• Korean Journal of Soil Science and Fertilizer
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• v.46 no.2
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• pp.117-121
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• 2013

Single or co-inoculation of phosphate solubilizing bacterial and fungal strains (Burkholderia anthina and Aspergillus awamori respectively) was performed separately to assess their synergistic and antagonistic interactions and the potential to be used as bio-inoculants. Co-inoculation was found to release the highest content of soluble phosphorus (1253 ${\mu}g\;ml^{-1}$) into the medium, followed by single inoculation of fungal strain (1214 ${\mu}g\;ml^{-1}$) and bacterial strain (997 ${\mu}g\;ml^{-1}$). However, there was no significant difference between single inoculation of fungal strain and co-inoculation of fungal and bacterial strain in terms of the phosphorous release. The highest pH reduction, organic acid production and glucose consumption were observed in the sole A. awamori inoculated culture medium. According to the plant growth promotion bioassays, co-inoculation of the microbial strains resulted in 21% and 43% higher shoot and root growth of the mung bean seedlings respectively as compared to the respective controls. Therefore, co-inoculation of B. anthina and A. awamori showed better performance in stimulating plant growth than that in inoculation of each strain alone. However, assessment period of the present study being short, we recommend in engaging further experimentation under field conditions in order to test the suitability of the strains to be used as bio-inoculants.

• Mycobiology
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• v.42 no.4
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• pp.331-338
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• 2014

To examine the effects of water stress and Cenangium ferruginosum (CF) on the fungal endophytic community of needles of Pinus koraiensis (PK), fungal endophytes isolated from the needles of 5-year-old PK seedlings were compared before and after exposure to water stress conditions and artificial inoculation with CF ascospores. Artificial CF inoculation was successfully confirmed using PCR with CF-specific primers (CfF and CfR). For comparison of the degree of water deficit in water-stressed and control groups of PK seedlings infected with CF, the water saturation deficit and water potential were measured. Lower water potential estimates were found in the water-stressed seedlings than in the control group. The fungal endophytes isolated from the second-year needles of non-water-stressed seedlings before and after CF inoculation revealed that primary saprobes were approximately 30% and 71.7%, respectively, and the remaining endophytes were rot fungi or pathogens. Sixty days after CF inoculation, diverse fungal endophytes in the first-year needles were isolated from the water-stressed seedlings. However, some fungal endophytes isolated from the non-water-stressed seedlings were also identified. Fungal endophytes in the second-year needles of the water-stressed and non-water-stressed seedlings were approximately 8% and 71.7% of saprobes, respectively, and the remaining endophytes were rot fungi or pathogens. On the basis of the results, we conclude that water deficit and CF can have an effect on fungal endophytic communities in the needles of PK seedlings.

• The Plant Pathology Journal
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• v.16 no.5
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• pp.247-251
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• 2000

Latent infections of healthy-appearing oaks of Hypoxylon atropunctatum complicates field studies by interfering with inoculation experiments to follow pathogenesis, fungal development and reproduction of this canker rot fungus. Mutants with unique and easily scorable phenotypes would be useful for inoculation studies. There is a broad range in the capacity of wild-type isolates to utilize nitrate as a sole nitrogen sources. Several types of nitrate-nonutilization mutants (nit1, Nit3, NitM) were selected from nitrate-utilizing wild-type isolates. Also, a few mutants of Hypoxylon atropunctatum were selected that could only grow poorly on basal medium supplemented with various nitrogen sources and even on yeast extract agar. These unknown mutants need to be characterized further. Nit mutants of Hypoxylon atropunctatum were readily selected, grew well and were recovered after inoculation into oak stems. These results suggest that nit mutants could be useful for inoculation studies in trees that contain latent infections.

• Korean journal of applied entomology
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• v.40 no.1
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• pp.51-58
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• 2001

