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Soil is home to microbiota with diverse metabolic activities. These microorganisms play vital roles in many ecological processes. Thus, the assessment of microbial functional diversity is an important quality indicator of soil ecosystems. In this study, we collected soil samples from three distinct forest habitats, i.e., an agroforest, a primary forest (PF), and a secondary forest, within the Angat Watershed Reservation in Bulacan, Northern Philippines. Community-level physiological profiling (CLPP) was done with the BIOLOG EcoPlate™ to analyze the responses of the soil microbial communities from the three forest habitats in the absence or presence of antibiotics. The BIOLOG EcoPlate represents 31 utilizable carbon sources. Based on the CLPP analysis, soil samples from the PF showed significantly higher utilization of most carbon sources than the other forest types (p < 0.05). Thus, less disturbed forest types constitute more functionally diverse microbial communities. The presence of antibiotics significantly decreased the carbon utilization patterns of the soil microbial communities (p < 0.05), indicating the possible use of CLPP in monitoring contamination in soil. [ABSTRACT FROM AUTHOR]

The rapid emergence of resistant bacteria is occurring worldwide, endangering the efficacy of antibiotics. Hence, there is a need to search for new sources of antibiotics that either exhibit novel structures or express a new mechanism of action. The lichen Usnea, with its wide range of unique, biologically potent secondary metabolites, may solve this problem. In this study, Usnea species were collected in the Northern Philippines, identified through combined morphological and biochemical characterization, and tested for antimicrobial activities against the multidrug-resistant ESKAPE pathogens, i.e., Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter cloacae, two standard antibiotic-sensitive test bacteria, and a yeast. A total of 46 lichen specimens were collected and later identified as Usnea baileyi (10), U. diffracta (10), U. glabrata (12), U. longissima (4), and U. rubicunda (10). The results show that the crude extracts of the Usnea species exhibited promising in vitro inhibitory activities against standard antibiotic-sensitive (E. faecalis ATCC 29212) and multidrug-resistant (methicillin-resistant S. aureus and E. faecalis) Gram-positive bacteria. Additionally, lichen compounds of representative specimens per species were identified and profiled using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The detection of lichen acids (LA) via HPLC showed the presence of 24 peaks of lichen acids. TLC-bioautography identified the bioactive lichen acids as alectronic acid, connorstictic acid, consalazinic acid, diffractaic acid, echinocarpic acid, erythrin acid, galbinic acid, hypoconstictic acid, hyposalazinic acid, hypostictic acid, lobaric acid, menegazzaic acid, micareic acid, pannarin, salazinic acid, stictic acid, and usnic acid. Our study highlighted the wide spectrum of opportunities for using lichens for the discovery of potential antimicrobial agents. [ABSTRACT FROM AUTHOR]

Protists affect soil microbiome composition and functioning, potentially increasing litter decomposition and plant growth. Yet, the role of different protist species, individually or in combinations, in regulating microbial-mediated litter decomposition remains unknown, as well as if these interactions feedback to plant growth. Using a full-factorial design of three protist species combinations with bacterial and fungal communities, we found that only one protist species reduced litter decomposition by 19%, with other species and combinations not affecting litter decomposition. Despite a positive correlation between plant growth and litter decomposition, we did not observe protist-induced changes in plant growth. Overall, our results highlight that protists affect litter decomposition in a species-specific manner, including reducing litter decomposition. • Only 1 of 3 protist species and any combinations reduced litter decomposition. • Protist presence did not affect plant growth. • Litter decomposition positively correlated with plant growth. [ABSTRACT FROM AUTHOR]

Citizen science is a research collaboration between scientists and volunteers who provide data for education, conservation, and environmental protection. Volunteers, often the locals in the area, provide data on species occurrence while researchers perform distribution mapping or other data analysis. Social networking sites including Facebook, Twitter, and Flickr can be used as platforms for the public to share their photos of species and for scientists to aid in identification. In this article, we show how social media groups can be used to generate data on species distribution of myxomycetes. [ABSTRACT FROM AUTHOR]

