• 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.

• 한국미생물·생명공학회지
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• 제25권1호
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• pp.30-36
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• 1997

To genetically breed powerful multifunctional antagonistic bacteria, the urease gene of alkalophilic Bacillus pasteurii was transferred into Bacillus subtilis YB-70 which had been selected as a powerful biocontrol agent against root-rotting fungus Fusarium solani. Urease gene was inserted into the HindIII site of pGB215-110 and designated pGU266. The plasmid pGU266 containing urease gene was introduced into the B. subtilis YB-70 by alkali cation transformation system and the urease gene was very stably expressed in the transformant of B. subtilis YB-70(pGU266). The optimal conditions for the transfomation were also evaluated. From the in vitro antibiosis tests against F. solani, the antifungal activity of B. subtilis YB-70 containing urease gene was much efficient than that of the non-transformed strain. Genetic improvement of B. subtilis YB-70 by transfer of urease gene for the efficient control seemed to be responsible for enhanced plant growth and biocontrol efficacy by combining its astibiotic action and ammonia producing ability.

• Journal of Microbiology
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• 제33권4호
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• pp.295-301
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• 1995

An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

• Mycobiology
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• 제49권1호
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• pp.69-77
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• 2021

The application of antagonistic fungi for plant protection has attracted considerable interest because they may potentially replace the use of chemical pesticides. Antipathogenic activities confirmed in volatile organic compounds (VOCs) from microorganisms have potential to serve as biocontrol agents against pre- and post-harvest diseases. In the present study, we investigated Galactomyces fungi isolated from rotten leaves and the rhizosphere of cherry tomato (Lycopersicon esculentum var. cerasiforme). VOCs produced by Galactomyces fungi negatively affected the growth of phytopathogenic fungi and the survival of nematodes. Mycelial growths of all nine examined phytopathogenic fungi were inhibited on agar plate, although the inhibition was more intense in Athelia rolfsii JYC2163 and Cladosporium cladosporioides JYC2144 and relatively moderate in Fusarium sp. JYC2145. VOCs also efficiently suppressed the spore germination and mycelial growth of A. rolfsii JYC2163 on tomatoes. The soil nematode Caenorhabditis elegans exhibited higher mortality in 24 h in the presence of VOCs. These results suggest the broad-spectrum activity of Galactomyces fungi against various plant pathogens and the potential to use VOCs from Galactomyces as biocontrol agents.

• 환경생물
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• 제21권3호
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• pp.292-296
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• 2003

균귄 곰팡이(arbuscular mycorrhizal fungus, AMF)의 하나인 Glomus manihot을 이용하여 고추의 성장과 무기이온 흡수 및 고추의 치명적 질병인 역병 방제에 미치는 효과를 조사한 결과 매우 유의적인 결과를 얻을 수 있었다. AMF 균주 처리에 의해 질소, 인산, 칼리의 흡수와 초기 중량이 증가되었음을 파종 2주만에 확인할 수 있었으며 고추 품종에 따른 차이는 크게 나타나지 않았다. 이는 Glomus manihot이 고추의 발아와 동시에 부리에 공생하게 되어 나타나는 결과였으며 따라서 파종 시기에 맞춰 AMF를 투입하는 것이 중용한 것으로 판단된다. 야외에서 6주간 경작한 결과, 중량은 AMF와 불용성 인산을 동시에 처리한 결과 40％ 가까운 증가 효과를 나타내었다. 이러한 결과는 불용성 인산을 사용 가능케 하는 AMF의 특징에 의한 것으로 토양 내 문제점이 되고 있는 인산 과용에 의한 불용성 인산염의 제거에도 매우 좋은 효과를 나타내리라 판단되었다. 또한 Phyto-phtohra capsici에 의한 고추 역병에 대해서도 Glomus manihot의 접종이 그 피해를 경감시킬 수 있는 것으로 나타났다. 따라서 고추 품종에 상관없이 파종 초기에 Glomus manihot을 접종한다면 고추의 성장과 질병 방제효과를 통해 보다 좋은 품질의 고추를 생산할 수 있을 것으로 판단되었다.

• The Plant Pathology Journal
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• 제25권2호
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• pp.165-171
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• 2009

Biological control activity of Acremonium strictum BCP, a mycoparasite on Botrytis cinerea, was examined against six plant diseases such as rice blast, rice sheath blight, cucumber gray mold, tomato late blight, wheat leaf rust, and barley powdery mildew in growth chambers. The spore suspension of strain BCP showed strong control activities against five plant diseases except against wheat leaf rust. On the other hand, the culture filtrate of A. strictum BCP was effective in controlling only cucumber gray mold and barley powdery mildew. Further in vivo biocontrol activities of A. strictum BCP against tomato gray mold were investigated under greenhouse conditions. Control efficacy of the fungus on tomato gray mold increased in a concentration-dependent manner. Treatment of more than $1{\times}10^6$ spores/ml significantly controlled the disease both in tomato seedlings and in adult plants. The high disease control activity was obtained from protective application of the strain BCP, whereas the curative application did not control the disease. Foliar infections of B. cinerea were controlled with $1{\times}10^8$ spores/ml of A. strictum BCP applied up to 7 days before inoculation. In a commercial greenhouse, application of A. strictum BCP exhibited the similar control efficacy with fungicide procymidone (recommended rate, $500{\mu}g/ml$) against strawberry gray mold. These results indicate that A. strictum BCP could be developed as a biofungicide for Botrytis diseases under greenhouse conditions.

