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The Transformation and Protein Expression of the Edible Mushroom Stropharia rugosoannulata Protoplasts by Agrobacterium-tumefaciens-Mediated Transformation

Summary

Researchers successfully developed a method to genetically modify the edible mushroom Stropharia rugosoannulata using Agrobacterium tumefaciens bacteria. This technique allows scientists to insert and express foreign genes in the mushroom, providing tools to study how specific genes control mushroom growth and the production of health-promoting compounds. The study demonstrates that both artificial and natural resistance markers can be used to identify successfully transformed mushrooms, offering a foundation for improving mushroom cultivation and breeding.

Background

Stropharia rugosoannulata is an edible cultivated mushroom with nutritional value and efficient cellulolytic enzymatic systems. However, the lack of genetic tools has severely limited investigation of its molecular mechanisms and functional genomic research, constraining precision breeding efforts.

Objective

To establish an Agrobacterium-tumefaciens-mediated genetic transformation (ATMT) system for S. rugosoannulata protoplasts and demonstrate expression of exogenous genes (mCherry, hph, GUS) and endogenous mutant genes (SDI) for development of molecular genetic tools.

Results

Successfully achieved stable expression of exogenous genes (mCherry, hph, GUS) and endogenous mutant SDI gene in S. rugosoannulata. Transformation efficiency was approximately 1.17‰ with hph resistance and 1.56‰ with mSDI resistance. T-DNA flanking sequences were characterized and integration sites confirmed.

Conclusion

The established ATMT system is feasible and effective for S. rugosoannulata genetic transformation, providing essential genetic tools for investigating target gene functions and molecular mechanisms. This system enables future research on gene deletion, overexpression, silencing, and precision breeding improvements.
  • Published in:Journal of Fungi,
  • Study Type:Experimental Research,
  • Source: 10.3390/jof11090674; PMID: 41003220
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