• The Plant Pathology Journal
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• v.19 no.2
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• pp.79-84
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• 2003

Sclerotinia rot occurred severely on some vegetable crops grown in Namyangju, Yangpyung, and Yangiu areas in Korea in 2001-2002. The crops infected with Scterotinia sp. were Adenophora remotiflora, Armoracia lapathfolia, Angelica acutiloba, Angelica archangelica, Anthriscus sylvestris, Aster tataricus, Beta vulgaris var. cicla, Brassica campestris var. marinosa, Brassica juncea var. laciniata, Chicholium intybus, Lactuca indica var. dracoglossa, Lactuca sativa var. oak-leaf, Petroselinum crispum, and Phyteuma japonicum. The fungus associated with the disease was identified as Sclerotinia sclerotiorum, based on the morphological characteristics of the pathogen. The symptoms were water-soaked spots that enlarged later and became a watery soft rot. Infected parts became yellow and then turned brown, followed by death of the whole plant. White mycelia developed on the upper petioles and leaves and on the soil where these plant parts lay. Then black sclerotia in variable size and shape formed from the mycelial mass. Pathogenicity of the fungus was proven by artificially inoculating each crop. This is the first report of Sclerotinia rot on the listed vegetable crops in Korea.

• Journal of Applied Biological Chemistry
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• v.44 no.2
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• pp.77-83
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• 2001

For defining the possibilities of the commercial mass liquid culture of Cordyceps sinensis, the pharmacological activities of mycelia were analyzed. The mycelium of C. sinensis consists of carbohydrate (5.1%) and fat (1.3%), and contains a low content of protein (0.7%) and ash (0.5%), and 92.4% moisture. The molecular sugar ratio of carbohydrate was composed mainly of glucose, mannose (1.0 : 0.9), in addition a small amount of galactose and arabinose (0.2 : 0.1). The cellular materials of mycelia were fractionated into ethylacetate (EA), MeOH (M) and hot-water extract fraction (HW). HW fraction showed the most potent intestinal immune system modulating activity, anti-coagulant activity, and anti-complementary activity, and M fraction had the inhibition activity of radical generation as effective as genistine. These results reveal that the mycelium of liquid cultured C. sinensis showed pharmacological activities and could be used for commercial purpose.

• Mycobiology
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• v.29 no.3
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• pp.135-141
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• 2001

Differential display of reverse transcription(DDRT)-PCR was conducted to have a profile of the differentially expressed genes during the formation of fruiting body of Lentinus edodes. The lines of L. edodes(ImHyup-1) employed were cultivated in the artificial blocks of sawdust, and the fruiting body was induced from the mycelia or the mass protruded from the brown surface of the sawdust blocks. RNAs were prepared from the four different developmental stages; mycelial, primordial, and stipes and pileus of fruiting body. The fragments of cDNA were synthesized from the combinations of the arbitrary primers and 3' one anchored Oligo-dT primer. Twelve combinations using the primers have been tested, and among them nineteen bands were identified as differentially expressed. Those genes were further analyzed by DNA sequencing and followed by homology search. Characterization of one clone was conducted as a preliminary data and more are under investigation.

• The Korean Journal of Mycology
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• v.44 no.3
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• pp.166-170
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• 2016

Ectomycorrhizal fungi are associated with plants roots and acquire significant amounts of nitrogen sources from the soil. For artificial cultivation, mass production of ectomycorrhizal fungi in liquid media is required. We studied the edible ectomycorrhizal mushrooms Hygrophorus russula, Ramaria fumigata, Sarcodon aspratus, and Tricholoma matsutake. All strains except S. aspratus NIFoS 2031 grew generally well on modified Melin-Norkran's (MMN) medium compared to on other media. All strains analyzed in this study showed significantly higher growth on organic nitrogen. Specifically, two strains of H. russula significantly responded to both tryptone and neopeptone media. Among different species and strains, there were clear differences in the capacity to grow on animal-based organic nitrogen sources.

• Research in Plant Disease
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• v.15 no.3
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• pp.222-229
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• 2009

A Pseudomonas strain SG3 producing biosurfactant and showing antifungal and insecticidal activities was isolated from agricultural soil severely contaminated with machine oils. The antagonistic bacterium inhibited mycelial growth of all of the tested fungal pathogens. The fermentation broth of SG3 also effectively suppressed the development of various plant diseases including rice blast, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew and red pepper anthracnose. An antifungal substance was isolated from the fermentation broth of SG3 by ethyl acetate partitioning, silica gel column chromatography and preparative HPLC under the guide of bioassay. The chemical structure of the antifungal substance was determined to be rhamnolipid B by mass and NMR spectral analyses. The antifungal biosurfactant showed a potent in vivo antifungal activity against gray mold and late blight on tomato plants. In addition, rhamnolipid B inhibited mycelial growth of B. cinerea causing tomato gray mold and zoospore germination and mycelial growth of P. infestans causing tomato late blight. Pseudomonas sp. SG3 producing rhamnolipid B could be used as a new biocontrol agent for the control of plant diseases occurring on tomato plants.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.37-37
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• 2015

