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.

Hot Springs ; Tracheophyta

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

Through studies on morphological characteristics, distribution and ecological habitat described in ancient literatures, Osmunda japonica is believed to the original plant of Chinese herbal medicine Cyrtomium Rhizome. Meanwhile, analysis is also made on causes for descriptions that do not comply with characteristics of O. japonica such as toxicity, flowers and fruits, illustrations, indumentums and flakes as well as appearance of other original plants. It is suggested to list O. japonica as the only original plant of Cyrtomium Rhizome and distinguish it from other frequently seen medical plants. Separated studies are also conducted for pesticide effect and active ingredients of Dryoteris Crassrhizomae Rhizome and Woodwardia japonica that show better effects.
Drugs, Chinese Herbal ; chemistry ; Dryopteridaceae ; chemistry ; Ferns ; chemistry ; Medicine, Chinese Traditional ; methods

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

To study the chemical constituents from the roots of Chirita longgangensis var. hongyao. The methanol extract was isolated and purified by silica gel, Sephadex LH-20 and preparative HPLC. Their structures were elucidated by MS and spectral data (1H, 13C-NMR). Seven compounds were isolated and identified as plantainoside A (1), plantainoside B (2), calcedarioside C (3), calcedarioside D (4), platyphylloside (5), hirsutanonol (6), and hirsutanonol-5-O-beta-D-glucopyranoside (7). Compounds 5-7 were isolated for the first time from the family Gesneriaceae.
Drugs, Chinese Herbal ; chemistry ; Organic Chemicals ; analysis ; Plant Roots ; chemistry ; Tracheophyta ; chemistry

OBJECTIVETo explore the identity and phylogenetic relationships among the three medicinally important species of Sphaerostephanos from South India using isozymic profile.

METHODSThe young fronds were homogenized with 3.5 mL of ice-cold homogenizing buffer in a pre-chilled pestle and mortar. The supernatant was subjected to electrophoresis as described by Anbalagan poly acrylamide gel electrophoresis. Staining solutions for isoperoxidase was prepared as per Smila method for the detection of isoenzymes.

RESULTSA total of six different bands in five different positions with different molecular weight/Rf values and four active zones have been observed in the isoperoxidase enzyme system of Sphaerostephanos. Only one band with MW/Rf 0.399 is common to two different species i.e. Sphaerostephanos arbuscula (S. arbuscula) and Sphaerostephanos unitus (S. unitus). Among the remaining four bands, two bands (Rf. 0.23, 0.47) are present in Sphaerostephanos subtruncatus (S. subtruncatus) and one distinct band has been observed individually in S. arbuscula (Rf. 0.507) and S. unitus (Rf. 0.56).

CONCLUSIONSThe present preliminary molecular study through isozymic analysis shows the identity of all the three species and the present results confirm distinctness of these three species based on macro-micromorphology, phytochemistry and cytology.

Electrophoresis, Polyacrylamide Gel ; India ; Isoenzymes ; analysis ; Peroxidases ; analysis ; Plants, Medicinal ; classification ; enzymology ; Staining and Labeling ; Tracheophyta ; classification ; enzymology

OBJECTIVETo prepare and evaluate the injectable temperature responsive in situ gel for sustained release of Glabrous Sarcandra Herb extract in vitro.

METHODThe novel temperature responsive poloxamer 407 was used as the carrier material of Glabrous Sarcandra Herb extract sustained release injection. The release of Glabrous Sarcandra Herb extract from poloxamer 407 based in situ gel in the phosphate buffer (pH 7. 6) was studied at 37 degrees C under agitation. HPLC method was used for the determination of Glabrous Sarcandra Herb extract.

RESULTThe best prescription was composed of by 16% P-407, 6% P-188, 0.2% HPMC, 0.5% benzil alcohol and 4% (g mL(-1)) Glabrous Sarcandra Herb extract. The optimal gelation temperature was around 33 degrees C. The data of release in vitro were analyzed according to the theoretical model of Korsmeyer-Peppas.

CONCLUSIONThe preparation method of the injectable thermosensitive in situ gel of Glabrous Sarcandra Herb extract is appropriate. The release in vitro can reach the expecting request.

Ferns ; chemistry ; Plant Extracts ; analysis ; Poloxamer ; chemistry ; Temperature

OBJECTIVETo study sesquiterpenoids of Coniogramme maxima.

METHODChemical constituents were separated by chromatography and their structures were identified according to physicochemical property and spectrum data.

RESULTFifteen compounds were separated by chromatography technique. Their structures were determined by spectral data, including 10 sesquiterpenoids as (3S)-pteroside D (1), epi-pterosin L (2), pterosin D (3), onitin (4), pterosin Z (5), onitisin (6), onitisin-glucopyranoside (7), onitin-15-O-beta-D-glucopyranoside (8), (2S,3R)-pterosin-L-2'-O-beta-D-glucopyranoside (9) and (3R)-peterosin D-3-O-beta-D-glucopyranoside (10). The other compounds were uracil (11), 3,4-dihydroxybenzaldehyde (12), 5-hydroxymethyl-2-furancarboxaldehyde (13), beta-sitosterol (14) and daucosterol (15).

CONCLUSIONThe above 15 compounds are separated from C. maxima for the first time, including 9 compounds being first separated from genus Coniogramme.

Ferns ; chemistry ; Indans ; chemistry ; Indenes ; chemistry ; Sesquiterpenes ; chemistry ; Sitosterols ; chemistry

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

The aim of this review was to explore the pharmacological activity of early tracheophytes (pteridophytes) as an alternative medicine for treating human ailments. As the first vascular plants, pteridophytes (aka, ferns and fern allies) are an ancient lineage, and human beings have been exploring and using taxa from this lineage for over 2000 years because of their beneficial properties. We have documented the medicinal uses of pteridophytes belonging to thirty different families. The lycophyte Selaginella sp. was shown in earlier studies to have multiple pharmacological activity, such as antioxidant, anti-inflammatory, anti-cancer, antidiabetic, antiviral, antimicrobial, and anti-Alzheimer properties. Among all the pteridophytes examined, taxa from the Pteridaceae, Polypodiaceae, and Adiantaceae exhibited significant medicinal activity. Based on our review, many pteridophytes have properties that could be used in alternative medicine for treatment of various human illnesses. Biotechnological tools can be used to preserve and even improve their bioactive molecules for the preparation of medicines against illness. Even though several studies have reported medicinal uses of ferns, the possible bioactive compounds of several pteridophytes have not been identified. Furthermore, their optimal dosage level and treatment strategies still need to be determined. Finally, the future direction of pteridophyte research is discussed.
Anti-Infective Agents/pharmacology* ; Anti-Inflammatory Agents/pharmacology* ; Antineoplastic Agents, Phytogenic/pharmacology* ; Antioxidants/pharmacology* ; Humans ; Phytochemicals/pharmacology* ; Phytotherapy ; Plant Extracts/pharmacology* ; Tracheophyta/chemistry*