• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.27-27
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• 2016

The ascomycete fungus Fusarium graminearum is the most common pathogen of Fusarium head blight (FHB), a devastating disease for major cereal crops worldwide. FHB causes significant crop losses by reducing grain yield and quality as well as contaminating cereals with trichothecenes and zearalenone (ZEA) that pose a serious threat to animal health and food safety. ZEA is a causative agent of hyperestrogenic syndrome in mammals and can result in reproductive disorders in farm animals. In F. graminearum, the ZEA biosynthetic cluster is composed of four genes, PKS4, PKS13, ZEB1, and ZEB2, which encode a reducing polyketide synthase, a nonreducing polyketide synthase, an isoamyl alcohol oxidase, and a transcription factor, respectively. Although it is known that ZEB2 primarily acts as a regulator of ZEA biosynthetic cluster genes, the mechanism underlying this regulation remains undetermined. In this study, two isoforms (ZEB2L and ZEB2S) from the ZEB2 gene in F. graminearum were characterized. It was revealed that ZEB2L contains a basic leucine zipper (bZIP) DNA-binding domain at the N-terminus, whereas ZEB2S is an N-terminally truncated form of ZEB2L that lacks the bZIP domain. Interestingly, ZEA triggered the induction of both ZEB2L and ZEB2S transcription. In ZEA producing condition, the expression of ZEB2S transcripts via alternative promoter usage was directly or indirectly initiated by ZEA. Physical interaction between ZEB2L and ZEB2L as well as between ZEB2L and ZEB2S was observed in the nucleus. The ZEB2S-ZEB2S interaction was detected in both the cytosol and the nucleus. ZEB2L-ZEB2L oligomers activated ZEA biosynthetic cluster genes, including ZEB2L. ZEB2S inhibited ZEB2L transcription by forming ZEB2L-ZEB2S heterodimers, which reduced the DNA-binding activity of ZEB2L. This study provides insight into the autoregulation of ZEB2 expression by alternative promoter usage and a feedback loop during ZEA production.

• 한국균학회소식:학술대회논문집
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• 2018.05a
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• pp.44-44
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• 2018

Fusarium head blight (FHB) caused by infection with Fusarium graminearum leads to enormous losses to crop growers, and may contaminate grains with a number of Fusarium mycotoxins that pose serious risks to human and animal health. Antagonistic bacteria that are used to prevent FHB offer attractive alternatives or supplements to synthetic fungicides for controlling FHB without the negative effects of chemical management. Out of 500 bacterial strains isolated from soil, Bacillus amyloliquefaciens JCK-12 showed strong antifungal activity and was considered a potential source for control strategies to reduce FHB. B. amyloliquefaciens JCK-12 produces several cyclic lipopeptides (CLPs) including iturin A, fengycin, and surfactin. Iturin A inhibits spore germination of F. graminearum. Fengycin or surfactin alone did not display any inhibitory activity against spore germination at concentrations less than 30 ug/ml, but a mixture of iturin A, fengycin, and surfactin showed a remarkable synergistic inhibitory effect on F. graminearum spore germination. The fermentation broth and formulation of B. amyloliquefaciens JCK-12 strain reduced the disease incidence of FHB in wheat. Furthermore, co-application of B. amyloliquefaciens JCK-12 and chemical fungicides resulted in synergistic in vitro antifungal effects and significant disease control efficacy against FHB under greenhouse and field conditions, suggesting that B. amyloliquefaciens JCK-12 has a strong chemosensitizing effect. The synergistic antifungal effect of B. amyloliquefaciens JCK-12 and chemical fungicides in combination may result from the cell wall damage and altered cell membrane permeability in the phytopathogenic fungi caused by the CLP mixtures and subsequent increased sensitivity of F. graminearum to fungicides. In addition, B. amyloliquefaciens JCK-12 showed the potential to reduce trichothecenes mycotoxin production. The results of this study indicate that B. amyloliquefaciens JCK-12 could be used as an available biocontrol agent or as a chemosensitizer to chemical fungicides for controlling FHB disease and as a strategy for preventing the contamination of harvested crops with mycotoxins.

