• Title/Summary/Keyword: antagonistic bacteria

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Cultivation Conditions for Mass Production of an Antagonistic Bacterium Stenotrophomonas maltophilia BW-13 (길항세균 Stenotrophomonas maltophilia BW-13의 대량배양을 위한 최적 배양조건)

  • Park Jong-Young;Kim Han-Woo;Kim Hyun-Ju;Chun Ok-Ju;Jung Soon-Je;Choi Woobong;Lee Seon-Woo;Moon Byung-Ju
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.158-161
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    • 2005
  • Stenotrophomonas maltophilia BW-13 is a potent biocontrol agent to control crisphead lettuce bottom rot caused by Rhizoctonia solani. To define the optimum conditions for the mass production of the S. maltophilia BW-13, we have investigated optimum culture conditions and effects of various carbon sources on the bacterial growth. The optimum initial pH and temperature were determined as pH $6.0\~7.0 and $35^{\circ}C$, respectively. For the selection of effective carbon source for the mass production, we tested the low molecular carbon sources such as sucrose, glucose, lactose, maltose, manose and the high molecular carbon source such as dough conditioner, rice bran, corn starch, sweet potato starch. As the results, the addition of dough conditioner in a basal medium ($1.25\%\;K_{2}HPO_4,\;0.38\%\;KH_{2}PO_4,\;0.01\%\;MgSO_4{\cdot}7H_{2}O,\;0.5\%\;Yeast extract$) was able to achieve higher cell density and the antifungal activity than others. Therefore, the basal medium containing $3\%$ dough conditioner (named as dough conditioner medium) was finally selected the optimized media for the mass production of BW-13 strain.

Characterization of the Antagonistic Activity against Lactobacillus plantarum and Induction of Bacteriocin Production (김치로부터 Lactobacillus plantarum 생육저해 박테리오신 생산균주의 분리 및 박테리오신 생산의 유도효과)

  • Yang, Eun-Ju;Chang, Ji-Yoon;Lee, Hyong-Joo;Kim, Jeong-Hwan;Chung, Dae-Kyun;Lee, Jong-Hoon;Chang, Hae-Choon
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.311-318
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    • 2002
  • A new bacteriocin producing lactic acid bacteria having antagonistic activity against Lactobacillus plantarum, was isolated from Kimchi. It was identified as Leuconostoc mesenteroides, and designated as Leuconostoc mesenteroides B7. The bacteriocin from Leuconostoc mesenteroides B7 named as bacteriocin B7 was stable in the pH range $2.5{\sim}9.5$. Bacteriocin B7 was active over a wide temperature range from $4^{\circ}C$ to $120^{\circ}C$. It was inactivated by proteinase K, trypsin, ${\alpha}-chymotrypsin$, and protease treatments indicating its proteinous nature. Tricine-SDS-PAGE of the purified bacteriocin B7 showed the presence of a single band, having a molecular mass of about 3,500 dalton. Mixed culture of the producer and the indicator, Lb. plantarum KFRI 464 or Lb. delbruekii KFRI 347, increased production of bacteriocin B7. This result suggested the presence of bacteriocin inducing factor in the indicator strain. The inducing factor was localized in cell debris and intracellular faction of the indicator cell, Lb. plantarum KFRI 464. Treatment of the inducing factor with proteinase K destroyed inducing activity. This result strongly suggested that the inducing factor is a protein.

Antifungal and Proteolytic Activity and Auxin Formation of Bacterial Strains Isolated from Highland Forest Soils of Halla Mountain (한라산 고지대 토양에서 분리한 미생물의 항균 및 단백질분해 활성, 오옥신 생산 특성)

