• Journal of Ginseng Research
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• v.44 no.1
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• pp.161-167
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• 2020

Background: The ascomycete fungi Cylindrocarpon destructans (Cd) and Fusarium solani (Fs) cause ginseng root rot and significantly reduce the quality and yield of ginseng. Cd produces the secondary metabolite radicicol, which targets the molecular chaperone Hsp90. Fs is resistant to radicicol, whereas other fungal genera associated with ginseng disease are sensitive to it. Radicicol resistance mechanisms have not yet been elucidated. Methods: Transcriptome analyses of Fs and Cd mycelia treated with or without radicicol were conducted using RNA-seq. All of the differentially expressed genes (DEGs) were functionally annotated using the Fusarium graminearum transcript database. In addition, deletions of two transporter genes identified by RNA-seq were created to confirm their contributions to radicicol resistance. Results: Treatment with radicicol resulted in upregulation of chitin synthase and cell wall integrity genes in Fs and upregulation of nicotinamide adenine dinucleotide dehydrogenase and sugar transporter genes in Cd. Genes encoding an ATP-binding cassette transporter, an aflatoxin efflux pump, ammonium permease 1 (mep1), and nitrilase were differentially expressed in both Fs and Cd. Among these four genes, only the ABC transporter was upregulated in both Fs and Cd. The aflatoxin efflux pump and mep1 were upregulated in Cd, but downregulated in Fs, whereas nitrilase was downregulated in both Fs and Cd. Conclusion: The transcriptome analyses suggested radicicol resistance pathways, and deletions of the transporter genes indicated that they contribute to radicicol resistance.

• Mycobiology
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• v.48 no.5
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• pp.383-391
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• 2020

Use of nanoparticles (NPs) in several commercial products has led to emergence of novel contaminants of air, soil and water bodies. The NPs may exhibit greater ecotoxicity due to nano-scale dependent properties over their bulk counterparts. The present investigation explores the effect of in vitro supplementation of TiO₂, silica and silver NPs on radial growth and ultrastructural changes in the hyphae and spores of two mushroom genera, Ganoderma lucidum and Volvariella volvaceae. A concentration dependent decrease in radial growth on NP amended potato dextrose agar medium was recorded. However, in comparison to control, there was decrease in radial diameter on supplementation with TiO₂ NPs while an increase was recorded for silica and silver NPs amendments as compared to their bulk salts at same concentrations after 48 h of incubation. Optical microscopy studies showed decrease in the number of spores while increase in spore diameter and thinning of hyphal diameter on NPs supplementation. Scanning electron microscopy analysis of fungal growth showed presence of deflated and oblong spores in two fruiting strains of Ganoderma while Volvariella exhibited decreased sporulation. Further, hyphal thinning and branching was recorded in response to NP amendments in both the test mushrooms. Enhancement of protein content was observed on NP compared to bulk supplementation for all cultures, concentrations and hours of incubation except for TiO₂ NPs. Likewise, bulk and NP supplementations (at 100 mg L^-1) resulted in enhanced laccase activity with occurrence of laccase specific protein bands on SDS-PAGE analysis.

• The Plant Pathology Journal
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• v.37 no.1
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• pp.24-35
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• 2021

Blue mold caused by Penicillium expansum is one of the most significant postharvest diseases of apples. Some microorganisms associated with the surface of ripening apples possess the ability to inhibit the growth of P. expansum. However, the existing literature about their colonization in the stages before ripening is not explored in depth. This study aims to characterize the antagonistic capacity of bacterial populations from five fruit development stages of 'Royal Gala' apples. The results have shown that the density of the bacterial populations decreases throughout the ripening stages of fruit (from 1.0 × 105 to 1.1 × 101 cfu/㎠). A total of 25 bacterial morphotypes (corresponding to five genera identified by 16S RNA) were differentiated in which Bacillus stood out as a predominant genus. In the in vitro antagonism tests, 10 Bacillus strains (40%) inhibited the mycelial growth of P. expansum from 30.1% to 60.1%, while in fruit bioassays, the same strains reduced the fruit rot ranging from 12% to 66%. Moreover, the bacterial strains with antagonistic activity increased in the ripening fruit stage. B. subtilis subsp. spiziennii M24 obtained the highest antagonistic activity (66.9% of rot reduction). The matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis revealed that bacteria with antagonistic activity produce anti-fungal lipopeptides from iturin and fengycin families.

