Penicillium ulleungdoense sp. nov. from Ulleung Island in Korea

In a study of the fungal diversity on Ulleung Island in Korea, three novel strains of Penicillium were isolated. Different sites on Ulleung Island were selected for collecting endophytic fungi, and three endophytic fungal strains showed unique morphological characteristics. DNA sequence of the internal transcribed spacer, β-tubulin, calmodulin, and RNA polymerase II second largest subunit regions of the strains were analyzed and they showed unique taxonomic position from the other species of Penicillium section Sclerotiora. The new strains were named Penicillium ulleungdoense sp. nov. As the novel endophytic Penicillium taxa were discovered in a unique environment, the data could be meaningful for understanding the geographical distribution of Ascomycetes on Ulleung Island.

Biodegradative Activities of Fungal Strains Isolated from Terrestrial Environments in Korea

Polylactic acid (PLA) and polycaprolactone (PCL) are commercially available bioplastics that are exploited worldwide, and both are biodegradable. The PLA and PCL polymer-degrading activity of 30 fungal strains that were isolated from terrestrial environments were screened based on the formation of a clear zone around fungal colonies on agar plates containing emulsified PLA or PCL. Among them, five strains yielded positive results of biodegradation.Strains Korean Agricultural Culture Collection (KACC) 83034BP and KNUF-20-PPH03 exhibited PCL degradation; two other strains, KACC 83035BP and KNUF-20-PDG05, degraded PLA; and the fifth strain, KACC 83036BP, biodegraded both tested plastics. Based on phylogenetic analyses using various combinations of the sequences of internal transcribed spacer (ITS) regions, RPB2, LSU, CAL, and b-TUB genes, the above-mentioned strains were identified as Apiotrichum porosum, Penicillium samsonianum, Talaromyces pinophilus, Purpureocillium lilacinum, and Fusicolla acetilerea, respectively. Based on our knowledge, this is the first report on (i) plastic biodegraders among Apiotrichum and Fusicolla species, (ii) the capability of T. pinophilus to degrade biodegradable plastics, (iii) the biodegradative activity of P. samsonianumagainst PCL, and (iv) the accurate identification of P. lilacinum as a PLA biodegrader. Further studies should be conducted to determine how the fungal species can be utilized in Korea.

Geographical Isolation and Root-Associated Fungi in the Marine Terrains:A Step Toward Establishing a Strategy for Acquiring Unique Microbial Resources

This study aimed to understand whether the geo-ecological segregation of native plant species affects the root-associated fungal community. Rhizoplane (RP) and rhizosphere (RS) fungal microbiota of Sedum takesimense native to three geographically segregated coastal regions (volcanic ocean islands) were analyzed using culture-independent methods: 568,507 quality sequences, 1399 operational taxonomic units, five phyla, and 181 genera were obtained. Across all regions, significant differences in the phyla distribution and ratio were confirmed. The Chao’s richness value was greater for RS than for RP, and this variance coincided with the number of genera. In contrast, the dominance of specific genera in the RS (Simpson value) was lower than the RP at all sites. The taxonomic identity of most fungal species (95%) closely interacting with the common host plant was different. Meanwhile, a considerable number of RP only residing fungal genera were thought to have close interdependency on their host halophyte. Among these, Metarhizium was the sole genus common to all sites. These suggest that the relationship between potential symbiotic fungi and their host halophyte species evolved with a regional dependency, in the same halophyte species, and of the same natural habitat (volcanic islands); further, the fungal community differenced in distinct geographical regions. Importantly, geographical segregation should be accounted for in national culture collections, based on taxonomical uniqueness.

Four Unrecorded Aspergillus Species from the Rhizosphere Soil in South Korea

The genus Aspergillus is commonly isolated from various marine and terrestrial environments;however, only a few species have been studied in rhizosphere soil. As part of the Korean indigenous fungal excavation project, we investigated fungal diversity from rhizosphere soil, focusing on Aspergillus species. A total of 13 strains were isolated from the rhizosphere soil of three different plants. Based on phylogenetic analysis of β-tubulin and calmodulin and morphological characteristics, we identified five Aspergillus species. A. calidoustus and A. pseudodeflectus were commonly isolated from the rhizosphere soil. Four species were confirmed as unrecorded species in Korea: A. calidoustus, A. dimorphicus, A. germanicus, and A. pseudodeflecuts. The detailed morphological descriptions of these unrecorded species are provided.