Nickel contamination is a serious environmental issue that requires immediate action. In this study, 23 strains of Trichoderma were isolated from terrestrial and marine environments and identified using a polyphasic approach of morphological characterization and ITS gene sequence analysis. The Trichoderma strains were tested for their tolerance and biosorption of nickel. Our results showed the growth of all Trichoderma strains on Trichoderma Selective Medium (TSM) with 50-1200-ppm nickel, indicating their tolerance of this heavy metal even at a relatively high concentration. Six Trichoderma strains (three isolated from terrestrial substrates and three from marine substates) had the highest radial growth on TSM with 50-ppm Ni. Among these fungal isolates, Trichoderma asperellum (S03) isolated from soil exhibited the best growth after 2 days of incubation. For the biosorption of nickel, the accumulation or uptake efficiency by the six selected Trichoderma was determined in Potato Dextrose Broth (PDB) supplemented with 50-ppm Ni using a Flame Atomic Absorption Spectrophotometer (AAS). The percent uptake efficiency of the three strains of T. asperellum (S03, S08, and LL14) was computed to be up to 66%, while Trichoderma virens (SG18 and SF22) and Trichoderma inhamatum (MW25) achieved up to 68% uptake efficiency. Observation of the Trichoderma strains with Scanning Electron Microscopy (SEM) before and after the absorption of nickel showed very minimal damage on the hyphal and conidial surface morphology, but changes in the colonial characteristics were observed. Our study highlighted the potential of terrestrial and marine strains of Trichoderma for the bioremediation of nickel pollution. [ABSTRACT FROM AUTHOR]

The culture-based approach was used to characterize the fungal endophytes associated with the coralloid roots of the endemic Cycas debaoensis and Cycas fairylakea from various population sites in China. We aim to determine if the assemblages of fungal endophytes inside these endemic plant hosts are distinct and could be explored for bioprospecting. The isolation method yielded a total of 284 culturable fungal strains. Identification based on the analysis of the internal transcribed spacer (ITS) rDNA showed that they belonged to two phyla, five classes, eight orders and 22 families. At least 33 known genera and 62 different species were confirmed based on >97% ITS sequence similarity. The most frequent and observed core taxa in the two host species regardless of their population origin were Talaromyces, Penicillium, Fusarium, Pochonia and Gliocladiopsis. Seventy percent was a rare component of the fungal communities with only one or two recorded isolates. Contrary to common notions, diversity and fungal richness were significantly higher in C. debaoensis and C. fairylakea collected from a botanical garden, while the lowest was observed in C. debaoensis from a natural habitat; this provides evidence that garden management, and to a minor extent, ex-situ conservation practice, could influence fungal endophyte communities. We further selected nineteen fungal isolates and screened for their antagonistic activities via a co-cultivation approach against the phytopathogens, Diaporthe sp. and Colletotrichum sp. Among these, five isolates with high ITS similarity matches with Hypoxylon vinosupulvinatum (GD019, 99.61%), Penicillium sp. (BD022, 100%), Penicillifer diparietisporus (GD008, 99.46%), Clonostachys rogersoniana (BF024, 99.46%) and C. rosea (BF011, 99.1%), which showed exceptional antagonistic activities against the phytopathogenic fungi with a significant inhibition rate of 70–80%. Taken together, our data presented the first and most comprehensive molecular work on culturable fungal endophytes associated with the coralloid roots of cycads. Our study also demonstrated that about 5% of fungal endophytes were not detected by the high-throughput sequencing approach, implying the equal importance of a culture-dependent approach to study fungal communities of cycads. We further highlighted the potential role of endemic and rare plants to discover and isolate unique plant-associated fungal taxa with excellent biocontrol properties. [ABSTRACT FROM AUTHOR]

Endolichenic fungi (ELF; asymptomatic microorganisms living inside healthy lichen thalli) were isolated from three Usnea species, namely U. baileyi, U. bismolliuscula and U. pectinata, collected near the town of Sagada, Philippines. A total of 101 ELF were recovered representing 12 genera (classes Sordariomycetes and Eurotiomycetes), with the genera Nemania (50.5%, 51 isolates) and Xylaria (22.8%, 23 isolates) being the most abundant. Comparative analyses on the antimicrobial activities of lichens and ELF revealed that lichen crude extracts were effective against the Gram-positive bacterium Staphylococcus aureus and the yeast Candida albicans, while ELF crude extracts were effective against S. aureus, C. albicans and the Gram-negative bacterium Escherichia coli. The broad-spectrum nature of ELF has provided medicinal and industrial advantages over the slow-growing lichens as shown on their respective bioactivities. Extracts from ELF also had a higher total flavonoid content (TFC; 6.29-85.69 mg QE/g of extract) and stronger antioxidant activities (IC50: 0.57-19.63 mg/ml) than lichen-derived extracts. Although only culturable ELF were identified, this study provides the first evaluation of the diversity and bioactivities of culturable ELF from fruticose lichens of the genus Usnea in the Philippines. [ABSTRACT FROM AUTHOR]

