Identification of the Pol Gene as a Species-Specific Diagnostic Marker for Qualitative and Quantitative PCR Detection of Tricholoma matsutake.

Luying Shan,Yinjiao Li,Wentao Xu,Shi Zheng,Dazhou Wang,Ying Shang

doi:10.3390/molecules24030455

Abstract

Tricholoma matsutake is a rare, precious, and wild edible fungus that could not be cultivated artificially until now. This situation has given way to the introduction of fake T. matsutake commodities to the mushroom market. Among the methods used to detect food adulteration, amplification of species-specific diagnostic marker is particularly important and accurate. In this study, the Pol gene is reported as a species-specific diagnostic marker to identify three T. matsutake varieties and 10 other types of edible mushrooms through qualitative and quantitative PCR. The PCR results did not reveal variations in the amplified region, and the detection limits of qualitative and quantitative PCR were found to be 8 ng and 32 pg, respectively. Southern blot showed that the Pol gene exists as a single copy in the T. matsutake genome. The method that produced the purest DNA of T. matsutake in this study was also determined, and the high-concentration salt precipitation method was confirmed to be the most suitable among the methods tested. The assay proposed in this work is applicable not only to the detection of raw materials but also to the examination of processed products containing T. matsutake.

Highlights

Tricholoma matsutake is an ectomycorrhizal agaricomycete predominantly associated with pines and oaks
These results indicate that the Pol gene did not show the sequence variation among the T. matsutake varieties studied
Lentinula edodes, Pleurotus eryngii,Sichuan, Flammulina velutipes, and Pleurotus varieties strains collected from the provinces of Yunnan, and Jilin, China, were kindly ostreatus

Summary

Introduction

Tricholoma matsutake is an ectomycorrhizal agaricomycete predominantly associated with pines and oaks. It is a commercially valuable edible mushroom [1] with great significance, because of its delicate flavor and because of its diverse biological properties [2], which include multiple immunostimulatory, hematopoietic, antineoplastic, antimutation, and antioxidation activities [3]. The growing number of T. matsutake consumers has steadily expanded market demands for this mushroom. Wild T. matsutake and its products are in a short supply in the market, and, as such, its economic value has risen sharply. To address demands for the mushroom and reap higher profits, merchants frequently pass off counterfeit or adulterated T. matsutake products as genuine items, for example, sliced dried Agaricus blazei [5].

Methods

Results

Conclusion