Biodegradative Activities of Fungal Strains Isolated from Terrestrial Environments in Korea

Polylactic acid (PLA) and polycaprolactone (PCL) are commercially available bioplastics that are exploited worldwide, and both are biodegradable. The PLA and PCL polymer-degrading activity of 30 fungal strains that were isolated from terrestrial environments were screened based on the formation of a clear zone around fungal colonies on agar plates containing emulsified PLA or PCL. Among them, five strains yielded positive results of biodegradation.Strains Korean Agricultural Culture Collection (KACC) 83034BP and KNUF-20-PPH03 exhibited PCL degradation; two other strains, KACC 83035BP and KNUF-20-PDG05, degraded PLA; and the fifth strain, KACC 83036BP, biodegraded both tested plastics. Based on phylogenetic analyses using various combinations of the sequences of internal transcribed spacer (ITS) regions, RPB2, LSU, CAL, and b-TUB genes, the above-mentioned strains were identified as Apiotrichum porosum, Penicillium samsonianum, Talaromyces pinophilus, Purpureocillium lilacinum, and Fusicolla acetilerea, respectively. Based on our knowledge, this is the first report on (i) plastic biodegraders among Apiotrichum and Fusicolla species, (ii) the capability of T. pinophilus to degrade biodegradable plastics, (iii) the biodegradative activity of P. samsonianumagainst PCL, and (iv) the accurate identification of P. lilacinum as a PLA biodegrader. Further studies should be conducted to determine how the fungal species can be utilized in Korea.

Geographical Isolation and Root-Associated Fungi in the Marine Terrains:A Step Toward Establishing a Strategy for Acquiring Unique Microbial Resources

This study aimed to understand whether the geo-ecological segregation of native plant species affects the root-associated fungal community. Rhizoplane (RP) and rhizosphere (RS) fungal microbiota of Sedum takesimense native to three geographically segregated coastal regions (volcanic ocean islands) were analyzed using culture-independent methods: 568,507 quality sequences, 1399 operational taxonomic units, five phyla, and 181 genera were obtained. Across all regions, significant differences in the phyla distribution and ratio were confirmed. The Chao’s richness value was greater for RS than for RP, and this variance coincided with the number of genera. In contrast, the dominance of specific genera in the RS (Simpson value) was lower than the RP at all sites. The taxonomic identity of most fungal species (95%) closely interacting with the common host plant was different. Meanwhile, a considerable number of RP only residing fungal genera were thought to have close interdependency on their host halophyte. Among these, Metarhizium was the sole genus common to all sites. These suggest that the relationship between potential symbiotic fungi and their host halophyte species evolved with a regional dependency, in the same halophyte species, and of the same natural habitat (volcanic islands); further, the fungal community differenced in distinct geographical regions. Importantly, geographical segregation should be accounted for in national culture collections, based on taxonomical uniqueness.

Four Unrecorded Aspergillus Species from the Rhizosphere Soil in South Korea

The genus Aspergillus is commonly isolated from various marine and terrestrial environments;however, only a few species have been studied in rhizosphere soil. As part of the Korean indigenous fungal excavation project, we investigated fungal diversity from rhizosphere soil, focusing on Aspergillus species. A total of 13 strains were isolated from the rhizosphere soil of three different plants. Based on phylogenetic analysis of β-tubulin and calmodulin and morphological characteristics, we identified five Aspergillus species. A. calidoustus and A. pseudodeflectus were commonly isolated from the rhizosphere soil. Four species were confirmed as unrecorded species in Korea: A. calidoustus, A. dimorphicus, A. germanicus, and A. pseudodeflecuts. The detailed morphological descriptions of these unrecorded species are provided.

