• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.41-41
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• 2015

Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

• The Korean Journal of Mycology
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• v.39 no.2
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• pp.91-98
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• 2011

Oudemansiella mucida, an edible and medicinal mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to produce antifungal substances to inhibit the mycelial growth and spore germination of the plant pathogenic fungi. To produce good amount of antifungal substances from culture media, the optimal culture conditions of O. mucida were investigated. The most favorable conditions for the mycelial growth were $25^{\circ}C$ and pH 5 in potato dextrose agar. The most favorable carbon and nitrogen sources promoting mycelial growth were maltose and calcium nitrate, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplemented to the basal medium as a carbon source. The optimal mycelial growth of O. mucida was found in the Hennerberg medium. The crude extract from submerged culture of potato dextrose broth exhibited inhibition of mycelial growth of Colletotrichum acutatum, Botrytis cinerea and Pyricularia oryzae but, fungicidal activity is not good enough to compared with commercially available fungicides tested. Therefore, the antifungal substances extracted from submerged culture of O. mucida might have a potential to be used for biocontrol agent of fungal diseases of plants.

• Journal of Applied Biological Chemistry
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• v.55 no.2
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• pp.79-83
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• 2012

It has been known that the proteome profiles in the period of growth and development of rice are changed by the growth conditions including temperature, soil, and fertilization. In this study, the proteome profiles of Odae polished white rice grown in Chulwon and Chilgog were compared on 2-dimensional(D) gels. The differentially expressed proteins were selected from the 112 identified total proteins and classified into functional groups. The most significantly differentially expressed proteins were stress responsive proteins; Ent-kaur-16-ene synthase, which is responsible for synthesizing a plant hormone gibberellin, was expressed in Chulwon rice and heat shock proteins were in Chilgog rice, respectively. Xylanase inhibitor protein, which inhibits the enzyme xylanase produced by pathogenic fungi and Bacilli, was expressed significantly high in Chilgog rice grown at high temperature. Differential expressions of transporter proteins were observed both in Chulwon and Chilgog rice. Regarding the facts that Chilgog rice contained relatively higher amount of proteins than Chulwon rice and Chulwon rice showed large number of proteins were differentially expressed, it can be concluded that different cultivation conditions could change the protein expression profiles in rice in various ways, including elevation of protein amount or differential expressions of specific proteins, etc. The results suggest that the characteristics of the profiles of the proteome in the polished white rice are definitely changed by the environmental factors including high temperature. The results can be utilized for the development of the proper cultivation conditions for the production of high quality rice with good palatability.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.135-141
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• 2006

We isolated a bacterium which produces antifungal substances from the Sanktpeterburg soils at Russia. The iso-lated strain was identified as Brevibacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Brevibacillus sp. KMU-391 produced maximum level of antifungal substances under incubation aerobically at $30^{\circ}C$ for 48 hours in trypticase soybean broth containing 1.0% sucrose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 7.0. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against Didymella bryoniae by dry cell weight. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Botrytis cinerea KACC 40573, Botrytis fabae KACC 40962, Colletotrichum gloeosporioides KACC 40804, Colletotrichum orbiculare KACC 40808, Didymella bryoniae KACC 40669, Fusarium graminearum KACC 41040, Fusarium oxysporum KACC 40037, Fusarium oxysporum KACC 40052, Fusarium oxysporum f, sp. radicis-Iycopersici KACC 40537, Fusarium oxysporum KACC 40902, Monosporascus cannonballus KACC 40940, Phytophthora camvibora KACC 40160, Rhizoctonia solani AG-1(IA) KACC 40101, Rhizoctonia solani AG-4 KACC 40142, and Scleotinia scleotiorum KACC 41065 by agar diffusion method.

