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Objective To establish a method for qualitative determination of Tricholoma matsutake ingredients by a TaqMan real-time fluorescence polymerase chain reaction (PCR), and identify the conformity of Tricholoma matsutake and its products with their labels by DNA barcoding technology. Methods Taking the pol gene of Tricholoma matsutake as the target gene, specific primers and probes were designed to study and analyze their specificity, sensitivity and repeatability. The established TaqMan real-time PCR method of Tricholoma matsutake was used to detect different types of commercially available samples and identify their authenticity. Results The established method was used for real-time PCR detection, and there was no cross-reactivity between Tricholoma matsutake and 32 kinds of other edible fungi, animals and plants, indicading strong specificity of the method. The sensitivity was 0.01% Tricholoma matsutake and 0.01 ng/μL Tricholoma matsutake genomic DNA. The authenticity test of 160 commercially available Tricholoma matsutake and their products showed that there was counterfeiting of dried Tricholoma matsutake. The test results did not match the labels at a rate of 36.70%. DNA barcoding technology was used to further identify the species of counterfeit matsutake mushrooms, with species attributes of Stropharia rugosoannulata, Tricholoma giganteum and Tricholoma bakamatsutake. The non-compliance rates with labels in the authenticity testing resultes of other collected matsutake products were 23.70%-60.00%, respectively. Conclusion This method has high sensitivity and strong specificity, which can quickly and accurately identify the authenticity and compliance of Tricholoma matsutake ingredients.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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