Fungal Clusters and Their Uniqueness in Geographically Segregated Wetlands: A Step Forward to Marsh Conservation for a Wealth of Future Fungal Resources

Here, we investigated fungal microbiota in the understory root layer of representative well-conserved geographically segregated natural wetlands in the Korean Peninsula. We obtained 574,143 quality fungal sequences in total from soil samples in three wetlands, which were classified into 563 operational taxonomic units (OTU), 5 phyla, 84 genera. Soil texture, total nitrogen, organic carbon, pH, and electrical conductivity of soil were variable between geographical sites. We found significant differences in fungal phyla distribution and ratio, as well as genera variation and richness between the wetlands. Diversity was greater in the Jangdo islands wetland than in the other sites (Chao richness/Shannon/Simpson’s for wetland of the Jangdo islands: 283/6.45/0.97 > wetland of the Mt. Gariwang primeval forest: 169/1.17/0.22 > wetland of the Hanbando geology: 145/4.85/0.91), and this variance corresponded to the confirmed number of fungal genera or OTUs (wetlands of Jangdo islands: 42/283 > of Mt. Gariwang primeval forest: 32/169 > of the Hanbando geology: 25/145). To assess the uniqueness of the understory root layer fungus taxa, we analyzed fungal genera distribution. We found that the percentage of fungal genera common to two or three wetland sites was relatively low at 32.3%, while fungal genera unique to each wetland site was 67.7% of the total number of identified fungal species. The Jangdo island wetland had higher fungal diversity than did the other sites and showed the highest level of uniqueness among fungal genera (Is. Jangdo wetland: 34.5% > wetland of Mt. Gariwang primeval forest: 28.6% > wetland of the Hanbando geology: 16.7%).

A New Species of Thelonectria and a New Record of Cephalotrichum hinnuleum from Gunwi and Ulleungdo in Korea

Three fungal strains belonging to the class Sordariomycetes were isolated from soils collected from Gyeongsangbuk-do in Korea. They were identified as Cephalotrichum hinnuleum (UD CT 1-3-3 and KNU-19GWF1) and Thelonectria chlamydospora sp. nov. (UD ST 1-2-1).T. chlamydospora sp. nov. was morphologically identical to T. truncata, but its specific macroconidial dimensions, lower number of septations, and chlamydospore diameter render it distinct from the strains of the genus Thelonectria. The strains UD CT 1-3-3 and KNU-19GWF1 were developed flat, velvety to felty, and golden gray to brown-gray after 14 days of incubation at 25℃ on PDA. These strains were produced polyblastic conidiogenous cells and conidia were pale brown to brown, smooth, thin-walled, subglobose to ellipsoidal, arranged in chains, and the diameters of 6.7–9.0 ×3.7–5.1 μm. The strains were also confirmed by using the multi-locus genes using internal transcribed spacer (ITS) regions, partial large subunit (LSU), translation elongation factor 1α(TEF1-α), b-tubulin (TUB2), and actin (ACT) genes. This is the discovery of T. chlamydospora sp. nov. and Cephalotrichum hinnuleum, a new record from Korea.

Penicillium from Rhizosphere Soil in Terrestrial and Coastal Environments in South Korea

Penicillium, the most common genus plays an important ecological role in various terrestrial and marine environments. However, only a few species have been reported from rhizosphere soil. As part of a project to excavate Korean indigenous fungi, we investigated rhizosphere soil of six plants in the forest (terrestrial habitat) and sand dunes (coastal habitat) and focused on discovering Penicillium species. A total of 64 strains were isolated and identified as 26 Penicillium species in nine sections based on morphological characteristics and the sequence analysis of b-tubulin and calmodulin. Although this is a small-scale study in a limited rhizosphere soil, eight unrecorded species and four potential new species have been identified. In addition, most Penicillium species from rhizosphere soil were unique to each plant. Penicillium halotolerans, P. scabrosum, P. samsonianum, P. jejuense, and P. janczewskii were commonly isolated from rhizosphere soil. Eight Penicillium species, P. aurantioviolaceum, P. bissettii, P. cairnsense, P. halotolerans, P. kananaskense, P. ortum, P. radiatolobatum, and P. verhagenii were recorded for the first time in Korea. Here, we provide the detailed morphological description of these unrecorded species.

