• Mycobiology
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• v.49 no.4
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• pp.385-395
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• 2021

Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.

• 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.

• Mycobiology
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• v.51 no.6
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• pp.401-409
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• 2023

Nigrospora (Xylariales, Apiosporaceae) consists of species of terrestrial plant endophytes and pathogens. Nigrospora has also been reported in marine environments such as mangroves, sea fans, and macroalgae. However, limited research has been conducted on Nigrospora associated with macroalgae. Here, we isolated Nigrospora species from three types of algae (brown, green, and red algae) from Korean islands (Chuja, Jeju, and Ulleung) based on phylogenetic analyses of multigenetic markers: the internal transcribed spacers (ITS), beta-tubulin (BenA), and translation elongation factor 1 (TEF1-α). A total of 17 Nigrospora strains were isolated from macroalgae and identified as nine distinct species. The majority of Nigrospora species (seven) were found on brown algae, followed by red algae (three), and then green algae (two). To our understanding, this study represents the first account of N. cooperae, N. covidalis, N. guilinensis, N. lacticolonia, N. osmanthi, N. pyriformis, and N. rubi occurring in marine environments. Additionally, this study provides the first report of the occurrence of N. cooperae, N. covidalis, N. guilinensis, N. lacticolonia, and N. osmanthi in South Korea. This study will provide valuable insights for future research exploring the functions of fungi in macroalgal communities.

• Mycobiology
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• v.51 no.4
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• pp.230-238
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• 2023

Glabridin is a well-known active isoflavone found in the root of licorice (Glycyrrhiza glabra L.) that possess a wide range of biological activity. Plant cells, hairy roots, and fungal endophytes cultures are the most important alternative methods for plant resources conservation and sustainable production of natural compounds, which has received much attention in recent decades. In the present study, an efficient culture condition was optimized for the biomass accumulation and glabridin production from fungal endophyte Aspergillus eucalypticola SBU-11AE isolated from licorice root. Type of culture medium, range of pH, and licorice root extract (as an elicitor) were tested. The results showed that the highest and lowest biomass production was observed on PCB medium (6.43 ± 0.32 g/l) and peptone malt (5.85 + 0.11 g/l), respectively. The medium culture PCB was produced the highest level of glabridin (7.26 ± 0.44 mg/l), while the lowest level (4.47 ± 0.02 mg/l) was obtained from the medium peptone malt. The highest biomass (8.51 ± 0.43 g/l) and glabridin (8.30 ± 0.51 mg/l) production were observed from the PCB medium adjusted with pH = 6, while the lowest value of both traits was obtained from the same medium with pH = 7. The highest production of total glabridin (10.85 ± 0.84 mg/l) was also obtained from the culture medium treated with 100 mg/l of the plant root extract. This information can be interestingly used for the commercialization of glabridin production for further industrial applications.

• Journal of Microbiology and Biotechnology
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• v.34 no.7
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• pp.1452-1463
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• 2024

Fungi generate different metabolites some of which are intrinsically bioactive and could therefore serve as templates for drug development. In the current study, six endophytic fungi namely Aspergillus flavus, Aspergillus tubigenesis, Aspergillus oryzae, Penicillium oxalicum, Aspergillus niger, and Aspergillus brasiliensis were isolated and identified from the medicinal plant, Silybum marianum. These endophytic fungi were identified through intra transcribed sequence (ITS) gene sequencing. The bioactive potentials of fungal extracts were investigated using several bioassays such as antibacterial activity by well-diffusion, MIC, MBC, anti-biofilm, antioxidant, and haemolysis. The Pseudomonas aeruginosa PAO1 was used to determine the antibiofilm activity. The ethyl acetate extract of Aspergillus flavus showed strong to moderate efficacy against Staphylococcus aureus, Escherichia coli, P. aeruginosa, and Bacillus spizizenii. Aspergillus flavus and Aspergillus brasiliensis exhibited significant antibiofilm activity with IC₅₀ at 4.02 and 3.63 mg/ml, while A. flavus exhibited maximum antioxidant activity of 50.8%. Based on HPLC, LC-MS, and NMR experiments kojic acid (1) and carbamic acid (methylene-4, 1-phenylene) bis-dimethyl ester (2) were identified from A. flavus. Kojic acid exhibited DPPH free radical scavenging activity with an IC₅₀ value of 99.3 ㎍/ml and moderate activity against ovarian teratocarcinoma (CH1), colon carcinoma (SW480), and non-small cell lung cancer (A549) cell lines. These findings suggest that endophytic fungi are able to produce promising bioactive compounds which deserve further investigation.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1100-1107
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• 2012

