First Report of Rust Caused by Puccinia sasicola on Corylopsis coreana in South Korea.

Korean winter hazel (Corylopsis coreana) is an endemic species of the South Korea (Seo et al. 2016; Kim et al. 2021), which is cultivated as an ornamental plant in this country, but also in China and Japan (Yoon et al. 2016). In July 2022, typical symptoms of a rust disease were observed on C. coreana at Jirisan National Park (35°22'07.7"N 127°34'57.7"E) in Namwon, South Korea. Spermogonia were epiphyllous, densely grouped, pale brown or orange-yellow, round, and 0.23 - 0.38 × 0.19 - 0.41 mm in size. Aecia were hypophyllous, mostly densely grouped, yellow or pale orange, resembling small wart-like galls, and 0.04 - 0.06 × 0.89 - 1.68 mm in size. Aeciospores were hyaline, mostly angularly globose, ellipsoid or oblong-ellipsoid, and 17.8 - 25.2 × 15 - 26.5 µm (average 19.2 × 19.1 µm; n=50) in size. Aeciospore walls were echinulate-verrucose, and 1.1 - 2.2 µm (average 1.7 µm; n=50) in thickness. In December 2022, dark brown telia were observed on the lower surface of Sasa borealis leaves near C. coreana. Telia were mostly scattered but often compacted, brown to dark brown, round, and 1.5 - 1.95 × 1.24 - 1.55 mm in size. Teliospores were either one- or two-celled with a long tapering apex, and light brown to brown in color. One-celled teliospores were globose, and 95.1 - 186.5 × 20.5 - 36.4 µm (average 136.4 × 27.7 µm; n=50) in size. Two-celled teliospores were ellipsoid-cylindrical, and 111.4 - 180.3 × 13.5 - 32.6 µm (average 149 × 21.1 µm; n=50) in size. Side walls of teliospores were golden and 2.2 - 5.5 µm thick (average 3.5 µm; n=50), and pedicels were hyaline, measuring 150 - 300 µm long. Uredinial stage was not observed. Disease symptomology and pathogen morphology were mostly consistent with that of Puccinia sasicola reported in Japan (Hino. 1955). For phylogenetic analysis, genomic DNA was extracted from the aeciospores collected from C. coreana and the teliospores collected from S. borealis. The internal transcribed spacer (ITS) rDNA and the large subunit (LSU) rDNA regions were amplified using ITS5u/ITS4rust (Pfunder and Schürch 2001) and LRust1R/LRust3 (Beenken et al. 2012) primers, respectively. Both sequences were identical for the spores collected from the two different hosts. The sequences were deposited in GenBank (PP171665, PP174211 [ITS], PP171709, PP174356 [LSU]). A GenBank BLAST search revealed 89.53% and 96.78% similarity with Puccinia kusanoi (KX610657) and Puccinia sp. (MT7298241) for ITS and LSU sequences, respectively. In maximum-likelihood phylogenetic analysis of ITS and LSU sequences, the isolates from C. coreana and S. borealis formed a separate clade from other Puccinia species. To test Koch's postulates, leaf disks with telia from S. borealis were directly attached to the adaxial surface of six healthy C. coreana leaves with tape. As controls, healthy S. borealis leaf disks were attached to the adaxial surface of six C. coreana leaves. After four weeks, four inoculated leaves developed small yellow wart-like galls on the abaxial surface, while the control leaves remained symptom-free. The isolates obtained from the inoculated leaves had identical sequences to the original isolate. There are no publicly available sequences for P. sasicola, nor did we find any sequences that match our Puccinia samples. Nevertheless, based on morphological characteristics and life cycle, our isolates closely matched with the previous description of P. sasicola by Hino (1955). To our knowledge, this is the first report of P. sasicola causing leaf rust in C. coreana in South Korea.

Exploring fine-scale assembly of ectomycorrhizal fungal communities through phylogenetic and spatial distribution analyses.