• The Plant Pathology Journal
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• 제34권2호
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• pp.126-138
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• 2018

Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

• Journal of Microbiology and Biotechnology
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• 제24권1호
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• pp.1-7
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• 2014

The possibility of using hyphomycete fungi as suitable biocontrol agents against greenhouse whitefly has led to the isolation of various insect pathogenic fungi. Among them is Beauveria bassiana, one of the most studied entomopathogenic fungi. The objective of this study was to use B. bassiana M130 as an insecticidal agent against the greenhouse whitefly. M130 isolated from infected insects is known to be a biocontrol agent against greenhouse whitefly. Phylogenetic classification of M130 was determined according to its morphological features and 18S rRNA sequence analysis. M130 was identified as B. bassiana M130 and showed chitinase (342.28 units/ml) and protease (461.70 units/ml) activities, which were involved in the invasion of the host through the outer cuticle layer, thus killing them. The insecticidal activity was 55.2% in petri-dish test, 84.6% in pot test, and 45.3% in field test. The results of this study indicate that B. bassiana has potential as a biological agent for the control of greenhouse whitefly to replace chemical pesticides.

• 한국미생물·생명공학회지
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• 제44권4호
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• pp.540-549
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• 2016

토양에 존재하는 식물의 잔사는 식물 병원균의 확산 및 월동을 위한 자원으로써 역할을 수행 할 수 있다. 본 연구에서는 식물 잔사인 밀짚에서 식물 병원균인 Fusarium solani의 군락 형성에 대한 생물학적 방제균인 Trichoderma harzianum의 억제 효과를 조사하였다. 두 종류의 토양 수분 조건(-50, -500 kPa)의 멸균하지 않은 토양에서 연구를 수행하였다. T. harzianum ThzID1-M3와 F. solani를 밀짚과 함께 토양에 첨가 한 후 ThzID1-M3, Trichoderma spp. 및 F. solani를 quantitative real-time PCR을 사용하여 21일 동안 관찰하였다. 모든 토양 수분 조건에서 ThzID1-M3는 F. solani의 밀짚 점유를 감소하고(p < 0.05), 반면에 F. SOLANI는 ThzID1-M3 및 Trichoderma spp.의 밀짚 점유를 감소시켰다(p < 0.05). 이 결과는 F. solani와 ThzID1-M3 사이의 경쟁적 억제를 보여주고 있다. 높은 수분 조건의 토양(-50 kPa)에서 ThzID1-M3 및 Trichoderma spp.는 더 높은 밀짚의 점유를 보여주었다(p < 0.05), 반면에 F. solani의 밀짚 점유는 두 수분조건에서 차이가 없었다. 따라서 ThzID1-M3의 병원균 억제 효과는 높은 수분 조건의 토양에서 더 컸다(p < 0.05). 토양내 식물 잔사를 점유할 때 발생되는 ThzID1-M3와 F. solani 사이의 강력한 경쟁 관계는 생물학적 방제 균인 T. harzianum가 작물 시스템에서 식물 병원균의 생존과 증식을 감소시킬 수 있음을 시사한다.

• The Plant Pathology Journal
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• 제27권1호
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• pp.78-84
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• 2011

Antagonistic rhizobacteria, more specifically fluorescent pseudomonads and certain species of Bacillus, are known as biocontrol agents of fungal root diseases of agronomic crops. In this study, 144 bacteria were isolated from cucumber rhizosphere and screened as potential biological control agents against Phytophthora drechsleri, the causal agent of cucumber root rot, in vitro condition. Non-volatile compounds of 23 isolates showed noticeable inhibition zone (> 30%) against P. drechsleri, whereas volatile compounds of 7 isolates could prevent more than 30% of the mycelial growth of the fungus. All promising isolates, except of Pseudomonas flourescens V69, promoted significantly plant growth under in vitro condition. P. flourescens CV69 and V11 exhibited the highest colonization on the root. Results of the greenhouse studies showed that a reduction in disease incidence by use of some strains, and particularly use of strains CV6 and V11 as a soil treatment, exhibited a reduction in disease incidence so that suppressed disease by 85.71 and 69.39% respectively. Pseudomonas flourescens CV6 significantly suppressed disease in comparison to Ridomil fungicide. The use of mixture bacterial strains in the soil inoculated by the fungus resulting in falling down the most of the plants which didn't show significant difference with infected control soils without bacteria.