New Pimarane Diterpenoids Isolated from EtOAc-Extract of Apiospora arundinis Culture Medium Show Antibenign Prostatic Hyperplasia Potential.

Three new pimarane diterpenoids, libertellenones U-W (1-3), together with libertellenone C (4) and myrocin A (5) were isolated from an EtOAc-extract of Apiospora arundinis culture medium. The chemical structures of the new compounds were elucidated using MS, NMR, and CD spectroscopic data. Benign prostatic hyperplasia (BPH), the abnormal and pathological proliferation of epithelial and stromal cells in prostatic tissues, is a common disease in middle-aged and elderly men. In this study, the anti-BPH effects of myrocin A (5) were evaluated using BPH-1 and WPMY-1 cells. Treatment with myrocin A (5) exerted antiproliferative effects in BPH-1 and dihydrotestosterone (DHT)-stimulated WPMY-1 cells. In BPH, treatment with myrocin A (5) significantly suppressed the mRNA levels of androgen receptor (AR) and its downstream targets nuclear receptor coactivator 1 (NCOA1), proliferating cell nuclear antigen (PCNA) and kallikrein-related peptidase 3 (KLK3). Additionally, DHT-stimulated WPMY-1 cells demonstrated an upregulated mRNA levels of AR, NCOA1, PCNA, and KLK3. However, treatment with myrocin A (5) resulted in suppression of the mRNA levels. Moreover, myrocin A (5) docked computationally into the binding site of the androgen receptor (-5.5 kcal/mol).

Novel Sarcoscypha Species from National Parks in Korea: Sarcoscypha humida sp. nov.

Sarcoscypha (Sarcoscyphaceae, Pezizales) is a saprobic fungus characterized by the cup or disc-shaped blight red apothecium and oblong to ellipsoid ascospores. The 18 species of Sarcoscypha were known to occur in Europe, North America, and tropical Asia. However, up to date, only two Sarcoscypha species have been reported in Korea. In this study, novel Sarcoscypha specimens were collected from Juwangsan, Odaesan, and Taebaeksan National Parks from September to October in Korea. This species is well distinguished from other Sarcoscypha species according to the molecular and phylogenetic analysis based on internal transcribed spacer (ITS) region. Here, we provided detailed descriptions with illustrations and a phylogenetic tree to report our specimens as novel Sarcoscypha species.

Notes of Five Wood-Decaying Fungi from Juwangsan National Park in Korea.

Wood-decaying fungi are essential decomposers in forest ecosystems. They decompose wood substrates by producing various lignocellulolytic enzymes, which have significant industrial and medical applications. A survey was conducted at the Juwangsan National Park from 2018 to 2019 to determine the diversity of macrofungi in Korea. Five previously unrecorded wood-decaying polyporoid and corticioid fungi were identified among the collected specimens: Eichleriella sinensis, Hymenochaete anomala, Hyphoderma subsetigerum, Lyomyces orientalis, and Pseudowrightoporia crassihypha. These species were identified based on morphological, molecular, and phylogenetic analyses of the internal transcribed spacer (ITS) and nuclear large subunit rDNA (nLSU) region. In this study, we provide detailed macro- and micro-morphological figures with phylogenetic trees to support the discovery of five new species in Korea.

Diversity of the Bambusicolous Fungus Apiospora in Korea: Discovery of New Apiospora Species.

Many Apiospora species have been isolated from bamboo plants - to date, 34 bambusicolous Apiospora species have been recorded. They are known as saprophytes, endophytes, and plant pathogens. In this study, 242 bambusicolous Apiospora were isolated from various bamboo materials (branches, culms, leaves, roots, and shoots) and examined using DNA sequence similarity based on the internal transcribed spacer, 28S large subunit ribosomal RNA gene, translation elongation factor 1-alpha, and beta-tubulin regions. Nine Apiospora species (Ap. arundinis, Ap. camelliae-sinensis, Ap. hysterina, Ap. lageniformis sp. nov., Ap. paraphaeosperma, Ap. pseudohyphopodii sp. nov., Ap. rasikravindrae, Ap. saccharicola, and Ap. sargassi) were identified via molecular analysis. Moreover, the highest diversity of Apiospora was found in culms, and the most abundant species was Ap. arundinis. Among the nine Apiospora species, two (Ap. hysterina and Ap. paraphaeosperma) were unrecorded in Korea, and the other two species (Ap. lageniformis sp. nov. and Ap. pseudohyphopodii sp. nov.) were potentially novel species. Here, we describe the diversity of bambusicolous Apiospora species in bamboo organs, construct a multi-locus phylogenetic tree, and delineate morphological features of new bambusicolous Apiospora in Korea.

