• Mycobiology
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• v.50 no.6
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• pp.457-466
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• 2022

Epicoccum nigrum is a saprophytic or endophytic fungus that is found worldwide. Because of the antagonist effects of E. nigrum on many plant pathogens, current studies on E. nigrum have focused on the development of biological control agents and the utilization of its various metabolites. In this study, E. nigrum was collected from a wheat field, and its genetic diversity was analyzed. Phylogenetic analyses identified 63 isolates of E. nigrum divided into seven groups, indicating a wide genetic diversity. Isolates antagonized the wheat pathogen Fusarium graminearum, and reduced disease symptoms caused by F. graminearum in wheat coleoptiles. Moreover, pretreatment of wheat coleoptiles with E. nigrum induced the upregulation of pathogen-related (PR) genes, PR1, PR2, PR3, PR5, PR9, and PR10 in wheat coleoptiles responding to F. graminearum invasion. Overall, this study indicates that E. nigrum isolates can be used as biological pathogen inhibitors applied in wheat fields.

• Journal of Applied Biological Chemistry
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• v.65 no.3
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• pp.159-166
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• 2022

Epicoccum nigrum produces epipyrone A (orevactaene), a yellow polyketide pigment. Its biosynthetic gene cluster was previously characterized in E. nigrum KACC 40642. The YES liquid culture of this strain revealed high-level production of epicoccamide (EPC), with an identity that was determined using liquid chromatography-mass spectrometry analysis and molecular mass search using the SuperNatural database V2 webserver. The production of EPC was further confirmed by compound isolation and nuclear magnetic resonance spectroscopy. EPC is a highly reduced polyketide with tetramic acid and mannosyl moieties. The EPC structure guided us to localize the hypothetical EPC biosynthetic gene cluster (BGC) in E. nigrum ICMP 19927 genome sequence. The BGC contains genes encoding highly reducing (HR)-fungal polyketide synthase (fPKS)-nonribosomal peptide synthetase (NRPS), glycosyltransferase (GT), enoylreductase, cytochrome P450, and N-methyltrasnferase. Targeted inactivation of the HR-fPKS-NRPS and GT genes abolished EPC production, supporting the successful localization of EPC BGC. This study provides a platform to explore the hidden biological activities of EPC, a bolaamphiphilic compound.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.225-236
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• 2021

The epipyrone (EPN) biosynthetic gene cluster of Epicoccum nigrum is composed of epnC, epnB, and epnA, which encode cytochrome P450 oxidase, glycosyltransferase, and highly reducing polyketide synthase, respectively. Gene inactivation mutants for epnA, epnB, and epnC were previously generated, and it was found that all of them were incapable of producing EPN and any of its related compounds. It was also reported that epnB inactivation abolished epnA transcription, generating ΔepnAB. This study shows that the introduction of native epnC readily restored EPN production in ΔepnC, suggesting that epnC is essential for polyketide release from EpnA and implies that EpnC works during the polyketide chain assembly of EpnA. Introduction of epnC promoter-epnA restored EPN production in ΔepnA. The ΔepnB genotype was prepared by introducing the epnA expression vector into ΔepnAB, and it was found that the resulting recombinant strain did not produce any EPN-related compounds. A canonical epnB inactivation strain was also generated by deleting its 5'-end. At the deletion point, an Aspergllus nidulans gpdA promoter was inserted to ensure the transcription of epnA, which is located downstream of epnB. Examination of the metabolite profile of the resulting ΔepnB mutant via LC-mass spectrometry verified that no EPN-related compound was produced in this strain. This substantiates that C-glycosylation by EpnB is a prerequisite for the release of EpnA-tethered product. In conclusion, it is proposed that cytochrome P450 oxidase and glycosyltransferase work in concert with polyketide synthase to generate EPN without the occurrence of any free intermediates.

• The Korean Journal of Mycology
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• v.40 no.3
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• pp.136-140
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• 2012

Leaves of Lindera obtusiloba were collected from four sites in Gangwon Province, Korea. Endophytic fungi were isolated from the leaves and identified using ITS sequences of rDNA. Total twelve species belonging of endophytic fungi were identified; Alternaria alternata, Annulohypoxylon annulatum, Creosphaeria sassafras, Diaporthe eres, Discosia sp., Epicoccum nigrum, Glomerella acutata, Glomerella cingulata, Paraconiothyrium brasiliense, Pestalotiopsis neglecta, Phomopsis amygdali, Xylaria sp. The endophytic fungus, Phomopsis amygdali, was the most dominant species isolated from L. obtusiloba in this study and the fungal diversities varied in the different sites.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.276-284
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• 2008

A lactic acid bacterium having antifungal activity was isolated from kimchi. It was identified as Lactobacillus plantarum based on its morphological and biochemical properties, and 16S rRNA sequence, and designated as Lb. plantarum AF1. This isolate inhibited the growth of Aspergillus flavus ATCC 22546, A. fumigatus ATCC 96918, A. petrakii PF-1, A. ochraceus PF-2, A. nidulans PF-3, Epicoccum nigrum KF-1, and Cladosporium gossypiicola KF-2 under a dual culture overlay assay. Also, the antimicrobial activity was found to be active against various species of Gram-positive and Gram-negative bacteria. The antifungal activity was found to be stable after heat ($121^{\circ}C$, 15 min) and proteolytic enzyme treatment, but it was unstable over pH 5.0. The antifungal compound(s) was estimated to have a low molecular mass (below 3,000 Da).