Aims Anthracnose caused by Colletotrichum species is one of the most devastating diseases of fruits and crops. We isolated and identified an antifungal compound from the mushroom Coprinus comatus and investigated its inhibitory potential against anthracnose disease-causing fungi with the goal of discovering natural products that can suppress anthracnose-caused plant disease. Methods and results The culture filtrate of C. comatus was subjected to a bioassay-guided isolation of antifungal compounds. The active compound was identified as orsellinaldehyde (2,4-dihydroxy-6-methylbenzaldehyde) based on mass spectroscopy and nuclear magnetic resonance analyses. Orsellinaldehyde displayed broad-spectrum inhibitory activity against different plant pathogenic fungi. Among the tested Colletotrichum species, it exhibited the lowest IC50 values on conidial germination and germ tube elongation of Colletotrichum orbiculare. The compound also showed remarkable inhibitory activity against Colletotrichum gloeosporiodes. The staining of Colletotrichum conidia with fluorescein diacetate and propidium iodide demonstrated that the compound is fungicidal. The postharvest in - vivo detached fruit assay indicated that orsellinaldehyde suppressed anthracnose lesion symptoms on mango and cucumber fruits caused by C. gloeosporioides and C. orbiculare , respectively. Conclusions Orsellinaldehyde was identified as a potent antifungal compound from the culture filtrate of C. comatus. The inhibitory and fungicidal activities of orsellinaldehyde against different Colletotrichum species indicate its potential as a fungicide for protecting various fruits against anthracnose disease-causing fungi. [ABSTRACT FROM AUTHOR]

This study investigates the impact of water quality, specifically elevated phosphate and zinc content, on the diversity and functional properties of mangrove fungal endophytes in two distinct mangrove forests. Mangrove plant performance is directly related to the presence of fungal leaf endophytes as these fungi could enhance plant health, resilience, and adaptability under stressed environmental conditions. Two distinct mangrove forest sites, one non-disturbed (ND) and one disturbed by aquaculture practices (D), were assessed for differences in water quality parameters. We further analyzed the fungal endophyte diversity associated with the leaves of a target host mangrove, Rhizophora mucronata Lamk., with the aim to elucidate whether fungal diversity and functional traits are linked to disturbances brought about by aquaculture practices and to characterize functional traits of selected fungal isolates with respect to phosphate (PO4) and zinc (Zn) solubilization. Contrary to expectations, the disturbed site exhibited a higher fungal diversity, challenging assumptions about the relationship between contamination and fungal community dynamics. Water quality, as determined by nutrient and mineral levels, emerged as a crucial factor in shaping both microbial community compositions in the phyllosphere of mangroves. From both sites, we isolated 188 fungal endophytes, with the ND site hosting a higher number of isolates and a greater colonization rate. While taxonomic diversity marginally differed (ND: 28 species, D: 29 species), the Shannon (H' = 3.19) and FAI (FA = 20.86) indices revealed a statistically significant increase in species diversity for fungal endophytes in the disturbed mangrove site as compared to the non-disturbed area (H' = 3.10, FAI = 13.08). Our chosen mangrove fungal endophytes exhibited remarkable phosphate solubilization capabilities even at elevated concentrations, particularly those derived from the disturbed site. Despite their proficiency in solubilizing zinc across a wide range of concentrations, a significant impact on their mycelial growth was noted, underscoring a crucial aspect of their functional dynamics. Our findings revealed a nuanced trade-off between mycelial growth and enzymatic production in fungal endophytes from ostensibly less contaminated sites, highlighting the relationship between nutrient availability and microbial activities. These insights provide a foundation for understanding the impact of anthropogenic pressures, specifically nutrient pollution, on mangrove-associated fungal endophytes. [ABSTRACT FROM AUTHOR]

