Fungal communities in Nelumbinis semen characterized by high-throughput sequencing

For a long period, Nelumbinis semen has been widely used as a medicinal and edible product. However, it is susceptible to contamination with toxigenic fungi and aflatoxins during the growth, collection, transportation, and storage processes, causing serious health threats to humans and huge economic losses. Effectively monitoring the fungal communities is of great importance to avoid aflatoxins contamination in Nelumbinis semen. High-throughput sequencing (HTS) is a new technology to evaluate fungal communities so as to overcome the limitations of the traditional cultural ways. In this study, the ITS2 based Illumina-MiSeq platform was developed to evaluate the fungal communities in normal and moldy Nelumbinis semen by using the HTS technology. Two different primer pairs were introduced to compare their performance in amplifying the target gene. The primer pair that produced more reads was selected to analyze the results. In all the nine tested Nelumbinis semen samples, 2 phyla, 5 classes, 6 orders, 8 families, 9 genera and 4 species were detected. A total of 9 genera were spotted, of which, Aspergillus (0.04%-72.93%) and Rhizopus (0.002%-48.12%) were the most dominant. ANOISM analysis showed no significant differences in the normal and moldy groups. The use of HTS technology can detect the fungal communities in complex Nelumbinis semen samples, providing an early warning for toxigenic fungi and aflatoxins contamination to warrant their quality and safety.