Ectomycorrhizal fungi (EMF) form symbiotic relationship with the roots of host plants. EMF communities are composed of highly diverse species; however, how they are assembled has been a long-standing question. In this study, we investigated from a phylogenetic perspective how EMF communities assemble on Pinus densiflora seedlings at different spatial scales (i.e., seedling scale and root tip scale). P. densiflora seedlings were collected from different habitats (i.e., disturbed areas and mature forests), and their EMF communities were investigated by morphotype sequencing and next-generation sequencing (NGS). To infer assembly mechanisms, phylogenetic relatedness within the community (i.e., phylogenetic structure) was estimated and spatial distribution of EMF root tips was analyzed. The EMF communities on pine seedlings were largely different between the two habitats. Phylogenetically restricted lineages (Amphinema, /suillus-rhizopogon) were abundant in the disturbed areas, whereas species from diverse lineages were abundant in the mature forests (Russula, Sebacina, /tomentella-thelephora, etc.). In the disturbed areas, phylogenetically similar EMF species were aggregated at the seedling scale, suggesting that disturbance acts as a powerful abiotic filter. However, phylogenetically similar species were spatially segregated from each other at the root tip scale, indicating limiting similarity. In the mature forest seedlings, no distinct phylogenetic signals were detected at both seedling and root tip scale. Collectively, our results suggest that limiting similarity may be an important assembly mechanism at the root tip scale and that assembly mechanisms can vary across habitats and spatial scales.

Fungal Catastrophe of a Specimen Room: Just One Week is Enough to Eradicate Traces of Thousands of Animals.

Indoor fungi obtain carbon sources from natural sources and even recalcitrant biodegradable materials, such as plastics and synthetic dye. Their vigorous activity may have negative consequences, such as structural damage to building materials or the destruction of precious cultural materials. The animal specimen room of the Seoul National University stocked 36,000 animal resources that had been well-maintained for over 80 years. Due to abandonment without the management of temperature and humidity during the rainy summer season, many stuffed animal specimens had been heavily colonized by fungi. To investigate the fungal species responsible for the destruction of the historical specimens, we isolated fungi from the stuffed animal specimens and identified them at the species level based on morphology and molecular analysis of the ß-tubulin (BenA) gene. A total of 365 strains were isolated and identified as 26 species in Aspergillus (10 spp.), Penicillium (14 spp.), and Talaromyces (2 spp.). Penicillium brocae and Aspergillus sydowii were isolated from most sections of the animal specimens and have damaged the feathers and beaks of valuable specimens. Our findings indicate that within a week of mismanagement, it takes only a few fungal species to wipe out the decades of history of animal diversity. The important lesson here is to prevent this catastrophe from occurring again through a continued interest, not to put all previous efforts to waste.

Diversity and Dynamics of Marine Arenicolous Fungi in Three Seasides of the Korean Peninsula.

Various arenicolous fungal species have been detected from the beach sand in the coastal area. However, little has been revealed regarding their distribution and dynamics. To investigate the overall diversity of marine arenicolous fungi (MAFs) in Korea and whether the composition of MAFs is affected by ocean currents, we isolated and analyzed the fungal community from the western, southern, and eastern seasides of the Korean Peninsula. In total, 603 strains were isolated and identified as 259 species based on appropriate molecular markers for each genus (ITS, BenA, CaM, tef1, and act). The composition of MAFs showed differences among the seasides. Our results indicate that many MAFs inhabit the beach sand on the Korean Peninsula, and the composition of MAFs is also affected by ocean currents flowing along each coast.

Mycorrhizal Fungal Diversity Associated with Six Understudied Ectomycorrhizal Trees in the Republic of Korea.

