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.     

The first report of two species of <Emphasis Type="Italic">Polyporus</Emphasis> (Polyporaceae,Basidiomycota) from South Korea

Based on morphological examination, two species of Polyporus, P. dictyopus, and P. tuberaster, were identified, which constitutes the first record of these species in South Korea. To confirm their affinity within the genus Polyporus, the phylogenetic relationships of Polyporus and allied genera were established from nuclear large subunit ribosomal DNA (nLSU rDNA) sequences, and a morphological diagnostic key is presented to clarify the Korean species of Polyporus.     

Report on a New Truffle Species,Tuber koreanum sp. nov., from Korea

The truffle and ectomycorrhizal roots formed by Tuber sp. were collected from the rhizosphere of Quercus aliena in Korea. The morphological characteristics of the ascoma, and molecular phylogenetic analysis using sequences from the internal transcribed spacer (ITS) and large subunit (LSU) of ribosomal DNA, translation elongation factor 1-alpha (TEF), and RNA polymerase second largest subunit (RPB2) regions confirmed the distinct morphology of the truffle. This truffle belongs to a monophyletic clade among the other Tuber species in the phylogeny. This study describes the truffle, Tuber koreanum, as a new species reported from Korea.     

COI DNA barcoding for molecular identification of the genus Aphidius (Braconidae: Aphidiinae) in South Korea,with diagnoses of two newly recorded species

The genus Aphidius is one of the most commercially important species in greenhouse for controlling pests, especially aphids. Although 13 Aphidius species had been recorded in South Korea, it was not carefully surveyed yet. In this study, we surveyed and collected 8 Aphidius spp. including two unrecorded species. Using the DNA barcode region of mitochondrial cytochrome c oxidase subunit I (COI), sequences were newly generated for 16 molecular operational taxonomic units (MOTUs) of 8 species, except for 15 duplicates, in addition of 58 MOTUs of 18 species referred from GenBank for comparison and identification, which were reconstructed in a barcode phylogeny with Neighbor-Joining method. Then, to define each species, both Automatic Barcode Gap Discovery (ABGD) and Bayesian Poisson Tree Processes (bPTP) analyses were performed. As results, although most of Aphidius spp. could be identified by clustering into a clade, we recognized that three Aphdius species were placed in a clade, of which each species was not separated as an independent species. Our data suggested that those species demonstrating a species complex were able to be hypothesized as one species. In addition, diagnoses and illustrations of the two newly recorded species in South Korea were provided.     

A New record of four <Emphasis Type="Italic">Penicillium</Emphasis> species isolated from <Emphasis Type="Italic">Agarum clathratum</Emphasis> in Korea

Agarum clathratum, brown algae, play important ecological roles in marine ecosystem, but can cause secondary environment pollution when they pile up on the beach. In order to resolve the environment problem by A. clathratum, we focus to isolate and identify Penicillium because many species are well known to produce extracellular enzymes. A total of 32 Penicillium strains were isolated from A. clathratum samples that collected from 13 sites along the mid-east coast of Korea in summer. They were identified based on morphological characters and phylogenetic analysis using β-tubulin DNA sequences as well as a combined dataset of β-tubulin and calmodulin. A total of 32 strains were isolated and they were identified to 13 Penicillium species. The commonly isolated species were Penicillium citrinum, P. roseomaculatum, and Penicillium sp. Among 13 Penicillium species, four species – P. bilaiae, P. cremeogriseum, P. madriti, and P. roseomaculatum – have not been previously recorded in Korea. For these four new species records to Korea, we provide morphological characteristics of each strain.     

