• Title/Summary/Keyword: Effective biocontrol formulation

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Isolation and Identification of Burkholderia pyrrocinia CH-67 to Control Tomato Leaf Mold and Damping-off on Crisphead Lettuce and Tomato

  • Lee, Kwang-Youll;Kong, Hyun-Gi;Choi, Ki-Hyuck;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.59-67
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    • 2011
  • A bacterial strain CH-67 which exhibits antagonism towards several plant pathogenic fungi such as Botrytis cinerea, Fulvia fulva, Rhizoctonia solani, Sclerotinia sclerotiorum, Colletotrichum sp. and Phytophthora sp. was isolated from forest soil by a chitin-baiting method. This strain was identified as Burkholderia cepacia complex (Bcc) and belonging to genomovar IX (Burkholderia pyrrocinia) by colony morphology, biochemical traits and molecular method like 16S rRNA and recA gene analysis. This strain was used to develop a bio-fungicide for the control of tomato leaf mold caused by Fulvia fulva. Various formulations of B. pyrrocinia CH-67 were prepared using fermentation cultures of the bacterium in rice oil medium. The result of pot experiments led to selection of the wettable powder formulation CH67-C containing modified starch as the best formulation for the control of tomato leaf mold. CH67-C, at 100-fold dilution, showed a control value of 85% against tomato leaf mold. Its disease control efficacy was not significantly different from that of the chemical fungicide triflumidazole. B. pyrrocinia CH-67 was also effective in controlling damping-off caused by Rhizoctonia solani PY-1 in crisphead lettuce and tomato plants. CH67-C formulation was recognized as a cell-free formulation since B. pyrrocinia CH-67 was all lethal during formulation process. This study provides an effective biocontrol formulation of biofungicide using B. pyrrocinia CH-67 to control tomato leaf mold and damping-off crisphead lettuce and tomato.

Field Sanitation and Foliar Application of Streptomyces padanus PMS-702 for the Control of Rice Sheath Blight

  • Yang, Chia-Jung;Huang, Tzu-Pi;Huang, Jenn-Wen
    • The Plant Pathology Journal
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    • v.37 no.1
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    • pp.57-71
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    • 2021
  • Rice sheath blight (ShB), caused by Rhizoctonia solani Kühn AG1-IA, is one of the destructive rice diseases worldwide. The aims of this study were to develop biocontrol strategies focusing on field sanitation and foliar application with a biocontrol agent for ShB management. Streptomyces padanus PMS-702 showed a great antagonistic activity against R. solani. Fungichromin produced by S. padanus PMS-702, at 3.07 mg/l inhibited 50% mycelial growth, caused leakage of cytoplasm, and inhibited the formation of infection structures of R. solani. Fungichromin could reach to 802 mg/l when S. padanus PMS-702 was cultured in MACC broth for 6 days. Addition of 0.5% S. padanus PMS-702 broth into soil decreased the survival rate of the pathogen compared to the control. Soil amended with 0.5% S. padanus broth and 0.5% tea seed pomace resulted in the death of R. solani mycelia in the infested rice straws, and the germination of sclerotia was inhibited 21 days after treatment. Greenhouse trials revealed that S. padanus cultured in soybean meal-glucose (SMGC-2) medium after mixing with different surfactants could enhance its efficacy for inhibiting the pathogen. Of six surfactants tested, the addition of 2% tea saponin was the most effective in suppressing the pathogen. S. padanus broth after being fermented in SMGC-2, mixed with 2% tea saponin, diluted 100 fold, and sprayed onto rice plants significantly reduced ShB disease severity. Thus, S. padanus PMS-702 is an effective biocontrol agent. The efficacy of S. padanus PMS-702 for disease control could be improved through formulation.

Seed Coating for the Application of Biocontrol Agent Bacillus subtilis YBL-7 against Phytopathogens (길항세균 Bacillus subtilis YBL-7 건조포자체의 종자피막화에 의한 생물학적 방제)

  • 장종원;김상달
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.243-248
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    • 1995
  • Agrochemicals for the plant-disease control are criticized severely for causing environmental pollution and residual problems, and consequently microbial disease control agents are expected to be safer and more economical for sustainable agriculture. Treatment of biological control agents to seed requires the use of effective delivery systems that allow full expression of the benefical qualities of the bioprotectant. For the activation and establishment of bioprotectant around the plant seed which are able protect the seeds and seedlings from pathogen attack, the optimal liquid coating formulation was obtained using 2% sodium carboxymethyl cellulose (binder), 20% sesame dregs (solid particulate material), and dried spore of Bacillus subtilis YBL-7 (bioprotectants, 10 mg/g of seed). Suppressive of root rot was demonstrated in pot trials with coated kidney bean (Phaseolus vulgaris L.) seeds. Coated seeds with B. subtilis YBL-7 spore in F. solani-infested soil reduced disease incidence by 85% to 90% after 30 days.