Heterocyte-forming cyanobacteria form symbiotic relationships with several lineages of plants. Here, twenty (20) strains of endosymbiotic cyanobacteria (cyanobionts) with Nostoc-like morphologies were isolated from the highly specialized coralloid roots of five host species in Cycadales—Cycas debaoensis, C. fairylakea, C. elongata, Ceratozamia robusta, and Macrozamia moorei. Molecular phylogeny based on the 16S rRNA gene placed these strains into seven different taxa within the Nostocaceae, specifically under the genera Desmonostoc and Dendronalium. The percent dissimilarity and unique patterns in the secondary structures of the D1-D1′, Box-B, V2, and V3 helices, which were based on the 16S–23S rRNA internal transcribed spacer (ITS) regions, supported three distinct species in Desmonostoc. These three morphologically distinct novel species are described in this report: Desmonostoc debaoense sp. nov., Desmonostoc meilinense sp. nov., and Desmonostoc xianhuense sp. nov. Other investigated strains were phylogenetically identified as members of the recently discovered genus Dendronalium and represent the first report of association of that genus with cycads. Our findings suggest that the order Cycadales hosts diverse species of cyanobionts in their coralloid roots and that many potential unreported or novel taxa are present in cycads occurring in their natural habitat and await discovery. [ABSTRACT FROM AUTHOR]

Carbon utilization by the marine Dendryphiella species, D. arenaria and D. salina, was investigated to detect differences in utilization and traits associated with their adaptation to the marine habitat. Fifty-four strains were isolated world-wide and tested for the utilization of various carbon sources using BIOLOG phenotype MicroArray (PM) and for the production of extracellular enzymes on solid culture media and on API ZYM assay strips. PM analysis showed that the fastest growth occurred on several monosaccharides and amino acids, 2-keto-d-gluconic acid, succinamide and turanose. Some polyols were poor carbon sources. However, the two species differed in their utilization rates of carbon sources, forming three major clusters: two separate clusters for D. arenaria and D. salina and a third cluster in which strains of the two species formed separate subclades that correlated with geographic origin. Several carbon sources were also found useful in differentiating the two speices. Dendryphiella salina did not utilize xylitol and quinic acid, whereas D. arenaria grew well on these substrates. The latter failed to grow on sorbitol and grew slowly on mannitol, both were good substrates for the former. There were also no qualitative differences between the extracellular enzymes produced, although laccase and peroxidase activities were confined only to some strains. The physiological similarities exhibited by the two species support the close relationship between D. arenaria and D. salina. [ABSTRACT FROM AUTHOR]

Antimicrobial compounds were purified from culture filtrates from 2 edible Pleurotus species. Using a bioassay-guided fractionation of the culture filtrate extracts, 3 compounds (1-3) were obtained from Pleurotus ostreatus, and another compound (4) was obtained from Pleurotus pulmonarius . Spectroscopic analysis revealed that 1-3 was identified as 5,7-dimethoxyphthalide, 4,6-dimethoxyphthalide, and cheimonophyllon E, respectively, while 4 were identified as pleuroton A. The minimum inhibitory concentration and minimum bactericidal concentration of these compounds were determined against 6 pathogenic bacterial species, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter cloacae. Compounds 2 and 4 were inhibitory against all tested bacteria, while 1 and 4 were inhibitory against 3 and 2 species, respectively. In addition, 1-4 inhibited tyrosinase, with IC50 values of 0.10-0.30 mg/mL, and α-glucosidase, with IC50 values of 0.12-0.54 mg/mL. However, their antioxidant capacities were marginal. [ABSTRACT FROM AUTHOR]

The excessive use of chemical pesticides in agricultural fields for controlling plant pathogenic microorganisms harms human health, the environment, and other beneficial microorganisms in the soil and plants. To address this challenge, it is essential to isolate and discover bioactive compounds from biological resources that could inhibit plant pathogenic microorganisms. In this study, the culture filtrate of the edible mushroom Pleurotus ostreatus was subjected to bioassay-guided isolation, and two phthalide derivatives--4,6-dimethoxyphthalide (1) and 5,7-dimethoxyphthalide (2)--were identified, along with an oxindole compound--3-hydroxy-3-methyloxindole (3). The inhibitory activities of the three compounds were evaluated against four fungal and five bacterial pathogens. Remarkably, 1 and 2 exhibited the lowest IC50 values against the conidial germination and germ tube elongation of the rice blast fungus Pyricularia oryzae. However, their effectiveness against bacterial pathogens was relatively low. The (S) and (R)-enantiomers of 3-hydroxy-3-methyloxindole showed different activities against plant fungal pathogens and bacterial plant pathogens. [ABSTRACT FROM AUTHOR]