Seven Unrecorded Indigenous Fungi from Mudeungsan National Park in Korea.

Fungi act as important decomposers in the forest environment. They recycle essential nutrients, promote plant growth through mycorrhizal relationships, and act as food for small animals. Samples of 265 indigenous fungal species were collected from Mudeungsan National Park in 2020. These species were identified based on morphological, molecular, and phylogenetic analyses using the internal transcribed spacer (ITS), nuclear large subunit rRNA (LSU), and RNA polymerase II second largest subunit (rpb2) regions. Subsequently, seven species were identified as unrecorded species in Korea: Cordyceps cicadae, Dentocorticium bicolor, Hymenochaete nanospora, Physisporinus crataegi, Rigidoporus piceicola, Russula raoultii, and Scutellinia crinita. This study reveals their detailed macro- and microscopic morphological characteristics with phylogenetic trees to report them as unrecorded species in Korea.

Two Unrecorded Apiospora Species Isolated from Marine Substrates in Korea with Eight New Combinations (A. piptatheri and A. rasikravindrae).

Although Apiospora Sacc. has previously been considered a sexual morph of Arthrinium species on the basis of phylogenetic, morphological, and ecological diagnoses, a recent study delimited these as different species. Recently, 14 species, including eight new species, of marine Arthrinium have been reported from Korea. Six known species have previously been renamed as species in the genus Apiospora (A. arundinis, A. marii, A. piptatheri, A. rasikravindrae, A. sacchari, and A. saccharicola). However, the eight new species of marine Arthrinium (Ar. agari, Ar. arctoscopi, Ar. fermenti, Ar. koreanum, Ar. marinum, Ar. pusillispermum, Ar. sargassi, and Ar. taeanense) are yet to be studied, and thus the taxonomic status of these species remains to be clarified. In this study, we conducted phylogenetic analyses using the internal transcribed spacer, 28S large subunit ribosomal RNA gene, translation elongation factor 1-alpha, and beta-tubulin regions to confirm the phylogenetic position of these eight species. Based on these analyses, we re-identified the eight Arthrinium species as new combinations in Apiospora. Additionally, among the six known Apiospora species, two (A. piptatheri and A. rasikravindrae) have not previously been recorded in Korea. On the basis of morphological and molecular analyses, we report these as new species in Korea. Herein, we present scanning electron micrographs detailing the morphologies of these species, along with phylogenetic trees and detailed descriptions.

Chemical constituents from basidiomycete Basidioradulum radula culture medium and their cytotoxic effect on human prostate cancer DU-145 cells.

Eight new naphtho[1,2-c]furan derivatives (1-8) along with six known analogues (9-14) were isolated from culture medium of the basidiomycete Basidioradulum radula. The structures of these compounds were identified using spectroscopic analysis, and their absolute configurations were resolved using X-ray diffraction, ECD, and VCD. Compounds 7 and 14 inhibited the cell viability of human prostate cancer DU-145 cells with IC50 values of 7.54 ± 0.03 µM and 5.04 ± 0.03 µM, respectively. At 8 µM, compounds 7 and 14 increased the percentage of apoptotic cells and upregulated the protein expression related to the apoptosis caspase pathways in DU-145 cells. Furthermore, the hallmarks of cells undergoing apoptosis, such as chromatin condensation, were also observed at this concentration. However, compound 7 and 14 showed no effect on the proliferation of splenocytes isolated from cyclophosphamide-induce immunosuppressed mice.

The genus Arthrinium (Ascomycota, Sordariomycetes, Apiosporaceae) from marine habitats from Korea, with eight new species.

