• 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.

• 미생물학회지
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• 제55권3호
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• pp.226-233
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• 2019

Ochratoxin A (OTA)는 주로 Aspergillus가 생성하는 진균 독소의 하나로 저장 곡물의 흔한 오염물질로 인간과 가축의 건강을 위협하고 있다. 본 연구의 목적은 분리 세균인 Bacillus subtilis AF13과 Streptomyces shenzhenensis YR226의 OTA 생성 Aspergillus 균주들에 대한 생장과 OTA 생성 저해능 조사이다. OTA 생성 세 균주들의 균사 생장과 포자 형성에 대한 항진균활성은 한천 평판에서 AF13과 YR226 균주와의 공동배양에 의해 조사되었다. AF13과 YR226은 10일 배양 중 진균 집락 직경을 각각 77.58%와 78.48% 감소시켰으며, 두 균주 모두 포자 형성을 99%까지 저해하였다. YR226은 또한 세 진균 균주의 포자 발아도 91% 이상 감소시켰다. Yeast extract sucrose 배지에서 Aspergillus와 AF13 또는 YR226 균주를 동시배양하였을 때 세 종류 진균 모두 균사체 생장과 OTA 생성이 감소하였다. 특히 AF13은 A. alutaceus의 균사체 생장을 완전히 저해하고 OTA 생성은 99% 감소시켰으며, YR226은 균사체 생장과 독소 생성을 99%까지 저해하였다. AF13과 YR226이 생성하는 항진균물질에는 siderophore, chitinase, protease, ${\beta}$-1,3-glucanase와 biosurfactant가 포함된다. 이 결과는 AF13과 YR226이 독소생성 진균으로부터 가치있는 농작물을 보호하는 생물학적 방법으로 이용될 수 있으며, 따라서 농업과 사료산업에서 경제적 피해를 감소시킬 수 있을 것이다.

• 한국식물병리학회지
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• 제14권3호
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• pp.278-285
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• 1998

Effects of chitosan on growth of tomato plant, and suppression of Fusaruim wilt caused by Fusarium oxysporum f. sp. lycopersici and late blight casued by Phytophthora infestans, were examined. Both late blight and fusarium wilt were suppressed by spray and irrigation of chitosan, respectively. Inhibition of mycelial growth was not greatly affected by molecular size of chitosan but, concentration dependent effects was observed. Ninty percent of P. infestans and 80% of F. oxysporum f. sp. lycopersici of mycelial growth was inhibited by 1,000 ppm of chitosan (MW 30,000~50,000) when amended in plate media. Induction of defense-related gene expression in plant by chitosan treatments were observed when chitosan treated tobacco and tomato RNA samples were hybridized with several defense-related genes as probes. The results revealed that $\beta$-1,3-glucanase and chitinase genes were strongly induced, while pathogenesis-related protein-1, 3-hydroxy-3-methylglutaryl coenzyme A reductase, anionic peroxidase, phenylalanine ammonia lyase genes were weakly induced by chitosan treatment. These results suggest that chitosan have dual effects on these host-pathogen interactions. Possible roles of chitosan in suppression of tomato diseases by inhibition of mycelial growth and activation of plant defense responses are discussed.

• Mycobiology
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• 제43권3호
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• pp.311-318
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• 2015

Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding ${\beta}$-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.

• The Plant Pathology Journal
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• 제28권3호
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• pp.282-289
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• 2012

The root rot of pepper (Capsicum annuum L.) caused by Phytophthora capsici is one of the most important diseases affecting this crop worldwide. This work presents the evaluation of the capacity of Streptomyces griseus H7602 to protect pepper plants against Phytophthora capsici and establishes its role as a biocontrol agent. In this study, we isolated an actinomycete strain H7602 from rhizosphere soil, identified it as Streptomyces griseus by 16S rRNA analysis and demonstrated its antifungal activity against various plant pathogens including P. capsici. H7602 produced lytic emzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase and protease. In addition, crude extract from H7602 also exhibited destructive activity toward P. capsici hyphae. In the pot trial, results showed the protective effect of H7602 against pepper from P. capsici. Application of H7602 culture suspension reduced 47.35% of root mortality and enhanced growth of pepper plants for 56.37% in fresh root and 17.56% g in fresh shoot as compared to control, resulting in greater protection to pepper plants against P. capsici infestation. Additionally, the enzymatic activities, chitinase and ${\beta}$-1,3-glucanase, were higher in rhizosphere soil and roots of pepper plants treated with H7602 than other treated plants. Therefore, our results indicated a clear potential of S. griseus H7602 to be used for biocontrol of root rot disease caused by P. capsici in pepper.

• BMB Reports
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• 제34권4호
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• pp.310-315
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• 2001

The uneven distribution of acidic and basic chitinases in different parts of rice seed, and also the characterization of hull-specific chitinases, are reported here. After extraction of chitinases from polished rice, bran, and rice hulls, the chitinases were separated into acidic and basic fractions, according to their behavior on an anion exchanger column. Both fractions from different parts of rice seed showed characteristic activity bands on SDS-PAGE that contained 0.01% glycol chitin. The basic chitinases from rice hulls were further purified using chitin affinity chromatography. The chitinase, specific to rice hulls (RHBC), was 88-fold purified with a 1.3% yield. RHBC has an apparent molecular weight of 22.2 kDa on SDS-PAGE. The optimal pH and temperature were 4.0 and $35^{\circ}C$, respectively. With [$^3H$]chitin as a substrate, RHBC has $V_{max}$ of 13.51 mg/mg protein/hr and $K_m$ of 1.36 mg/ml. This enzyme was an endochitinase devoid of ${\beta}$-1,3-glucanase, lysozyme, and chitosanase activities.

