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Identification of matB used as an endogenous reference gene for the qualitative and real-time quantitative polymerase chain reaction detection of Lentinus edodes

Summary

This study identifies the matB gene as a reliable genetic marker for detecting shiitake mushrooms (Lentinus edodes) in food products. Researchers developed a simple DNA test that can identify L. edodes in processed foods where it might be fraudulently substituted for more expensive wild mushrooms. The test is highly sensitive, detecting DNA at extremely low concentrations, and works on both raw mushrooms and processed products. This method provides food manufacturers and regulators with an effective tool to prevent mushroom fraud and ensure food authenticity.

Background

Lentinus edodes is a widely cultivated medicinal and edible mushroom accounting for approximately 25% of world mushroom production. Food adulteration, particularly disguising L. edodes as expensive wild edible mushrooms in processed foods, is an increasing concern. Traditional morphological and physiochemical identification methods are inadequate for processed products that have undergone complex treatments.

Objective

To identify and validate the matB gene as an endogenous reference gene for species-specific detection of L. edodes using qualitative and real-time quantitative PCR. To establish a convenient and accurate method for detecting L. edodes products and adulteration in wild edible mushroom products.

Results

The matB gene showed no homology with other mushroom species and amplified only L. edodes DNA in qualitative PCR with no amplification in 18 non-L. edodes species. Conventional PCR detected DNA at concentrations as low as 1 ng/µl. Real-time quantitative PCR achieved a detection limit of 16 pg/µl with a linear relationship (R² = 0.999) between DNA quantity and Ct values. Application to processed products successfully detected L. edodes in L. edodes sauce but not in Pleurotus eryngii sauce.

Conclusion

The matB gene is an ideal endogenous reference gene for L. edodes detection with high species specificity and excellent sensitivity (16 pg/µl). The established qualitative and quantitative PCR methods are suitable for detecting L. edodes in both raw materials and processed products, providing a convenient and accurate approach for preventing food adulteration in wild edible mushroom products.
  • Published in:Food Science & Nutrition,
  • Study Type:Experimental Study,
  • Source: 10.1002/fsn3.2860
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