Molecular identification and phylogenetic analysis of economically important acaroid mites (Acari: Astigmata: Acaroidea) in Korea

Molecular diagnosis is highly valuable for the species identification of microscopic mites. Here, we collected some economically important mites of the superfamily Acaroidea from various stored products in Korea. Those nucleotide sequences of ribosomal second internal transcribed spacer (ITS2) and the mitochondrial cytochrome oxidase subunit I (COI) regions were determined by PCR using universial primers. In nucleotide sequence comparison at GenBank database, seven species including Rhizoglyphus robini, R. echinopus, Sancassania sp. Acarus siro, Tyrophagus putrescentiae, T. similis in Acaridae and Suidasia medanensis in Suidasiidae were identified. Particularly, COI sequences of R. robini, R. echinopus, Sancassania sp. and T. similis were firstly determined. Our results suggest that the phylogenetic relationships inferred from the ITS2 region, rather than COI region, were similar to those derived based on their morphological classification. Our study provides molecular information for the identification and phylogenetic relationship of acaroid mites in Korea.     

Re-Identification on Korean Penicillium Sequences in GenBank Collected by Software GenMine

Penicillium species have been actively studied in various fields, and many new and unrecorded species continue to be reported in Korea. Moreover, unidentified and misidentified Korean Penicillium species still exist in GenBank. Therefore, it is necessary to revise the Korean Penicillium inventory based on accurate identification. We collected Korean Penicillium nucleotide sequence records from GenBank using the newly developed software, GenMine, and re-identified Korean Penicillium based on the maximum likelihood trees. A total of 1681 Korean Penicillium GenBank nucleotide sequence records were collected from GenBank. In these records, 1208 strains with four major genes (Internal Transcribed Spacer rDNA region, β-tubulin, Calmodulin and RNA polymerase II) were selected for Penicillium re-identification. Among 1208 strains, 927 were identified, 82 were identified as other genera, the rest remained undetermined due to low phylogenetic resolution. Identified strains consisted of 206 Penicillium species, including 156 recorded species and 50 new species candidates. However, 37 species recorded in the national list of species in Korea were not found in GenBank. Further studies on the presence or absence of these species are required through literature investigation, additional sampling, and sequencing. Our study can be the basis for updating the Korean Penicillium inventory.     

Revisiting Rhytisma lonicericola: Morphological Characterization and Molecular Phylogenetic Analysis

Rhytisma lonicericola was identified as a tar spot fungus on Lonicera sp. in 1902, and has since been recorded on several species of Lonicera in China, Japan, and Korea. Most of the previous records of R. lonicericola have been based on a list of disease occurrences in the absence of any formal morphological identification or molecular analyses. Using six newly obtained specimens collected in the past 2 years, we confirmed the tar spot fungus found on L. japonica in Korea as R. lonicericola based on morphological examinations and molecular phylogenetic analyses. This fungus was distinguished from R. xylostei, another tar spot fungus on Lonicera, by ascospore size and geographical distributions. We present detailed mycological information and, for the first time, DNA sequence data useful for the identification of R. lonicericola.     

A novel Trichoderma species isolated from soil in Guizhou, T. guizhouense

A new species of Trichoderma, Trichoderma guizhouense, isolated from soil in Guizhou Province, is described based on morphology and phylogenetic analyses. This species exhibits characteristic Trichoderma morphology but is distinct from related species based on characters of phialides and conidia. Two DNA markers, the translation elongation factor 1 alpha (tef1) and RNA polymerase II subunit b (rpb2) were used for phylogenetic analyses. The correlation between morphological and molecular-based clustering demonstrated two studied isolates are a new species.     

First Report of Six Macrofungi from Daecheongdo and Socheongdo Islands,Korea

Daecheongdo and Socheongdo Islands are located in the West Sea of Korea, 210 km away from land, and are military border areas very close to North Korea, making them difficult to access. Although the ecosystem of the islands is relatively well preserved due to the lack of accessibility, research on fungi of the regions is insufficient. Therefore, we aimed to investigate indigenous fungi in these geographically and geopolitically constrained regions. A survey of the indigenous fungal diversity of the islands was conducted in 2018. All specimens were identified at the species level based on morphological and molecular analyses. Among them, six macrofungi—namely, Agaricus menieri, Crepidotus praecipuus, Dichomitus squalens, Hortiboletus amygdalinus, Melanoleuca friesii, and Trametes lactinea—were not previously reported in Korea. Considering that the proportion of unrecorded species is high in the survey area and period as well as the number of samples collected, similar research on adjacent islands may be necessary.     