• The Korean Journal of Pesticide Science
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• v.5 no.3
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• pp.26-35
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• 2001

In order to search potent antifungal substances from domestic plants, 40 plants cultivated in Korea were collected. After extracting with methanol (MeOH) and concentrating to dryness, the MeOH extracts were screened for in vivo antifungal activity against six plant diseases at a concentration of $2000{\mu}g/mL$. Fourteen extracts showed disease-controlling activity more than 90% against at least one of the 6 plant diseases tested; eight, seven, and three extracts controlled more than 90% the development of rice blast, tomato late blight, and wheat leaf rust, respectively. However, none of the extracts exhibited in vivo antifungal activity more than 90% against rice sheath blight, tomato gray mold, and barley powdery mildew. From the MeOH extracts of Angelica gigas and A. dahurica showing potent controlling activity against rice blast, 1 and 2 antifungal substances, respectively, were isolated by solvent partitioning and column chromatography. The three compounds were identified to be coumarins, namely, decursin, imperatorin, and isoimperatorin, by mass spectrometry and NMR spectroscopy. They were examined for in vitro and in vivo antifungal activities together with umbelliferone (7-bydroxycournarin) and scopoletin (6-methoxy-7-hydroxycoumarin) containing a free hydroxyl group at position 7 to investigate the structure-activity relationship. In vitro, most of 50% growth inhibitory concentrations ($IC_{50}$) were over $200{\mu}g/mL$, indicating that they have relatively weak antifungal activity. The antifungal activity of decursin and scopoletin, containing cyclic alkoxy groups instead of free hydroxyl group at position 7, was stronger than umbelliferone and scopoletin. Especially, decursin and imperatorin showed potent antifungal activities against Pythium ultimum and Magnaporthe grisea, respectively, with $IC_{50}$ values less than $25{\mu}g/mL$. In vivo, decursin and imperatorin showed potent antifungal activity against rice blast, whereas other coumarins hardly controlled the development of 6 plant diseases tested. These results suggest that the antifungal activity of 7-hydroxycoumarin derivative is substantially increased when the hydroxyl group at position 7 is protected by a stable cyclic alkoxy grouping.

• The Korean Journal of Pesticide Science
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• v.11 no.4
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• pp.331-338
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• 2007

The major fungal diseases which effecting garlic storage are blue mold and dry rot, caused by Penicillium hirsutum and Fusarium oxysporum, respectively. In order to reduce the damage by the pathogenic fungi, here we report the effects of 11 fungicides tested to reduce spoilage during storage of garlics. In the in vitro antimicrobial activity test, the fungicides, diphenylamine, prochloraz and tebuconazole showed 0.3, 2.2, and 1.3 nun inhibition zone to F. oxysporium, and cyprodinil, diphenylamine, fenbuconazole, hexaconazole, penconazole, prochloraz, propiconazole, pyrimethanil and tebuconazole exhibited 0.2, 2.4, 0.8, 0.4, 1.2, 1.5, 1.2, 0.4 and 1.5 mm to P. hirsutum, respectively. To test the in vivo control effect, when the diphenylamine, prochloraz, and tebuconazole were treated by standard concentration, the fungal mycelium of F. oxysporium started to grow 5 days after inoculation, and 80, 63.3 and 83.3% of the inoculated cloves are infected 11 days after inoculation. When the tebuconazole were treated by standard concentration, the P. hirsutum was completely inhibited the growth of the fungi. In case of diphenylamine, penconazole and propiconazole treatment, the P. hirsutum was observed 7 days after inoculation and $20{\sim}23.3%$ of the cloves were infected 11 days after inoculation. When cyprodinil, prochloraz and pyrimethanil were treated, pathogens occurred 5 days after inoculation and $60{\sim}100%$ of the cloves infected 11 days after inoculation. Three fungicides such as diphenylamine, prochloraz and tebuconazole also suppressed remarkably the infection and growth of F. oxysporium and P. hirsutum on garlic when both of the pathogens are inoculated after the garlic cloves were dipped for 10 min in the suspension of each agrochemical. Overall, diphenylamine, prochloraz and tebuconazole showed effective control efficacy on dry rot and blue mold There was significant correlation between in vitro and in vivo assay in diphenylamine and prochloraz to F. oxysporum and cyprodinil, prochloraz and pyrimethanil to P. hirsutum.

