Abstract
Abstract
Mushroom poisoning contributes significantly to global foodborne diseases and related fatalities. Amanita mushrooms frequently cause such poisonings; however, identifying these toxic species is challenging due to the unavailability of fresh and intact samples. It is often necessary to analyze residues, vomitus, or stomach extracts to obtain DNA sequences for the identification of species responsible for causing food poisoning. This usually proves challenging to obtain usable DNA sequences that can be analyzed using conventional molecular biology techniques. Therefore, this study aimed to develop a DNA mini-barcoding method for the identification of Amanita species. Following the evaluation and optimization of universal primers for DNA mini-barcoding in Amanita mushrooms, we found that the internal transcribed spacer (ITS) gene sequence primer ITS-a was the most suitable DNA barcode primer for identifying Amanita species. Forty-three Amanita samples were subsequently amplified and sequenced. The sequences obtained were analyzed for intra- and inter-species genetic distances, and a phylogenetic tree was constructed. The findings indicated that the designed primers had strong universality among the Amanita samples and could accurately identify the target gene fragment with a length of 290 bp. Notably, the DNA mini-barcode accurately identified the 43 Amanita samples, demonstrating high consistency with the conventional DNA barcode. Furthermore, it effectively identified DNA from digested samples. In summary, this DNA mini-barcode is a promising tool for detecting accidental ingestion of toxic Amanita mushrooms. It may be used as an optimal barcode for species identification and traceability in events of Amanita-induced mushroom poisoning.
Key points
• Development of a DNA mini-barcoding method for Amanita species identification without fresh samples.
• The ITS-a primer set was optimized for robust universality in Amanita samples.
• The mini-barcode is suitable for screening toxic mushroom species in mushroom poisoning cases.
Graphical Abstract
Funder
Key Technologies Research and Development Program
Publisher
Springer Science and Business Media LLC
Reference48 articles.
1. Badotti F, de Oliveira FS, Garcia CF, Vaz ABM, Fonseca PLC, Nahum LA, Oliveira G, Góes-Neto A (2017) Effectiveness of ITS and sub-regions as DNA barcode markers for the identification of Basidiomycota (Fungi). BMC Microbiol 17(1):42
2. Bagley JC, de Aquino PDPU, Breitman MF, Langeani F, Colli GR (2019) DNA barcode and minibarcode identification of freshwater fishes from Cerrado headwater streams in Central Brazil. J Fish Biol 95(4):1046–1060
3. Dadpour B, Tajoddini S, Rajabi M, Afshari R (2017) Mushroom poisoning in the northeast of Iran; a retrospective 6-year epidemiologic study. Emergency 5(1):e23
4. Diaz JH (2018) Amatoxin-containing mushroom poisonings: species, toxidromes, treatments, and outcomes. Wilderness Environ Med 29(1):111–118
5. Dieffenbach CW, Lowe TMJ, Dveksler GS (1993) General concepts for PCR primer design. PCR Methods Appl 3(3):S30–S37