• The Korean Journal of Mycology
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• v.49 no.4
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• pp.455-467
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• 2021

Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH₄)₂SO₄ after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

• Mycobiology
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• v.50 no.5
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• pp.389-398
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• 2022

Endophytic fungi are promising sources for the production of podophyllotoxin-an important anticancer compound, replacing depleted medical plants. In this study, the endophytes associated with Dysosma difformis-an ethnomedicinal plant species were isolated to explore novel sources of podophyllotoxin. Fifty-three endophytic fungi were isolated and identified by morphological observation and ITS-based rDNA sequencing, assigning them to 27 genera in 3 divisions. Fusarium was found the most prevalent genus with a colonization frequency of 11.11%, followed by Trametes (9.26%) and Penicillium (7.41%). Phylogenetic trees were constructed for the endophytic fungi community in two collection sites, Ha Giang and Lai Chau, revealing the adaptation of the species to the specific tissues and habitats. Cytotoxic activity of endophytic fungal extracts was investigated on cancer cell lines such as SK-LU-1, HL-60, and HepG2, demonstrating strong anti-cancer activity of six isolates belonging to Penicillium, Trametes, Purpureocillium, Aspergillus, and Ganoderma with IC₅₀ value of lower than 10 ㎍/mL. The presence of podophyllotoxin was indicated in Penicillium, Trametes, Aspergillus and for the first time in Purpureocillium and Ganoderma via high-performance liquid chromatography, which implied them as a potential source of this anticancer compound.

• The Korean Journal of Mycology
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• v.24 no.1 s.76
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• pp.56-66
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• 1996

Ecological variations of Arbuscular mycorrhizal (AM) fungi were determined from the soils collected from different sites of the polluted and unpolluted areas related to the soil pollutions. Average 24.5 spores $(per\;20g)^{-1}$ soil of AM fungal spore were counted from the 32 sites of soils collected from On-san (polluted), whereas average 4.1 spores $(per\;20g)^{-1}$ soil from the 18 sites of those from Mt. Kwanak (polluted); Average 23.6 spores $(per\;20g)^{-1}$ soil of AM fungal spore were also counted from the 30 sites of soils collected from Chung-mu (unpolluted), whereas average 15.8 spores $(per\;20g)^{-1}$ soil from the 14 sites of those from Mt. Chungwang (unpolluted).The spores of AM fungi were the species of Glomus, Gigaspora, Acaulospora and Scutellospora. Among the above four genera, the species of Glomus were observed to be more abundant than the other genera in the soils collected from On-san, Chung-mu and Mt. Chungwang whereas the species of Gigaspora in those from Mt. Kwan-ak. The parameters of soils measured showed some variations between the polluted and unpolluted areas; 12.9 to 16.4% in the soil moisture, 5.6 to 8.3% in the organic matter and 4.3 to 5.7 at soil pH (polluted to unpolluted areas, respectively). The soils collected, thereby, appeared to be more strongly acidic and also lower in the contents of soil moisture or organic matter at the polluted area than unpolluted area. Based on the ecological criteria, the species richness or species diversity had significant differences (p<0.05) between polluted and unpolluted area. The spore density of genus Glomus or Gigaspora was significantly different (p<0.05) among the soils of three different plant vegetations (conifer plants, broad leaf plants, and grass plants). Also, there were significant differences (p<0.05) in the species evenness or species diversity among the soils referring to three different plant vegetations. There was a direct relationship $(r^2=0.38)$ between soil moisture and organic matter measured from 94 soil samples. Since there was a direct relationship $(r^2=0.22)$ between organic matter and total spores, it seems to be likely to presume that mycorrhizal spores can be increased in proportion to enhanced organic matters in soils. The species richness or species diversity was inclined to increase in proportion to enhanced soil pH and total spores in soils.

• The Korean Journal of Mycology
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• v.15 no.1
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• pp.48-69
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• 1987

To understand host ranges and host specificity of ectomycorrhizal fungi and fungus specificity of host tree species, higher fungi from pure forest stands of following twelve tree species were collected for six years from 1981 to 1986: Pinus densiflora, P. rigida, P. koraiensis, Picea koraiensis, Larix leptolepis, Abies holophylla, Populus $alba{\times}glandulosa$, Castanea crenata, Quercus aliena, Q. acutissima, Q. mongolica, and Betula platyphylla. A total of 196 ectomycorrhizal species with 8 varieties in 48 genera were identified. Thirty-one species with one variety belonged to Russula, 22 species with 5 varieties to Amanita, 18 species with one variety to Lactarius. Fungi belonging to above three gepera, Laccaria, and Cantharellus had a relatively wide host range. Laccaria laccata was collected under all the 12 tree species, Amanita vaginata group under 11 tree species, Laccaria amethystina and Russula foetens under 10 tree species, Lactarius gerardii and Russula sororia under 9 tree species, and Amanita agglutinata, Cantharellus cibarius, Russula bella, and R. virescens under 8 tree species. Above 10 fungal species may be classified to have a wide host range. Following fungi were collected under 4 to 5 tree species of both conifers and broad-leaved trees: Amanita citrina, Boletus bicolor, B. erythropus, Lactarius piperatus, L. subzonar­ius, and Russula pseudodelica. Above fungi may be classified to have an intermediate host range. Chroogomphus, Gomphidius, Rhizopogon, and Suillus were collected only under Pinaceae and may be classified to have a narrow host range. Particularly Suillus grevillei was collected only under Larix leptolepis, and appeared to have high host specificity. A total of 83 ectomycorrhizal species were collected under Abies holophylla, 66 species under Pinus densiflora, 50 species under Pinus rigida, 49 species under Pinus koraiensis, 46 species under Quercus aliena, and 23 fungal species under Larix leptolepis. Particularly, Larix leptolepis was associated with different fungal flora, with fewer species of Amanita, Lactarius and Russula, and more species of Laccaria. Most host tree species surveyerl in the present study appeared to have low fungus specificity for ectomycorrhizal formation.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.134-140
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• 2012