Penicillium from Rhizosphere Soil in Terrestrial and Coastal Environments in South Korea

Penicillium, the most common genus plays an important ecological role in various terrestrial and marine environments. However, only a few species have been reported from rhizosphere soil. As part of a project to excavate Korean indigenous fungi, we investigated rhizosphere soil of six plants in the forest (terrestrial habitat) and sand dunes (coastal habitat) and focused on discovering Penicillium species. A total of 64 strains were isolated and identified as 26 Penicillium species in nine sections based on morphological characteristics and the sequence analysis of b-tubulin and calmodulin. Although this is a small-scale study in a limited rhizosphere soil, eight unrecorded species and four potential new species have been identified. In addition, most Penicillium species from rhizosphere soil were unique to each plant. Penicillium halotolerans, P. scabrosum, P. samsonianum, P. jejuense, and P. janczewskii were commonly isolated from rhizosphere soil. Eight Penicillium species, P. aurantioviolaceum, P. bissettii, P. cairnsense, P. halotolerans, P. kananaskense, P. ortum, P. radiatolobatum, and P. verhagenii were recorded for the first time in Korea. Here, we provide the detailed morphological description of these unrecorded species.

Foodborne Infectious Diseases Mediated by Inappropriate Infection Control in Food Service Businesses and Relevant Countermeasures in Korea

OBJECTIVES: The objective of this review is to propose an appropriate course of action for improving the guidelines followed by food handlers for control of infection. For this purpose, previous epidemiological reports related to acute gastroenteritis in food service businesses mediated by food handlers were intensively analyzed. METHODS: Relevant studies were identified in international databases. We selected eligible papers reporting foodborne infectious disease outbreaks. Among primary literature collection, the abstract of each article was investigated to find cases that absolutely identified a causative factor to be food handlers’ inappropriate infection control and the taxon of causative microbial agents by epidemiological methodologies. Information about the sites (type of food business) where the outbreaks occurred was investigated. RESULTS: A wide variety of causative microbial agents has been investigated, using several epidemiological methods. These agents have shown diverse propagation pathways based on their own molecular pathogenesis, physiology, taxonomy, and etiology. CONCLUSION: Depending on etiology, transmission, propagation, and microbiological traits, we can predict the transmission characteristics of pathogens in food preparation areas. The infected food workers have a somewhat different ecological place in infection epidemiology as compared to the general population. However, the current Korean Food Safety Act cannot propose detailed guidelines. Therefore, different methodologies have to be made available to prevent further infections.

Distribution, Characterization, and Diversity of the Endophytic Fungal Communities on Korean Seacoasts Showing Contrasting Geographic Conditions

This study analyzed the distribution of endophytic fungi in 3 coastal environments with different climatic, geographical, and geological characteristics: the volcanic islands of Dokdo, the East Sea, and the West Sea of Korea. The isolated fungal endophytes were characterized and analyzed with respect to the characteristics of their host environments. For this purpose, we selected common native coastal halophyte communities from three regions. Molecular identification of the fungal endophytes showed clear differences among the sampling sites and halophyte host species. Isolates were also characterized by growth at specific salinities or pH gradients, with reference to previous geographical, geological, and climate studies. Unlike the East Sea or West Sea isolates, some Dokdo Islands isolates showed endurable traits with growth in high salinity, and many showed growth under extremely alkaline conditions. A smaller proportion of West Sea coast isolates tolerate compared to the East Sea or Dokdo Islands isolates. These results suggest that these unique fungal biota developed through a close interaction between the host halophyte and their environment, even within the same halophyte species. Therefore, this study proposes the application of specific fungal resources for restoring sand dunes and salt-damaged agricultural lands and industrialization of halophytic plants.