Mycorrhizal fungi are key components of forest ecosystems and play essential roles in host health. The host specificity of mycorrhizal fungi is variable and the mycorrhizal fungi composition for the dominant tree species is largely known but remains unknown for the less common tree species. In this study, we collected soil samples from the roots of six understudied ectomycorrhizal tree species from a preserved natural park in the Republic of Korea over four seasons to investigate the host specificity of mycorrhizal fungi in multiple tree species, considering the abiotic factors. We evaluated the mycorrhizal fungal composition in each tree species using a metabarcoding approach. Our results revealed that each host tree species harbored unique mycorrhizal communities, despite close localization. Most mycorrhizal taxa belonged to ectomycorrhizal fungi, but a small proportion of ericoid mycorrhizal fungi and arbuscular mycorrhizal fungi were also detected. While common mycorrhizal fungi were shared between the plant species at the genus or higher taxonomic level, we found high host specificity at the species/OTU (operational taxonomic unit) level. Moreover, the effects of the seasons and soil properties on the mycorrhizal communities differed by tree species. Our results indicate that mycorrhizal fungi feature host-specificity at lower taxonomic levels.

Taxonomic study of Collybiopsis (Omphalotaceae, Agaricales) in the Republic of Korea with seven new species.

Collybiopsis is a genus of the gymnopoid/marasmioid complex of the family Omphalotaceae. The classification system of Collybiopsis has recently undergone large changes through molecular approaches. The new classification system has not been applied for Collybiopsis in the Republic of Korea, and general research on this genus was also lacking. In this study, we analyzed the Collybiopsis species in the Republic of Korea by assessing all gymnopoid/marasmioid specimens collected nationwide for ten years by combining morphological approaches and multilocus (ITS + nrLSU) phylogenetic analysis. We thus confirmed that 16 species of Collybiopsis are present in the Republic of Korea, including two previously unreported species (Co.nonnulla and Co.dichroa) and seven new species (Co.albicantipes sp. nov., Co.clavicystidiata sp. nov., Co.fulva sp. nov., Co.orientisubnuda sp. nov., Co.subumbilicata sp. nov., Co.undulata sp. nov., and Co.vellerea sp. nov.). A thorough examination of the Collybiopsis suggested that it is difficult to distinguish or identify the species based on morphological characteristics only; a combined molecular approach is needed for accurate identification. The Collybiopsis database of the Republic of Korea is updated, and information on the new species is provided. Five new combinations from Marasmiellus to Collybiopsis are also proposed (Co.istanbulensis comb. nov., Co.koreana comb. nov., Co.omphalodes comb. nov., Co.pseudomphalodes comb. nov., and Co.ramuliciola comb. nov.).

Five Previously Unrecorded Fungal Species Isolated from Marine Plastic Wastes in South Korea.

Plastic wastes have a negative impact on marine environments; however, they can be used as carbon sources and habitats by certain microbes. Microbes in the marine plastisphere can migrate worldwide through the ocean and cause serious environmental problems when they encounter suitable environments. Therefore, efforts to investigate the microbes inhabiting the marine plastisphere are increasing. In the present study, fungal strains were isolated from plastic wastes buried in Korean sea sands and mudflats and identified using molecular and morphological analyses. Five species were identified that were previously unrecorded from South Korea: Cladosporium funiculosum, Neosetophoma poaceicola, Neosetophoma rosigena, Parasarocladium gamsii, and Trichoderma fomiticola. Their molecular phylogenies and morphological characteristics are described in this study.

Different patterns of belowground fungal diversity along altitudinal gradients with respect to microhabitat and guild types.

Fungi are key components of belowground ecosystems with various ecological roles in forests. Although the changes in the richness and composition of belowground fungi across altitudinal gradients have been widely reported, only a few studies have focused on the microhabitat types along altitudinal gradients. Here, we analysed the effect of altitude on the ectomycorrhizal and non-ectomycorrhizal fungal communities in belowground microhabitats. We collected root and soil samples from 16 Pinus densiflora forests at various altitudes across Korea, and measured the soil properties as potential factors. Fungal communities were analysed by high-throughput sequencing of the internal transcribed spacer 2 (ITS2) region. We found that altitude negatively affected the species richness of root-inhabiting fungi but did not influence that of soil-inhabiting fungi. In addition, the composition of ectomycorrhizal (ECM) fungi was less influenced by altitude than non-ECM fungi. Most of the soil properties did not show a significant relationship with altitude, but the effect of soil properties was different across microhabitat types and ecological roles of fungi. Our results reveal that microhabitat types and altitudinal gradients differently affect the richness and composition of fungal communities associated with P. densiflora, providing a better understanding of plant-associated fungal communities.

