• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.490-496
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• 1998

Aspergillus oryzae producing high proteolytic enzyme was isolated from soybean koji and named tentatively A. oryzae O-1. A. oryzae U-1 was obtained by mutation of A. oryzae O-1 with ultraviolet (UV)-irradiation and produced 14 times higher pretense activity compared with A. oryzae O-1. A. oryzae E-1 was acquired by treatment of A. oryzae U-1 with 0.5 M ethylmethanesulfonate (EMS) for 6 min at 3$0^{\circ}C$ and produced 39 times higher proteolytic activity than A. oryzae O-1. With protoplast fusion between A. oryzae O-1 and A. oryzee E-1 in the presence of polyethylenegylcol (PEG)-CaCl$_2$, proteolytic activity was increased to 82 times compared to A. oryzae O-1, and the fusant was named A. oryzae PF. The activities of the cultures containing proteolytic enzymes produced by the strains were determined to be 0.23 U/$m\ell$ for A. oryzae O-1, 3.29 U/$m\ell$ for A. oryzae U-1, 8.91 U/$m\ell$ for A. oryzae E-1, and 19.0 U/$m\ell$ for A. oryzae PF.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.229-233
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• 2004

The bacterial leaf blight, which is caused by Xantho-monas oryzae pv. oryzae, is the most damaging and intractable disease of rice. To identify the genes involved in the virulence mechanism of transposon TnS complex, which possesses a linearized transposon and transposase, was successfully introduced into X. oryzae pv. oryzae by electroporation. The transposon mutants were selected and confirm the presence of transposition in X. oryzae pv. oryzae by the PCR amplification of transposon fragments and the Southern hybridization using these mutants. Furthermore, transposon insertion sites in the mutant bacterial chromosome were deter-mined by direct genomic DNA sequencing using transposon-specific primers with ABI 3100 Genetic Analyzer. Efficiency of transposition was influenced mostly by the competence status of X. oryzae pv. oryzae cells and the conditions of electroporation. These results indicated that the insertion mutagenesis strategy could be applied to define function of uncharacterized genes in X. oryzae pv. oryzae.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.218-223
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• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

• Korean Journal of Agricultural Science
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• v.11 no.1
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• pp.8-24
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• 1984

In this study, investigated the effects of four strains belonging to Aspergillus oryzae on the quality of Kochujang. In the Koji and Kochujang making, investigated the difference of enzyme production of each strain, the change of each component and color during the aging, and tested the sensory taste. The results obtained were as follows; 1. Protease activity (acid, neutral) in wheat flour Koji was high in the case of Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type. The strain Aspergillus $oryzae-S_1$ showed maximum activity after two days of Koji making, while the strain Aspergillus oryzae - M showed low activity till two days, but showed maximum activity after three days-four days of Koji making. 2. In $\alpha$-amylase activity, strain Aspergillus $oryzae-S_1$, Aspergillus $oryzae-S_2$ and Aspergillus oryzae - M showed high activity after two days of Koji making. Aspergillus oryzae-NB strain showed slower ${\alpha}$ - amylase activity than that strains. 3. In glucoamylase activity, all strain tested showed high activity after three days of Koji making, but st rain Aspergillus oryzae - NB showed slower activity than ot - hers. 4. In protease activity (acid, neutral) during the aging of Kochujang, strain Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type showed higher activity than that of long stalked type. 5. Amino type nitrogen contents during the aging of Kochujang was very higher in the case of strains Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type than other strains, and each contents was 315mg% and 337mg% after aged for ninty days. 6. The results that analysed free sugar of Kochujang aged for ninty days with HPLC were; glucose 5.84~7.13%, fructose 4.13~5.00%, rhamnose 0.91~1.04%, maltose 0.72~0.92% and presence of xylose was recognized. 7. The results that analysed alcohols of Kochujang aged for ninty days with gas chromatography were; ethanol 1.51~1.78%, n-propyl alcohol 1.13~1.20mg%, iso-amyl alcohol 3.5~4.4mg%. 8. In the sensory test of Kochujang aged for sixty days and for ninty days, the case of strains Aspergillus oryzae-M and Aspergillus $oryzae-S_1$ of short stalked type showed good taste, while the case of strains Aspergillus $oryzae-S_2$ and Aspergillus oryzae-NB of long stalked type showed good flavor and color.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.188-194
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• 1993

Anastomosis types and hyphal interactions in culture among different location and field isolates of Rhizoctonia solani AG-1(IA), R. oryzae and R. oryzae-sativae were examined. In the pairings of R. solani AG-1(IA) isolates, cytoplasmic fusion only occurred in the self-anastomoses, and non-cytoplasmic fusion occurred in the other combinations. In the pairings of R. oryzae isolates, cytoplasmic fusion occurred in six combinations between different location isolates and in two combinations between different field isolates from the same locations as well as in the self-anastomoses. In that case, four isolates of the fungus reciprocally made the cytoplasmic fusion. In the pairings of R. oryzae-sativae isolates, only non-cytoplasmic fusion occurred among the different location and field isolates, in which cytoplasmic fusion also occurred in the self-anastomoses. When non-cytoplasmic fusion isolates(NCFIs) of R. solani AG-1(IA) were opposed on PDA, a killing zone developed between the NCFls paired after incubation. The killing zone also developed between the NCFls of R. oryzae paired. No killing zone developed between the cytoplasmic fusion isolates(CFIs) of R. oryzae, in which mycelia of the CFIs intermingled with each other without formation of any demarcation line. An entangled zone instead of the killing zone developed between the NCFIs of R. oryzae-sativae.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.256-264
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• 2004