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

• The Korean Journal of Mycology
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• v.21 no.2
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• pp.94-105
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• 1993

The genus Cordyceps known as an insect parasite forms a sclerotium in insect bodies and then produces perithecia on the single or multiple stromata produced from sclerotium. Collected Cordyceps were identified into 5 species: Cordyceps militaris, C. nutans, Cordyceps sphecocephala, Isaria japonica, and Torrubiella sp. The fruit bodies of Cordyceps in petri-dish cover were fixed by tape and put the lid on water agar plates to isolate these collected Cordyceps. The germinated spores were transferred from water agar to Potato dextrose agar(PDA) after six hours. Mycelial growth of C. nutans and C. militaris was the most successful on Hamada media and was also good on Complete media and PDA. Mannose as a carbon source was good for two species and Glutamic acid as a nitrogen source was satisfactory to C. militaris and Asparagine gave a good result to C. nutans. C. militaris and C. nutans showed similar mycelial growth rate on the media that contained thiamine-HCI, biotine or nicotinic acid as a vitamine. When conidia of C. nutans were inoculated to insects, mortality was high in Artogeia napi L, Hemiptera, Plutella xylostella and 50% in Orthoptera, 12% in Acantholyda posticalise M, but not Agelastica coerulea B. in Aphididae, C. nutans was collected from only Hemiptera in nature, but killing effect on other insects was proved. Mycelial growth and fruit-body formation were good on the media that consist of rice powder 5g, wheat flour 5g, water 100ml, but formed fruit-body was not complete stromata but a mass of conidia according to results of observing microscope.

• Journal of Life Science
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• v.11 no.3
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• pp.235-241
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• 2001

To select the sntagonistic bacteria against B. cinerea, isolates were screened from the eggplant leaves and rhizosphere soils in the eggplnat fields in the greenhouse. W1 and P99 isolates were selected by the inhibition of mycelial growth of B. cinerea E12 in vitro test. These isolates, W1 and P99, were identified as Bacillus subtilis and Pseudomonas putida, respectively, by the Bergeys manual and API systems, For the formulation of the antagonistic bacteria, the media for the mass production were prepared with biji(soybean curd residues) or soybean flour. B. subtilis W1 or P. putida P99 was mass cultured in biji broth or soybean flour extrect broth and then soybean flour, corn starch flour, rice glutinous flour and biji flour as high molecular substrates were added. These mixtures were dried, grinded and formulated as brofungicides of wettable powder type. The assess the control effect of biofungicides against the infection of B. cinerea, six types of formulations were assayed at the pot culturing with eggplant in the greenhouse. According to the results, there were no significant differences among the formulation methods. However, P99S or PppB formulated with P. putida P99 showed the highest control values as 90.4% and 96.1%, respectively. Then. BSB or BSD formulated whit B. subtilis W1 were 80.8% and 83.0%, respectively. There afforementioned values were more effective than that of chemical fungicide. Ipro W.P which showed as 72.6%.

• KSBB Journal
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• v.25 no.2
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• pp.155-166
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• 2010

Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.

• KSBB Journal
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• v.11 no.1
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• pp.22-29
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• 1996

Immobilized bioprocess was carried out for continuous production of cyclosporin A (CyA) produced intracellularly as a secondary metabolite by a filamentous fungus, Tolypocladium inflatum. Immobilization procedure for entrapping conidiospores of the producer was significantly simplified by use of a modified immobilization technique. A newly-designed immobilized perfusion reactor system (IPRS) showed good process benefits as demonstrated by the role of the high density immobilized cells as an efficient biomass generator, continuously supplying highly active CyA-producing free cells (1.0g/$\ell$/hr) even at very high dilution rate ($0.1hr^{-1}$). IPRS bioprocess was possible since efficient decantor system developed in our laboratory separated the sloughed-off free cells from the immobilized biomass effectively, thus overcoming wash-out phenomenon frequently encountered in continuous free cell cultures. Furthermore the released-free cells remaining in the bulk solution did not appear to cause substrate mass transfer limitation which was often experienced in suspended mycelial fungal cell fermentations. The primary reason for this was that the suspension broth of the IPRS mainly consisted of roundshaped short mycelial fragments and conidiospores, still remaining Newtonian even at high cell density. In parallel with IPRS bioprocess development, other key factors to be considered necessarily for significant increase in CyA productivity would be strain improvement and medium optimization for the immobilized cells.