Identification of pyrGUsed as an Endogenous Reference Gene in Qualitative and Real‐Time Quantitative PCR Detection of Pleurotus ostreatus

• Autores: Shi Zheng, Luying Shan, Yongliang Zhuang, Ying Shang
• Localización: Journal of food science, ISSN 0022-1147, Vol. 83, Nº 3, 2018, págs. 750-755
• Idioma: inglés
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• Resumen

  • As a well‐known edible fungus rich in nutrients, Pleurotus ostreatushas been used as an alternative to expensive wild edible fungi. Specifically, the fact that using P. ostreatusinstead of other expensive wild edible fungi has damaged the rights and interests of consumers. Among the existing methods for detection of food adulteration, the amplification of endogenous reference gene is the most accurate method. However, an ideal endogenous reference gene for P. ostreatushas yet to be developed. In this study, a DNA extraction method for P. ostreatuswas optimized, and pyrGwas selected as a species‐specific gene through sequence alignment. This gene was subsequently subjected to qualitative and quantitative Polymerase Chain Reaction (PCR) assays with 3 different P. ostreatusvarieties and 7 other species. A low detection limit of 5 pg/μL was obtained by TaqMan quantitative PCR, and no pyrGamplification product was observed in the 7 other species. No allelic variation was detected in P. ostreatusvarieties. These experiments confirmed that pyrGwas an ideal endogenous reference gene for the qualitative and real‐time quantitative PCR detection of P. ostreatus. This method was also suitable for the examination of processed P. ostreatussamples and determination of adulteration in wild mushrooms. The pyrGgene was chosen as an ideal endogenous reference gene for the qualitative and real‐time quantitative PCR detection of P. ostreatus, and the detection limit was 5 pg/μL for the quantification. This method is used not only for raw materials but also for processed P. ostreatusproducts and other processed mushroom foods.