• Mycobiology
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• 제35권2호
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• pp.54-61
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• 2007

Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

• The Plant Pathology Journal
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• 제26권4호
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• pp.394-401
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• 2010

Resin flow, symptom development, and lignin biosynthesis in response to wounding and fungal inoculation were investigated in Pinus rigida and Pinus densiflora. The two-year-old seedling stems were subjected to three types of treatments: (i) wounding without inoculation, (ii) wound-inoculation with a conidial suspension of Fusarium circinatum, and (iii) pre-wounding woundinoculation with the fungus 20 days after the initial wounding. Resin flow from wounding sites was more evident in P. rigida than P. densiflora in all treatments. The wound-inoculation with the fungus induced almost two-fold higher levels of resin flow than the other treatments in both species. The pre-wounding woundinoculation appeared to result in a decrease in pitch canker development in the two pine species. Some reductions in disease severity were observed in the prewounding wound-inoculated P. rigida, showing a mean disease severity of less than 85%, compared with approximately 100% in the wound-inoculated stems. Disease severity was approximately 50% in the woundinoculated P. densiflora, whereas 10% in the pre-wounding wound-inoculated stems. Higher amounts of lignin were found from bark (ca. 40%) than from xylem (ca. 30%). The wound-inoculated bark and the pre-wounding wound-inoculated bark exhibited higher amounts of lignin among the other treatments. These results suggest that the wound-inoculation apparently prompt the increase in resin flow and lignin biosynthesis from the two pine species, and the prior wounding may be involved in decreased disease severity against the further invasion of F. circinatum.

• 한국잠사곤충학회지
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• 제44권2호
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• pp.74-81
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• 2002

The optimal conditions for mycelial growth of P. linteus ASI 26011 were 25-30$^{\circ}C$ and pH 6.0, respectively. The mycelial growth of P. linteus was excellent on MCM medium. In case of carbon sources, the mycelial growth of P. linteus was best on the culture media that were contained with sucrose, mannose and glucose. Potassium nitrate and sodium nitrate were good for the mycelial growth of P. linteus as a nitrogen source. For comparison of the mycelial colonization of P. linteus on logs, several techniques of inoculation were tested; the sterilized short log inoculation, drilling inoculation and log-end sandwich inoculation. The mycelial colonization of P. linteus on logs was good in the treatment of sterilized short log inoculation, but poor in the traditional methods such as drilling inoculation and log-end sandwich. The initial mycelial growth and the full mycelial colonization of P. linteus were the best on 20 cm logs under the condition of 42% of moisture content in log. Also the initial mycelial growth of P. linteus was accelerated over 12 hours of sterilization. Burying method of logs after 5-6 months of incubation was the best for formation of basidiocarp of P. linteus. The formation of fruiting body of P. linteus was quite good in the cultivation house at the 31-35$^{\circ}C$ and over 96% of relative humidity.

• 한국주조공학회지
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• 제20권1호
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• pp.46-56
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• 2000

• IMMUNE NETWORK
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• 제6권4호
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• pp.185-191
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• 2006

Background: Based on outstanding progresses in animal experiments, vaccines for some human tumors have been developed. However, clinical effects of these vaccines have been far below than expected. This discrepancy might come from differences between animal models and human patients with respect to immunocompetency. The immune status of mice after tumor inoculation has not been well studied, which make us cautious in interpreting and applying the results from mice to human. We evaluated cell-mediated immune responses in mice after tumor cell inoculation. Methods: Mice were inoculated with TA3Ha, CT26, or 4T1. Delayed-type hypersensitivity (DTH) responses were induced 2-4 weeks after inoculation using 2,4-dinitro-1-fluorobenzene as an antigen. The relationships between the severity of DTH responses and the duration of tumor inoculation or the size of tumor mass were analyzed. Results: In T A3Ha groups, DTH response was elevated 2 weeks after inoculation, but depressed after 4 weeks, compared to the control group. When analyzed based on the sizes of tumor masses elicited, DTH responses were inversely related to the mass size, especially in those greater than 10 mm in diameter. In CT26 groups, while the duration after inoculation did not affect the severity of DTH responses, those with large mass showed depressed responses regardless the duration of inoculation. 4T1 cells grew so slowly that the size of tumor mass was small even 4 weeks after inoculation, and this group showed much higher DTH responses compared to that of tumor-free group. Conclusion: At least in an experimental setting where tumor model was induced by inoculating tumor cell lines into immunologically competent mice, the host immune response was elevated in early stage, and then depressed in late stage when the mass grew over a critical size.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.69.2-70
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• 2003

For the selection and breeding of chestnut varieties resistant to the chestnut blight fungus Cryphonectria parasitica, disease resistance and susceptibility of 28 varieties widely planted and growing in Korea were evaluated by artificial inoculation of a pathogenic fungus. For this experiment, a typical virulent strain (KCPC-19) was selected. Artificial inoculation was conducted into all varieties by using two different materials and methods, i.e., bark and wood tissue sections in the laboratory and living trees in the field. In the bark and wood tissue section method, the size of necrotic area and canker development on chestnut varieties were examined and compared 4 days after inoculation. There were wide variations of chestnut varieties in disease resistance and susceptibility against chestnut blight fungus, but 3 varieties, Daebo！, Ishizuchi, and Sandae, were shown to be relatively resistant to the disease with the necrotic area of 0.95-1.03 cm2, while Arima was the most susceptible with the size of 2.0 cm2. In the living tree inoculation examined 5 weeks after inoculation, 3 varieties, Daebo, Ishizuchi, and Riheiguri, showed the higher resistance, but Tono 2 did the highest susceptibility among tested varieties.

