• Mycobiology
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• 제33권3호
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• pp.131-136
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• 2005

Infection structures were observed at the penetration sites on the leaves of cucumber plants inoculated with Colletotrichum orbiculare using a fluorescence microscope. The cucumber plants were previously drenched with suspension of bacterial strains Pseudomonas putida or Micrococcus luteus. The plants pre-inoculated with both bacterial strains were resistant against anthracnose after inoculation with C. orbiculare. To investigate the resistance mechanism by both bacterial strains, the surface of infected leaves was observed at the different time after challenge inoculation. At 3 days after inoculation there were no differences in the germination and appressorium formation of conidia of C. orbiculare as well as in the callose formation of the plants between both bacteria pre-inoculated and non-treated. At 5 days, the germination and appressorium formation of the fungal conidia were, however, significantly decreased on the leaves of plants pre-inoculated with M. luteus at the concentration with $1.0{\times}10^7\;cfu/ml$. Furthermore, callose formation of plants cells at the penetration sites was apparently increased. In contrast, there were no defense reactions of the plants at the concentration with $1.0{\times}10^6\;cfu/ml$ of M. luteus. Similarly, inoculation P. putida caused no plant resistance at the low concentration, whereas increase of callose formation was observed at the higher concentration. The results of this study suggest that the resistant mechanisms might be differently expressed by the concentration of pre-treatment with bacterial suspension.

• 한국균학회지
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• 제47권2호
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• pp.131-142
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• 2019

법주사와 선암사는 다른 지리적 조건에 위치하고 있으며 그에 따라 건물의 구조와 배치에도 차이를 보인다. 또한 지역적 기후 차이가 뚜렷한 것으로 나타나 두 곳의 실내 외 공기질에 분포하는 부유 진균의 농도와 다양성을 2년 동안 계절별로 비교하고자 하였다. 그 결과, 전반적으로 선암사의 실내 외 환경에서 부유 진균의 농도가 법주사보다 높았지만 실내의 경우, 종 다양성은 낮았으며 전각들간의 농도 편차는 컸다. 실내의 경우, 법주사에는 총 54속 173점의 진균이, 선암사에서는 총 49속 162점의 진균이 분리되었고 외기 환경의 경우, 법주사에서는 33속 80점의 진균이, 선암사에서는 39속 745점의 진균이 분리되어 법주사는 실내에서, 선암사는 외기 환경에 더 다양한 진균이 분포하였다. 두 곳의 공기질에는 자낭균류가 90% 내외로 가장 높은 비율을 차지하였고 담자균류와 접합균류 순으로 구성되어 있었으며 담자균류의 경우, 실내보다는 외기 환경에 더 많이 분포하였다. 두 곳 모두 다른 계절에 비해 4월 봄철과 가을철에 부유 진균의 농도가 높은 것으로 나타났으며 법주사는 특히 가을철에, 선암사는 4월 봄철에 더 높았다. 자낭균류인 Cladosporium 속은 모든 시기와 지점에서 검출되었으며 특히, 가을철 공기질에서 우점하였다. 또한, 비가 내려 대기가 습해진 공기질에는 담자균류의 번식이 급증하였는데, 이러한 대기 조건에 놓일때, 선암사가 법주사보다 생물피해에 더 노출되기 쉬운 환경에 접해있는 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제4권2호
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• pp.134-140
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• 1994

An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and $\beta$-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50$^{\circ}C$ and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50$^{\circ}C$. The enzyme was significantly inhibited by metal compounds such as $HgCl_2$, but was stimulated by $CoCl_2$. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.

• 아시안잔디학회지
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• 제15권1호
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• pp.1-8
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• 2001

Antifungal activity of 20 chemicals registered to turfgrass diseases was evaluated. Among the chemicals, iprodione, benomyl, iprodione+thiram, pencycuron+tebuconazole, hexaconazole, and iprodione+thiophanate-methyl exhibited high antifungal activity to the dollar spot fungus. All isolates were greatly inhibited by the chemicals at the concentration over 32ppm($\mu\textrm{g}$/ml). However, sensitivity of the isolates to chemicals was varied at the lower concentration as 8 and 16 ppm as follows. The isolate originated from Ora golf course was resistant to iprodione and iprodione+thiophanate methyl, Gonjiam isolate to iprodion, Youngpyung isolate to iprodione+thiram, and Dogo isolate to iprodione+thiram, pencycuron+tebuconazole, and tebuconazole. It was found from this study that the varied chemical resistance among the isolates was positively related to the application time of the chemical in the golf course. Consequently, when a chemical was applied more often times than the others to the golf course, the fungal isolates originate from the field showed higher resistance to the former chemical. Effects of the chemicals on control of the dollar spot was evaluated in the field. All tested chemicals revealed over 70% disease control efficacy, however, mepronil+propiconazole was the best showing 83% control efficacy and followed by fenarimo, iprodione, terbuconazole, thiram, and thiophanate-methyl.

• 생약학회지
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• 제40권4호
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• pp.357-364
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• 2009

Phospholipids are crucially important in a cell membrane function and could thereby influence antibiotic susceptibility. In order to investigate the antifungal mechanism the total lipid was extracted from C. albicans treated with citronellol or thymol in concentration of their minimum inhibiting concentration and the changes in phospholipids composition were analyzed using ketoconazole as control. The cell growth and total lipid synthesis in cell walls of C. albicans were inhibited by treatment with citronellol. The levels of total lipids were decreased by 35.85% compared to the control. They also showed a significant decrease in the contents of phospholipid, phosphatidylcholine(PC), phosphatidyl ethanolamine(PE) and phosphatidylinositol(PI). As the result of GC assay for total fatty acid methyl esters of PC, PE and PI in C. albicans treated with citronellol, it was found that the major fatty acid composed of three phospholipid were palmitic acid, stearic acid and oleic acid. Moreover, the pattern of the fatty acid compositions of PC, PE and PI were changed by the oil. Based on the results, the anti-Candida mechanism of citronellol or thymol might be closely associated with disrupting the permeability barriers of the fungal cell wall composition or construction.