Entomopathogenic fungus, Paecilomyces japonicus, has been commercially used as medicinal purpose . The silkworm, Bombyx mori, as an optimal host for the fungi, has been selected and used for the production of the fungal fruit bodies. In current method, newly molted fifth instal larvae should be exposed to the adverse stress environment of high temperature (3$0^{\circ}C$), high relative humidity ( 90%), and starvation for 24h for better fungal inoculation to the host insects. In this study, an alternative method using chemical agent, dexamethasone (DEX: an eicosanoid biosynthesis inhibitor), was tried to get the immunodepressive effect on the larvae to elevate the inoculation rate of the fungi to the silkworm without any harsh rearing environment. DEX (100$\mu\textrm{g}$) showed significantly synergistic effect on the hemocyte lethality of the fungus, and was effective to decrease cellular immune responses measured by the number of hemocyte microaggregation and phenoloxidase activity of the fifth instar larvae in response to the fungal injection. A detergent of 0.05% Triton-X was effective to increase the in- oculation rate of the fungi to the larvae and used in all fungal spraying solutions. Without any environ- mental stress treatment, only DEX (100$\mu\textrm{g}$) injection to the fifth instar larvae followed by the fungal spray was effective to get the inoculation rate equivalent to the current fungal spray method requiring harsh rearing environment. These results suggest that the inoculation of P. japonicus can be elevated by the help of DEX and that the silkworms use eicosanoids to elicit cellular immune response against fungal pathogen.

• Mycobiology
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• v.50 no.6
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• pp.475-486
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• 2022

The antifungal activity of thymol against Aspergillus awamori F23 and Botrytis aclada F15 in onions was examined through direct treatment with amended media and gaseous treatment with I-plates (plastic plates containing central partitions). The protective and curative control efficacy of thymol was examined 24 h before and after the inoculation of onion bulbs with the fungal isolates. Mycelial growth, sporulation, and spore germination of the isolates were inhibited on potato dextrose agar amended with various concentrations of thymol or acetic acid (positive control). Overall, thymol produced a stronger inhibitory effect on the mycelial growth and development of the isolates than acetic acid. Following gaseous treatment in I-plates, mycelial growth, sporulation, and spore germination of the isolates were inhibited at higher concentrations of thymol or acetic acid; however, acetic acid showed a little effect on the sporulation and spore germination of the isolates. Following the treatment of onion bulbs with 1000 mg L^-1 of thymol 24 h before and after fungal inoculation, lesion diameter was greatly reduced compared with that following treatment with 0.5% ethanol (solvent control). Onion bulbs sprayed with thymol 24 h before fungal inoculation generally showed reduced lesion diameters by isolate F23 but not in isolate F15 compared with those sprayed 24 h after fungal inoculation. Collectively, thymol effectively inhibited the growth and development of A. awamori and B. aclada on amended media and in I-plates. In addition, spraying or fumigation of thymol is more desirable for effectively controlling these postharvest fungal pathogens during long-term storage conditions.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.4
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• pp.565-574
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• 2011

The development of satisfactory alternatives for supplying the nutrients needed by crops could decrease the problems associated with conventional NPK chemical fertilizers. In this study, the effects of bacterial and fungal effective microorganisms (EM) on the growth of Chinese cabbage (Brassica rapa) were evaluated. This investigation was carried out parrallel with conventional NPK chemical fertilizer and a commercial sold microbial fertilizer to compare between each of their effect. Sterile water and molasses were served as controls. Azotobacter chroococcum effect also was studied either alone or in combination with the effective microorganisms on the growth parameters. In contrast to the bacterial EM, the fungal EM alone without A. chroococcum had a more stimulating effect than fungal EM combined with A. chroococcum. Results showed that seedling inoculation significantly enhanced B. rapa growth. Shoot dry and fresh weight, and leaf length and width significantly were increased by both bacterial and fungal inoculation. The results indicated that the NPK chemical fertilizer deteriorates the microflora inhabiting the soil, while the effective microorganisms either fungal or bacterial ones increased the microbial density significantly. This study implies that both of fungal and bacterial EM are effective for the improvement of the Chinese cabbage growth and enhance the microorganisms in soil. The results showed antagonism occurred between A. chroococcum and each of Penicillium sp and Trichoderma sp in both agar and plant assays. The data were statistically analyzed by ANOVA and Dunnett test.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.392-399
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• 2008

Detached chili pepper fruits were inoculated with the conidial suspension of Colletotrichum acutatum JC-24 simultaneously (simultaneous inoculation, SI) and at delayed time (delayed inoculation, DI) after wounding with (delayed wound inoculation, DWI) or without additional wounding (delayed non-wound inoculation, DNI) at the inoculation time. Disease severity was significantly lowered by DNI, compared to SI. By DNI, the disease reduction rates were proportional with the length of delayed time, and greater at the high temperature range (18, 23 and $28^{\circ}$) than at the low temperature ($13^{\circ}$) tested. DWI was also effective in reducing the disease severity especially at 18oC; however, its effectiveness was lower than for DNI. In light microscopy, parenchyma cells at the wounding sites were modified structurally, initially forming new cell walls crossing cytoplasm, enlarged with multiple periclinal cell divisions, and finally layered like wound periderms. In DWI, the above structural modifications occurred, showing the restriction of the fungal invasion by the cell walls in enlarged modified cells, while no definite cellular modifications were found with proliferation of fungal hyphae in SI. Sclerenchyma-like cells with thickened cell walls were proliferated around the wounding sites, which were partially dissolved by DWI, probably leading to some disease development. All of these results suggest that the decline of the anthracnose disease in pepper fruit by the delayed inoculations may be derived from the structural modifications related to the healing processes of the previous wound inflicted on the tissues.

