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Highly Efficient Electroporation-Mediated Transformation into Edible Mushroom Flammulina velutipes

Summary

This research developed an improved method for introducing new genes into the edible mushroom Flammulina velutipes (also known as enoki mushroom). The technique uses electrical pulses to transfer genetic material into mushroom cells with high success rates. This advancement has important implications for mushroom research and cultivation. Impacts on everyday life: • Could lead to improved varieties of edible mushrooms with better growth characteristics • May help develop mushrooms with enhanced nutritional content • Could contribute to more efficient and productive mushroom farming methods • Enables better understanding of mushroom biology for improved cultivation practices • May lead to development of mushrooms with novel beneficial properties

Background

Flammulina velutipes is a widely cultivated edible mushroom, especially in Asia, with limited genetic information available. Its genome is approximately 33 Mb organized into 8 chromosomes. Transformation technology is essential for studying gene functions, but existing methods often have low efficiency. The flammutoxin (FTX) gene produces a cardiotoxic and cytolytic polypeptide specifically expressed during fruiting body formation.

Objective

To develop a highly efficient transformation method for Flammulina velutipes using protoplasts and to study the integration of the flammutoxin gene through electroporation-mediated transformation.

Results

The highest transformation efficiency was achieved with an electric pulse of 1.25 kV/cm, yielding 177 transformants/µg of DNA in 1 × 107 protoplasts. PCR and Southern blot analysis confirmed that single copies of the plasmid DNA were inserted at different locations in the F. velutipes genome through non-homologous recombination. The transformed traits remained stable through multiple generations.

Conclusion

The study established a highly efficient electroporation-mediated transformation system for F. velutipes that could serve as a valuable tool for gene function analysis. While homologous recombination with the ftx target gene was not achieved, the method proved effective for stable transformation and random gene disruption studies.
  • Published in:Mycobiology,
  • Study Type:Laboratory Research,
  • Source: 10.4489/MYCO.2010.38.4.331
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