Bacterial endophytes may be important factors in plant growth and ecologically relevant functions in rice. Using pyrosequencing technology, we analyzed the composition of endophytic bacterial communities that colonized the roots of rice cultivated in long-term fertilized (APK) and non-fertilized (NF) paddy soils. A total of 1,900 reads were obtained from 2 samples. All sequences were classified into 177 OTUs (APK sample) or 72 OTUs (NF sample) at a 97% similarity cut-off. Twenty-two OTUs were shared between the 2 samples, and these were also the most dominant OTUs in both samples. Proteobacteria was the most dominant phylum with 90.2%, followed by Actinobacteria (7.1%) and Bacteroidetes (1.1%). Furthermore, Pseudomonas was the most abundant genus in both samples. We observed clear differences in the structure of the endophytic bacterial community structure between the 2 samples. Notably, the distributions of Alphaproteobacteria and Gammaproteobacteria were markedly different. The diversity index of the APK sample was higher than that of the NF sample. These findings showed that the endophytic bacterial community of rice roots was affected by the presence of fertilizers in the rice field soil.

• Korean Journal of Medicinal Crop Science
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• v.13 no.1
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• pp.1-5
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• 2005

While the rusty-colored root is common in ginsengs culture and, often results in a severe economic loss, the major factors have not been found. This study was focused on the determination of a potential relationship between rusty root and endophytic bacteria. The number of endophytes was $9.6\;{\times}\;10^1{\sim}1.5\;{\times}\;10^2\;cfu/g$ fw in normal ginseng roots compared to $3.7\;{\times}\;10^6{\sim}5.1\;{\times}\;10^7\;cfu/g$ fw in rusty ones. Of 31 isolates from rusty ginseng roots, twenty-four isolates repeatedly induced severe to moderate rust on root while seven isolates induced slight rust. The bacteria responsible for rusty ginseng roots were mainly Gram negative aerobic. Rust inducing bacteria were identified as Agrobacterium tumefaciens, A. rhizogenes, Burkholderia phenazinium, Ensifer adharens, Lysobacter gummosus, Microbacterium luteolum, M. oxydans, Pseudomonas marginalis, P. veronii, Pseudomonas sp., Rhizobium leguminosarum, R. tropica, Rhodococcus erythropolis, Rh. globerulus, Variovorax paradoxus on the basis of bacteriological characters and 16S rDNA sequences analysis. The results in this study strongly suggested that the rusty ginseng roots were produced by infection and growth of endophytic bacteria.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.152-164
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• 2015

Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.893-902
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• 2011

Endophytic fungi are little known for their role in gibberellins (GAs) synthesis and abiotic stress resistance in crop plants. We isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (CF) on the GAs biosynthesis mutant rice line - Waito-C. CF bioassay showed that endophyte GMH-1B significantly promoted the growth of Waito-C compared with controls. GMH-1B was identified as Penicillium minioluteum LHL09 on the basis of ITS regions rDNA sequence homology and phylogenetic analyses. GC/MS-SIM analysis of CF of P. minioluteum revealed the presence of bioactive $GA_4$ and $GA_7$. In endophyte-soybean plant interaction, P. minioluteum association significantly promoted growth characteristics (shoot length, shoot fresh and dry biomasses, chlorophyll content, and leaf area) and nitrogen assimilation, with and without sodium chloride (NaCl)-induced salinity (70 and 140 mM) stress, as compared with control. Field-emission scanning electron microcopy showed active colonization of endophyte with host plants before and after stress treatments. In response to salinity stress, low endogenous abscisic acid and high salicylic acid accumulation in endophyte-associated plants elucidated the stress mitigation by P. minioluteum. The endophytic fungal symbiosis of P. minioluteum also increased the daidzein and genistein contents in the soybean as compared with control plants, under salt stress. Thus, P. minioluteum ameliorated the adverse effects of abiotic salinity stress and rescued soybean plant growth by influencing biosynthesis of the plant's hormones and flavonoids.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.13-13
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• 2016

Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.