Karst forest represents a distinct landscape with highly alkaline soil and limestone rocks. This specialized topography supports many unique species of plants and animals. Thus, documenting species in this area is important for any biodiversity research. In this study, a field survey was conducted to assess abundance, diversity, and distribution of myxomycetes in a karst forest within Quezon National Park, Philippines. Fruiting bodies were collected in addition to decaying substrates (e.g., aerial leaves and ground leaf litter) and twigs for culture in moist chambers. A total of 35 species from 16 genera was identified. The majority of these species occurred only rarely. Myxomycete communities between aerial and ground litter had the highest level of similarity based on their species composition and corresponding relative abundance. This study documented diversity of myxomycetes from the lowland karst landscape in the Philippines and now serves as baseline information for investigating plasmodial slime molds in Quezon National Park. [ABSTRACT FROM AUTHOR]

Ice-ice disease in cultivated seaweeds is often associated with environmental stress and infection by pathogenic marine bacteria. No studies have associated the disease with marine fungi. Our study aimed to isolate marine fungi from cultivated Kappaphycus species and assess their ability to induce the disease. Kappaphycus alvarezii and K. striatum were collected from Calatagan, Batangas, Philippines. Following washing with sterile artificial seawater and inoculation into culture media, 18 morphospecies of marine-derived fungi (MDF) were isolated. Fungal diversity (Hs=2.4) in infected seaweeds was higher than in healthy specimens. K. striatum (orange variety) had the highest incidence of MDF with 67 isolates, while K. striatum (green variety) had the lowest incidence with only 17 isolates. The ability of MDF to produce carrageenolytic and cellulolytic enzymes and utilize algal components was also tested. Of the 18 MDF selected, three had carrageenolytic activity and ten had cellulolytic activity. Most isolates utilized carrageenan, agar, and cellulose. Among the 10 MDF assayed for their ability to induce ice-ice disease, three isolates ( Aspergillus ochraceus, A. terreus and Phoma sp.) induced ice-ice disease symptoms (thallus bleaching) in healthy, non-axenic cultures of K. alvarezii. [ABSTRACT FROM AUTHOR]

Cycads have developed a complex root system categorized either as normal or coralloid roots. Past literatures revealed that a great diversity of key microbes is associated with these roots. This recent study aims to comprehensively determine the diversity and community structure of bacteria and fungi associated with the roots of two Cycas spp. endemic to China, Cycas debaoensis Zhong & Chen and Cycas fairylakea D.Y. Wang using high-throughput amplicon sequencing of the full-length 16S rRNA (V1-V9 hypervariable) and short fragment ITS region. The total DNA from 12 root samples were extracted, amplified, sequenced, and analyzed. Resulting sequences were clustered into 61 bacteria and 2128 fungal OTUs. Analysis of community structure revealed that the coralloid roots were dominated mostly by the nitrogen-fixer Nostocaceae but also contain other non-diazotrophic bacteria. The sequencing of entire 16S rRNA gene identified four different strains of cyanobacteria under the heterocystous genera Nostoc and Desmonostoc. Meanwhile, the top bacterial families in normal roots were Xanthobacteraceae, Burkholderiaceae, and Bacillaceae. Moreover, a diverse fungal community was also found in the roots of cycads and the predominating families were Ophiocordycipitaceae, Nectriaceae, Bionectriaceae, and Trichocomaceae. Our results demonstrated that bacterial diversity in normal roots of C. fairylakea is higher in richness and abundance than C. debaoensis. On the other hand, a slight difference, albeit insignificant, was noted for the diversity of fungi among root types and host species as the number of shared taxa is relatively high (67%). Our results suggested that diverse microbes are present in roots of cycads which potentially interact together to support cycads survival. Our study provided additional knowledge on the microbial diversity and composition in cycads and thus expanding our current knowledge on cycad-microbe association. Our study also considered the possible impact of ex situ conservation on cyanobiont community of cycads. [ABSTRACT FROM AUTHOR]

A total of nine endolichenic fungi were isolated from the foliose lichen Parmotrema rampoddense (Nyl.) Hale. Of the nine endolichenic fungi, three taxa (Fusarium proliferatum, Nemania primolutea, Daldinia eschsholtzii) showed antibacterial activities as determined by the disk diffusion assay against ESKAPE bacterial pathogens. Fusarium proliferatum gave the most active fungal extract with zone of inhibition values of 15 mm and 19 mm against E. faecalis and S. aureus, respectively. Further chromatographic purification of the F. proliferatum ethyl acetate extract led to the isolation and identification of bis(2-ethylhexyl)terephthalate (1), acetyl tributyl citrate (2), and fusarubin (3). Acetyl tributyl citrate (2) exhibited moderate antibacterial activity against Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. [ABSTRACT FROM AUTHOR]