Species of Arthrinium are well-known plant pathogens, endophytes, or saprobes found in various terrestrial habitats. Although several species have been isolated from marine environments and their remarkable biological activities have been reported, marine Arthrinium species remain poorly understood. In this study, the diversity of this group was evaluated based on material from Korea, using morphological characterization and molecular analyses with the internal transcribed spacer (ITS) region, ß-tubulin (TUB), and translation elongation factor 1-alpha (TEF). A total of 41 Arthrinium strains were isolated from eight coastal sites which represented 14 species. Eight of these are described as new to science with detailed descriptions.

Genomic Analysis and Assessment of Melanin Synthesis in Amorphotheca resinae KUC3009.

This study reports the draft genome of Amorphotheca resinae KUC30009, a fungal isolate with promising industrial-scale melanin production potential. The mechanisms for melanin or melanin-related pigment formation of this strain were examined through bioinformatic and biochemical strategies. The 30.11 Mb genome of A. resinae contains 9638 predicted genes. Genomic-based discovery analyses identified 14 biosynthetic gene clusters (BGCs) associated with secondary metabolite production. Moreover, genes encoding a specific type 1 polyketide synthase and 4-hydroxynaphthalene reductase were identified and predicted to produce intermediate metabolites of dihydroxy naphthalene (DHN)-melanin biosynthesis pathway, but not to DHN-melanin. These findings were further supported by the detection of increased flaviolin concentrations in mycelia and almost unchanged morphologies of the culture grown with tricyclazole. Apart from this, the formation of melanin in the culture filtrate appeared to depend on the laccase-like activity of multi-copper oxidases. Simultaneously, concentrations of nitrogen-containing sources decreased when the melanin formed in the media. Interestingly, melanin formation in the culture fluid was proportional to laccase-like activity. Based on these findings, we proposed novel strategies for the enhancement of melanin production in culture filtrates. Therefore, our study established a theoretical and methodological basis for synthesizing pigments from fungal isolates using genomic- and biochemical-based approaches.

Environmental drivers affecting the bacterial community of intertidal sediments in the Yellow Sea.

Intertidal flats, as transition zones where terrestrial and marine ecosystems meet, provide unique environments and play an important role in marine ecosystems. In particular, the environmental characteristics of tidal marshes show are different than those of bare flats, especially in the rhizosphere. However, unlike the rhizosphere in terrestrial ecosystems, the rhizosphere of plants in tidal marsh areas and the associated microbial community have been the focus of very little research. Thus, this study investigated the diversity and variation in bacterial communities in the rhizosphere of a Phragmites australis and Suaeda japonica and along the sediment depths. High-throughput sequencing was performed by amplifying the 16S rRNA gene of environmental DNA extracted from sediment cores, and indicator species were identified with respect to the vegetation type and sediment depth. The most abundant phylum was Proteobacteria, followed by Chloroflexi, Bacteroidetes, Acidobacteria, and Firmicutes. In general, the results indicated that not only vegetation type and sediment depth themselves but also their interaction resulted in significant differences among the bacterial communities. The envfit results revealed that the environmental variables of sediment, such as mud content, organic matter, total organic carbon, and total nitrogen, had significant effects on the bacterial community structure. The indicator species varied depending on the vegetation type and sediment depth, showing significant correlations with certain selected environmental variables, but were fundamentally related to the rhizosphere. Overall, this study revealed the key factors that determine the bacterial community structure in tidal marshes and the indicator species according to vegetation type in the little studied rhizosphere of the intertidal ecosystem.

Production and characterization of melanin pigments derived from Amorphotheca resinae.

As melanin has emerged as functional pigment with cosmetic, health and food applications, the demand for the pigments is expected to increase. However, the conventional sources (e.g. mushroom, hair, and wool) of melanin production entail pigments inside the substrates which requires the costly extraction procedures, leading to inappropriate scalable production. In this study, we screened 102 of fungal isolates for their ability to produce melanin in the supernatant and selected the only Amorphotheca resinae as a promising candidate. In the peptone yeast extract glucose broth, A. resinae produced the melanin rapidly during the autolysis phase of growth, reaching up 4.5 g/L within 14 days. Structural characterization of the purified melanin from A. resinae was carried out by using elemental analysis, electron paramagnetic resonance, 13C solid-state nuclear magnetic resonance spectroscopy, and pyrolysis-gas chromatography-mass spectrometry in comparison with the standard melanins. The results indicate that the structural properties of A. resinae melanin is similar to the eumelanin which has a wide range of industrial uses. For example, the purified melanin from A. resinae has the potent antioxidant activities as a result of free radical scavenging assays. Consequently, A. resinae KUC3009 can be a promising candidate for scalable production of industrially applicable melanin.