  • Kim, Tack-Soo;Ko, Min-Jung;Lee, Se-Weon;Han, Ji-Hee;Park, Kyung-Seok;Park, Jin-Woo
    • The Korean Journal of Pesticide Science
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    • v.15 no.4
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    • pp.495-501
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    • 2011
  • Bacterial strains were isolated from forest soils of Halla mountain, Jeju island in Korea. The soil samples were collected at each altitude of 100m from 1,000 m above sea level. Total 398 strains were isolated and tested for their physiological characteristics of antagonistic and proteolytic activities, and auxin production. Among the isolates, 172 strains were selected as antifungal strains showing antagonistic activity against at least one of 8 plant fungal pathogens (Alternaria alternata, Botrytis cinerea, Collectotrichum acutatum, Fusarium oxysporum, Phytophthora capsici, Pythium ultimum and Sclerotinia sclerotiorum). In addition 203 strains for proteolytic activity and 26 strains for auxin production were characterized for further study. Je28-4 (Rhodococcus sp.) were showed 80% of control value against tomato gray mold in vivo. Thus, it is suggested that soil bacteria isolated from forest soils of Halla mountain can be important sources of bioactive compounds for improving plant growth or promising biocontrol agents.

Biological Control of Stem Rot of Pepper caused by Sclerotium rolfsii using by Bacillus amyloliquefaciens KBC1009 (길항세균 Bacillus amyloliquefaciens KBC1009를 이용한 고추 흰비단병의 생물학적 방제)

  • Kang, Jae-Gon;Lee, Young-Ui;Park, Jeong-chan;Jeong, Yoon-Woo;Park, Chang-Seuk;Kang, Hoon-Serg
    • Journal of agriculture & life science
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    • v.50 no.4
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    • pp.27-34
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    • 2016
  • Sclerotium rolfsii is a well known broad host range soil borne plant pathogenic fungus and caused serious damage to various vegetable crops. To develop an effective biological control agent for S. rolfsii, an isolate which showed strong inhibitory effect on the mycelial growth of S. rolfsii was selected among the antagonistic bacterial isolates collected from vinyl-house soil. The bacterial isolate was identified as Bacillus amyloliquefaciens KBC1009 based on the morphological, physiological characteristics and by 16S rRNA sequence analysis. The growth conditions for B. amyloliquefaciens KBC1009 were optimized in LB media(pH7) by culturing at 30℃ for 72 hrs. Glucose and yeast extract were confirmed as the best carbon and nitrogen sources, respectively. In order to test the inhibitory effect of B. amyloliquefaciens KBC1009 to stem rot of pepper, green house experiment was conducted. Drench of 1/500 diluted bacterial suspension of B. amyloliquefaciens KBC1009(5×108 cfu/ml) to each pepper plant 3 times with 10 days interval showed 66.7% control effectiveness. These results suggest that B. amyloliquefaciens KBC1009 is one of promising biocontrol agent to control stem rot caused by Sclerotium rolfsii.

Biological Control of White Rot in Apple Using Bacillus spp. (Bacillus spp.를 이용한 사과 겹무늬썩음병의 생물학적 방제)

  • Ha-Kyoung Lee;Jong-Hwan Shin;Seong-Chan Lee;You-Kyoung Han
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.390-398
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    • 2023
  • Apple white rot, caused by Botryosphaeria dothidea, is one of the important diseases in Korea. B. dothidea can cause pre- and postharvest decay on apple fruit as well as canker and dieback of apple trees. In this study, we isolated bacteria from the trunk of apple trees and tested their antagonistic activity against B. dothidea. Five bacterial isolates (23-168, 23-169, 23-170, 23-172, and 23-173) were selected that were most effective at inhibiting the mycelial growth of the pathogens. The isolate 23-172 was identified as Bacillus amyloliquefaciens and four isolates 23-168, 23-169, 23-170, and 23-173 were identified as Bacillus velezensis by RNA polymerase beta subunit (rpoB) and DNA gyraseA subunit (gyrA) gene sequencing. All isolates showed strong antagonistic activity against B. dothidiea as well as Colletotrichum fructicola and Diaporthe eres. All isolates exhibited cellulolytic, proteolytic and phosphate solubilizing activities. In particular, two isolates 23-168, 23-169 were shown to significantly reduce the size of white rot lesions in pretreated apple fruits. These results will provide the basis for the development of a fungicide alternative for the control of white rot of apple.