A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.873-878
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• 2002

In order to industrialize traditional Meju-tasting Koji, the characteristics of Koji manufactured with bacteria and fungi found in traditional Meju were investigated. Bacillus subtilis B-4 and Aspergillus oryzae F-5 showing high enzyme activities including those of amylase and protease were used. L-value of Koji manufactured with B. subtilis B-4 had darker color and higher enzyme production than A. oryzae F-5 made one. B. subtilis B-4 made Koji showed higher enzyme production and sensony evaluation score than A. oryzae F-5 Koji. A. oryzae F-5 Koji showed superior color to B. subtilis B-4 Koji. Activity in color, capacity of enzyme production, viable cell count, and sensory evaluation of water activity controlled Koji was superior to the uncontrolled one.

• Korean Journal of Microbiology
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• v.3 no.2
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• pp.9-14
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• 1965

The enzyme-producing potentials of industrially important strains of Aspergillus spp. were studied. Irradiation of three original isolates of Aspergillus oryzae to ultra-violet rays resulted in the production of mutants which differed from the parent riboflavin and vitamin $B_{12}$ in culture media. 1. Irradition three strains of Aspergillus oryzae to ultraviolet light produced mutants and two strains of them were selected for soy sauce brewing. 2. The two strains are the physiological mutants of Aspergillus oryzae. Both were found to have superior enzyme activity to their relatives. 3. Aspergillus oryzae UV-induced mutant 172-722 and 569-713 were more powerful than others in the production of riboflavin and vitamin $B_{12}$. The enzyme activity of these strain were high and decreased only slightly even in 20 percent solution of NaCl. 4. Aspergillus oryzae UV-induced mutant 172-722 had more powerful protease producibility in wheat bran media than in modified Czapek's solution. On the contrary, Aspergillus oryzae UV-induced mutant 569-713 had more powerful producibility of saccharogenic and dextrinogenic amylase in modified Czapek's solution than in mold bran. 5. Aspergillus oryzae UV-induced mutant 172-722 formed the spore rapidly and Aspergillus oryzae UV-induced mutant 569-713 did ordinarily. 6. It is found from the results that Aspergillus oryzae UV-induced mutant 172-722 is valuable material for the manufacture of soy sauce because of its high protease activity in 20 percent solution of NaCl. Aspergillus oryzae UV-induced mutant 569-713 is suitable for soy bean mash and for fermented red pepper sauce for its high saccharogenic and dextrinogenic amylase activity in 20 percent solution of sodium chloride.

• Korean Journal of Microbiology
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• v.6 no.3
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• pp.81-86
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• 1968

1. Amylolytic enzyme activities of Asp. oryzae and Asp. oryzae var. fulvus using the glucose as the carbon sources increased remarkably according to the decrease of the residual sugars. 2. The amylase productions of Asp. oryzae and Asp. oryzae var. fulvus were increased and enhanced when the organisms lave belen cultured in modified Koji media containing maltose as adaptive substrate. However, being devoid of maltose the level of amylase activities were lower and the begining of the production was prolonged. 3. The effects of C-sources on the amylase production of them were observed. The level of amylase activity varied with C-sources and their concentrations Marked increase of amylase production was afforded by starch and maltose. The effects of citric acid and tartaric acid were little or nothing. 4. Using the sucrose and lactose as the adaptive substrates both strains show the maximum amylolytic enzyme activities at the 3% concentrations of those sugars.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.506-513
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• 2010

Plant growth promoting ability of Methylobacterium oryzae CBMB20 was evaluated under different levels of organic fertilizer application on red pepper plants in a pot experiment. Oil cake as an organic N fertilizer was applied at the rates of 70, 85, 100 and 120% of the conventional recommended level. Each treatment was further treated with or without M. oryzae CBMB20 inoculation. The recommended amount of compost for red pepper was added in all the treatments. Results revealed that plant height, dry biomass and fruit yield were enhanced in increasing order as the rate of fertilization increased. Overall plant growth was improved due to the inoculation of M. oryzae CBMB20 and red pepper fruit yield was also increased by 10-35% in the plants inoculated with M. oryzae CBMB20 at different rates of organic fert1izer application. Total methylotrophic bacterial population in rhizosphere soil measured at the time of harvest was significantly higher in M. oryzae CBMB20 inoculated treatments. The growth promoting effect of M. oryzae CBMB20 found in red pepper could be due to the effective colonization of the bacteria in the rhizosphere and its ability of enhancing nutrient availability and producing plant growth hormones. With the plant growth promoting effect of M. oryzae CBMB20, the rate of organic fertilizer application can be reduced without any significant decreases in biomass production and yield of red pepper.