• The Plant Pathology Journal
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• v.40 no.2
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• pp.218-224
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• 2024

Plants are treasure trove of novel compounds that have potential for antifungal chemicals and drugs. In our previous study, we had screened plant extracts obtained from more than eight hundred plant materials collected in Korea, and found that butanol fraction of the Actinostemma lobatum were most potent in suppressing growth of diverse fungal pathogens of plants. Here in this study, we describe further analysis of the butanol fraction, and summarize the results of subsequent antifungal activity test for the sub-fractions against a selected set of plant pathogenic fungi. This line of analyses allowed us to identify the sub-fractions that could account for a significant proportion of observed antifungal activity of initial butanol fraction from A. lobatum. Further analysis of these sub-fractions and determination of structure would provide the shortlist for novel compounds that can be a lead to new agrochemicals.

• The Korean Journal of Pesticide Science
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• v.18 no.4
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• pp.383-395
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• 2014

When we investigated seed infestation by fungal pathogens from 51 varieties in 9 crops, the contamination rate of rice and sesame seeds was high. Therefore, to control seed-borne diseases, we obtained extracts from commercial products of Kimchi, Gochujang, Doenjang, Ganjang, Makgeolli and Tohajut and their suppressive effects against seed-borne diseases were studied. In addition, bacterial strains were screened to control rice seed-borne diseases in vitro and in vivo. Among forty food extracts, eleven food-extracts suppressed incidence of seedling rots in vitro and five food extracts increased 8-33% of healthy seedling in the greenhouse. Among 218 isolates from 40 fermented foods, 43 isolates showed high antifungal activity against seven fungal pathogens. When we tested 43 isolates for the reduction of rice seed borne disease, 32 isolates were able to reduce the rice seed borne disease. Among 32 isolates, 17 isolates reduced significantly seedling rot and increased healthy seedlings, the other isolates except for Kc4-2 and Mkl 2-2 increased shoot emergence and the percentage of healthy plants. Thirty isolates with high antifungal activity and suppressive effect against rice seedling rots were identified by 16S rRNA sequencing. Twenty one of thirty isolates were identified as Bacillus spp. Three isolates from Makgeolli were identified as Saccharomyces cerevisiae. B. amyloliquifaciens were isolated from six Korean traditional fermented foods except for Ganjang. B. amyloliquifaciens were majority in the effective bacterial population of Gochujang and Jutgal. Relatively diverse Bacillus species including B. subtilis, B. pumilus, and B. amyloliquificiens were isolated from Kimchi. The selected effective microorganisms from Korean fermented foods founded to be effective for controlling seed-borne diseases of rice in vitro and in the greenhouse. We think that Korean fermented foods and their useful microorganisms can be used as biocontrol agents for suppressing rice seed-borne diseases based on above described results.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• Journal of Applied Biological Chemistry
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• v.60 no.2
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• pp.149-153
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• 2017

Contamination and growth of Trichoderma, a green mold, on the oak log and wooden chip or sawdust media can severely inhibit the growth of oak mushroom. Chemicals including pesticides and antibiotics are generally not allowed for the control of green mold disease during mushroom cultivation. In this study, bacterial pathogens causing blotch disease on the oyster mushrooms were isolated and their peptide toxins were purified for the control of green mold disease. Strains of Pseudomonas tolaasii secret various peptide toxins, tolaasin and its structural analogues, having antifungal activities. These peptides have shown no effects on the growth of oak mushrooms. When the peptide toxins were applied to the green mold, Trichoderma harzianum H1, they inhibited the growth of green molds. Among the 20 strains of peptide-forming P. tolaasii, strong, moderate, and weak antifungal activities were measured from 8, 5, and 7 strains, respectively. During oak mushroom cultivation, bacterial culture supernatants containing the peptide toxins were sprayed on the aerial mycelia of green molds grown on the surface of sawdust media. The culture supernatants were able to suppress the fungal growth of green molds while no effect was observed on the mushroom growth and production. They changed the color of molds from white aerial mycelium into yellowish dried scab, representing the powerful anti-fungal and sterilization activities of peptide toxins.

• Korean Journal of Environmental Biology
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• v.20 no.2
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• pp.130-135
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• 2002

Species of Genus Trichoderma are commercially applied as biological control agents against fungal Pathogens. A powerful biocontrol agent, Trichoderma sp. SJG-99721 was isolated from 305 isolates by morphological characters, chitinase activities and antifungal activities against Phytophthora capsiei. The isolate was identified as Trichoderma harzianum from various features such as growth rate at $27^\circ{C}$, significant growth ratio of $27^\circ{C}$ to $17^\circ{C}$, amount of aerial mycelium, types of branching: system, and disposition patterns of phialide and phialospore. Trichoderma harzianum SJG-99721 have been shown to act as a powerful biological agent against fungal phytopathogens; Botrytis cinerea, Rhizoctonia solani, Phytophthora cryptogea, Phytophthora capsiei, Sclerotinia sclerotiorum, Mycoshaerella melonis, Alternaria sotani, Fusarium oxysporum, Collectotrichum gloesporioodes, Alternaria alternata, Phythium ultimum, Phytophthora drechsleri, Pyricularia grisea.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1654-1663
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• 2013

