Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i>
Anthracnose is one of the most widespread and destructive diseases in grapes. Grape anthracnose can be caused by various <i>Colletotrichum</i> species, such as <i>Colletotrichum gloeosporioides</i> and <i>Colletotrichum cuspidosporium</i>. In recent years, <i&g...
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MDPI AG
2023-04-01
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| author | Shendan Yu Jing Wang Rongyao Chai Haiping Qiu Ziqi Lu Zhen Zhang Lin Li Jiaoyu Wang Guochang Sun |
| author_facet | Shendan Yu Jing Wang Rongyao Chai Haiping Qiu Ziqi Lu Zhen Zhang Lin Li Jiaoyu Wang Guochang Sun |
| author_sort | Shendan Yu |
| collection | DOAJ |
| description | Anthracnose is one of the most widespread and destructive diseases in grapes. Grape anthracnose can be caused by various <i>Colletotrichum</i> species, such as <i>Colletotrichum gloeosporioides</i> and <i>Colletotrichum cuspidosporium</i>. In recent years, <i>Colletotrichum aenigma</i> was reported as a causal agent of Grape anthracnose in China and South Korea. Peroxisome is an important organelle in eukaryotes, which plays a very important role in the growth, development, and pathogenicity of several plant-pathogenic fungal species i, but it has not been reported in <i>C. aenigma.</i> In this work, the peroxisome of <i>C. aenigma</i> was labeled with a fluorescent protein, using green fluorescent protein (GFP) and red fluorescent protein (DsRED and mCherry) as reporter genes. Via <i>Agrobacterium tumefaciens</i>-mediated transformation (<i>At</i>MT), two fluorescent fusion vectors to mark the peroxisomes, with GFP and DsRED, respectively, were introduced into a wild-type strain of <i>C. aenigma</i>. In the transformants, bright dots of green or red fluorescence in hyphae and spores could be seen in the strains labeled peroxisome. The nuclei labeled by the same method showed bright round fluorescent spots. In addition, we also combined fluorescent protein labeling with chemical staining to show the localization more clearly. The ideal peroxisome and nuclear fluorescence-labeled <i>C. aenigma strain</i> was obtained, which provided a reference for the study of its growth, development, and pathogenicity. |
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| last_indexed | 2024-03-11T04:51:31Z |
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| spelling | doaj.art-ee9ce672033944e59cb7697124e1b1822023-11-17T19:58:56ZengMDPI AGJournal of Fungi2309-608X2023-04-019449310.3390/jof9040493Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i>Shendan Yu0Jing Wang1Rongyao Chai2Haiping Qiu3Ziqi Lu4Zhen Zhang5Lin Li6Jiaoyu Wang7Guochang Sun8College of Life Sciences, Zhejiang Normal University, Jinhua 321004, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaCollege of Advanced Agricultural Sciences, Zhejiang Agriculture and Forestry University, Hangzhou 310007, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaAnthracnose is one of the most widespread and destructive diseases in grapes. Grape anthracnose can be caused by various <i>Colletotrichum</i> species, such as <i>Colletotrichum gloeosporioides</i> and <i>Colletotrichum cuspidosporium</i>. In recent years, <i>Colletotrichum aenigma</i> was reported as a causal agent of Grape anthracnose in China and South Korea. Peroxisome is an important organelle in eukaryotes, which plays a very important role in the growth, development, and pathogenicity of several plant-pathogenic fungal species i, but it has not been reported in <i>C. aenigma.</i> In this work, the peroxisome of <i>C. aenigma</i> was labeled with a fluorescent protein, using green fluorescent protein (GFP) and red fluorescent protein (DsRED and mCherry) as reporter genes. Via <i>Agrobacterium tumefaciens</i>-mediated transformation (<i>At</i>MT), two fluorescent fusion vectors to mark the peroxisomes, with GFP and DsRED, respectively, were introduced into a wild-type strain of <i>C. aenigma</i>. In the transformants, bright dots of green or red fluorescence in hyphae and spores could be seen in the strains labeled peroxisome. The nuclei labeled by the same method showed bright round fluorescent spots. In addition, we also combined fluorescent protein labeling with chemical staining to show the localization more clearly. The ideal peroxisome and nuclear fluorescence-labeled <i>C. aenigma strain</i> was obtained, which provided a reference for the study of its growth, development, and pathogenicity.https://www.mdpi.com/2309-608X/9/4/493<i>Colletotrichum aenigma</i>fluorescent protein labelingperoxisomenucleus |
| spellingShingle | Shendan Yu Jing Wang Rongyao Chai Haiping Qiu Ziqi Lu Zhen Zhang Lin Li Jiaoyu Wang Guochang Sun Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> Journal of Fungi <i>Colletotrichum aenigma</i> fluorescent protein labeling peroxisome nucleus |
| title | Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> |
| title_full | Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> |
| title_fullStr | Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> |
| title_full_unstemmed | Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> |
| title_short | Fluorescent Labeling of Peroxisome and Nuclear in <i>Colletotrichum aenigma</i> |
| title_sort | fluorescent labeling of peroxisome and nuclear in i colletotrichum aenigma i |
| topic | <i>Colletotrichum aenigma</i> fluorescent protein labeling peroxisome nucleus |
| url | https://www.mdpi.com/2309-608X/9/4/493 |
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