Suppressive Mechanism of Soil-borne Disease Development and its Practical Application -Isolation and Identification of Species of Trichoderma Antagonistic to Soil diseases and its activities in the Rhizosphere- (토양병의 발병억제 기작과 그 실용성 -길항성 Trichoderma spp.의 분리, 동정 및 근권내 활동-)

  • Kim, S.I.;Shim, J.O.;Shin, H.S.;Choi, H.J.;Lee, M.W.
    • The Korean Journal of Mycology
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    • v.20 no.4
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    • pp.337-346
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    • 1992
  • Trichoderma spp. are an effective control agent for damping-off or other plant diseases. The interaction between. T. hamatum and Rhizoctonia solani on the rhizosphere or surface soil were examined to assess the possible roles of antibiosis or competition in the mechanisms of biological control agents as a basic research. In a proportional comparison, total bacteria, fungi, actinomycetes and Trichoderma spp were 65%, 8.8%, 25.9% and 0.28% respectively in their distribution in the soil. Among Trichoderma spp isolated, the 5 species of Trichoderma spp were indentified as T. koninggii, T. pseudokoninggii, T. aureoviridi, T. hamatum and T. viride respectively. In a mycoparasitic test, one isolate of T. hamatum strain Tr-5 showed an enzymatic ability to break fungal hyphae into piecies and infected on the R. solani hyphae showing a parasitism. Spore germination of the all isolates of Trichoderma spp showed a 1.7-7.3% of germination in natural soil conditions, but the percentage was high in sterile soil indicating all the natural soil were fungistatic on conidia of Trichoderma spp. In rhizosphere competent assay in pea plant, the antagonistic T. hamatum, T. viride, T. koninggii, T. pseudokoninggii showed a colonizing upper soil depth in rhizosphere around 1-3 cm in root zone, but the colonizing ability was much reduced along the deeper the soil depth. Propagule density was decreased in deeper the soil layer. Disease development rate treated alone with plant pathogens, Fusarium solani, Rhizoctonia solani, Cylindrocarpon destructans increased, but disease incidence rate reduced in treatment with combinations with antagonistic T. hamatum strain Tr-5.

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Biological Control Activity of Two Isolates of Pseudomonas fluorescens against Rice Sheath Blight

  • Choi Gyung-Ja;Kim Jin-Cheol;Park Eun-Jin;Choi Yong-Ho;Jang Kyoung-Soo;Lim He-Kyoung;Cho Kwang-Yun;Lee Seon-Woo
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.289-294
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    • 2006
  • Two isolates of mucous bacteria, mc75 and pc78, were isolated from fungal culture plate as culture contaminants with an interesting swarming motility. Both isolates were identified as Pseudomonas fluorescens based on microscopy, biochemical analysis, Biolog test and DNA sequence analysis of the 16S rRNA gene. Both strains have the exactly the same 16S rRNA gene sequences, and yet their biological control activity were not identical each other. In vitro analysis of antagonistic activity of two isolates against several plant pathogenic fungi indicated that both produced diffusible and volatile antifungal compounds of unknown identities. Treatment of the bacterial culture of P. fluorescens pc78 and its culture filtrate exhibited a strong biological control activity against rice sheath blight in vivo among six plant diseases tested. More effective disease control activity was obtained from treatment of bacterial culture than that of culture filtrate. Therefore, in addition to antifungal compound and siderophore production, other traits such as biofilm formation and swarming motility on plant surface may contribute to the biological control activity of P.fluorescens pc78 and mc75.

Control Effect of Stenotrophomonas maltophilia BW-13 strain to the lettuce Bottom rot

  • Park, Jong-Young;Kim, Hyun-Ju;Bak, Joung-Woo;Lee, Kwang-Youll;Jun, Ok-Ju;Lee, Jin-Woo;Jung, Soon-Je;Moon, Byung-Ju
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.103.1-103
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    • 2003
  • An antagonistic bacteria, Stenotrophomonas maitophilia BW-13 strain which was effectively inhibited mycerial growth of Bottom rot pathogen, Rhizoctonia solani PY-1 strain was isolated from the rhizosphere of the lettuce in Uiryeong-Gun, Gyeongsangnam-Do from 2002 to 2003. For the biological control, the most suitable inoculum and its density of pathogen, PY-1 strain ware tested prior biological control test, For the pathogenicity test, A inoculum (wheat bran)sawdust+rice bran+PDB) showing disease incidence of 100% was selected as the most suitable inoculum, which showed more effective than B inoculum (sawdust+rice bran+DW) and mycelial disc. also, In selection of the amount of inoculum (40g, 50g, 60g, 70g, 80g), most suitable amount of inoculum of pathogen determined as 40g showing disease incidence of 80%. For the selection of effective microorganism to control bottom rot on lettuce, about 200 isolates were isolated from the diseased soil and lettuce leaves, and examined their antifungal activity to the pathogen on PDA. As the pots assay, BW-13 strain showed the highest control value as 90%, and followed by R-13 and R-26 strain as 80% and 60%, respectively. Selected BW-13 isolates identified as 5. maltophilia (GeneBank accession no. AJ293473.1, 99%) by 16S rRNA sequencing. This is the first report on the biological control using by S. maltophilia to the bottom rot pathogen, Rhizoctonia solani PY-1 strain.

