Identification and poisoning diagnosis of Aconitum materials using a genus-specific nucleotide signature

• Published in: Ecotoxicology and environmental safety Vol. 237; p. 113539
• Main Authors: Wang, Gang, Liu, Yang, Bai, Xuanjiao, Cao, Pei, Pang, Xiaohui, Han, Jianping
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.06.2022; Elsevier
• Subjects: Aconitum; Aconitum - genetics; Alkaloids; Food poisoning; Genome, Chloroplast; High-Throughput Nucleotide Sequencing; High-throughput sequencing; Identification; Nucleotide signature; Nucleotides; Simulated gastric fluid digestion; Nucleotide signature; Aconitum; Identification; High-throughput sequencing; Food poisoning; Simulated gastric fluid digestion
• ISSN: 0147-6513; 1090-2414
• DOI: 10.1016/j.ecoenv.2022.113539

Aconitum genus generally contains hypertoxic alkaloids. Poisoning incidents due to the improper ingestion of Aconitum materials frequently occur around the world. DNA barcoding is considered as a powerful tool for species identification, but complete sequences of conventional DNA barcodes are sometimes unattainable from food and highly processed products due to severe DNA degradation. Therefore, a shorter molecular marker will be more profitable for the authentication and poisoning diagnosis of Aconitum materials. In this study, 1246 psbA-trnH sequences and chloroplast genomes representing 183 taxa of Aconitum were collected, and a 23-bp nucleotide signature unique to Aconitum genus (5′-TATATGAGTCATTGAAGTTGCAG-3′) was developed. The nucleotide signature was conserved and universal within Aconitum while divergent among other genera. The specific molecular signature was then successfully applied to the detection of processed Aconitum ingredients. To further evaluate the application potential of nucleotide signature in completely unknown mixture samples, boiled food mixtures, containing different ratios of Aconitum materials, were sequenced by high-throughput sequencing technology. The results showed that the nucleotide signature sequence could be directly extracted from raw sequencing data, even at a low DNA concentration of 0.2 ng/µl. Consequently, the 23-bp genus-specific nucleotide signature represents a significant step forward in the use of DNA barcoding to identify processed samples and food mixtures with degraded DNA. This study undoubtedly provides a new perspective and strong support for the identification and detection of Aconitum-containing products, which can be further introduced to the diagnosis of food poisoning. •A 23-bp genus-specific nucleotide signature was innovatively developed for toxic Aconitum genus.•The nucleotide signature was more efficient in the identification of deeply processed materials than conventional DNA barcodes.•A combination of nucleotide signature and high-throughput sequencing was proposed for the detection of unknown mixtures.•This method is of great potential in the diagnosis of food poisoning.