• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.11-11
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• 2014

Fungal pathogens have huge impact on health and economic wellbeing of human by causing life-threatening mycoses in immune-compromised patients or by destroying crop plants. A key determinant of fungal pathogenesis is their ability to undergo developmental change in response to host or environmental factors. Genetic pathways that regulate such morphological transitions and adaptation are therefore extensively studied during the last few decades. Given that epigenetic as well as genetic components play pivotal roles in development of plants and mammals, contribution of microbial epigenetic counterparts to this morphogenetic process is intriguing yet nearly unappreciated question to date. To bridge this gap in our knowledge, we set out to investigate histone modifications among epigenetic mechanisms that possibly regulate fungal adaptation and processes involved in pathogenesis of a model plant pathogenic fungus, Magnaporthe oryzae. M. oryzae is a causal agent of rice blast disease, which destroys 10 to 30% of the rice crop annually. Since the rice is the staple food for more than half of human population, the disease is a major threat to global food security. In addition to the socioeconomic impact of the disease it causes, the fungus is genetically tractable and can undergo well-defined morphological transitions including asexual spore production and appressorium (a specialized infection structure) formation in vitro, making it a model to study fungal development and pathogenicity. For functional and comparative analysis of histone modifications, a web-based database (dbHiMo) was constructed to archive and analyze histone modifying enzymes from eukaryotic species whose genome sequences are available. Histone modifying enzymes were identified applying a search pipeline built upon profile hidden Markov model (HMM) to proteomes. The database incorporates 22,169 histone-modifying enzymes identified from 342 species including 214 fungal, 33 plants, and 77 metazoan species. The dbHiMo provides users with web-based personalized data browsing and analysis tools, supporting comparative and evolutionary genomics. Based on the database entries, functional analysis of genes encoding histone acetyltransferases and histone demethylases is under way. Here I provide examples of such analyses that show how histone acetylation and methylation is implicated in regulating important aspects of fungal pathogenesis. Current analysis of histone modifying enzymes will be followed by ChIP-Seq and RNA-seq experiments to pinpoint the genes that are controlled by particular histone modifications. We anticipate that our work will provide not only the significant advances in our understanding of epigenetic mechanisms operating in microbial eukaryotes but also basis to expand our perspective on regulation of development in fungal pathogens.

• Molecules and Cells
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• v.27 no.5
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• pp.563-570
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• 2009

We previously isolated the OsCBT gene, which encodes a calmodulin (CaM)-binding protein, from a rice expression library constructed from fungal elicitor-treated rice suspension cells. In order to understand the function of OsCBT in rice, we isolated and characterized a T-DNA insertion mutant allele named oscbt-1. The oscbt-1 mutant exhibits reduced levels of OsCBT transcripts and no significant morphological changes compared to wild-type plant although the growth of the mutant is stunted. However, oscbt-1 mutants showed significant resistance to two major rice pathogens. The growth of the rice blast fungus Magnaporthe grisea, as well as the bacterial pathogen Xanthomonas oryzae pv. oryzae was significantly suppressed in oscbt-1 plants. Histochemical analysis indicated that the hypersensitive-response was induced in the oscbt-1 mutant in response to compatible strains of fungal pathogens. OsCBT expression was induced upon challenge with fungal elicitor. We also observed significant increase in the level of pathogenesis-related genes in the oscbt-1 mutant even under pathogen-free condition. Taken together, the results support an idea that OsCBT might act as a negative regulator on plant defense.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.805-810
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• 2000

An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

• The Plant Pathology Journal
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• v.30 no.4
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• pp.367-374
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• 2014

Genomes contain a large number of unique genes which have not been found in other species. Although the origin of such "orphan" genes remains unclear, they are thought to be involved in species-specific adaptive processes. Here, we analyzed seven orphan genes (MoSPC1 to MoSPC7) prioritized based on in planta expressed sequence tag data in the rice blast fungus, Magnaporthe oryzae. Expression analysis using qRT-PCR confirmed the expression of four genes (MoSPC1, MoSPC2, MoSPC3 and MoSPC7) during plant infection. However, individual deletion mutants of these four genes did not differ from the wild-type strain for all phenotypes examined, including pathogenicity. The length, GC contents, codon adaptation index and expression during mycelial growth of the four genes suggest that these genes formed during the evolutionary history of M. oryzae. Synteny analyses using closely related fungal species corroborated the notion that these genes evolved de novo in the M. oryzae genome. In this report, we discuss our inability to detect phenotypic changes in the four deletion mutants. Based on these results, the four orphan genes may be products of de novo gene birth processes, and their adaptive potential is in the course of being tested for retention or extinction through natural selection.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.305-313
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• 2020