Rice straw is closely associated with meju fermentation and it is generally known that the rice straw provides meju with many kinds of microorganisms. In order to elucidate the origin of meju fungi, the fungal diversity of rice straw was examined. Rice straw was collected from 12 Jang factories where meju are produced, and were incubated under nine different conditions by altering the media (MEA, DRBC, and DG18), and temperature ($15^{\circ}C$, $25^{\circ}C$, and $35^{\circ}C$). In total, 937 strains were isolated and identified as belonging to 39 genera and 103 species. Among these, Aspergillus, Cladosporium, Eurotium, Fusarium, and Penicillium were the dominant genera. Fusarium asiaticum (56.3%), Cladosporium cladosporioides (48.6%), Aspergillus tubingensis (37.5%), A. oryzae (31.9%), Eurotium repens (27.1%), and E. chevalieri (25.0%) were frequently isolated from the rice straw obtained from many factories. Twelve genera and 40 species of fungi that were isolated in the rice straw in this study were also isolated from meju. Specifically, A. oryzae, C. cladosporioides, E. chevalieri, E. repens, F. asiaticum, and Penicillium polonicum (11.8%), which are abundant species in meju, were also isolated frequently from rice straw. C. cladosporioides, F. asiaticum, and P. polonicum, which are abundant in the low temperature fermentation process of meju fermentation, were frequently isolated from rice straw incubated at $15^{\circ}C$ and $25^{\circ}C$, whereas A. oryzae, E. repens, and E. chevalieri, which are abundant in the high temperature fermentation process of meju fermentation, were frequently isolated from rice straw incubated at $25^{\circ}C$ and $35^{\circ}C$. This suggests that the mycobiota of rice straw has a large influence in the mycobiota of meju. The influence of fungi on the rice straw as feed and silage for livestock, and as plant pathogens for rice, are discussed as well.

• 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.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.34-34
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• 2014

Filamentous fungi of the genus Colletotrichum (teleomorph, Glomerella) are considered major plant pathogens worldwide. Cereals, legumes, vegetables, and fruit trees may be seriously affected by this pathogen (1). Colletotrichum species cause typical disease symptoms known as anthracnoses, characterized by sunken necrotic tissue, where orange conidial masses are produced. Anthracnose appears in both developing and mature plant tissues (2). We investigated disease occurrence in apple orchards from 2013 to 2014 in northern Gyeongbuk province, Korea. Typical anthracnose with advanced symptoms was observed in all apple orchards studied. Of late, static fruit spot symptoms are being observed in apple orchards. A small lesion, which does not expand further and remains static until the harvesting season, is observed at the beginning of fruit growth period. In our study, static symptoms, together with the typical symptoms, were observed on apples. The isolated fungus was tested for pathogenicity on cv. 'Fuji apple' (fully ripe fruits, unripe fruits, and cross-section of fruits) by inoculating the fruits with a conidial suspension ($10^5$ conidia/ml). In apple inoculated with typical anthracnose fungus, the anthracnose symptoms progressed, and dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. However, in apple inoculated with fungi causing static symptoms, the size of the spots did not increase. Interestingly, the shape and size of the conidia and the shape of the appressoria of both types of fungi were found to be similar. The conidia of the two types of fungi were straight and cylindrical, with an obtuse apex. The culture and morphological characteristics of the conidia were similar to those of C. gloeosporioides (5). The conidia of C. gloeosporioides germinate and form appressoria in response to chemical signals such as host surface wax and the fruitripening hormone ethylene (3). In this study, the spores started to germinate 4 h after incubation with an ethephon suspension. Then, the germ tubes began to swell, and subsequently, differentiation into appressoria with dark thick walls was completed by 8 h. In advanced symptoms, fungal spores of virtually all the appressoria formed primary hyphae within 16 h. However, in the static-symptom fungus spores, no primary hyphae formed by 16 h. The two types of isolates exhibited different growth rates on medium containing apple pectin, Na polypectate, or glucose as the sole carbon. Static-symptom fungi had a >10% reduction in growth (apple pectin, 14.9%; Na polypectate, 27.7%; glucose, 10.4%). The fungal isolates were also genetically characterized by sequencing. ITS regions of rDNA, chitin synthase 1 (CHS1), actin (ACT), and ${\beta}$-tubulin (${\beta}t$) were amplified from isolates using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, ACT-512F and ACT-783R, and T1 and ${\beta}t2$ (5), respectively. The resulting sequences showed 100% identity with sequences of C. gloeosporioides at KC493156, and the sequence of the ${\beta}$t gene showed 100% identity with C. gloeosporioides at JX009557.1. Therefore, sequence data from the four loci studied proves that the isolated pathogen is C. gloeosporioides. We also performed random amplified polymorphic DNA-PCR, which showed clearly differentiated subgroups of C. gloeosporioides genotypes. The clustering of these groups was highly related to the symptom types of the individual strains.

• The Korean Journal of Mycology
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• v.41 no.3
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• pp.192-196
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• 2013

Fungal development is largely affected by many environmental factors. In a model filamentous fungus Aspergillus nidulans, asexual development is promoted by exposure of light, presence of salt and non-fermentable sugars. In other hand, sexual development is largely induced by absence of light, fermentable sugars and hypoxic condition. Also, some important genes including veA and nsdD play positive roles in activating sexual development. Here, we reported that the effect of high concentration of $CO_2$ on developmental decision in A. nidulans. When wild-type $veA^+$ strain was cultured in normal condition, sexual and asexual development occurred in balanced manner. However, high concentration of $CO_2$ (~5%) strongly activated sexual development and inhibited asexual development. Furthermore, this $CO_2$ effect was controlled by the veA or nsdD gene. High $CO_2$ culture of $veA^-$ or $nsdD^-$ mutant didn't activate sexual development, suggesting that the activation of sexual development induced by high $CO_2$ cannot overcome the genetic requirement of sexual development such as veA or nsdD. Since 5% $CO_2$ is an important condition for human pathogenic fungi for surviving and adapting in human body, this developmental pattern of A. nidulans affected by $CO_2$ concentration may provide interesting clues for comparative study with human fungal pathogens including Aspergillus fumigatus.

• Journal of Life Science
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• v.31 no.1
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• pp.83-89
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• 2021

Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.