Trichoderma mienum sp. nov., isolated from mushroom farms in Japan

During an investigation of Hypocrea/Trichoderma species inhabiting mushroom bedlogs, we found five strains of an undescribed species from a culture collection. These were analyzed using a combined approach, including morphology of holomorph, cultural studies, and phylogenetic analyses of the rRNA gene cluster of the internal transcribed spacer region, translation elongation factor 1-α, and RNA polymerase subunit II gene sequences. Distinctive morphological characters include stromata with green ascospores produced on potato dextrose agar medium, and Gliocladium-like to irregularly Verticillium-like conidiophores. In phylogenetic analyses, this species belongs to the Semiorbis clade, but its morphological characteristics do not match the other members of this clade. Based on morphological observations and phylogenetic analyses, we describe this as a new species, Trichoderma mienum, representing its Hypocrea teleomorph and Trichoderma anamorph.     

Morphometrics of the final instar exuviae of five cicada species occurring in urban areas of central Korea

Cicadas produce loud calling songs for mate attraction that can be a nuisance to city dwellers in Korea. Exuviae of final instars can be used to estimate population density. We investigated the morphological characteristics of final instar exuviae for five cicada species that are abundant in central Korea: Cryptotympana atrata, Hyalessa fuscata, Graptopsaltria nigrofuscata, Meimuna opalifera, and Meimuna mongolica. The characters analyzed were pro-mesonotum length, pronotum width, pronotum length, head width, abdominal circumference, body length, wing length, and femoral tooth angle. The results of a general linear model showed that species and sex were significant for morphological characters, and the result of post hoc analyses revealed that the mean values of most of the morphological characters measured were significantly different among the five cicada species. Three groups, Cryptotympana atrata, H. fuscata/G. nigrofuscata, and M. opalifera/M. mongolica, were distinctively identified based on most morphological characters. The distributions of femoral tooth angles did not overlap between H. fuscata and G. nigrofuscata, and the distributions of abdominal circumference did not overlap between M. opalifera and M. mongolica. Thus, the exuviae of all five cicada species could be easily distinguished based on morphological characters. We also provide a morphological key for the exuviae of all five cicada species in Korea.     

Molecular and morphological characterization of a poorly known marine ciliate, Myoschiston duplicatum precht 1935: implications for phylogenetic relationships between three morphologically similar genera -- Zoothamnium, Myoschiston, and Zoothamnopsis (Ciliophora, Peritrichia, Zoothamniidae)

We studied the morphology and molecular phylogeny of Myoschiston duplicatum, a peritrich ciliate that has been recorded as an epibiont of crustaceans, but which we also identified on marine algae from Korea. The important morphological characteristics revealed by silver staining of Myoschiston species have not been described because they are rarely collected. Using morphological methods, we redescribed the type species of the genus, Myoschiston duplicatum, and provided an improved diagnosis of Myoschiston. In addition, the coding regions for nuclear small subunit (SSU) rRNA and internal transcribed spacer 1‐5.8S‐internal transcribed spacer 2 sequences were sequenced. Phylogenetic analyses that included available SSU rDNA sequences of peritrichs from GenBank strongly supported a position of M. duplicatum within the family Zoothamniidae. In addition, phylogenetic analyses were performed with single datasets (ITS1‐5.8S‐ITS2) and combined datasets (SSU rDNA + ITS1‐5.8S‐ITS2) to explore further the phylogenetic relationship in the family Zoothamniidae between the three morphologically similar genera—Zoothamnium, Myoschiston, and Zoothamnopsis.     

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island,Korea

There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.