• 대한수의학회지
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• 제33권2호
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• pp.241-247
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• 1993

Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

• Journal of Microbiology
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• 제45권1호
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• pp.15-20
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• 2007

The majority of soil microorganisms can derive ethylene from L-methionine (L-MET), while some rhizobacteria can hydrolyze 1-aminocyclopropane-1-carboxylate (ACC) due to their ACC-deaminase activity. In this study, three strains having either ACC-deaminase activity (Pseudomonas putida biotype A, $A_7$), or the ability to produce ethylene from L-MET (Acinetobacter calcoaceticus, $M_9$) or both (Pseudomonas fluorescens, $AM_3$) were used for inoculation. The highly ethylene specific bioassay of a classical 'triple' response in pea seedlings was used to investigate the effect of the inoculation with the rhizobacteria in the presence of 10 mM ACC or L-MET. The exogenous application of ACC had a concentration-dependent effect on the etiolated pea seedlings in creating the classical 'triple' response. The inoculation with P. putida diluted the effect of ACC, which was most likely due to its ACC-deaminase activity. Similarly, the application of $Co^{2+}$ reduced the ACC-imposed effect on etiolated pea seedlings. In contrast, the inoculation of A. calcoaceticus or P. fluorescens in the presence of L-MET caused a stronger classical 'triple' response in etiolated pea seedlings; most likely by producing ethylene from L-MET. This is the first study, to our knowledge, reporting on the comparative effect of rhizobacteria capable of utilizing ACC vs L-MET on etiolated pea seedlings.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.938-945
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• 2018

Salinity stress is an important environmental problem that adversely affects crop production by reducing plant growth. The impacts of rhizobacterial strains to alleviate salinity stress on the germination of Lactuca sativa and Raphanus sativus seeds were assessed using different concentrations of NaCl. Plant growth-promoting rhizobacteria (PGPR) strains were also examined to improve the early germination of Chinese cabbage seeds under normal conditions. Lactobacillus sp. and P. putida inoculation showed higher radicle lengths compared with non-inoculated radish (Raphanus sativus) seeds. LAP mix inoculation increased the radicle length of lettuce (Lactuca sativa) seedlings by 2.0 and 0.5 cm at salinity stress of 50 and 100 mM NaCl concentration, respectively. Inoculation by Azotobacter chroococcum significantly increased the plumule and radicle lengths of germinated seeds compared with non-inoculated control. A. chroococcum increased the radicle length relative to the uninoculated seeds by 4.0, 1.0, and 1.5 cm at 50, 100, and 150 mM NaCl concentration, respectively. LAP mix inoculation significantly improved the radicle length in germinated radish seeds by 7.5, 1.3, 1.2, and 0.6 cm under salinity stress of 50, 100, 150, and 200 mM NaCl concentration, respectively. These results of this study showed that PGPR could be helpful to mitigate the salinity stress of different plants at the time of germination.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.1971-1981
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• 2018

In this study, we investigated the altered enzymatic activities and metabolite profiles of koji fermented using varying permutations of Aspergillus oryzae and/or Bacillus amyloliquefaciens. Notably, the protease and ${\beta}$-glucosidase activities were manifold increased in co-inoculated (CO) koji samples (co-inoculation of A. oryzae and B. amyloliquefaciens). Furthermore, gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling indicates that levels of amino acids, organic acids, sugars, sugar alcohols, fatty acids, nucleosides, and vitamins were distinctly higher in CO, SA (sequential inoculation of A. oryzae, followed by B. amyloliquefaciens), and SB (sequential inoculation of B. amyloliquefaciens, followed by A. oryzae). The multivariate principal component analysis (PCA) plot based on GC-MS datasets indicated a clustered pattern for MA and MB (koji samples inoculated either with A. oryzae or B. amyloliquefaciens) across PC2 (20.0%). In contrast, the CO, SA, and SB metabolite profiles displayed segregated patterns across PLS1 (22.2%) and PLS2 (21.1%) in the partial least-square discriminant analysis (PLS-DA) model. Intriguingly, the observed disparity in the levels of primary metabolites was engendered largely by higher relative levels of sugars and sugar alcohols in MA, SA, and CO koji samples, which was commensurate with the relative amylase activities in respective samples. Collectively, the present study emphasizes the utility of integrated biochemical and metabolomic approaches for achieving the optimal permutation of fermentative inocula for industrial koji preparation.