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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 beneficial fungus called Penicillium sclerotiorum by removing its cell wall and introducing new genes. They added a glowing green marker (GFP) to track the fungus as it colonizes tomato plant roots. The study shows that enzymatic treatment of seeds significantly improves how well the fungus attaches to roots, potentially helping plants grow better while revealing how the genetic modification affects the fungus’s internal chemistry.

Background

Penicillium sclerotiorum is an important fungal species with biotechnological applications. Protoplast-based transformation is a vital tool for genetic studies in fungi, but no protoplast method previously existed for P. sclerotiorum-scaumcx01.

Objective

To develop and optimize an efficient protoplast isolation, regeneration, and transformation system for P. sclerotiorum-scaumcx01, and to investigate metabolomic changes and root colonization dynamics following GFP tagging.

Results

Optimal protoplast yield of 6.72 × 10⁶ cells/mL was achieved after 12h enzymatic digestion at 28°C using combined enzymes and 1M MgSO₄. PEG-mediated transformation resulted in stable GFP expression over five generations. Metabolomic analysis revealed significant shifts in glycerophospholipid metabolism. GFP-tagged strain successfully colonized tomato roots, particularly in cortical tissues after enzymatic pretreatment.

Conclusion

An efficient protoplast transformation system for P. sclerotiorum-scaumcx01 was successfully established, enabling stable genetic modification and GFP tagging. The study demonstrates metabolic impacts of genetic modifications and reveals enzymatic seed treatment as an effective method for enhancing plant-fungal interactions.
  • Published in:Journal of Fungi,
  • Study Type:Experimental Research,
  • Source: 40422719
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