• The Plant Pathology Journal
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• v.15 no.3
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• pp.144-151
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• 1999

Chitinase activities from Shumard oak tissues were determined to study changes in chitinase activities related to water stress. The enzyme extracted in sodium acetate buffer (0.1M, pH 4.5) was assayed by a colorimetric method. In addition, the fungal hyphae of Hypoxylon atropunctatum in xylem tissues of oak were observed through scanning electron microscopy. The enzyme in oak tissues was mainly endochitinase, and optimum pH for enzyme activity was 5. Specific chitinase activities from both of stems held under high relative humidity (ranges of 0.63-1.11 pKatal/$\mu\textrm{g}$ of protein) and stems held under low relative humidity (ranges of 0.41-0.99 pKatal/$\mu\textrm{g}$ of protein) were significantly increased following fungal inoculation with H. atropunctatum. However, there was no significant difference in chitinase activities between tissues held under high and low humidities, which might be due to fungal chitinase. Scanning electron microscopy showed holes in fungal hyphae in the xylem tissues of stems held under high humidity but not in the stems held under ow humidity, suggesting that hyphae might be hydrolyzed by plant hydolases such as chitinase.

• The Plant Pathology Journal
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• v.17 no.1
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• pp.45-51
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• 2001

A root rot/vine decline disease occurred naturally on bottle gourd-stocked watermelon, melon, oriental melon and squash grown in greenhouses, but not on these plants grown in fields. Self-rooted watermelon, cucumber, pumpkin and luffa were also proven to be hosts of the pathogen by artificial inoculation in this experiment. The pathogen was identified as Monosporascus cannonballus by comparing microscopic characteristics of fungal structures with those of previously identified fungal strains. Our field investigations showed that the temperature and electric conductivity of soil in infected greenhouses were higher and the soil moisture content was lower than in noninfected greenhouses. To investigate soil-environmental factors affecting disease development, greenhouse trials and inoculation experiments were conducted. The host plants inoculated and grown under conditions of high soil temperature and electrical conductivity ($35\pm2^{\circ}$, 3.2-3.5 mS) and with low soil moisture content (pF 3.0-4.5) were most severely damaged by the fungal disease. Since plants growing in greenhouses ae usually exposed to such environmental conditions, this may be the reason why the monosporascus root rot/vine decline disease has occurred only on cucurbits cultivated in greenhouses but not in field conditions.

• The Plant Pathology Journal
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• v.19 no.2
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• pp.85-91
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• 2003

Pre-treatment with DL-3-aminobutyric acid (BABA) in the cucumber plants caused the decrease of disease severity after inoculation with anthracnose pathogen Colletotrichum orbiculare. In this study, ultrastructures of the vascular bundle and the infection structures in the leaves of BABA-treated as well as untreated cucumber plants were observed after inoculation with the anthracnose pathogen by electron microscopy. The ultrastructures of vascular bundle in the leaves of BABA-treated plants were similar to those of the untreated plants except plasmodesmata. In the BABA-treated plants, the plasmodesmata were more numerous than in the untreated plants, suggesting that the BABA treatment may cause the active transfer of metabolites through the vascular bundle. In the leaves of untreated plants, the fungal hyphae were spread widely in the plant tissues at 5 days after pathogen inoculation. Most cellular organelles in the hyphae were intact, indicating a compatible interaction between the plant and the parasite. In contrast, in the leaves of BABA pre-treated plants the growth of most hyphae was restricted to the epidermal cell layer at 5 days after inoculation. Most hyphae cytoplasm and nucleoplasm was electron dense or the intracellular organelles were degenerated. The cell walls of some plant cells became thick at the site adjacent to the intercellular hyphae, indicating a mechanical defense reaction of the plant cells against the fungal attack. Furthermore, hypersensitive reaction (HR) of the epidermal cells was often observed, in which the intracellular hyphae were degenerated. Based on these results it is suggested that BABA causes the enhancement of defense mechanisms in the cucumber plants such as cell wall apposition or HR against the invasion of C. orbiculare.