Asian dust storms originating in the arid desert of China and Mongolia usually occur from late winter through spring, and more than one million tons of dust per year is transported to the Korean Peninsula by the prevalent westerly winds. We supposed that these dust particles could include bioaerosols and act as carriers of microorganisms. In order to clarify the dynamics of microorganisms moving with these particles, the concentration and composition of microorganisms associated with settled particles were compared between samples collected during Asian dust events and those under non-dust periods. From February to April 2008, settled dust particles were collected at one location in Ulsan using rainfall meter of 200 mm diameter. During this period, there was one Asian dust event in Ulsan. The bacterial concentrations were higher in samples collected during Asian dust event than those under non-dust period, whereas fungal concentrations were rather similar regardless of the Asian dust event. We analyzed 16S rRNA gene sequences of 45 bacterial isolates obtained from the settled particle samples. These isolates belonged to either genus Bacillus or genus Streptococcus and were tentatively identified as B. amyloliquefaciens, B. aryabhattai, B. atrophaeus, B. licheniformis, B. megaterium, B. methylotrophicus, B. pumilus, B. sonorensis, B. subtlis, B. vallismortis, S. epidermidis, and S. succinus. In cases of fungal isolates, genera such as Mucor, Alternaria, Cladosporium, and Aspergillus were tentatively identified from samples collected at both Asian dust and non-Asian dust periods. It appears that endospore-forming bacteria such as Bacillus sp. rather than fungal spores are more likely to be associated with Asian dust particles.

• Journal of Sericultural and Entomological Science
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• v.47 no.1
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• pp.18-30
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• 2005

Morphological characteristics of Cordyceps species and its allies collected in Korea were clarified. Through the survey conducted from June 1999 to October 2002 in 19 mountains in Korea, 667 samples of entomogenous fungi were collected. Cordyceps and its allies of 17 species of 5 genera were identified as Cordyceps gracilioides, C. japonica, C. longissima, C. martialis, C. militaris, C. myrmecophila, C. nutans, C. pruinosa, C. sphecocephala, C. tricentri, Hirsutella nutans, Paecilomyces cicadae, P. farinosus, P. tenuipes, Paecilomyces sp., Shimizuomyces paradoxa, Tilachlidiopsis nigra. The fungi with insect hosts have been collected mainly in the place of shade or mosses near brooks and streams that had high humidity. Overall the frequenct of fungal infection in natural ecosystem was relatively low as few as 10 collections per each species. However, many species were found in terms of the few number of colleciton sites with seasonal limitations. Occurrence of the fungi in Jeju island remote from inland of the Korean peninsula were diverse in their species due to the varied weather of vertical distribution following the altitude. Three most common species were C. nutans, P. tenuipes and C. militaris, mainly found early in August when the relative humidity and temperature were high, of which C. nutans occupied the highest frequency consisting of 65% in total collections. Neither variation in ascomata arrangement in stromata nor developement of secondary spores was recognizable, while the number, shape and colour of stromata varied with insect hosts and weather conditions.

• Microbiology and Biotechnology Letters
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• v.43 no.2
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• pp.142-149
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• 2015

Bioaerosols are comprised of particles 0.02-100 μm in size that originate in natural and artificial environments, and as a result of human activities. They consist of microorganisms including viruses, bacteria, fungi, and protozoa; fungal spores; microbial toxins; pollen; plant or animal material; expectorated liquid from humans; and glucans (peptidoglycan and β-glucan). Bioaerosols can cause respiratory and other diseases in humans and animals. In this study, bioaerosol samples acquired from agricultural sources, livestock, a sewage treatment plant, a beach, and a pristine area were analyzed to identify and phylogenetically characterize culturable microorganisms. The isolated bacteria exhibited regional differences, with different species dominating. However, Bacillus cereus was isolated in all samples, with a total of 31 strains isolated from all areas, and Acinetobacter baumannii was isolated from an indoor poultry farm. In addition, bacteria determined to be of novel genus or species of the genera Domibacillus, Chryceobacterium, Nocardioides and family Comamonadaceae were isolated from the agricultural, livestock and beach environments.