Two dibenzo-α-pyrones, botrallin (1) and TMC-264 (2) were preparatively separated from crude ethyl acetate extract of the endophytic fungus Hyalodendriella sp. Ponipodef12, which was isolated from the hybrid 'Neva' of Populus deltoides Marsh × P. nigra L. using a combination of high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC. Botrallin (1) with 74.73% of purity and TMC-264 (2) with 82.29% of purity were obtained through HSCCC by employing a solvent system containing n-hexane-ethyl acetate-methanol-water at a volume ratio of 1.2:1.0:0.9:1.0. It was the first time for TMC-264 (2) to be isolated from this fungus. TMC-264 (2) showed strong antimicrobial and antinematodal activity, and botrallin (1) exhibited moderate inhibitory activity on acetylcholinesterase.

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A total of 57 endophytic fungal isolates were obtained from the roots of Salvia miltiorrhiza Bunge (Lamiaceae). Fourteen (14) distinct isolates were selected for further taxonomical identification by morphological traits and internal transcribed spacer (ITS) rRNA gene sequence analysis. Twelve (12) genera were identified among which Alternaria and Fusarium were dominants. Eight endophytic fungi (that is, Pleosporales sp. Samif02, Leptosphaeria sp. Samif03, Peyronellaea glomerata Samif04, Xylomelasma sp. Samif07, Bionectria ochroleuca Samif08, Sarocladium kiliense Samif11, Petriella setifera Samif13 and Cadophora sp. Samif14) were separated as the endophytic fungi from S. miltiorrhiza for the first time. Most of the fungal isolates were observed to have antibacterial activity that suggests antibacterial compounds mainly exist in mycelia. The ethyl acetate extracts of Alternaria sp. Samif01, Xylomelasma sp. Samif07, Fusarium redolens Samif09, Sarocladium kiliense Samif11 and Petriella setifera Samif13 were also observed to have antifungal activity. Among the isolates, Alternaria sp. Samif01 and Sarocladium kiliense Samif11 were found to have strong antibacterial and antifungal activities. The results indicate that there is a diversity of the endophytic fungi from S. miltiorrhiza, and these endophytic fungi could be an excellent resource for searching natural antimicrobial compounds.

High-speed counter-current chromatography (HSCCC) was applied for the first time for the preparative separation of spirobisnaphthalenes from a crude extract of the endophytic fungus Berkleasmium sp. Dzf12, associated with the medicinal plant Dioscorea zingiberensis. Six spirobisnaphthalenes were successfully separated by HSCCC with a two-phase solvent system composed of n-hexane-chloroform-methanol-water (1.5:3.0:2.5:2.0, v/v). About 18.0 mg of diepoxin κ (1), 245.7 mg of palmarumycin C 13 (2), 42.4 mg of palmarumycin C 16 (3), 42.2 mg of palmarumycin C 15 (4), 32.6 mg of diepoxin δ (5), and 22.3 mg of diepoxin γ (6) with purities of 56.82, 71.39, 76.57, 75.86, 91.01 and 82.48%, respectively, as determined by high-performance liquid chromatography (HPLC), were obtained from 500 mg of the crude extract in a one-step elution within 7 h of separation procedure by HSCCC. The purified spirobisnaphthalenes were further structurally characterized by means of physicochemical and spectrometric analysis.

The effects of three polysaccharides; exopolysaccharide, water-extracted mycelial polysacharide, and sodium hydroxide-extracted mycelial polysaccharide (EPS, WPS and SPS) and their corresponding oligosaccharides; oligosaccharide obtained by hydrolysis of EPS, oligosaccharide obtained by hydrolysis of WPS and oligosaccharide obtained by hydrolysis of SPS (EOS, WOS and SOS) as the elicitors prepared from endophytic fungus Berkleasmium sp. Dzf12 on growth and diosgenin accumulation in cell and seedling cultures of Dioscorea zingiberensis were investigated. Among all the elicitation treatments, EOS generated the most satisfactory effect on enhancing diosgenin production in both cell and seedling cultures. The highest diosgenin yield (2.89 mg/L) in the cell cultures treated with EOS at 20 mg/L was achieved, which was 6.88-fold of control (0.42 mg/L). Meanwhile, when the seedlings were treated with EOS at 40 mg/L, the maximum diosgenin yield (14.68 mg/L) was achieved, which was 4.60-fold as the control (3.19 mg/L). Oligosaccharides WOS and SOS also showed obvious effects on enhancing diosgenin accumulation in the cell and seedling cultures, which were stronger than their corresponding polysaccharides WPS and SPS. The diosgenin yield in the cell cultures separately treated with WOS at 40 mg/L or SOS at 20 mg/L reached the maximum, which were 4.27-and 2.86-fold of control, respectively. For the seedlings treated with WOS or SOS, the maximum diosgenin yield was 8.43 mg/L, which was 2.64-fold of control. The results indicate that the polysaccharides and oligosaccharides as the elicitors from Berkleasmium sp. Dzf12 could be applied in D. zingibereneis cell and seedling cultures to produce diosgenin.