Background. The growing resistance of microorganisms to antimicrobial agents is a pressing public health issue. Bioprospecting efforts have mainly focused on well-known environments such as soil and animal gut in search for microorganisms with antibiotic production or antimicrobial activity, or terrestrial ecosystems for endemic plants with bioactive compounds. However, microbial communities thriving in stressed environments such as hot springs, are potential sources of metabolites that can be screened for antimicrobial activity. There is a need for research on bioprospecting of fungi as potential sources of antimicrobials.

Objectives. The study aimed to test the antimicrobial activity of endophytic and rhizospheric fungi associated with soft fern (Christella spp.) and Cinderella weed (Synederella nodiflora) inhabiting a hot spring in Los Baños, Laguna, Philippines.

Methods. A total of 23 endophytic and rhizospheric fungi isolated from soft fern and Cinderella weed were purified and phenotypically identified. These isolates were subjected to agar well diffusion and agar plug diffusion methods as preliminary assays for antimicrobial activity against Bacillus subtilis var. spizizenii (ATCC® 6633™), Staphylococcus  aureus (ATCC® 25923™), four multi-antibiotic resistant Escherichia coli (OT11, OT16, OT18, OT22), and Cladosporium  cladosporioides. Based on the results of the preliminary screening, ethyl-acetate extracts of selected fungal isolates were subjected to broth microdilution assay to determine the minimum inhibitory concentrations (MICs) for antibacterial activity, as well as poisoned food technique to determine the percent mycelial inhibition for antifungal activity. The nearest phylogenetic affiliations of fungal isolates with higher antimicrobial activities were determined.

Results. Ten rhizospheric fungal isolates from Cinderella weed and seven rhizospheric and six endophytic fungal isolates from soft fern were phenotypically identified as Aspergillus, Coniothyrium, Fusarium, Penicillium, Talaromyces,  and Trichoderma species. Ethyl acetate extracts from endophytic fungal isolates UL1 (Trichoderma sp.) and UL2 (Trichoderma sp.) and rhizospheric fungal isolates UR1 (Trichoderma sp.) and UR3 (Trichoderma sp.) showed activity against the test bacteria at 128-256 ?g/mL concentrations. Isolates UL1, UL2, and UR3, which exhibited higher antibacterial activities, were sequenced and confirmed to be most phylogenetically related to Trichoderma  virens. Eleven fungal isolates belonging to Aspergillus spp., Coniothyrium spp., Fusarium spp., Penicillium spp., and Talaromyces spp. demonstrated antagonism against C. cladosporioides. The rhizospheric fungal isolate FCRU4 (Talaromyces sp.), from where ethyl acetate extracts were recovered for testing mycelial inhibition, was confirmed to be most phylogenetically related to Talaromyces islandicus.

Conclusion. Endophytic and rhizospheric fungi asso ciated with Cinderella weed (Synedrella nodiflora) and soft fern (Christella sp.) from a hot spring in Los Baños, Laguna, Philippines have antimicrobial activity.

A new phloroglucinol was isolated from 50% ethanol extract of Dryopteris fragrans by silica gel column chromatography, Sephadex LH-20 gel column chromatography, thin-layer chromatography(TLC), and preparative liquid column chromatography. On the basis of MS, ~1H-NMR, ~(13)C-NMR, and reference materials, compound 1 was identified as 2,5-cyclohexadien-1-one, 2-{[2,6-dihydroxy-4-methoxy-3-methyl-5-(1-isobutyl)phenyl]methyl}-3,5-dihydroxy-4,4-dimethyl-6-(1-oxobutyl)(1), and named disaspidin BB. Compound 1 was evaluated for its antibacterial activity. The experimental results showed that compared with the commonly used topical antibiotics erythromycin or mupirocin, disaspidin BB exhibited significant antibacterial activities against Staphylococcus epidermidis(SEP), S. haemolyticus(SHA), and methicillin-resistant S. aureus(MRSA)(P<0.05). Additionally, disaspidin BB was sensitive to ceftazidime-resistant SEP1-SEP4, SHA5-SHA7, MRSA8, and MRSA9. The MIC values of disaspidin BB against SEP and SHA were 1.67-2.71 μg·mL~(-1) and 10.00-33.33 μg·mL~(-1) respectively. Disaspidin BB has good antibacterial activities and deserves development as a new anti-infective drug for external use.
Anti-Bacterial Agents/pharmacology* ; Dryopteris ; Methicillin-Resistant Staphylococcus aureus ; Microbial Sensitivity Tests ; Phloroglucinol/pharmacology* ; Plant Extracts/pharmacology*