Note of Five Unrecorded Mushrooms Including Three Rare Species on Mount Juwang in Korea.

The Mount Juwang was designated as a national park in 1976 because of its unique bedrock geology. Although Juwang National Park has maintained its natural ecosystem well, few macrofungal surveys have been carried out. As a part of a project supported by the National Institute of Biological Resources (NIBR) for the discovery of indigenous fungal species, we surveyed the mushrooms in Juwang National Park from 2018 to 2019. The macrofungi were identified based on morphological and molecular analyses. Among these fungi, five specimens were identified as species previously unrecorded in Korea: Calocybe decolorata, Crepidotus brunnescens, Mycena pearsoniana, Psathyrella phegophila, and P. sulcatotuberculosa. Three of these species are known to be rare in the world: Crepidotus brunnescens, P. phegophila, and P. sulcatotuberculosa. In this study, we provide detailed morphological descriptions of the five unrecorded species from Mt. Juwang in Korea.

Recovery of the benthic bacterial community in coastal abandoned saltern requires over 35 years: A comparative case study in the Yellow Sea.

Salt is an essential nutrient for humans, and salterns exist worldwide. Although the construction of salterns has stopped and typical salterns are now mostly abandoned, there has been no research on the ecological recovery of the abandoned salterns. Here, we analyzed the bacterial diversity and community structure in three pairs of abandoned salterns that have undergone 1-35 years of natural restoration and tidal flats to determine the recovery time and process. Partial 16S rRNA sequences were amplified and sequenced to investigate the biodiversity and structure of the bacterial community in sediments collected from abandoned salterns and adjacent natural tidal flats (viz., controls) in the Yellow Sea. The most abundant microorganisms across locations were found to be members of Proteobacteria, ranging from 45 to 72%, which was also a crucial taxon in the bacterial recovery process. The benthic bacterial community of the salterns showed time-dependent recovery, as demonstrated by the similarity between the salterns and controls. Indeed, dissimilarities between bacterial communities were significant for the saltern that had been abandoned for one year, according to ANOSIM (R = 1.0, p < 0.01). The genera that were determined to contribute to the dissimilarity exhibited a significant correlation with the sedimentary phosphorus concentration. The dataset generally supported that the indigenous benthic bacterial community in an altered marine environment might require a considerable time to return to a natural status. Meanwhile, a delay between the recovery of the physicochemical environment and biological component was evidenced, which seemed to influence the recovery time in a site-specific manner. Overall, the present study provided new insight and understanding of the recovery of the benthic bacterial community in abandoned salterns in terms of recovery time and the associated process.

Fungal Diversity in Intertidal Mudflats and Abandoned Solar Salterns as a Source for Biological Resources.

Intertidal zones are unique environments that are known to be ecological hot spots. In this study, sediments were collected from mudflats and decommissioned salterns on three islands in the Yellow Sea of South Korea. The diversity analysis targeted both isolates and unculturable fungi via Illumina sequencing, and the natural recovery of the abandoned salterns was assessed. The phylogeny and bioactivities of the fungal isolates were investigated. The community analysis showed that the abandoned saltern in Yongyudo has not recovered to a mudflat, while the other salterns have almost recovered. The results suggested that a period of more than 35 years may be required to return abandoned salterns to mudflats via natural restoration. Gigasporales sp. and Umbelopsis sp. were selected as the indicators of mudflats. Among the 53 isolates, 18 appeared to be candidate novel species, and 28 exhibited bioactivity. Phoma sp., Cladosporium sphaerospermum, Penicillium sp. and Pseudeurotium bakeri, and Aspergillus urmiensis showed antioxidant, tyrosinase inhibition, antifungal, and quorum-sensing inhibition activities, respectively, which has not been reported previously. This study provides reliable fungal diversity information for mudflats and abandoned salterns and shows that they are highly valuable for bioprospecting not only for novel microorganisms but also for novel bioactive compounds.

Importance of functional diversity in assessing the recovery of the microbial community after the Hebei Spirit oil spill in Korea.