Investigation of the Fungal Diversity of the Federated States of Micronesia and the Construction of an Updated Fungal Inventory.

The Federated States of Micronesia (FSM) is an island country in the western Pacific and is a known biodiversity hotspot. However, a relatively small number of fungi (236 species) have been reported till July 2021. Since fungi play major ecological roles in ecosystems, we investigated the fungal diversity of FSM from various sources over 2016 and 2017 and constructed a local fungal inventory, which also included the previously reported species. Fruiting bodies were collected from various host trees and fungal strains were isolated from marine and terrestrial environments. A total of 99 species, of which 78 were newly reported in the FSM, were identified at the species level using a combination of molecular and morphological approaches. Many fungal species were specific to the environment, host, or source. Upon construction of the fungal inventory, 314 species were confirmed to reside in the FSM. This inventory will serve as an important basis for monitoring fungal diversity and identifying novel biological resources in FSM.

Determination of Diversity, Distribution and Host Specificity of Korean Laccaria Using Four Approaches.

The genus Laccaria (Hydnangiaceae, Agaricales) plays an important role in forest ecosystems as an ectomycorrhizal fungus, contributing to nutrient cycles through symbiosis with many types of trees. Though understanding Laccaria diversity and distribution patterns, as well as its association with host plants, is fundamental to constructing a balanced plant diversity and conducting effective forest management, previous studies have not been effective in accurately investigating, as they relied heavily on specimen collection alone. To investigate the true diversity and distribution pattern of Laccaria species and determine their host types, we used four different approaches: specimen-based analysis, open database search (ODS), NGS analysis, and species-specific PCR (SSP). As a result, 14 Laccaria species have been confirmed in Korea. Results regarding the species distribution pattern were different between specimen-based analysis and SSP. However, when both were integrated, the exact distribution pattern of each Laccaria species was determined. In addition, the SSP revealed that many Laccaria species have a wide range of host types. This study shows that using these four different approaches is useful in determining the diversity, distribution, and host of ECM fungi. Furthermore, results obtained for Laccaria will serve as a baseline to help understand the role of ECM fungi in forest management in response to climate change.

Two New Species of Laccaria (Agaricales, Basidiomycota) from Korea.

Species of Laccaria (Hydnangiaceae, Agaricales, and Basidiomycota) are well-known ectomycorrhizal symbionts of a broad range of hosts. Laccaria species are characterized by brown, orange, or purple colored basidiocarps, and globose or oblong, echinulate and multinucleate basidiospores. While some Laccaria species are easily identified at the species level using only the morphological characteristics, others are hard to distinguish at the species level due to small differences in morphology. Heretofore, ten Laccaria species have been reported in Korea. While studying the fungal diversity in the National Parks of Korea, two new Laccaria species were discovered. Species identification was done based on molecular analyses (ITS, 28S rDNA, rpb2, and tef1), then were confirmed by their corresponding morphologies. The two newly discovered Laccaria species are proposed here as Laccaria macrobasidia and Laccaria griseolilacina. The unique morphological characters of L. macrobasidia that distinguish it from its closely related species are orange-brown colored basidiocarp, long basidia and the absence of cheilocystidia. L. griseolilacina is characterized by a light grayish lavender-colored pileus and the absence of cheilocystidia. Two new species are described and illustrated in the present paper.

Influence of Season and Soil Properties on Fungal Communities of Neighboring Climax Forests (Carpinus cordata and Fraxinus rhynchophylla).

