang Sheng Xiang Weiheng Ren Yina Jiang Ling Xu Pinkuan Zhu
author_facet	Xue Chen Chuanxi Zhu Yantao Na Dandan Ren Chenghua Zhang Yifan He Yiwen Wang Sheng Xiang Weiheng Ren Yina Jiang Ling Xu Pinkuan Zhu
author_sort	Xue Chen
title	Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
title_short	Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
title_full	Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
title_fullStr	Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
title_full_unstemmed	Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>
title_sort	compartmentalization of melanin biosynthetic enzymes contributes to self-defense against intermediate compound scytalone in <italic toggle="yes">botrytis cinerea</italic>
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/3168d767944c473e8d6b35cf2dc8e69f
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Compartmentalization of Melanin Biosynthetic Enzymes Contributes to Self-Defense against Intermediate Compound Scytalone in <italic toggle="yes">Botrytis cinerea</italic>

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