Identification of a Huperzine A-producing endophytic fungus from Phlegmariurus taxifolius

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].…”

Microbial acetylcholinesterase inhibitors for Alzheimer’s therapy: recent trends on extraction, detection, irradiation-assisted production improvement and nano-structured drug delivery

“…Finally, ethanol (70%) for 1 min and 1% available chlorine for 3 min were also investigated (Zaki et al 2019). After surface tissue sterilization, the plant parts were cut into small segments and directly cultured on a suitable media; potato dextrose agar (PDA) was mostly chosen, which was enriched by streptomycin, 60 μg/ml, and ampicillin, 100 μg/ml, to avoid bacterial contamination (Zhu et al 2010;Wang et al 2011a, b;Zhang et al 2015a;Zaki et al 2019;Cruz-Miranda et al 2019). Shu et al (2014) and Dong et al (2014) used Murashige and Skoog (MS) media for endophytes' isolation.…”

“…The chloroform extracts were evaporated, and the obtained residue was dissolved in 1 mL methanol. Zhu et al (2010) and Cruz-Miranda et al (2019) used sequential extraction. Primarily, the dried cells underwent extraction with 75% ethanol for 30 min in a 40°C ultrasonic bath.…”

“…Methanol: water (85:15) (Zhu et al 2010;Wang et al 2011a;Zaki et al 2019). Ammonium acetate (80 mM, or 0.1 M, pH 6.0): methanol (7:3 or 6:4 or 36: Confirmatory analyses combining both liquid chromatography (LC) and mass analysis by mass spectrometry (MS) using LC/MS and UPLC/MS analyses were also performed, where the alkaloid extract underwent chromatographic separation using a suitable run solvent as methanoic acid (0.1%): acetonitrile (9: 1 v/v) at a flow rate of 1.0 mL/min (Su and Yang 2015), 10 mM ammonium acetate (pH 3.5) and methanol with 5: 56: 56% gradient elution (T: 0: 20: 30 min) (Shu et al 2014), methanol: 0.2% aqueous formic acid (50: 50 v/v), flow rate of 0.15 mL/min, 308 nm as a detecting wavelength, and 5 μL injected volume (Zhang et al 2015a), and acetonitrile: 0.025% formic acid 15: 85 v/v, 0.25 mL/ min as a flow rate (Cruz-Miranda et al 2019), then the elute of the chromatography was subjected to MS analysis.…”