• Journal of Microbiology and Biotechnology
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• 제13권1호
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• pp.57-63
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• 2003

Using fungal (Fusarium solani f. pisi) and bacterial (Pseudomonas mendocina) cutinases, the initial hydrolysis rate of p-nitrophenyl esters was systematically estimated for a wide range of enzyme and substrate concentrations using a 96-well microplate reader. Both cutinases exhibited a high substrate specificity; i.e. a high hydrolytic activity on p-nitrophenyl butyrate (PNB), yet extremely low activity on p-nitrophenyl palmitate (PNP). When compared to the hydrolysis of PNB and PNP by other hydrolases [lipases and esterases derived from different microbial sources, such as bacteria (Pseudomonas cepacia, Psedomonas furescens, Baciilus stearothermophilus), molds (Aspeillus niger, mucor miehei), and yeasts (Candida rugosa, Candida cylindracea)], the above substrate specificity would seem to be a unique characteristic of cutinases. Secondly, the hydrolytic activity of the cutinases on PNB appeared much faster than that of the other hydrolytic enzymes mentioned above. Furthermore, the current study proved that even when the cutinases were mixed with large amounts of other hydrolases (lipases or esterases), the Initial hydrolysis rate of PNB was determined only by the cutinase concentration for each PNB concentration. This property of cutinase activity would seem to result from a higher accessibility to the substrate PNB, compared with the other hydrolytic enzymes. Accordingly, these distinct properties of cutinases may be very useful in the rapid and easy isolation of various natural cutinases with different microbial sources, each of which may provide a novel industrial application with a specific enzymatic function.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1065-1070
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• 2017

This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtained as a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of $3.1{\mu}g/ml$. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to $70^{\circ}C$, but its anti-candidal activity was affected at pH 2.

• Journal of Microbiology and Biotechnology
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• 제30권7호
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• pp.967-973
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• 2020

The fungal cell wall is a major target of antifungals. In this study, we report the antifungal activity of an ethanol extract from Aucklandia lappa against Candida albicans. We found that the extract caused cell wall injury by decreasing chitin synthesis or assembly and (1,3)-β-D-glucan synthesis. A sorbitol protection assay demonstrated that the minimum inhibitory concentration (MIC) of the A. lappa extract against C. albicans cells increased eight-fold from 0.78 to 6.24 mg/ml in 72 h. Cell aggregates, which indicate damage to the cell wall or membrane, were commonly observed in the A. lappatreated C. albicans cells through microscopic analysis. In addition, the relative fluorescence intensities of the C. albicans cells incubated with the A. lappa extract for 3, 5, and 6 h were 92.1, 84.6, and 79.8%, respectively, compared to the controls, estimated by Calcofluor White binding assay. This result indicates that chitin content was reduced by the A. lappa treatment. Furthermore, synthesis of (1,3)-β-D-glucan polymers was inhibited to 84.3, 79.7, and 70.2% of that of the control treatment following incubation of C. albicans microsomes with the A. lappa extract at a final concentration equal to its MIC, 2× MIC, and 4× MIC, respectively. These findings suggest that the A. lappa ethanol extract may aid the development of a new antifungal to successfully control Candidaassociated disease.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1690-1698
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• 2014

Partial purification of glucoamylase from solid-state fermentation culture was, firstly, investigated by reverse micellar extraction (RME). To avoid back extraction problems, the glucoamylase was kept in the original aqueous phase, while the other undesired proteins/enzymes were moved to the reverse micellar organic phase. The individual and interaction effects of main factors (i.e., pH and NaCl concentration in the aqueous phase, and concentration of sodium bis-2-ethyl-hexyl-sulfosuccinate (AOT) in the organic phase) were studied using response surface methodology. The optimum conditions for the maximum recovery of the enzyme were pH 2.75, 100 mM NaCl, and 200 mM AOT. Furthermore, the optimum organic to aqueous volume ratio ($V_{org}/V_{aq}$) and appropriate number of sequential extraction stages were 2 and 3, respectively. Finally, 60% of the undesired enzymes including proteases and xylanases were removed from the aqueous phase, while 140% of glucoamylase activity was recovered in the aqueous phase and the purification factor of glucoamylase was found to be 3.0-fold.

• 한국초지조사료학회지
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• 제32권4호
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• pp.325-334
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• 2012

It has been known that endophyte (Neotyphodium coenophialum) is beneficial to tall fescue (Schedonorus phoenix Scop.) because the mutualistic endophyte is able to confers tolerance against abiotic and biotic stresses to tall fescue. However, this fungal endophyte produces toxic alkaloid resulting in negative effects on animal performance. Recently, Non-toxic endophyte have been developed and inserted into tall fescue to avoid detrimental effect on animal but remaining positive influence on tall fescue. In order to keep this beneficial impact, it is essential to have endophyte infected tall fescue through vertical transmission from maternal plants to seeds. Little research has been carried out on endophyte transmission. To get basic information related to endophyte transmission, experiment was conducted to examine the effect of plant genotype on endophyte transmission under water stresses. Overall endophyte concentration in seeds was higher than that in panicles and endophyte concentration in seeds and panicles relied on plant. This study revealed that drought is not a critical component to control the endophyte transmission from maternal plants to seeds. Plant genotype is an important factor controlling the endophyte transmission from plant to seed.