This study intended to explore the effect and mechanism of total flavonoids of Drynariae Rhizoma in improving scopola-mine-induced learning and memory impairments in model mice. Ninety four-month-old Kunming(KM) mice were randomly divided into six groups. The ones in the model group and blank group were treated with intragastric administration of normal saline, while those in the medication groups separately received the total flavonoids of Drynariae Rhizoma, Kangnaoshuai Capsules, donepezil, as well as total flavonoids of Rhizoma Drynariae plus estrogen receptor(ER) blocker by gavage. The mouse model of learning and memory impairments was established via intraperitoneal injection of scopolamine. Following the measurement of mouse learning and memory abilities in Morris water maze test, the hippocampal ERβ expression was detected by immunohistochemistry, and the expression levels of ERβ and phosphorylated p38(p-p38) in the hippocampus and B-cell lymphoma 2(Bcl-2), Bcl-2-associated death promoter(Bad), and cysteinyl aspartate-specific protease-3(caspase-3) in the apoptotic system were assayed by Western blot. The contents of malondia-ldehyde(MDA), superoxide dismutase(SOD), and nitric oxide(NO) in the hippocampus were then determined using corresponding kits. Compared with the control group, the model group exhibited significantly prolonged incubation period, reduced frequency of cros-sing the platform, shortened residence time in the target quadrant, lowered ERβ, Bcl-2 and SOD activity in the hippocampus, and increased p-p38/p38, Bad, caspase-3, MDA, and NO. Compared with the model group, the total flavonoids of Rhizoma Drynariae increased the expression of ERβ and SOD in the hippocampus, down-regulated the expression of neuronal pro-apoptotic proteins, up-re-gulated the expression of anti-apoptotic proteins, and reduced p-p38/p38, MDA, and NO. The effects of total flavonoids of Drynariae Rhizoma on the above indexes were reversed by ER blocker. It has been proved that the total flavonoids of Drynariae Rhizoma obviously alleviate scopolamine-induced learning and memory impairments in mice, which may be achieved by regulating the neuronal apoptotic system and oxidative stress via the ER-p38 mitogen-activated protein kinase(ER-p38 MAPK) signaling pathway.
Animals ; Flavonoids ; Hippocampus ; Maze Learning ; Mice ; Polypodiaceae ; Receptors, Estrogen ; Scopolamine/toxicity* ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases/genetics*

Drynariae Rhizoma is warm in nature and bitter in taste, mainly acting on liver and kidney systems. It is a common Chinese herbal medicine for the treatment of fracture and bone injury. The chemical compositions of Drynariae Rhizoma mainly include flavonoids, triterpenoids, phenylpropanoids and lignans. At present, modern pharmacological and clinical studies have shown that Drynariae Rhizoma has the effects of anti osteoporosis, promoting fracture healing, kidney protection, anti-inflammatory, promoting tooth growth, preventing and treating aminoglycoside ototoxicity and lowering blood lipid. In addition, the toxicity evaluation experiment of Drynariae Rhizoma has also shown that it has no obvious toxic and side effects. Naringin is a kind of dihydroflavone in Drynariae Rhizoma. Many studies have shown that naringin and other total flavonoids play an important role in anti-osteoporosis, promoting fracture healing, anti-inflammation, promoting tooth growth and lowering blood lipid. In this study, the research progresses on chemical consti-tuents and pharmacological activities of Drynariae Rhizoma in recent years were reviewed, and some mechanisms of action were summarized, to provide references for the further research and development of Drynariae Rhizoma.
Drugs, Chinese Herbal/pharmacology* ; Flavonoids ; Humans ; Osteoporosis/drug therapy* ; Polypodiaceae ; Rhizome

Two phloroglucinol compounds(1-2) were isolated and purified from 95% ethanol extract of Dryopteris fragrans through various column chromatographies on silica gel, Sephadex LH-20, medium pressure column chromatography, and preparative HPLC. Their structures were elucidated as 2',4',6'-trihydroxy-5'-methyl acetate-3'-methyl-1'-butyrophenone(1) and aspidinol B(2) based on their chemical and physicochemical methods and spectroscopic data. Compound 1 is a new phloroglucinol compound named "dryofraginol".
Chromatography, High Pressure Liquid ; Dryopteris ; Ethanol ; Phloroglucinol ; Plant Extracts

Botany/history* ; Ferns ; History, 19th Century ; History, 20th Century ; Humans ; Portraits as Topic