• Mycobiology
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• 제45권4호
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• pp.409-420
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• 2017

Foliar sprays of three plant resistance inducers, including chitosan (CH), potassium sorbate (PS) ($C_6H_7kO_2$), and potassium bicarbonates (PB) ($KHCO_3$), were used for resistance inducing against Erysiphe cichoracearum DC (powdery mildew) infecting okra plants. Experiments under green house and field conditions showed that, the powdery mildew disease severity was significantly reduced with all tested treatments of CH, PS, and PB in comparison with untreated control. CH at 0.5% and 0.75% (w/v) plus PS at 1.0% and 2.0% and/or PB at 2.0% or 3.0% recorded as the most effective treatments. Moreover, the highest values of vegetative studies and yield were observed with such treatments. CH and potassium salts treatments reflected many compounds of defense singles which leading to the activation power defense system in okra plant. The highest records of reduction in powdery mildew were accompanied with increasing in total phenolic, protein content and increased the activity of polyphenol oxidase, peroxidase, chitinase, and ${\beta}$-1,3-glucanase in okra plants. Meanwhile, single treatments of CH, PS, and PB at high concentration (0.75%, 2.0%, and/or 3.0%) caused considerable effects. Therefore, application of CH and potassium salts as natural and chemical inducers by foliar methods can be used to control of powdery mildew disease at early stages of growth and led to a maximum fruit yield in okra plants.

• Mycobiology
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• 제32권1호
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• pp.47-53
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• 2004

Pseudomonas fluorescens 1-94 induced systemic resistance in chickpea against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri by the synthesis and accumulation of phenolic compounds, phenylalanine ammonia lyase(PAL) and pathogenesis related(PR) proteins(chitinase, $\beta$-1,3-glucanase and peroxidase). Time-course accumulation of these enzymes in chickpea plants inoculated with P. fluorescens was significantly(LSD, P=0.05) higher than control. Maximum activities of PR-proteins were recorded at 3 days after inoculation in all induced plants; thereafter, the activity decreased progressively. Five PR peroxidases detected in induced chickpea plants. Molecular mass of these purified peroxidases was 20, 29, 43, 66 and 97 kDa. Purified peroxidases showed antifungal activity against plant pathogenic fungi.

• 한국유기농업학회지
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• 제26권1호
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• pp.151-161
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• 2018

This study was designed to investigate the effects of multi-enzyme on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.1% multienzyme (Multi; mixture of ${\beta}-mannanase$, xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Frequency of diarrhea, levels of packed cell volume (PCV), white blood cells (WBC), immunoglobulins, cortisol, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), and C-reactive protein (CRP) were measured. Multi group tended to decrease (p<0.1) diarrhea frequency than CON group during 2 wk after weaning. Lower values of PCV on d 3 (p<0.05) and d 7 (p<0.1) were found in Multi group compared with CON group. There were no significant differences on WBC number and immunoglobulin (Ig) M and A between Multi and CON groups. However, Multi group tended to increase (p<0.1) Ig G on d 7 than CON group. Moreover, Multi group showed modulated immune responses, indicated by decreased levels of cortisol (p<0.05) on d 7 and 14, $TNF-{\alpha}$ on d 3 (p<0.05) and d 7 (p<0.10), $TGF-{\beta}$ on d 2 (p<0.05) and d 7 (p<0.10), and CRP (p<0.10) on d 3 and 7 after weaning compared with CON group. Consequently, inclusion of multi-enzyme in diets for weaned pigs improved gut health and modulated immune responses of weaned pigs.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1327-1336
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• 2014

Bacillus amyloliquefaciens strain NJZJSB3 has shown antagonism of several phytopathogens in vitro, especially Sclerotinia sclerotiorum. Both the broth culture and cell suspension of strain NJZJSB3 could completely protect the detached leaves of canola (Brassica napus) from S. sclerotiorum infection. In pot experiments, the application of strain NJZJSB3 cell suspension ($10^8CFU/ml$) decreased the disease incidence by 83.3%, a result similar to commercially available fungicide (Dimetachlone). In order to investigate the potential biocontrol mechanisms of strain NJZJSB3, the nonvolatile antifungal compounds it produces were identified as iturin homologs using HPLC-ESI-MS. Antifungal volatile organic compounds were identified by gas chromatography-mass spectrometry. The detected volatiles toluene, phenol, and benzothiazole showed antifungal effects against S. sclerotiorum in chemical control experiments. Strain NJZJSB3 also produced biofilm, siderophores and cell-wall-degrading enzymes (protease and ${\beta}$-1,3-glucanase). These results suggest that strain NJZJSB3 can be a tremendous potential agent for the biological control of sclerotinia stem rot.