• Journal of Microbiology
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• v.43 no.5
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• pp.417-423
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• 2005

In this study, the nematode-trapping fungus, Monacrosporium sphaeroides, was transformed with a plasmid harboring the hygromycin B phosphotransferase gene, via restriction enzyme-mediated integration (REMI). Frequencies of up to 94 transformants ${\mu}g^{-1}$ per linearized plasmid DNA were obtained by optimizing the PEG concentration, as well as the category and quantity of the added restriction enzyme. $90\%$ of the transformants were determined to be stable for drug resistance when 20 randomly selected transformants were tested. Southern analyses revealed that the transforming DNA was integrated into the M. sphaeroides genome either with or without rearrangement. Five mitotic stable mutant strains were obtained using this approach, all of which had been altered with regard to sporulation capacity and pathogenicity toward nematodes. Southern blot analyses of the five mutants revealed that foreign plasmid DNA had integrated into the genome. Three of the mutants, Tms2316, Tms3583 and Tms1536, exhibited integration at a single location, whereas the remaining two, Tms32 and Tms1913, manifested integration at double or multiple locations. Our results suggest that the transformation of M. sphaeroides via REMI will facilitate insertional mutagenesis, the functional analysis of a variety of genes, and the tagging or cloning of genes of interest.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1994.06a
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• pp.27-49
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• 1994

Root colonization of biocontrol agents via seed treatment was investigated and a compatible combination, Gliocladium virens G872B and Pseudomonas putida Pf3, in colonizing cucumber rhizosphere was confirmed through the study. Much higher number of fungal and bacterial propagules were detected when two isolates were inoculated together. The presence of Pf3 in root system was greatly helpful to G872B to colonize at root tip. The mechanism of this phenomenon is partially elucidated through the results of in vitro experiments and the observations of scanning electron and fluorescence microscope. Addition of Pf3 cells resulted earlier germination of G872B conidia and increased mycelial growth. And the more number of germinated conidia on seed coat, the more vigorous hypal streching and sporulation on the root surface were observed in coinoculated treatment. The propagules of G872B on the cucumber root when they were challenged against the pathogenic Fusarium oxysporum, were even higher than that of G872B treated alone, and the magnitude of such a difference was getting grater toward the root ip and the population of F. oxysporum on the root was reduced by seed inoculation of G872B. The rhizosphere competence was obviously reflected to disease suppression and plant growth promotion that induced by the given isolate. Green house experiments revealed that the combined treatment provided long-term disease suppression with greater rate and the larger amount of fruit yield than single treatments. Through this study the low temperature growing Pseudomonas fluorescens M45 and MC07 were evaluated to apply them to the winter crops in field or plastic film house. In vitro tests reveal that M45 and MC07 inhibited the mycelial growth of Pythium ultimum, Rhizoctona solani and Phytophthora capsici and enhanced growth of cucumber cotyledon in MS agar. This effect was more pronounced when the bacteria were incubated at 14$^{\circ}C$ than at 27$^{\circ}C$. And disease suppression and plant growth promotion in green house were also superior at low temperature condition. Seed treatment of M45 or soil treatment of MC07 brought successful control of damping-off and enhanced seedling growth of cucumber. The combined treatment of two isolates was more effective than any single treatment.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.202-210
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• 2022

Aflatoxin contamination in rice has been documented in a number of studies, and has a high incidence in Asian countries, and as such, there has been a growing interest in alternative biocontrol strategies to address this issue. In this study, 147 strains of yeasts and yeast-like fungi were screened for their potential to produce volatile organic compounds (VOCs) active against Aspergillus flavus strains that produce aflatoxin B₁ (AFB₁). Five strains within four different genera showed greater than 50% growth inhibition of some strains of A. flavus. These were Anthracocystis sp. DMKU-PAL124, Aureobasidium sp. DMKU-PAL120, Aureobasidium sp. DMKU-PAL144, Rhodotorula sp. DMKU-PAL99, and Solicococcus keelungensis DMKU-PAL84. VOCs produced by these microorganisms ranged from 4 to 14 compounds and included alcohols, alkenes, aromatics, esters and furans. The major VOCs produced by the closely related Aureobasidium strains were found to bedistinct. Moreover, 2-phenylethanol was the most abundant compound generated by Aureobasidium sp. DMKU-PAL120, while methyl benzeneacetate was the major compound emitted from Aureobasidium sp. DMKU-PAL144. On the other hand, 2-methyl-1-butanol and 3-methyl-1-butanol were significant compounds produced by the other three genera. These antagonists apparently inhibited A. flavus sporulation and mycelial development. Additionally, the reduction of the AFB1 in the fungal-contaminated rice grains was observed after co-incubation with these VOC-producing strains and ranged from 37.7 ± 8.3% to 60.3 ± 3.4%. Our findings suggest that these same microorganisms are promising biological control agents for use against aflatoxin-producing fungi in rice and other agricultural products.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.912-920
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• 2021