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Characterization of Bacillus luciferensis Strain KJ2C12 from Pepper Root, a Biocontrol Agent of Phytophthora Blight of Pepper

  • Kim, Hye-Sook;Sang, Mee-Kyung;Myung, Inn-Shik;Chun, Se-Chul;Kim, Ki-Deok
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.62-69
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    • 2009
  • In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P=0.05) reduced both final disease severity and areas under the disease progress curves of 5-week-old pepper plants inoculated with P. capsici compared to buffer-treated controls. As for the production of antibiotics, biofilms, biosurfactant, extracellular enzyme, HCN, and swarming activity, strain KJ2C12 produced an extracellular enzyme with protease activity, but no other productions or swarming activity. However, Escherichia coli produced weak biofilm only. Strain KJ2C12 could colonize pepper roots more effectively in a gnotobiotic system using sterile quartz sand compared to E. coli over 4 weeks after treatments. However, no bacterial populations were detected in 10 mM $MgSO_4$ buffer-treated controls. Strain KJ2C12 produced significantly higher microbial activity than the $MgSO_4$-treated control or E. coli over 4 weeks after treatments. Bacterial strain KJ2C12 was identified as Bacillus luciferensis based on morphological, physiological, and biochemical characteristics as well as FAME and 16S rRNA gene sequence analyses. In addition, these results suggested that B. luciferensis strain KJ2C12 could reduce Phytophthora blight of pepper by protecting infection courts through enhanced effective root colonization with protease production and an increase of soil microbial activity.

Biological Control of Bacterial Fruit Blotch of Watermelon Pathogen (Acidovorax citrulli) with Rhizosphere Associated Bacteria

  • Adhikari, Mahesh;Yadav, Dil Raj;Kim, Sang Woo;Um, Young Hyun;Kim, Hyun Seung;Lee, Seong Chan;Song, Jeong Young;Kim, Hong Gi;Lee, Youn Su
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.170-183
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    • 2017
  • Bacterial fruit blotch (BFB), which is caused by Acidovorax citrulli, is a serious threat to watermelon growers around the world. The present study was conducted to screen effective rhizobacterial isolates against 35 different A. citrulli isolates and determine their efficacy on BFB and growth parameters of watermelon. Two rhizobacterial isolates viz. Paenibacillus polymyxa (SN-22), Sinomonas atrocyanea (NSB-27) showed high inhibitory activity in the preliminary screening and were further evaluated for their effect on BFB and growth parameters of three different watermelon varieties under greenhouse conditions. The greenhouse experiment result revealed that SN-22 and NSB-27 significantly reduced BFB and had significant stimulatory effect on total chlorophyll content, plant height, total fresh weight and total dry weight compared to uninoculated plants across the tested three watermelon varieties. Analysis of the 16S ribosomal RNA (rRNA) sequences revealed that strains SN-22 belong to P. polymyxa and NSB-27 to S. atrocyanea with the bootstrap value of 99% and 98%, respectively. The isolates SN-22 and NSB-27 were tested for antagonistic and PGP traits. The result showed that the tested isolates produced siderophore, hydrolytic enzymes (protease and cellulose), chitinase, starch hydrolytic enzymes and they showed phosphate as well as zinc solubilizing capacity. This is the first report of P. polymyxa (SN-22) and S. atrocyanea (NSB-27) as biocontrol-plant growth promoting rhizobacteria on watermelon.