Cutaneous Microflora from Geographically Isolated Groups of Bradysia agrestis, an Insect Vector of Diverse Plant Pathogens

Larvae of Bradysia agrestis, an insect vector that transports plant pathogens, were sampled from geographically isolated regions in Korea to identify their cutaneous fungal and bacterial flora. Sampled areas were chosen within the distribution range of B. agrestis; each site was more than 91 km apart to ensure geographical segregation. We isolated 76 microbial (fungi and bacteria) strains (site 1, 29; site 2, 29; site 3, 18 strains) that were identified on the basis of morphological differences. Species identification was molecularly confirmed by determination of universal fungal internal transcribed spacer and bacterial 16S rRNA gene sequences in comparison to sequences in the EzTaxon database and the NCBI GenBank database, and their phylogenetic relationships were determined. The fungal isolates belonged to 2 phyla, 5 classes, and 7 genera; bacterial species belonged to 23 genera and 32 species. Microbial diversity differed significantly among the geographical groups with respect to Margalef's richness (3.9, 3.6, and 4.5), Menhinick's index (2.65, 2.46, and 3.30), Simpson's index (0.06, 0.12, and 0.01), and Shannon's index (2.50, 2.17, and 2.58). Although the microbial genera distribution or diversity values clearly varied among geographical groups, common genera were identified in all groups, including the fungal genus Cladosporium, and the bacterial genera Bacillus and Rhodococcus. According to classic principles of co-evolutionary relationship, these genera might have a closer association with their host insect vector B. agrestis than other genera identified. Some cutaneous bacterial genera (e.g., Pseudomonas) displaying weak interdependency with insect vectors may be hazardous to agricultural environments via mechanical transmission via B. agrestis. This study provides comprehensive information regarding the cutaneous microflora of B. agrestis, which can help in the control of such pests for crop management.

First Report of Leaf Spot Caused by Alternaria tenuissima on Black Chokeberry (Aronia melanocarpa) in Korea

In July 2015, diseased leaves of black chokeberry (Aronia melanocarpa) were observed in Danyang and Gochang, Korea. The symptoms appeared as circular or irregular brown leaf spots, from which Alternaria tenuissima was isolated. The isolates were cultured on potato dextrose agar, and their morphological characteristics were observed under a light microscope. The colonies were whitish to ash colored. The pathogenicity test on healthy black chokeberry leaves produced circular brown spots, in line with the original symptoms. Molecular analyses of the ITS, GPD, RPB2, and TEF genes were conducted to confirm the identity of the pathogen. The phylogeny of the multi-gene sequences indicated that the causal agent was A. tenuissima. This study is the first report of A. tenuissima leaf spot on black chokeberry (A. melanocarpa).

Aspergillus clavatus Y2H0002 as a New Endophytic Fungal Strain Producing Gibberellins Isolated from Nymphoides pe ltata in Fresh Water

Eighteen endophytic fungi with different colony morphologies were isolated from the roots of Nymphoides peltata growing in the Dalsung wetland. The fungal culture filtrates of the endophytic fungi were treated to Waito-c rice seedling to evaluate their plant growth-promoting activities. Culture filtrate of Y2H0002 fungal strain promoted the growth of the Waito-c rice seedlings. This strain was identified on the basis of sequences of the partial internal transcribed spacer region and the partial beta-tubulin gene. Upon chromatographic analysis of the culture filtrate of Y2H0002 strain, the gibberellins (GAs: GA1, GA3, and GA4) were detected and quantified. Molecular and morphological studies identified the Y2H0002 strain as belonging to Aspergillus clavatus. These results indicated that A. clavatus improves the growth of plants and produces various GAs, and may participate in the growth of plants under diverse environmental conditions.

Diversity of Endophytic Fungi Associated with the Roots of Four Aquatic Plants Inhabiting Two Wetlands in Korea

A total of 4 aquatic plants, Eleocharis kuroguwai Ohwi, Hydrocharis dubia Backer, Salvinia natans All., and Zizania latifolia Turcz., were sampled from representative two wetlands of South Korea. A total of 38 endophytic fungal strains were isolated from aquatic plants native to the Daepyeong wetland, and 27 strains were isolated from the Jilnal wetland. The internal transcribed spacer regions of fungal isolates were sequenced and a phylogenetic analysis was performed. In addition, endophytic fungal diversity from each wetland and host plant species was deduced. A total of 25 fungal genera were purely isolated, and 16 fungal genera were isolated from each of the two wetlands. Commonly isolated genera from both wetlands were Aspergillus, Cladosporium, Clonostachys, Fusarium, Leptosphaeria, Penicillium, and Talaromyces. This study revealed that fungal diversity varied with environmental conditions and by host plant in representative two wetlands.

