Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i>
The <i>Alternaria alternata</i> tangerine pathotype causes Alternaria brown spot, a devastating disease of susceptible tangerine varieties and their hybrids. <i>Alternaria citri</i> toxin (ACT) is the primary virulence factor, but the regulatory mechanisms governing ACT synth...
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2025-06-01
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| author | Jiejing Tang Hang Zhou Chen Jiao Hongye Li |
| author_facet | Jiejing Tang Hang Zhou Chen Jiao Hongye Li |
| author_sort | Jiejing Tang |
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| description | The <i>Alternaria alternata</i> tangerine pathotype causes Alternaria brown spot, a devastating disease of susceptible tangerine varieties and their hybrids. <i>Alternaria citri</i> toxin (ACT) is the primary virulence factor, but the regulatory mechanisms governing ACT synthesis remain unclear. Deubiquitinating enzymes maintain ubiquitination homeostasis and regulate fungal pathogenicity, yet their role in <i>A. alternata</i> remains unexplored. We characterized 13 ubiquitin-specific protease (UBP) family members in <i>A. alternata</i> tangerine pathotype. Six UBP genes (<i>Aaubp2</i>, <i>Aaubp3</i>, <i>Aaubp4</i>, <i>Aaubp6</i>, <i>Aaubp14</i>, and <i>Aaubp15</i>) regulated mycelial growth. <i>Aaubp14</i> deletion abolished sporulation, while mutations of <i>Aaubp3</i>, <i>Aaubp4</i>, <i>Aaubp6</i>, <i>Aaubp8</i>, and <i>Aaubp15</i> altered conidial morphology. qRT-PCR demonstrated distinct host-induced expression patterns among <i>Aaubp</i> genes. Pathogenicity tests showed that Δ<i>Aaubp6</i>, Δ<i>Aaubp14</i>, and Δ<i>Aaubp15</i> mutants failed to produce lesions on <i>Citrus reticulata</i> cv. Hongjv leaves. Moreover, <i>Aaubp14</i> deletion significantly suppressed ACT biosynthesis gene expression and blocked ACT production. Comparative proteomics showed Aaubp14 regulates ACT biosynthesis by modulating protein ubiquitination in metabolic pathways and controls pathogenicity via a complex network. Our findings elucidate <i>Aaubp</i> gene function in development and pathogenicity, particularly the Aaubp14-mediated regulation mechanism, providing insights into ubiquitination-mediated pathogenicity in phytopathogenic fungi. |
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| publishDate | 2025-06-01 |
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| spelling | doaj-art-cbd357e89f3b4889a7fc7a81a3329fb22025-07-25T13:27:19ZengMDPI AGJournal of Fungi2309-608X2025-06-0111749510.3390/jof11070495Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i>Jiejing Tang0Hang Zhou1Chen Jiao2Hongye Li3The Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, The Key Laboratory of Molecular Biology of Crop Pathogens and Insects of Ministry of Agriculture, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, ChinaThe Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Institute of Insect Science, Zhejiang University, Hangzhou 310058, ChinaThe Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, The Key Laboratory of Molecular Biology of Crop Pathogens and Insects of Ministry of Agriculture, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, ChinaThe Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, The Key Laboratory of Molecular Biology of Crop Pathogens and Insects of Ministry of Agriculture, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, ChinaThe <i>Alternaria alternata</i> tangerine pathotype causes Alternaria brown spot, a devastating disease of susceptible tangerine varieties and their hybrids. <i>Alternaria citri</i> toxin (ACT) is the primary virulence factor, but the regulatory mechanisms governing ACT synthesis remain unclear. Deubiquitinating enzymes maintain ubiquitination homeostasis and regulate fungal pathogenicity, yet their role in <i>A. alternata</i> remains unexplored. We characterized 13 ubiquitin-specific protease (UBP) family members in <i>A. alternata</i> tangerine pathotype. Six UBP genes (<i>Aaubp2</i>, <i>Aaubp3</i>, <i>Aaubp4</i>, <i>Aaubp6</i>, <i>Aaubp14</i>, and <i>Aaubp15</i>) regulated mycelial growth. <i>Aaubp14</i> deletion abolished sporulation, while mutations of <i>Aaubp3</i>, <i>Aaubp4</i>, <i>Aaubp6</i>, <i>Aaubp8</i>, and <i>Aaubp15</i> altered conidial morphology. qRT-PCR demonstrated distinct host-induced expression patterns among <i>Aaubp</i> genes. Pathogenicity tests showed that Δ<i>Aaubp6</i>, Δ<i>Aaubp14</i>, and Δ<i>Aaubp15</i> mutants failed to produce lesions on <i>Citrus reticulata</i> cv. Hongjv leaves. Moreover, <i>Aaubp14</i> deletion significantly suppressed ACT biosynthesis gene expression and blocked ACT production. Comparative proteomics showed Aaubp14 regulates ACT biosynthesis by modulating protein ubiquitination in metabolic pathways and controls pathogenicity via a complex network. Our findings elucidate <i>Aaubp</i> gene function in development and pathogenicity, particularly the Aaubp14-mediated regulation mechanism, providing insights into ubiquitination-mediated pathogenicity in phytopathogenic fungi.https://www.mdpi.com/2309-608X/11/7/495<i>Alternaria alternata</i> tangerine pathotypeubiquitin-specific proteaseACT biosynthesispathogenicityubiquitome |
| spellingShingle | Jiejing Tang Hang Zhou Chen Jiao Hongye Li Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> Journal of Fungi <i>Alternaria alternata</i> tangerine pathotype ubiquitin-specific protease ACT biosynthesis pathogenicity ubiquitome |
| title | Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> |
| title_full | Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> |
| title_fullStr | Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> |
| title_full_unstemmed | Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> |
| title_short | Functional Characterization of Deubiquitinase UBP Family and Proteomic Analysis of Aaubp14-Mediated Pathogenicity Mechanism in <i>Alternaria alternata</i> |
| title_sort | functional characterization of deubiquitinase ubp family and proteomic analysis of aaubp14 mediated pathogenicity mechanism in i alternaria alternata i |
| topic | <i>Alternaria alternata</i> tangerine pathotype ubiquitin-specific protease ACT biosynthesis pathogenicity ubiquitome |
| url | https://www.mdpi.com/2309-608X/11/7/495 |
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