SOS response is a conserved response to DNA damage in prokaryotes and is negatively regulated by LexA protein, which recognizes specifically an "SOS-box" motif present in the promoter region of SOS genes. Myxococcus xanthus DK1622 possesses a lexA gene, and while the deletion of lexA had no significant effect on either bacterial morphology, UV-C resistance, or sporulation, it did delay growth. UV-C radiation resulted in 651 upregulated genes in M. xanthus, including the typical SOS genes lexA, recA, uvrA, recN and so on, mostly enriched in the pathways of DNA replication and repair, secondary metabolism, and signal transduction. The UV-irradiated lexA mutant also showed the induced expression of SOS genes and these SOS genes enriched into a similar pathway profile to that of wild-type strain. Without irradiation treatment, the absence of LexA enhanced the expression of 122 genes that were not enriched in any pathway. Further analysis of the promoter sequence revealed that in the 122 genes, only the promoters of recA2, lexA and an operon composed of three genes (pafB, pafC and cyaA) had SOS box sequence to which the LexA protein is bound directly. These results update our current understanding of SOS response in M. xanthus and show that UV induces more genes involved in secondary metabolism and signal transduction in addition to DNA replication and repair; and while the canonical LexA-dependent regulation on SOS response has shrunk, only 5 SOS genes are directly repressed by LexA.

• The Korean Journal of Mycology
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• v.6 no.2
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• pp.1-10
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• 1978

In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

• The Korean Journal of Mycology
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• v.2 no.1
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• pp.1-6
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• 1974

1. The mycelial growth and sporulation of Mycogone perniciosa was compared on modified Czapek's media deficient in carbon source, nitrogen source, K,Mg, P or the heavy metal elements. The mycelial growth was significantly reduced in solution cultures lacking Mg, K or P and only a trace of growth occurred in solutions lacking carbon source or nitrogen source. Most sparse sporulation and smaller chlamydospores than on any of deficient agar media occurred on agar media dificient in carbon source or nitrogen source. 2. In both potato dextrose agar and malt extract solution, growth of the fungus was optimum at $25^{\circ}C$, and undetectable at $10^{\circ}C$ and $35^{\circ}C$. 3. Optimum pH for growth of this fungus was 7.0. 4. This fungus was killed in soil when exposed to $50^{\circ}C$ or higher for 20 minutes or more.

• The Korean Journal of Mycology
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• v.23 no.3 s.74
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• pp.246-256
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• 1995

Phenotypic characteristics, pathogenicity and fungicides resistance of fifty one isolates of Botrytis cinerea obtained from various host plants were observed and determined. The relationships between these characteristics were also investigated on the basis of isolation host plants. The isolates of B. cinerea varied in the capacity of sclerotia formation and sporulation. The pathogenicity of 44 isolates from tomato, cucumber, and strawberry was significantly stronger with 3.2 cm in average diameter of necrotic lesions on cucumber leaves than that of seven isolates from other host plants such as orange, gerbera, ginseng, kiwi, grape, pear and from butter with 1.8 cm in average diameter of necrotic lesions. Benomyl resistance of 12 isolates from tomato plants was much higher with the $EC_{50}$, 562 ppm than that of 19 isolates from various host plants. Diethofencarb resistance, however, of 11 isolates from strawberry plants was highest with the $EC_{50}$, 210 ppm among isolates from other host plants. Polygalacturonase activity varied among isolates in the range of 0 to 103 unit and that of isolates from tomato, cucumber and strawberry was slightly lower than that of isolates from other host plants. No significant relationship between pathogenicity and fungicides resistance, polygalacturonase activity was found among 51 isolates of B. cinerea. Isozyme patterns of polygalacturonase produced from two strongly and weakly pathogenic isolates (FC122, KC6) were slightly different depending upon carbon sources during cultivation.