Over 10 years after the Hebei Spirit oil spill (HSOS), the concentrations of pollutants, such as TPH and PAHs, in spilled crude oil have recovered to background levels, but in some areas, the environment has not fully recovered. In particular, PAHs were more resistant to degradation, and their persistence could have deleterious impacts on the sediment ecosystem. This study aimed to evaluate the microbial recovery of coastal sediments from the HSOS by analyzing the structure and diversity of the microbial community and its functional contribution to PAHs degradation. High-throughput sequencing on the MiSeq platform was conducted using tidal flat sediments collected in 2014 and 2016 from the area contaminated by the HSOS. The microbial recovery was evaluated by various diversity factors, including microbial composition and structure and functional diversity based on PICRUSt analysis. The abundance of microbial taxa associated with TPH degradation was higher in 2014 than that in 2016, but the taxa associated with PAHs degradation were similar between years. These results are consistent with the dynamics of microbes associated with the fate of pollutants, and they also showed similar tendency in functional profiles. That is, even if the pollutants are completely degraded, the microbial community has not yet completely recovered from the HSOS. The evaluation of microbial ecosystems in contaminated environments should consider both the fate of pollutants and the dynamics of microbial species that make functional contributions to the degradation of pollutants.

Diversity and Ecology of Marine Algicolous Arthrinium Species as a Source of Bioactive Natural Products.

In our previous study, all Arthrinium isolates from Sargassum sp. showed high bioactivities, but studies on marine Arthrinium spp. are insufficient. In this study, a phylogenetic analysis of 28 Arthrinium isolates from seaweeds and egg masses of Arctoscopus japonicus was conducted using internal transcribed spacers, nuclear large subunit rDNA, ß-tubulin, and translation elongation factor region sequences, and their bioactivities were investigated. They were analyzed as 15 species, and 11 of them were found to be new species. Most of the extracts exhibited radical-scavenging activity, and some showed antifungal activities, tyrosinase inhibition, and quorum sensing inhibition. It was implied that marine algicolous Arthrinium spp. support the regulation of reactive oxygen species in symbiotic algae and protect against pathogens and bacterial biofilm formation. The antioxidant from Arthrinium sp. 10 KUC21332 was separated by bioassay-guided isolation and identified to be gentisyl alcohol, and the antioxidant of Arthrinium saccharicola KUC21221 was identical. These results demonstrate that many unexploited Arthrinium species still exist in marine environments and that they are a great source of bioactive compounds.

New Report of Three Unrecorded Species in Trichoderma harzianum Species Complex in Korea.

The genus Trichoderma (Hypocreaceae, Ascomycota) consists of globally distributed fungi. Among them, T. harzianum, one of the most commonly collected Trichoderma species, had been known as a polyphyletic or aggregate species. However, a total of 19 species were determined from the polyphyletic groups of T. harzianum. Thus, we explored Korean "T. harzianum" specimens that were collected in 2013-2014. These specimens were re-examined based on a recent study with translate elongation factor 1-alpha (EF1α) sequences to reveal cryptic Trichoderma species in Korea. As a result, four different species, T. afroharzianum, T. atrobruneum, T. pyramidale, and T. harzianum, were identified. Except T. harzianum, the other three species have not been reported in Korea. In this work, we describe these species and provide figures.

Investigation of Filamentous Fungi Producing Safe, Functional Water-Soluble Pigments.

The production of water-soluble pigments by fungal strains indigenous to South Korea was investigated to find those that are highly productive in submerged culture. Among 113 candidates, 34 strains that colored the inoculated potato dextrose agar medium were selected. They were cultured in potato dextrose broth and extracted with ethanol. The productivity, functionality (radical-scavenging activities), and color information (CIELAB values) of the pigment extracts were measured. Five species produced intense yellowish pigments, and two produced intense reddish pigments that ranked the highest in terms of absorbance units produced per day. The pigment extracts of Penicillium miczynskii, Sanghuangporus baumii, Trichoderma sp. 1, and Trichoderma afroharzianum exhibited high radical-scavenging activity. However, the S. baumii extract showed moderate toxicity in the acute toxicity test, which limits the industrial application of this pigment. In conclusion, P. miczynskii KUC1721, Trichoderma sp. 1 KUC1716, and T. afroharzianum KUC21213 were the best fungal candidates to be industrial producers of safe, functional water-soluble pigments.