Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation

<i>Fusarium acuminatum</i> is recognized as the causative agent of root rot in many forestry and agricultural plants. In recent years, root rot and foliage blight caused by <i>F. acuminatum</i> have become widespread and severe in China, particularly affecting <i>Dianth...

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Main Authors: Fangyi Ju, Zhongqiang Qi, Jiajin Tan, Tingting Dai
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/12/12/2427
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author Fangyi Ju
Zhongqiang Qi
Jiajin Tan
Tingting Dai
author_facet Fangyi Ju
Zhongqiang Qi
Jiajin Tan
Tingting Dai
author_sort Fangyi Ju
collection DOAJ
description <i>Fusarium acuminatum</i> is recognized as the causative agent of root rot in many forestry and agricultural plants. In recent years, root rot and foliage blight caused by <i>F. acuminatum</i> have become widespread and severe in China, particularly affecting <i>Dianthus chinensis</i>. The infection mechanism of <i>F. acuminatum</i> remains a pressing area for research. A crucial approach to elucidating its pathogenic mechanisms involves the genetic modification of candidate genes, which necessitates effective transformation systems. Currently, protoplast-mediated transformation (PMT) serves as a valuable tool for studying plant-pathogen interactions and offers several advantages over conventional transformation methods. In this study, we employed the PMT technique to establish a transformation system for the <i>F. acuminatum</i> strain FDCY-5 due to its benefits such as ease of operation, low cost, high conversion efficiency, and broad applicability. We successfully developed a transformation system capable of producing abundant high-quality protoplasts from <i>F. acuminatum</i> and generating green fluorescent protein (GFP) transformants. To verify whether GFP was constitutively expressed, we utilized fluorescence microscopy alongside PCR technology. The results demonstrated that GFP was effectively transformed into the protoplasts of <i>F. acuminatum</i> and expressed successfully. The established protoplast transformation system for <i>F. acuminatum</i> provides a foundational platform for analyzing functional genes within infected host plants as well as understanding the molecular mechanisms underlying host plant infections by <i>F. acuminatum.</i>
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spelling doaj-art-6f9b9d9c06b84b90b0e86f62ce43d50a2024-12-27T14:41:07ZengMDPI AGMicroorganisms2076-26072024-11-011212242710.3390/microorganisms12122427Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic TransformationFangyi Ju0Zhongqiang Qi1Jiajin Tan2Tingting Dai3Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210000, ChinaInstitute of Plant Protection, Jiangsu Academy of Agricultural Science, Nanjing 210014, ChinaCo-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210000, ChinaCo-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210000, China<i>Fusarium acuminatum</i> is recognized as the causative agent of root rot in many forestry and agricultural plants. In recent years, root rot and foliage blight caused by <i>F. acuminatum</i> have become widespread and severe in China, particularly affecting <i>Dianthus chinensis</i>. The infection mechanism of <i>F. acuminatum</i> remains a pressing area for research. A crucial approach to elucidating its pathogenic mechanisms involves the genetic modification of candidate genes, which necessitates effective transformation systems. Currently, protoplast-mediated transformation (PMT) serves as a valuable tool for studying plant-pathogen interactions and offers several advantages over conventional transformation methods. In this study, we employed the PMT technique to establish a transformation system for the <i>F. acuminatum</i> strain FDCY-5 due to its benefits such as ease of operation, low cost, high conversion efficiency, and broad applicability. We successfully developed a transformation system capable of producing abundant high-quality protoplasts from <i>F. acuminatum</i> and generating green fluorescent protein (GFP) transformants. To verify whether GFP was constitutively expressed, we utilized fluorescence microscopy alongside PCR technology. The results demonstrated that GFP was effectively transformed into the protoplasts of <i>F. acuminatum</i> and expressed successfully. The established protoplast transformation system for <i>F. acuminatum</i> provides a foundational platform for analyzing functional genes within infected host plants as well as understanding the molecular mechanisms underlying host plant infections by <i>F. acuminatum.</i>https://www.mdpi.com/2076-2607/12/12/2427<i>F. acuminatum</i>transformationPMTGFP
spellingShingle Fangyi Ju
Zhongqiang Qi
Jiajin Tan
Tingting Dai
Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
Microorganisms
<i>F. acuminatum</i>
transformation
PMT
GFP
title Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
title_full Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
title_fullStr Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
title_full_unstemmed Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
title_short Development of Green Fluorescent Protein-Tagged Strains of <i>Fusarium acuminatum</i> via PEG-Mediated Genetic Transformation
title_sort development of green fluorescent protein tagged strains of i fusarium acuminatum i via peg mediated genetic transformation
topic <i>F. acuminatum</i>
transformation
PMT
GFP
url https://www.mdpi.com/2076-2607/12/12/2427
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