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Development of Green Fluorescent Protein-Tagged Strains of Fusarium acuminatum via PEG-Mediated Genetic Transformation

Summary

Scientists developed a method to genetically modify a harmful fungus called Fusarium acuminatum that causes root rot in plants like carnations. Using a technique that breaks down the fungal cell wall and uses a special chemical (PEG) to insert genes, they successfully added a green-glowing protein (GFP) marker to the fungus. This allows researchers to track where and how the fungus infects plants. The modified fungus still behaves normally, making it a useful tool for identifying which genes make the fungus dangerous, potentially leading to better disease control methods.

Background

Fusarium acuminatum is a causative agent of root rot in forestry and agricultural plants, with increasing severity in China particularly affecting Dianthus chinensis. Understanding the infection mechanisms of F. acuminatum requires effective genetic transformation systems. Protoplast-mediated transformation (PMT) offers advantages over conventional transformation methods including ease of operation, low cost, and high conversion efficiency.

Objective

The study aimed to establish an efficient PEG-mediated protoplast transformation system for F. acuminatum strain FDCY-5 to develop green fluorescent protein (GFP)-tagged strains. This system would provide a foundational platform for analyzing functional genes and understanding the molecular mechanisms underlying host plant infections by F. acuminatum.

Results

Thirty of 32 transformants successfully expressed the GFP gene with 93.75% amplification efficiency. GFP was stably expressed in both mycelia and conidia after multiple generations on selective plates. The transformant FaGFP-1 showed no significant differences from wild-type in growth rate, dry weight, spore production, stress responses, or pathogenicity.

Conclusion

An effective and reproducible PEG-mediated protoplast transformation system was successfully established for F. acuminatum. The system does not adversely affect fungal growth or pathogenicity when foreign genes are introduced, providing a reliable method for identifying genes related to pathogenicity and observing colonization dynamics in host plants.
  • Published in:Microorganisms,
  • Study Type:Experimental Research,
  • Source: PMID: 39770630, DOI: 10.3390/microorganisms12122427
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