Diversity and Plant Growth Promoting Capacity of Endophytic Fungi Associated with Halophytic Plants from the West Coast of Korea

Five halophytic plant species, Suaeda maritima, Limonium tetragonum, Suaeda australis, Phragmites australis, and Suaeda glauca Bunge, which are native to the Muan salt marsh of South Korea, were examined for fungal endophytes by sequencing the internal transcribed spacer (ITS) region containing ITS1, 5.8S rRNA, and ITS2. In total, 160 endophytic fungal strains were isolated and identified from the roots of the 5 plant species. Taxonomically, all 160 strains belonged to the phyla Ascomycota, Basidiomycota, and Zygomycota. The most dominant genus was Fusarium, followed by the genera Penicillium and Alternaria. Subsequently, using 5 statistical methods, the diversity indices of the endophytes were determined at genus level. Among these halophytic plants, P. australis was found to host the greatest diversity of endophytic fungi. Culture filtrates of endophytic fungi were treated to Waito-C rice seedlings for plant growth-promoting effects. The fungal strain Su-3-4-3 isolated from S. glauca Bunge provide the maximum plant length (20.1 cm) in comparison with wild-type Gibberella fujikuroi (19.6 cm). Consequently, chromatographic analysis of the culture filtrate of Su-3-4-3 showed the presence of physiologically active gibberellins, GA(1) (0.465 ng/mL), GA(3) (1.808 ng/mL) along with other physiologically inactive GA(9) (0.054 ng/mL) and GA(24) (0.044 ng/mL). The fungal isolate Su-3-4-3 was identified as Talaromyces pinophilus.

Culturable Fungal Endophytes Isolated from the Roots of Coastal Plants Inhabiting Korean East Coast

Twelve plant species were collected from the east coast of Korea to identify culturable endophytes present in their roots. The fungal internal transcribe spacer (ITS) region (ITS1-5.8SrRNA-ITS2) was used as a DNA barcode for identification of fungi. A total of 194 fungal strains were identified and categorized into 31 genera. The genus Penicillium accounted for the largest number of strains, followed by the genus Aspergillus. Furthermore, using 5 statistical methods, the diversity indices of the fungi were calculated at the genus level. After comprehensive evaluation, the endophytic fungal group from Phragmites australis ranked highest in diversity analyses. Several strains responsible for plant growth and survival (Penicillium citrinum, P. funiculosum, P. janthinellum, P. restrictum, and P. simplicissimum), were also identified. This study provides basic data on the sheds light on the symbiotic relationship between coastal plants and fungi.

X-linked Recessive Bulbospinal Muscular Atrophy (Kennedy's Syndrome): A case report

X-linked recessive bulbospinal muscular atrophy (Kennedy's syndrome) is a variant of the spinal muscular atrophies caused by mutation of androgen receptor gene on X-chromosome. A 69-year-old man had suffered from slowly progressive lower extremity weakness and gynecomastia. Muscle weakness was more severe in proximal muscles and showed symmetrical features. He had fascicular contraction on his face and tongue. All tendon reflexes were absent and pyramidal signs were not detected. Nerve conduction studies were normal except low amplitude of sensory nerve action potential in median nerve. Needle electromyography revealed widespread chronic denervation potentials in all sampling muscles of extremities, facial and tongue muscles. Histopathologic findings showed chronic denervation atrophy. DNA analysis showed abnormal expansion of CAG repeats in the androgen receptor gene and we confirmed this case as Kennedy's syndrome. If an adult patient has slowly progressive muscle weakness, bulbar symptoms and signs of male genital failure, DNA analysis should be taken to differentiate Kennedy syndrome from other motor neuron disease or myopathy.