As a well-known edible fungus rich in nutrients, Pleurotus ostreatus has been used as an alternative to expensive wild edible fungi. Specifically, the fact that using P. ostreatus instead 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. ostreatus has yet to be developed. In this study, a DNA extraction method for P. ostreatus was optimized, and pyrG was 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. ostreatus varieties and 7 other species. A low detection limit of 5 pg/μL was obtained by TaqMan quantitative PCR, and no pyrG amplification product was observed in the 7 other species. No allelic variation was detected in P. ostreatus varieties. These experiments confirmed that pyrG was 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. ostreatus samples and determination of adulteration in wild mushrooms. PRACTICAL APPLICATION The pyrG gene 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. ostreatus products and other processed mushroom foods.

中文翻译：

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在平菇定性和实时定量 PCR 检测中用作内源参考基因的 pyrG 的鉴定

作为著名的营养丰富的食用菌，平菇已被用作昂贵野生食用菌的替代品。具体而言，使用牡蛎代替其他价格昂贵的野生食用菌，损害了消费者的权益。在现有的食品掺假检测方法中，内参基因扩增是最准确的方法。然而，尚未开发出一种理想的 P. ostreatus 内源性参考基因。本研究优化了牡蛎的DNA提取方法，通过序列比对选择pyrG作为种特异性基因。随后对该基因进行了定性和定量聚合酶链式反应 (PCR) 测定，其中包括 3 个不同的 P. ostreatus 品种和 7 个其他物种。TaqMan 定量 PCR 检测限低至 5 pg/μL，其他 7 个物种均未观察到 pyrG 扩增产物。在 P. ostreatus 品种中未检测到等位基因变异。这些实验证实pyrG是一种理想的内源性参考基因，可用于P. ostreatus的定性和实时定量PCR检测。该方法也适用于加工平菇样品的检验和野生蘑菇中掺假的测定。实际应用 pyrG基因被选为理想的内参基因，用于P. ostreatus的定性和实时定量PCR检测，定量检测限为5 pg/μL。该方法不仅用于原料，还用于加工平菇产品和其他加工蘑菇食品。