Abstract: In this study, we collected a total of 38 samples of 27 Amanita species and extracted their genomic DNA. Universal primers were used to amplify the internal transcribed spacer (ITS), large ribosomal subunit (LSU), the second-largest subunit of RNA polymerase II (RPB2), and the β-tubulin gene sequences. Sanger bidirectional sequences were obtained, proofread and then submitted to the NCBI GenBank for sequence alignment to identify the species. We calculated the intra-species and inter-species Kimura-2-Parameter (K2P) genetic distance and constructed the phylogenetic tree. The results indicated that β-tubulin and ITS were more suitable than RPB2 and LSU for use in the identification of Amanita species. The combined use of β-tubulin and ITS could be recommended to identify Amanita species, providing early warning of foodborne poisoning caused by poisonous mushrooms. β-tubulin was shorter than LSU, ITS, and RPB2, being suitable for use in the analysis of highly-processed mushroom products and vomits after eating poisonous mushrooms by mistake. Thus, β-tubulin can be used as the optimal barcode to identify and trace Amanita species causing mushroom poisoning.

Key words: Amanita; DNA barcoding; species identification