开发针对iboH基因的PCR检测方法，以评估羊肚菌属中伊博滕酸/马西米醇生物合成潜力的分布情况

Abstract

Ibotenic acid and muscimol, synthesized by the ibotenic acid biosynthetic gene cluster (iboBGC) inAmanitaspecies, are two of the primary neurotoxins responsible for mushroom poisoning incidents worldwide. However, the molecular detection methods for these toxins remain underdeveloped. In this study, we designed a pair of universal PCR primers targetingiboH, a hydroxylase-encoding gene within theiboBGC, and developed a PCR assay with a detection limit of 10 pg of genomic DNA per reaction. Using the assay on 53Amanitaspecies, 21 out of the 23 species tested from sectionAmanitawere positive, whereas all 30 species from sects.Amidella(1),Caesareae(3),Phalloideae(6),Roanokenses(8),Strobiliformes(1),Vaginatae(4), andValidae(7) tested negative. We then integrated these PCR signals into our global phylogenetic analysis ofAmanitaand found thatiboHis restricted to a large clade within sect.Amanita. Amanita sinensis, commonly considered an edible species in China, was detected carryingiboH, indicating its potential to produce ibotenic acid/muscimol and pose associated health risks. In conclusion, this study establishes an effective universal PCR assay for detecting ibotenic acid/muscimol-producingAmanitaspecies and offer a reference for toxigenic mushroom recognition and food safety.