To clarify the difference of soil moisture characteristics between mixed broad leaf-conifer forest soil and artificial cultivation of Notopterygium incisum, the HYPROP system and the dew point potential meter were used to determine soil water retention curves(SWRC) for samples of two horizons(i.e. 2-7 cm, 10-15 cm). The basic physical and chemical properties of soil and its water characteristic parameters were also determined. The result showed as fllows:①The bulk density of mixed coniferous-broad leaf forest soil was between 0.33 and 0.52 g·cm~(-3), significantly lower than the corresponding value of field soil(1.01-1.18 g·cm~(-3))(P<0.05), While the organic matter content was significantly higher than the corresponding value of field soil(P<0.05). ②The saturated water content(θ_s), field water holding capacity(θ_(FC)) and Water that can be effectively utilized by plants(θ_(PAC)) of mixed coniferous-broadleaved forest soil were significantly higher than the corresponding value of field soil(P<0.05), while the retained water content(θ_r) value that cannot be effectively utilized by plants was significantly lower than that of field soil(P<0.05). ③The values of structural porosity(0.13-0.24 cm~3·cm~(-3)) and Matrix porosity(0.34-0.44 cm~3·cm~(-3)) of mixed coniferous-broadleaved forest soil were higher than the corresponding values of field soil. Therefore, with low bulk density and high content of organic matter, mixed coniferous-broadleaved forest soil can store more water in soil in the form of effective water to meet the needs of plants for water, thus possibly forming high quality medicinal materials of Notopterygii Rhizoma et Radix. In conclusion, the results of this study can provide theoretical basis guidance for soil structure improvement and water management to form high quality medicinal materials in the artificial cultivation of N. incisum.
Apiaceae ; China ; Forests ; Soil ; Tracheophyta ; Water ; analysis

It is of great significance to carry out scientific investigation of threatened species and assess their in-situ conservation status in order to guide the conservation and management of these species within a region. In this study, we explored and assessed the biodiversity and in-situ conservation status of the threatened medicinal vascular plants(TMVPs) in central China. The results showed that there were 276 TMVPs in central China, including 18 critically endangered(CR), 77 endangered(EN), and 181 vulnerable(VU) species. Of which, 222 TMVPs were distributed in 49 national nature reserves, with an in-situ conservation rate of 80.43%. And the in-situ conservation rate of CR, EN and NR species were 83.33%, 77.92% and 81.22%, respectively. The complementary algorithm was used to select the nine national nature reserves with the highest protection and complementary contribution to the TMVPs. The cumulative protection contribution rate of these nine nature reserves was 81.98%. Fifty-four TMVPs were distributed outside of national nature reserves, while mainly distributed within 10 hotspot counties including Badong county, Hubei province, Sangzhi county, Hunan province, Jianghua Yao Autonomous county, Hunan province and so on. In general, TMVPs are well protected by the national nature reserves in central China. However, there are still some conservation gaps. It is necessary to carry out further field investigation on the species with conservation gaps and identify the gap areas, so as to provide scientific guidance for the optimization of the conservation of TMVPs in this region.
Biodiversity ; China ; Conservation of Natural Resources ; Endangered Species ; Plants, Medicinal ; Tracheophyta

This study is based on the data analysis of medicinal plant resources and diversity collected from the fourth Chinese traditional medicine resource survey( pilot). Through the analysis of relevant data from 33 census pioneer plots in Guizhou province( area),a total of 265 families,1 432 genera and 5 296 species of medicinal resources were reported,including algae,fungi,lichens,mosses,a total of 43 genera and 35 families,57,48 families,120 genera and 453 species of ferns,gymnosperms 11 families,22 genera and 61 species,167 families,1 243 genera and 4 721 species of angiosperms,4 genera and 4 families four medicinal animals.Compared with the data related to the third survey of traditional Chinese medicine resources,the number of ferns,gymnosperms and angiosperms in the fourth survey has increased far more than that of the third survey. From the regional distribution of medicinal resources,the composition of the genus,the type of life,and the location of the medicine,the richness of the medicinal plant resources in Guizhou province is not only reflected in many types,but also in the variety of medicinal resources. These studies provide a scientific basis for vigorously developing the Chinese herbal medicine industry and the sustainably using medicinal plant resources in Guizhou province.
China ; Cycadopsida ; Drugs, Chinese Herbal ; Ferns ; Magnoliopsida ; Medicine, Chinese Traditional ; Plants, Medicinal ; classification

In 2017, small, elliptical, brownish purple spots on spears and ferns of asparagus were found in fields of Gangwon-do. The isolated fungal species was identified as an ascomycete Stemphylium vesicarium based on morphological characteristics and molecular phylogenic analyses including nucleotide sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and cytochrome b (cytb). A pathogenicity test revealed that S. vesicarium was the causal agent of purple spot disease on asparagus. The occurrence of purple spots caused by S. vesicarium on asparagus is the first report in Korea.
Ascomycota ; Base Sequence ; Cytochromes b ; Ferns ; Gangwon-do ; Korea ; Oxidoreductases ; Virulence