The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses
<i>Metarhizium anisopliae</i> is an entomopathogenic fungus which may enhance plant growth and resistance when acting as an endophyte in host plants. However, little is known about the protein interactions nor their activating mechanisms. Common in fungal extracellular membrane (CFEM) pr...
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/4/4037 |
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| author | Ni Cai Xiangqun Nong Rong Liu Mark Richard McNeill Guangjun Wang Zehua Zhang Xiongbing Tu |
| author_facet | Ni Cai Xiangqun Nong Rong Liu Mark Richard McNeill Guangjun Wang Zehua Zhang Xiongbing Tu |
| author_sort | Ni Cai |
| collection | DOAJ |
| description | <i>Metarhizium anisopliae</i> is an entomopathogenic fungus which may enhance plant growth and resistance when acting as an endophyte in host plants. However, little is known about the protein interactions nor their activating mechanisms. Common in fungal extracellular membrane (CFEM) proteins have been identified as plant immune regulators that suppress or activate plant resistance responses. Here, we identified a CFEM domain-containing protein, MaCFEM85, which was mainly localized in the plasma membrane. Yeast two-hybrid (Y2H), glutathione-S-transferase (GST) pull-down, and bimolecular fluorescence complementation assays demonstrated that MaCFEM85 interacted with the extracellular domain of a <i>Medicago sativa</i> (alfalfa) membrane protein, MsWAK16. Gene expression analyses showed that MaCFEM85 and MsWAK16 were significantly upregulated in <i>M. anisopliae</i> and <i>M. sativa</i>, respectively, from 12 to 60 h after co-inoculation. Additional yeast two-hybrid assays and amino acid site-specific mutation indicated that the CFEM domain and 52th cysteine specifically were required for the interaction of MaCFEM85 with MsWAK16. Defense function assays showed that JA was up-regulated, but <i>Botrytis cinerea</i> lesion size and <i>Myzus persicae</i> reproduction were suppressed by transient expression of MaCFEM85 and MsWAK16 in the model host plant <i>Nicotiana benthamiana</i>. Collectively, these results provide novel insights into the molecular mechanisms underlying interactions of <i>M. anisopliae</i> with host plants. |
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| last_indexed | 2024-03-11T08:41:01Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9f19bcbc8d7c46aba60a22629bbb515e2023-11-16T21:08:38ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01244403710.3390/ijms24044037The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant DefensesNi Cai0Xiangqun Nong1Rong Liu2Mark Richard McNeill3Guangjun Wang4Zehua Zhang5Xiongbing Tu6The State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaThe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaThe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaAgResearch, Lincoln Science Centre, Private Bag 4749, Canterbury 8140, New ZealandThe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaThe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaThe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China<i>Metarhizium anisopliae</i> is an entomopathogenic fungus which may enhance plant growth and resistance when acting as an endophyte in host plants. However, little is known about the protein interactions nor their activating mechanisms. Common in fungal extracellular membrane (CFEM) proteins have been identified as plant immune regulators that suppress or activate plant resistance responses. Here, we identified a CFEM domain-containing protein, MaCFEM85, which was mainly localized in the plasma membrane. Yeast two-hybrid (Y2H), glutathione-S-transferase (GST) pull-down, and bimolecular fluorescence complementation assays demonstrated that MaCFEM85 interacted with the extracellular domain of a <i>Medicago sativa</i> (alfalfa) membrane protein, MsWAK16. Gene expression analyses showed that MaCFEM85 and MsWAK16 were significantly upregulated in <i>M. anisopliae</i> and <i>M. sativa</i>, respectively, from 12 to 60 h after co-inoculation. Additional yeast two-hybrid assays and amino acid site-specific mutation indicated that the CFEM domain and 52th cysteine specifically were required for the interaction of MaCFEM85 with MsWAK16. Defense function assays showed that JA was up-regulated, but <i>Botrytis cinerea</i> lesion size and <i>Myzus persicae</i> reproduction were suppressed by transient expression of MaCFEM85 and MsWAK16 in the model host plant <i>Nicotiana benthamiana</i>. Collectively, these results provide novel insights into the molecular mechanisms underlying interactions of <i>M. anisopliae</i> with host plants.https://www.mdpi.com/1422-0067/24/4/4037wall-associated kinaseCFEMs<i>Metahizium anisopliae</i>plant immunity<i>Medicago sativa</i> |
| spellingShingle | Ni Cai Xiangqun Nong Rong Liu Mark Richard McNeill Guangjun Wang Zehua Zhang Xiongbing Tu The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses International Journal of Molecular Sciences wall-associated kinase CFEMs <i>Metahizium anisopliae</i> plant immunity <i>Medicago sativa</i> |
| title | The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses |
| title_full | The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses |
| title_fullStr | The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses |
| title_full_unstemmed | The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses |
| title_short | The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses |
| title_sort | conserved cysteine rich secretory protein macfem85 interacts with mswak16 to activate plant defenses |
| topic | wall-associated kinase CFEMs <i>Metahizium anisopliae</i> plant immunity <i>Medicago sativa</i> |
| url | https://www.mdpi.com/1422-0067/24/4/4037 |
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