• Research in Plant Disease
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• v.21 no.4
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• pp.280-289
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• 2015

Maize (Zea mays L.) is an economically important crop in worldwide. While the consumption of the maize is steadily increasing, the yield is decreasing due to continuous mono-cultivation and infection of soil-borne fungal pathogens such as Fusarium species. Recently, stalk rot disease in maize, caused by F. subglutinans and F. temperatum has been reported in Korea. In this study, we isolated bacterial isolates in rhizosphere soil of maize and subsequently tested for antagonistic activities against F. subglutinans and F. temperatum. A total of 1,357 bacterial strains were isolated from rhizosphere. Among them three bacterial isolates (GC02, GC07, GC08) were selected, based on antagonistic effects against Fusarium species. The isolates GC02 and GC07 were most efficient in inhibiting the mycelium growth of the pathogens. The three isolates GC02, GC07 and GC08 were identified as Bacillus methylotrophicus, B. amyloliquefaciens and B. thuringiensis using 16S rRNA sequence analysis, respectively. GC02 and GC07 bacterial suspensions were able to suppress over 80% conidial germination of the pathogens. GC02, GC07 and GC08 were capable of producing large quantities of protease enzymes, whereas the isolates GC07 and GC08 produced cellulase enzymes. The isolates GC02 and GC07 were more efficient in phosphate solubilization and siderophore production than GC08. Analysis of disease suppression revealed that GC07 was most effective in suppressing the disease development of stalk rot. It was also found that B. methylotrophicus GC02 and B. amyloliquefaciens GC07 have an ability to inhibit the growth of other plant pathogenic fungi. This study indicated B. methylotrophicus GC02 and B. amyloliquefaciens GC07 has potential for being used for the development of a biological control agent.

• The Korean Journal of Pesticide Science
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• v.8 no.4
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• pp.338-346
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• 2004

The methanol extract of Chloranthus japonicus roots effectively controlled the development of rice blast(Magnaporthe grisea), rice sheath blight(Corticium sasaki), tomato pay mold(Btrytis cinerea), tomato late blight(Phytophthora infestans), and wheat leaf rust(Puccinia recondita). From the methanol extract of C. japonicus roots, three antifungal substances were isolated. Their chemical structures were determined to be shizukanols B, C, and D mainly by mass and NMR spectral data. Among the three substances, shizukanol C showed the strongest inhibitory activity against mycelial growth of the plant pathogenic fungi tested; it completely inhibited mycelial growth of M. grisea. Colletotrichum gloeosporioides, and C. acutatum at concentrations of more than $12.5{\mu}g$/ and P. infestans at concentrations of more than $3.13{\mu}g/m\ell$. They also controlled effectively the development of rice blast and wheat leaf rust. On the other hand, they caused phytotoxic symptoms on barley leaves and inhibited the growth of duckweed (Lemna paucicostata) with $EC_{50}$ values of $30.0{\mu}g/m\ell$ for shizukanol B, $49.9{\mu}g/m\ell$ for shizukanol C, and $154{\mu}g/m\ell$ for shizukanol D. In addition, shizukanol C showed an insecticidal activity against brown planthopper (Nilaparavata lugens), peen peach aphid (Myzus persicae), diamond-back moth (Plutella xylostella), and tobacco cutworm (Spodoptera litura) of the 5 arthropod pests tested with mortality values of more than 60% at a concentration of $1,000{\mu}g/m\ell$.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.61 no.1
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• pp.9-16
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• 2016

To investigate the effect of soluble silicate zeolite dressing of the rice against bakanae disease, field trial in reclaimed land and in vitro were carried out. The coated rice seeds (SCS) which were dressed with the mixture of 25% silicic acids (binder), and the zeolite (coating powder). In wet direct seeding, uniform scattering of rice seeds on the soil surface and the better seedling establishment were shown in SCS treatment plots. The incidence of bakanae disease began from the mid tillering stage toward the heading stage. Around heading stage, the ratio of infected tillers reached its highest point by 9.9% in non-SCS treatment plots. While, in SCS treatment plots, the ratio of infected tillers was no more than 0.01%. The vitality of the pathogenic fungi of bakanae disease in the SCS and non-SCS samples were assessed. Samples were incubated for one week keeping proper humidity at $30^{\circ}C$ after inoculated with panicles of infected rice plants from experimental field plots. In non-SCS treatment, pinkish colonies were formed on the grain surface of panicle of infected plants, and mycelium, macro-conidia and micro-conidia were developed actively inside part of infected grain inoculated. While in SCS treatment, micro-conidia and mycelium were not survived and the growth of macro-conidia, mycelia were greatly inhibited and withered. Based on the results, it is concluded that the environmental friendly control of bakanae disease by use of SCS is possible and soluble silicate can be applied as agents for replacement of seed disinfection.