Host shifting and host expansion of fungal plant pathogens increases the rate of emergence of new pathogens and the incidence of disease in various crops, which threaten global food security. Magnaporthe species cause serious disease in rice, namely rice blast disease, as well as in many alternative hosts, including wheat, barley, and millet. A severe outbreak of wheat blast due to Magnaporthe oryzae occurred recently in Bangladesh, after the fungus was introduced from South America, causing great loss of yield. This outbreak of wheat blast is of growing concern, because it might spread to adjacent wheat-producing areas. Therefore, it is important to understand the host range and population structure of M. oryzae and related species for determining the evolutionary relationships among Magnaporthe species and for managing blast disease in the field. Here, we collected isolates of M. oryzae and related species from various Poaceae species, including crops and weeds surrounding rice fields, in Korea and determined their phylogenetic relationships and host species specificity. Internal transcribed spacer-mediated phylogenetic analysis revealed that M. oryzae and related species are classified into four groups primarily including isolates from rice, crabgrass, millet and tall fescue. Based on pathogenicity assays, M. oryzae and related species can infect different Poaceae hosts and move among hosts, suggesting the potential for host shifting and host expansion in nature. These results provide important information on the diversification of M. oryzae and related species with a broad range of Poaceae as hosts in crop fields.

• Research in Plant Disease
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• v.7 no.2
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• pp.86-92
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• 2001

This experiment was carried out to analyze the recent epidemic of rice blast in southern provinces of Korea in 1999 and 2000. Incidences of leaf blast and panicle blast in these years were 1.5 and 2.9 times greater than those in 1998, respectively. Cultivation areas of rice cultivars, Daesan, Ilmi and Dongan bred from Milyang 95 as a recurrent parent increased over the country from 11% in 1998 to 38.4% in 2000, which were grown in more than 85% of total paddy fields in Jeonnam province. Predominant race populations of Pyricularia grisea in the farmers' fields had been changed from KJ-301 to KI-1117 in 1999 and KI-1113 and KJ-105 in 2000. Distribution ratio of the KI-1117 race in 1998 in the southern provinces was 1.7%, but increased to 30% in 1999. Although the cultivars Daesan, Ilmi and Dongan have shown wide spectrum of resistance to many races including KJ-301, they were susceptible to such races as KI-1117, KI-1113 and, KJ-105. These three races could be further classified into two sub-races, KI-1117 and KI-1117a, KI-1113 and Kl-1113a, and KJ-105 and KJ-105a based on virulence to Daesan. The newly classfied races, Kl-1113a, Kl-1117a, and KJ-l05a were also pathogenic to Ilmi and Dongan. This indicates that rice blast epidemic in southern provinces in 1999 and 2000 resulted from the breakdown of the resistance of Daesan, Ilmi and Dongan by the rapid increase of virulent races KI-1117, KI-1113, KJ-105 and their new sub-races.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.413-417
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• 1998

Magnaporthe grisea, the casual agent of rice blast, requires formation of an appressorium, a dome-shaped and well melanized infection structure, to penetrate its host. Environmental cues that induce appressorium formation include hydrophobicity and hardness of contact surface and chemicals from its host. Artificial surfaces are widely used to induce appressorium formation, but frequencies of appressorium induction are not always consistent. To understand variable induction of appressorium formation in M. grisea, several factors were tested on GelBond. High levels of appressorium formation were induced over a wide range of temperature (20~3$0^{\circ}C$) and pH (4~7). spore age up to 3-week-old did not significantly affect appressorium formation, but only a few apressoria on GelBond. However, adenosine specifically inhibited appressorium formation. Adenosine inhibition of appressorium formation was restored by exogenous addition of cAMP. Germ tube tips of M. grisea maintained the ability to differentiate appressoria by chemical inducers on GelBond at least up to 16 h after conidia germination. These results suggest that environmental factors have little effect on the variable induction of appressorium formation on the artificial surface in M. grisea.

• The Plant Pathology Journal
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• v.34 no.2
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• pp.126-138
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• 2018

Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

• Molecules and Cells
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• v.23 no.2
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• pp.207-214
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• 2007

lant ${\beta}$-1, 3-glucanases are involved in plant defense and in development. Very little data are available on the expression of rice glucanases both in developmental tissues and under various stresses. In this study, we cloned and characterized twenty-seven rice ${\beta}$-1, 3-glucanases (OsGlu) from at total of 71 putative glucanases. The OsGlu genes were obtained by PCR from a cDNA library and were classified into seven groups (Group I to VII) according to their DNA or amino acid sequence homology. Analysis of the expression of the twenty-seven OsGlu genes by Northern blotting revealed that they were differentially expressed in different developmental tissues as well as in response to plant hormones, biotic stress, high salt etc. OsGlu11 and 27 in Group IV were clearly expressed only in stem and leaf and were also induced strongly by SA (5 mM), ABA ($200{\mu}M$), and M. grisea. OsGlu1, 10, 11, and 14 were induced earlier and to higher levels in incompatible M. grisea interaction than in compatible one. Taken together, our findings suggest that the twenty-seven rice OsGlu gene products play diverse roles not only in plant defense but also in hormonal responses and in development.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.09a
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• pp.194.2-194
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• 2002