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PEG-Mediated Protoplast Transformation of Penicillium sclerotiorum (scaumcx01): Metabolomic Shifts and Root Colonization Dynamics

Summary

Scientists developed a new method to genetically modify a fungus called Penicillium sclerotiorum by using protoplasts, which are fungal cells with their protective outer walls removed. They added a glowing green protein (GFP) to track the fungus and discovered that this modification changed how the fungus uses fats and lipids. When they treated tomato seeds with enzymes before exposing them to the modified fungus, it enhanced the fungus’s ability to colonize plant roots, potentially helping plants grow better.

Background

Penicillium sclerotiorum is an endophytic fungus with biotechnological applications in production of bioactive compounds and antibiotics. Protoplast-based transformation is a vital tool for genetic studies in fungi, but no protoplast method previously existed for P. sclerotiorum-scaumcx01. This study addresses the need for developing an efficient and reproducible protoplast transformation protocol for this fungal species.

Objective

To optimize protoplast isolation, regeneration, and transformation efficiency in P. sclerotiorum-scaumcx01 using PEG-mediated transformation. To investigate metabolomic shifts following GFP tagging and to demonstrate the colonization dynamics of GFP-labeled fungal strains on tomato roots.

Results

Highest protoplast yield of 6.72 × 10⁶ cells/mL was obtained after 12 h enzymatic digestion at 28°C with 1 M MgSO₄ as osmotic stabilizer, achieving 1.02% regeneration rate. PEG-mediated transformation successfully generated stable GFP-expressing transformants over five generations. Metabolomic analysis revealed significant changes in glycerophospholipid metabolism. GFP-tagged scaumcx01 successfully colonized tomato roots, particularly in cortical tissues, with enhanced colonization following enzymatic seed pretreatment.

Conclusion

An efficient protoplast transformation system for P. sclerotiorum-scaumcx01 was successfully established with stable GFP expression achieved. Genetic modification induced metabolic shifts particularly affecting lipid metabolism pathways. Enzymatic seed treatment enhanced fungal colonization of tomato roots, suggesting potential applications in plant-fungal interaction studies and agricultural enhancement.
  • Published in:Journal of Fungi,
  • Study Type:Experimental Study,
  • Source: 10.3390/jof11050386, PMID: 40422719
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