• Research in Plant Disease
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• v.10 no.4
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• pp.260-267
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• 2004

Fusarium spp. were isolated from the postharvest fruit rot of oriental melon fruits at commercial fruit markets in Korea during 2001 to 2003. The decayed fruits were covered with the fungal mycelia and eventually soft rotted. The disease started at the fruit stalk area, the calyx end of the fruit and skin of fruit. As the disease advanced, white to pinkish mycelia covered with the surface of decayed fruit. The cultural and morphological characteristic of Fusarium spp. were compared with descriptions of those reported previously, and identified as Fusarium equiseti, F. graminearum, F. moniliforme, F. proliferatum, F. sambucinum, and F. semitectum. Pathogenicity of the isolates was proved by artificial wound and unwound inoculation onto the healthy fruits. Two days after inoculation, aerial mycelia were noticed on the wound inocultion region of the fruit and developed soft rot symptoms. Although Fusarium spp. causing fruit rot disease in oriental melon have been reported in Korea, identification of the those species was not described. Therefore, this is the first report of Fusarium spp. causing postharvest fruit rot on oriental melon in Korea.

• Research in Plant Disease
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• v.25 no.4
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• pp.157-163
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• 2019

This study aimed to assess the incidence and distribution of toxigenic fungi in Korean oat. Toxigenic fungi were isolated from oat samples collected from 12 oat fields from heading to harvest in 2017 and 2018. A total of 745 fungal colonies were isolated based on morphology and identified using marker genes. About 92% of the fungal isolates were Fusarium spp. and others were Penicillium (5.9%) and Aspergillus (2.1%). Fusarium isolates comprised mostly of F. asiaticum (83.1%), followed by F. incarnatum (5.4%), F. proliferatum (3.5%), F. fujikuroi (2.8%), F. tricinctum species complex (FTSC) 11 (1.5%) and F. graminearum (1.0%). About 97% of F. asiaticum was nivalenol type, and 3-acetyl deoxynivalenol (3.2%) and 15-acetyl deoxynivalenol (0.4%) types also were found. Pathogenicity test of the selected Fusarium isolates revealed that F. asiaticum isolates have a wide range of virulence depending on the tested plants. F. graminearum and FTSC 11 isolates from blighted spikelets were the most virulent in naked oat. All Fusarium isolates (n=18) except one (FTSC 11) produced nivalenol (0.2-7.6 ㎍/g), deoxynivalenol (0.03-6.1 ㎍/g), and zearalenone (0.1-27.0 ㎍/g) on rice medium. This study is first report that F. asiaticum causes Fusarium head blight disease of oat in Korea. These findings demonstrate the dominance of F. asiaticum in oat agroecosystems as in rice, wheat and barley in Korea.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.407-413
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• 2016

Fusarium graminearum species complex (FGSC) causes Fusarium head blight in small grain cereals. To date, four species (F. graminearum, F. asiaticum, F. boothii, and F. meridionale ) belonging to FGSC frequently occur in Korean cereals. In addition, we first reported the occurrence of additional species (F. vorosii ) within FGSC, which was isolated from barley, corn, and rice in Korea. Phylogenetic analysis of the Fusarium isolates of this group using combined multigene sequences confirmed species identification. Moreover, the macroconidia produced by these isolates were morphologically similar to those of the F. vorosii holotype. Chemical analysis indicated that the F. vorosii isolates produced various trichothecenes such as nivalenol and deoxynivalenol with their acetyl derivatives along with zearalenone. Pathogenicity tests demonstrated that all of the F. vorosii isolates examined were pathogenic on barley, corn, and rice with variation in aggressiveness. This study is the first report of F. vorosii in Korean cereals, their pathogenicity towards barley and corn, and their ability to produce trichothecenes and zearalenone.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.123.2-124
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• 2003

Fusarium graminearum (telomorph：Gibberella zeae), an important fungal pathogen of cereal crops with ubiquitous geographic distribution, produces mycotoxins on diseased crops that has threaten human and animal health. Recently severe epidemics of scab diseases of barley and rice by this fungus occurred in Korea, causing serious economic losses. To determine genetic diversity of F. graminearum from rice in Korea, a total of 269 isolates were obtained from Southern part of Korea during 2001-2002. A phylogenetic tree of the isolates was constructed by using amplified fragment length polymorphism (AFLP). Population structure of the rice isolates consists of a single lineage (lineage 6). Frequency of female fertility among these Isolates was relatively low (37％) compared to that among lineage 7 isolates from Korean corn. PCR amplification using chemotype specific primers derived from Tri7 and Tri13 genes at the trichothecene biosynthesis gene cluster revealed that most isolates (260) were NIV chemotype；9 isolates were identified as DON chemotype by Tri13 but as either NIV chemotype or unknown by Tri7. The result of chemical analysis also supported the chemotype determination；all of the NIV chemotype isolates produced NIV, whereas the 9 isolates produce either DON or no toxin.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.85.2-86
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• 2003