• Korean Journal Plant Pathology
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• v.1 no.1
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• pp.79-84
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• 1985

Some characteristics of the resistance induced by Erysiphe graminis f. sp. hordei on near-isogenic barley leaves were evaluated. Inoculation of heat-killed incompatible inducer conidia did not protect barley leaves against compatible challenger race when the inducer inoculum was removed prior to inoculation of challenger. However, the colony formation of challenger race was greatly reduced by 87.1 to $91.2\%$ when the heat-killed inducer inoculum was not removed from the leaves. Although incompatible inducer conidia were removed before they penetrate the host cell, colony formation of challenger was markedly decreased without change in its infection type. After penetrating the host cell by inducer, however, a change in infection types occurred on the challenged leaves. Irrespective of compatibility of previously inoculated inducer on middle part of leaves, there was no reduction in colony formation of challenger race both on the adjacent acropetal and basipetal parts of the same leaves free of inducer inoculation. The colonies formed on the basipetal part by challenger race showed normal 4 type, whereas the infection type of colonies formed on the acropetal part was somewhat changed, thereby sporulation being reduced. The possibility of translocation of resistance-inducing factors was discussed.

• KSBB Journal
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• v.16 no.1
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• pp.30-35
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• 2001

High cell density culture of Bacillus thuringiensis was conducted in fed-batch culture and TCRC using a bioreactor incorporating ceramic membrane filter. Cell growth of B. thuringiensis in fed-batch culture increased linearly, which was well matched by the results of cell growth modeling. In spite of the slower growth rate during fed-batch culture, no spore formation was observed, which was contrary to the results of continuous culture. Changing culture mode to batch culture after fed-batch operation induced a 2.7$\times$$10^9$ CFU/mL spore concentration using a 300 g/L glucose feed concentration. In TCRC operation incorporating ceramic filter within the bioreactor, the effect of glucose feed concentrations on the cell growth and spore formation of B. thuringiensis was determined. A maximum cell concentration of 1.8$\times$$10^{10}$ CFU/ml, which corresponds to 82.6 g-cell/L, was obtained in the TCRC using a 50 g/L glucose feed concentration. In the TCRC, cell growth increased linearly and glucose concentration was limited, which agreed well with the results of cell growth modeling. No spore formation was observed except when 1 g/L of glucose was fed. Changing to batch culture induced a 1.2$\times$$10^{10}$ CFU/mL of spore concentration, which was the highest spore concentration obtained among the various culture modes examined. The optimal glucose feed rate was found to be 0.55 g-glucose/h.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.27 no.4
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• pp.361-370
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• 1982

The effects of climatic factors on organisms lire variable and complex, and it, however, can be interpreted in terms of those on the distribution and those on the population densities. The distribution of an organism may largely be determined by the temperatures, except some temporal organisms which are depended on the air mass movements. Population density of an organism is determined by various climatic factors, such as previous winter temperature, temperature of growing season and rainfall. The start of growing season of the rice plants has been shifted to earlier since last decade in Korea. This may mean that the overall climatic condition during the growing season might be considerably different from those in past years, and such a difference in climatic conditions might have close relation with the recent status of the diseases and insect pests through direct effects on the physiology and population dynamics of the organisms, as well as through on the biotic associations of the pest organisms. The white back planthopper and brown planthopper have become the key insect pests in Korea in recent years. They are migratory and have high reproductive pontentials and more generations than average residential insects. The synchronization of the migrants and physiological condition of the rice plants seems to be the important factors in relation to the recent outbreaks of these insects; the high reproductive rate can be obtained with the growth stage of rice being 30-50 days after transplanting. The modication of the microclimate associated with high plant density and some other introduced new cultural techniques also have some relation with the outbreak. The key diseases of the rice are the blast disease, sheath blight and the bacterial leaf blight. For the rice blast, the seedling blast and leaf blast during the early growing season and the neck blast, have become more serious, the former may be related to hotbed nursery and the later may be related to the high humidity in early August, and synchronization of the heading time which has been shifted to early part from middle or late part of August. In general, for the rice diseases, the development of the new races have been the most serious which are largely resulted from the introduction of the new varieties, but it also seems to be related with the prolonged periods of the favorable condition associated with the shifted growing seasons. In general, the diseases and insect pest problems have become much more variable and complex, and control measures should be based on the thorough knowledge of the ecology of the pest organisms, that is, effects of various environmental factors on the disease cycle; spore release, spore deposition, infection, colonization and sporulation of the disease organisms, and those on the development, reproductive potentials, dispersal, age specific responses of the insects. The well organized real-time pest management systems, such as alfalfa weevil management system developed at the Purdue University in U.S., is the prime importance for the implementation of the pest management principles.