Trees in forest ecosystems constantly interact with the soil fungal community, and this interaction plays a key role in nutrient cycling. The diversity of soil fungal communities is affected by both environmental factors and host tree species. We investigated the influence of both of these factors by examining the total fungal communities in the rhizospheric soil of climax tree species that have similar ecological roles (Carpinus cordata, an ectomycorrhizal [ECM] tree, and Fraxinus rhynchophylla, an arbuscular mycorrhizal [AM] tree) in temperate forests with continental climates of Mt. Jeombong, South Korea. Fungal communities were assessed by Illumina-MiSeq sequencing the internal transcribed spacer (ITS) region of environmental DNA, and comparing their environmental factors (season and soil properties). We found that soil fungi of the two forest types differed in terms of community structure and ecological guild composition. The total fungal community composition changed significantly with seasons and soil properties in the F. rhynchophylla forest, but not in the C. cordata forest. However, potassium and carbon were significantly correlated with fungal diversity in both forests, and a positive correlation was found only between symbiotrophs of C. cordata and the carbon to nitrogen (C/N) ratio. Thus, the effects of environmental factors on soil fungal communities depended on the host trees, but some factors were common in both forests. Our results indicate that individual tree species should be considered when anticipating how the fungal communities will respond to environmental change.

Successional Change of the Fungal Microbiome Pine Seedling Roots Inoculated With Tricholoma matsutake.

The pine mushroom (Tricholoma matsutake; Agaricales, Tricholomataceae) is an ectomycorrhizal fungus that produces a commercially valuable, edible mushrooms. Attempts to artificially cultivate T. matsutake has so far been unsuccessful. One method used to induce T. matsutake to produce fruiting bodies of in the wild is shiro (mycelial aggregations of T. matsutake) transplantation. In vitro ectomycorrhization of T. matsutake with seedlings of Pinus densiflora has been successful, but field trials showed limited production of fruiting bodies. Few studies have been done to test what happens after transplantation in the wild, whether T. matsutake persists on the pine seedling roots or gets replaced by other fungi. Here, we investigated the composition and the interaction of the root fungal microbiome of P. densiflora seedlings inoculated with T. matsutake over a 3 year period after field transplantation, using high-throughput sequencing. We found a decline of T. matsutake colonization on pine roots and succession of mycorrhizal fungi as P. densiflora seedlings grew. Early on, roots were colonized by fast-growing, saprotrophic Ascomycota, then later replaced by early stage ectomycorrhiza such as Wilcoxina. At the end, more competitive Suillus species dominated the host roots. Most of the major OTUs had negative or neutral correlation with T. matsutake, but several saprotrophic/plant pathogenic/mycoparasitic species in genera Fusarium, Oidiodendron, and Trichoderma had positive correlation with T. matsutake. Four keystone species were identified during succession; two species (Fusarium oxysporum, and F. trincintum) had a positive correlation with T. matsutake, while the other two had a negative correlation (Suillus granulatus, Cylindrocarpon pauciseptatum). These findings have important implications for further studies on the artificial cultivation of T. matsutake.

Seventeen Unrecorded Species from Gayasan National Park in Korea.

Macrofungi play important roles in forest ecology as wood decayers, symbionts, and pathogens of living trees. For the effective forest management, it is imperative to have a comprehensive overview of macrofungi diversity in specific areas. As a part of the National Institute of Biological Resources projects for discovering indigenous fungi in Korea, we collected macrofungi in Gayasan National Park from 2017 to 2018. These specimens were identified based on morphological characteristics and sequence analysis of internal transcribed spacer (ITS) or the nuclear large subunit rRNA (LSU) region. We discovered 17 macrofungi new to Korea: Butyrea japonica, Ceriporia nanlingensis, Coltricia weii, Coltriciella subglobosa, Crepidotus crocophyllus, Cylindrobasidium laeve, Fulvoderma scaurum, Laetiporus cremeiporus, Lentinellus castoreus, Leucogyrophana mollusca, Marasmius insolitus, Nidularia deformis, Phaeophlebiopsis peniophoroides, Phanerochaete angustocystidiata, Phlebiopsis pilatii, Postia coeruleivirens, and Tengioboletus fujianensis. We described their detailed morphological characteristics.