Fusarium graminearum is an important pathogen of cereal crops in many areas of the world causing head blight and ear rot of small grains. In addition to serious economic losses, this fungus produces mycotoxins, such as trichothecenes and zearalenone on diseased crops and has been a potential threat to human and animal health. To massively identify pathogenesis-related genes from F. graminearum, two representative strains (SCKO4 from rice and Z03643 from wheat) were mutagenized using restriction enzyme-mediated integration (REMI). In total, 20,DOD REMI transformants have been collected from the two strains. So far, 63 mutants for several traits involved in disease development such as virulence, mycotoxin production, and sporulation have been selected from 3,000 REMI transformants. Now, selected mutants of interest have being genetically analyzed using a newly developed outcross method (See Jungkwan Lee et al poster). In addition, cloning and characterization of genomic DNA regions flanking the insertional site in the genome of the mutants are in progress.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.349-353
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• 2011

The ascomycete fungus Fusarium graminearum is an important plant pathogen responsible for Fusarium head blight in small grains and ear rot on maize. This fungus also produces the estrogenic metabolite, zearalenone (ZEA) that causes estrogenic disorders in humans and animals. Previously, we developed a conditional gene expression system for this fungus using a ZEA-inducible promoter (Pzear). In the present study, four other estrogenic compounds, including ${\beta}$-estradiol, estriol, estrone, and secoisolariciresinol, were screened as possible substitutes for ZEA in this system. Among them, ${\beta}$-estradiol was able to successfully induce the expression of a gene controlled by Pzear, while estrone was only able to partially induce its expression but the other two compounds were not effective. In combination, these results demonstrate that ${\beta}$-estradiol can replace ZEA in this conditional gene expression system, thereby eliminating the need to use the more expensive reagent, ZEA, and facilitating high-throughput functional analyses of F. graminearum in future studies.

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.967-974
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• 2016

Zearalenone (ZEA) is an estrogenic mycotoxin that is produced by several Fusarium species, including Fusarium graminearum. One of the ZEA biosynthetic genes, ZEB2, encodes two isoforms of Zeb2 by alternative transcription, forming an activator (Zeb2L-Zeb2L homooligomer) and an inhibitor (Zeb2L-Zeb2S heterodimer) that directly regulate the ZEA biosynthetic genes in F. graminearum. Cyclic AMP-dependent protein kinase A (PKA) signaling regulates secondary metabolic processes in several filamentous fungi. In this study, we investigated the effects of the PKA signaling pathway on ZEA biosynthesis. Through functional analyses of PKA catalytic and regulatory subunits (CPKs and PKR), we found that the PKA pathway negatively regulates ZEA production. Genetic and biochemical evidence further demonstrated that the PKA pathway specifically represses ZEB2L transcription and also takes part in posttranscriptional regulation of ZEB2L during ZEA production. Our findings reveal the intriguing mechanism that the PKA pathway regulates secondary metabolite production by reprograming alternative transcription.

• 한국균학회소식:학술대회논문집
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• 2018.05a
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• pp.33-33
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• 2018

Air-borne plant pathogenic fungus Fusarium graminearum and seed-borne plant pathogenic bacterium Burkholderia glumae are cause similar disease symptoms in rice heads. Here we showed that two pathogens frequently co-isolated in rice heads and F. graminearum is resistant to toxoflavin produced by B. glumae while other fungal genera are sensitive to the toxin. We have tried to clarify the resistant mechanism of F. graminearum against toxoflavin and the ecological reason of co-existence of the two pathogens in rice. We found that F. graminearum carries resistance to toxoflavin as accumulating lipid in fungal cells. Co-cultivation of two pathogens resulted in increased conidia and enhanced chemical attraction and attachment of the bacterial cells to the fungal conidia. Bacteria physically attached to fungal conidia, which protected bacterium cells from UV light and allowed disease dispersal. Chemotaxis analysis showed that bacterial cells moved toward the fungal exudation compared to a control. Even enhanced the production of phytotoxic trichothecene by the fungal under presence of toxoflavin and disease severity on rice heads was significantly increased by co-inoculation rather than single inoculation. This study suggested that the undisclosed potentiality of air-born infection of bacteria using the fungal spores for survival and dispersal.