Jong-Hwan Shin

Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea.

Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

Characterization of the Maize Stalk Rot Pathogens Fusarium subglutinans and F. temperatum and the Effect of Fungicides on Their Mycelial Growth and Colony Formation

Maize is a socioeconomically important crop in many countries. Recently, a high incidence of stalk rot disease has been reported in several maize fields in Gangwon province. In this report, we show that maize stalk rot is associated with the fungal pathogens Fusarium subglutinans and F. temperatum. Since no fungicides are available to control these pathogens on maize plants, we selected six fungicides (tebuconazole, difenoconazole, fluquinconazole, azoxystrobin, prochloraz and kresoxim-methyl) and examined their effectiveness against the two pathogens. The in vitro antifungal effects of the six fungicides on mycelial growth and colony formation were investigated. Based on the inhibition of mycelial growth, the most toxic fungicide was tebuconazole with 50% effective concentrations (EC50) of <0.1 μg/ml and EC90 values of 0.9 μg/ml for both pathogens, while the least toxic fungicide was azoxystrobin with EC50 values of 0.7 and 0.5 μg/ml for F. subglutinans and F. temperatum, respectively, and EC90 values of >3,000 μg/ml for both pathogens. Based on the inhibition of colony formation by the two pathogens, kresoxim-methyl was the most toxic fungicide with complete inhibition of colony formation at concentrations of 0.1 and 0.01 μg/ml for F. subglutinans and F. temperatum, respectively, whereas azoxystrobin was the least toxic fungicide with complete inhibition of colony formation at concentrations >3,000 μg/ml for both pathogens.

Effectiveness of Different Classes of Fungicides on Botrytis cinerea Causing Gray Mold on Fruit and Vegetables.

Botrytis cinerea is a necrotrophic pathogen causing a major problem in the export and post-harvest of strawberries. Inappropriate use of fungicides leads to resistance among fungal pathogens. Therefore, it is necessary to evaluate the sensitivity of B. cinerea to various classes of fungicide and to determine the effectiveness of different concentrations of commonly used fungicides. We thus evaluated the effectiveness of six classes of fungicide in inhibiting the growth and development of this pathogen, namely, fludioxonil, iprodione, pyrimethanil, tebuconazole, fenpyrazamine, and boscalid. Fludioxonil was the most effective (EC50 < 0.1 μg/ml), and pyrimethanil was the least effective (EC50 = 50 μg/ml), at inhibiting the mycelial growth of B. cinerea. Fenpyrazamine and pyrimethanil showed relatively low effectiveness in inhibiting the germination and conidial production of B. cinerea. Our results are useful for the management of B. cinerea and as a basis for monitoring the sensitivity of B. cinerea strains to fungicides.

Antifungal Activity of Bacillus sp. GJ-1 Against Phytophthora capsici

Phytophthora capsici is one of major limiting factors in production of pepper and other important crops worldwide by causing foliage blight and rot on fruit and root. Increased demand for the replacement of fungicides has led to searching a promising strategy to control the fungal diseases. To meet eco-friendly agriculture practice, we isolated microorganisms and assessed their beneficial effects on plant health and disease control efficacy. A total of 360 bacterial strains were isolated from rhizosphere soil of healthy pepper plants, and categorized to 5 representative isolates based on colony morphology. Among the 5 bacterial strains (GJ-1, GJ-4, GJ-5, GJ-11, GJ-12), three bacterial strains (GJ-1, GJ-11, GJ-12) presented antifungal activity against P. capsici in an fungal inhibition assay. In phosphate solubilization and siderophore production, the strain GJ-1 was more effective than others. The strain GJ-1 was identified as Bacillus sp. using 16S rDNA analysis. Bacillus sp. GJ-1 was also found to be effective in inhibiting other plant pathogenic fungi, including Rhizoctonia solani, Pythium ultimum and Fusarium solani. Therefore, the Bacillus sp. GJ-1 can serve as a biological control agent against fungal plant pathogens.

Distinct roles of the YPEL gene family in development and pathogenicity in the ascomycete fungus Magnaporthe oryzae

Members of the Yippee-like (YPEL) gene family are highly conserved in eukaryotes and are homologous to the Drosophila yippee gene. In this study, we functionally characterized two YPEL-homologous genes, MoYPEL1 and MoYPEL2, in the rice blast pathogen Magnaporthe oryzae using the deletion mutants ΔMoypel1, ΔMoypel2, and ΔΔMoypel1,2. The MoYPEL1 deletion mutant was significantly defective in conidiation and unable to undergo appressorium development; however, deletion of MoYPEL2 resulted in a significant increase in conidiation and the abnormal development of two appressoria per conidium. These data demonstrate the opposite roles of each member of the YPEL gene family during the development of M. oryzae. The double mutant was phenotypically similar to the ΔMoypel1 mutant in conidiation, but similar to the ΔMoypel2 mutant in appressorium development. Subcellular localization of the MoYPEL1 protein was dynamic during appressorium development, while the MoYPEL2 protein consistently localized within the nuclei during developmental stages. Our studies indicate that the two YPEL gene family members play distinct roles in the developmental stages of M. oryzae, furthering our understanding of disease dissemination and development in fungi.

The Effect of Fungicides on Mycelial Growth and Conidial Germination of the Ginseng Root Rot Fungus, Cylindrocarpon destructans

Abstract Ginseng root rot caused by Cylindrocarpon destructans is the most destructive disease of ginseng. Six different fungicides (thiophanate-methyl, benomyl, prochloraz, mancozeb, azoxystrobin, and iprodione) were selected to evaluate the inhibitory effect on the mycelial growth and conidial germination of C. destructans isolates. Benomyl and prochloraz were found to be the most effective fungicides in inhibiting mycelial growth of all tested isolates, showing 64.7% to 100% inhibition at a concentration of 10 μg/mL, whereas thiophanate-methyl was the least effective fungicide, showing less than 50% inhibition even at a higher concentration of 100 μg/mL. The tested fungicides exhibited less than 20% inhibition of conidium germination at concentrations of 0.01, 0.1, and 1 μg/mL. However, the inhibition effect of mancozeb on condium germination of C. destructans was significantly increased to 92% to 99% at a higher concentration of 100 μg/mL, while the others still showed no higher than 30% inhibition.

Genome-wide analyses of DNA-binding proteins harboring AT-hook motifs and their functional roles in the rice blast pathogen, Magnaporthe oryzae

Transcriptional regulation is a complex process mediated by coordinated assembly complexes to ensure temporal and spatial gene expression. The AT-hook is a DNA-binding motif originally described in the high mobility group A of non-histone chromatin components. The AT-hook proteins bind to the minor groove of adenine–thymine (AT) rich regions of DNA and act as transcriptional cofactors coordinating nucleoproteins during transcriptional regulation. In this study, a genome-wide in silico analysis of AT-hook proteins was performed on the ascomycete plant pathogenic fungus, Magnaporthe oryzae. Quantitative real-time RT-PCR analysis revealed differential expression patterns of MoATH genes during development and plant infection. To increase our understanding of the functional role of AT-hook proteins in M. oryzae development and pathogenicity, a deletion mutant of MoATH10 was functionally characterized. Targeted deletion of MoATH10 significantly increased pigmentation and conidiation, indicating that MoATH10 is negatively involved in the regulation of pigmentation and conidiation in M. oryzae. Pathogenicity assays revealed that the ΔMoath10 mutant was less virulent. The reduced disease development of the ΔMoath10 mutant was due to a partial defect in invasive growth inside plant cells, but not appressorium-mediated penetration. These results suggest that MoATH10 is important for growth, development, and virulence in M. oryzae.