Macrolepiota in Korea: New Records and a New Species.

The genus Macrolepiota (Agaricales, Basidiomycota) is easy to recognize at the genus level because of big, fleshy basidiocarps with squamules covering the pileus; a single or double annulus; and big, thick-walled basidiospores with a germ pore. However, morphological identification is often unreliable in Macrolepiota due to similar morphological features among species. Due to the uncertainty of previous morphological identification in the genus Macrolepiota, it is necessary to re-examine Korean Macrolepiota using molecular data. We re-examined 34 Macrolepiota specimens collected from 2012 to 2018 in Korea using a reverse taxonomic approach, whereby species identification was first done based on the internal transcribed spacer (ITS) region analysis, followed by morphological confirmation. We identified the presence of four species: M. detersa, M. mastoidea, M. procera, and M. umbonata sp. nov. Two species (M. detersa and M. mastoidea) were previously unrecorded from Korea and M. umbonata is a new species. Detailed descriptions of all four species and taxonomic key are provided in this study. Macrolepiota procera and M. umbonata are distributed through the country, but M. detersa and M. mastoidea are distributed only in limited areas. According to our results, the combination of ITS locus and morphology proved to be a robust approach to evaluate the taxonomic status of Macrolepiota species in Korea. Additional surveys are needed to verify the species diversity and clarify their geographic distribution.

Diversity of fungi associated with roots of Calanthe orchid species in Korea.

While symbiotic fungi play a key role in the growth of endangered Calanthe orchid species, the relationship between fungal diversity and Calanthe species remains unclear. Here, we surveyed root associated fungal diversity of six Calanthe orchid species by sequencing the internal transcribed spacer (ITS) region using 454 pyrosequencing. Our results revealed that Paraboeremia and Coprinopsis are dominant fungal genera among Calanthe species. In terms of overall relative abundance, Paraboeremia was the most common fungal genus associated with Calanthe roots, followed by Coprinopsis. Overall fungal diversity showed a significant degree of variation depending on both location and Calanthe species. In terms of number of different fungal genera detected within Calanthe species, C. discolor had the most diverse fungal community, with 10 fungal genera detected. This study will contribute toward a better understanding of those fungi that are required for successful cultivation and conservation of Korean Calanthe species.

Three New Recorded Species of the Physalacriaceae on Ulleung Island, Korea.

Most known species in the Physalacriaceae are saprotrophs that grow on decaying leaves and wood, and approximately 21 genera in the Physalacriaceae have been reported worldwide. During an ongoing survey of indigenous fungi in Korea, four specimens belonging to the Physalacriaceae were collected on Ulleung Island. These specimens were identified as three species based on morphological characteristics and molecular analysis of rDNA-internal transcribed spacer sequences. Three species in three genera were shown to be new records in Korea: Hymenopellis orientalis, Paraxerula hongoi, and Ponticulomyces orientalis. The latter two are the first records of these genera in Korea. In this study, we provide detailed morphological descriptions of these species and describe their phylogenetic position within the Physalacriaceae.

Ten New Recorded Species of Macrofungi on Ulleung Island, Korea.

Ulleung Island is a biodiversity hotspot in South Korea. During a survey of indigenous fungal species from Ulleung Island conducted from 2015 to 2016, we discovered 10 unrecorded macrofungi in Korea. These macrofungi were identified to the species level using morphological features and phylogenetic analysis based on the internal transcribed spacer region: Deconica phyllogena, Mycena zephirus, Phaeomarasmius proximans, Phlebia radiata, Pluteus semibulbosus, Postia alni, Resinicium pinicola, Scytinostroma portentosum, Tricholomopsis flammula, and Tyromyces kmetii. We also provide detailed morphological descriptions for these 10 species.