Isolation and characterzation of endophytic Huperzine A-producing fungi from Phlegmariurus phlegmaria

Abstract

Based on special colony morphology, 340 endophytic fungi were isolated from the stems, leaves, and roots of Phlegmariurus phlegmaria. Among the 340 strains, eight, representative species (MY298, MJ216, MY237, MJ422, MY311, MY183, MJ484, and MY252) could produce HupA analyzed by UPLC-MS/MS (Ultra performance liquid chromatography), which compared with standard HupA under the same chromatographic conditions. And through the standard curve generated, among these HupA-producing fungi, the MY311 had highest content which was 40.53 µg/L. The extract of eight strains was examined by AChE inhibition activity assay in vitro and ELISA (enzyme-linked immunosorbent assay). These eight strains were found to have positive results for HupA production and MY311 extract obviously exhibited the highest inhibition activity. Furthermore, the genera of the eight fungi were identified based on the internal transcribed spacer sequence analysis. All eight distinctive genotypes were grouped into four fungal genera and six fungal species, however, MY311 and MY183 were Ceriporia lacerata and Hypoxylon investiens, which had never been reported as HupA-producing endophytic fungi.