Analysis of microbiomes and mycotoxins revealing the importance in the standardized storage of Aesculus wilsonii Rehd fruit

Aesculus wilsonii Rehd, a medicinal food homology plant, whose fruit are used to treat diseases such as edema and exudation of soft tissue. Nonetheless, the presence of microbial and mycotoxin contaminants in A. wilsonii fruit is still unclear. In this study, high-throughput sequencing and liquid chromatograph tandem-mass spectrometry were conducted to determine the diversity of fungal microbiome and mycotoxin contents in moldy A. wilsonii fruit. The prevalent fungi in A. wilsonii fruit included Fusarium, Colletotrichum, Dictyostelium, and Aspergillus. Furthermore, mycotoxin tests showed that all samples tested positive for ochratoxin A, with deoxynivalenol detected in 72.4 % of samples, zearalenone in 24.1 %, and T-2 toxin in 62.1 %, revealing these mycotoxins constituted the primary contaminants. Moreover, to monitor the mycotoxin-producing fungi, multiplex ligation-dependent probe amplification (MLPA) targeting key genes in toxin biosynthetic pathway of Fusarium and Aspergillus was established. and it can detect fungi with DNA content less than 20 ng. In conclusion, this study reported the microbiome diversity and the prevalence of fungi contamination in A. wilsonii fruit, while also established a rapid method for detecting toxigenic fungi. This work focused on analyzing and detecting microbiomes and mycotoxins in A. wilsonii fruit, providing a reference for the study of potentially harmful microorganisms in the storage of medicinal products.