Isolation of endophytic fungi and screening of Huperzine A–producing fungus from Huperzia serrata in Vietnam

Abstract

Huperzine A (HupA), a natural Lycopodium alkaloid derived from Huperzia serrata (Thunb. ex Murray) Trev. plants, is a highly active acetylcholinesterase inhibitor and a key compound used for treating Alzheimer’s disease (AD). Recently, HupA has been reported in various endophytic fungi isolated from H. serrata. In the present study, 153 endophytic fungi were isolated from healthy tissues of H. serrata collected from natural populations in Lam Dong province of Central Vietnam. The endophytic fungi were identified based on morphological characteristics and Internal Transcribed Spacer sequences. Among them, 34 strains were classified into seven genera belonging to Ascomycota, including Alternaria, Fusarium, Trichoderma, Penicillium, Paecilomyces, and Phoma, and eight strains belonging to the genus Mucor (Zygomycota). The other strains remained unidentified. According to the results of thin-layer chromatography and high-performance liquid chromatography, only one of the 153 strains, Penicillium sp. LDL4.4, could produce HupA, with a yield 1.38 mg l−1 (168.9 µg g−1 dried mycelium) when cultured in potato dextrose broth, which was considerably higher than that of other reported endophytic fungi. Such a fungus is a promising candidate and alternative to presently available HupA production techniques for treating AD and preventing further memory decline.

Illumina-based analysis yields new insights into the diversity and composition of endophytic fungi in cultivated Huperzia serrata

“…Currently, about 12 fungal genera have been reported to produce huperzine A [15][16][17][42][43][44][45][46][47], of which 7 genera were found in H. serrata in this study. Among them, Cladosporium, Trichoderma, and Fusarium with the relative abundance > 0.1% had the highest distribution in the stems, followed by the leaves and the roots, indicating that the endophytic fungi in the stems and the leaves were more involved in the synthesis of some secondary metabolites than those in the roots.…”

“…Previous studies examining the endophytic fungi of H. serrata and their secondary metabolic products revealed that they exhibited various biological activities, including antimicrobial activity, acetylcholinesterase inhibitory activity, nematocidal activity, and inhibition of nitric oxide production, among others [11][12][13][14]. In addition, some of endophytic fungi in H. serrata can produce huperzine A [15][16][17]. The distribution of some endophytic fungi in the host plants exhibited tissue specificity, which is an important influencing factor for accumulation of bioactive substances in different tissues [18,19].…”

Cannabis Microbiome and the Role of Endophytes in Modulating the Production of Secondary Metabolites: An Overview

“…The alkaloid drug Huperzine A (HupA) used to treat Alzheimer's disease is not only derived from the Huperzia serrata plant but also is produced and biosynthesized by the fungal endophyte Penicillium sp. LDL4.4 isolated from H. serrata [114]. In the legume Crotalaria (subfamily Fabaceae), the biosynthesis of pyrrolizidine alkaloids (antiherbivore, nematicide) depends on the nodulation by Bradyrhizobium sp.…”

Potential Fungi Isolated From Anti-biodegradable Chinese Medicine Residue to Degrade Lignocellulose

“…A suitable anti-biodegradable residue tissue block of 10 g was selected and washed with sterile normal saline three times, fungi were isolated by plant tissue separation ( Makhuvele et al, 2017 ; Thi Minh Le et al, 2019 ), the sample was repeated three times. Genomic DNA of the purified fungi was extracted using Ezup Column Fungal genomic DNA Extraction Kit (Qingke, Chongqing) and used as a template for internal transcribed spacer (ITS) region amplification ( Bellemain et al, 2010 ).…”