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Biological control of Gray Mold Rot of Perilla Caused by Botrytis cinerea II. Formulation of Antagonistic Bacteria and Its Control Effect (들깨 잿빛곰팡이병의 생물학적 방제 II. 미생물농약의 제조 및 그 방제효과)

  • Moon, Byung-Ju;Kim, Choul-Soung;Song, Ju-Hee;Kim, Ju-Hee;Lee, Jae-Pil;Park, Hyean-Cheal;Shin, Dong-Bum
    • Research in Plant Disease
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    • v.8 no.3
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    • pp.184-188
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    • 2002
  • An antagonistic bacteria, Bacillus licheniformis Nl strain which effectively inhibited mycelial growth of gray mold rot pathogen, Botrytis cinerea was isolated from the rhizosphere of perilla crop. Powder soy formulation by B. lichentfomis Nl strain as a biocontrol agent was developed far the first time and estimated its control effect on perilla leaves in this study. First of all, far the mass production of antifungal metabolites of B. lichentfomis Nl strain in flask liquid culture, the most effective carbon and nitrogen source were selected as glucose and tryp-tone, respectively, For the formulation, vegetative biomass of B. licheniformis Nl strain from 5-day-old liquid culture in nutrient broth added glucose and tryptone was mixed with soy flour, rice flour glucose, FeSo$_4$~7$H_2O$, and MnCl$_2$. 4$H_2O$, and dried and pulverized. In plastic house test, powder soy formulation effectually controlled gray mold rot as the control value of 93.1 %, was more effective than chemical fungicide, benomyl showing the control value of 86.1%. Thus, development of powder soy formulation of B. lichentfomis Nl will aid large-scale application of biological control in field trials.

Biological Control of Tea Anthracnose Using an Antagonistic Bacterium of Bacillus subtilis Isolated from Tea Leaves

  • Kim, Gyoung-Hee;Lim, Myoung-Taek;Hur, Jae-Seoun;Yum, Kyu-Jin;Koh, Young-Jin
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.99-102
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    • 2009
  • An antagonistic bacterium of Bacillus subtilis BD0310 against Colletotrichum theae-sinensis was isolated from the phylloplane of tea trees at a tea plantation in Korea. SC (suspension concentrate)-type biofungicide was formulated with the antagonist. Cell viability and antifungal activity of B. subtilis were maintained in the formulation more than 12 months at room temperature. The antagonist was sensitive only to copper sulfate among the chemical pesticides currently registered for tea trees in Korea. Greenhouse application demonstrated that the biofungicide controlled more effectively the disease in a protective mode than in a curative mode. Field trial showed that alternate applications of the biofungicide and chemical fungicide were more effective in controlling tea anthracnose than single application of the biofungicide or chemical fungicide with less use of chemicals. This study suggests that the biofungicide of B. subtilis 8D0310 is an effective method for biological control of anthracnose in tea plantations.

Biological Control of Perilla Sclerotinia Rot Caused by Sclerotinia sclerotiorum Using Bacillus megaterium N4. (Bacillus megaterium N4에 의한 들깨 균핵병 (Sclerotinia sclerotiorum)의 생물학적 방제)

  • 문병주;김현주;송주희;이광열;백정우;정순재
    • Journal of Life Science
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    • v.14 no.5
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    • pp.761-769
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    • 2004
  • This study was investigated the occurrence of sclerotinia rot caused by Sclerotinia sclerotiorum at the major perilla cultivating area, Gangdong-dong, Gangseo-gu, Busan in 1998. The incidence of this disease ranged from 8.1 to 28.3% at Gangdong-dong area during the growing seasons. Symptoms of the disease initially appeared damping-off of infected stems and soft-rot on the leaves of perilla. Under the relatively high humidity, abundant white mycelia of the pathogen formed on the lesion developed into black sclerotia later and the infected leaves were finally fell down. Sixteen isolates, Sl-S16, isolated from diseased lesions showing typical symptoms, and pathogenicity was tested using mycerlial disks. Among them, S2 isolate showing the most strong pathogenicity was selected and identified as Sclerotinia sclerotiorum on the basis of morphological and cultural characteristics. For biological control, an antagonistic bacteria, N4 isolate which effectively inhibited not only mycelial growth of S2 isolate but also suppress sclerotinia rot on the pot assay, was selected and identified as Bacillus megaterium according to Bergey's manual and API system., Wettable powder type, N4 formulation using B. megaterium N4 isolate was developed and estimated its control effect on perilla crops in a plastic house. As a results, N4 formulation which applied before 3 days inoculation of pathogen was effectually controlled Sclerotinia rot as the control value of 98.0%, was more effective than chemical fungicide, benomyl showing the control value of 78.0%. This is the first report of wettable powder formulation as a biocontrol agent using B. megaterium N4 against Sclerotinia rot caused by S. sclerotiorum on perilla.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Study on Medium Ingredient Composition for Enhancing Biomass Productionand Anti-potato Common Scab Activity of Streptomyces sp. A020645 as a BCA Candidate (생물제제(BCA) 후보균주인 Streptomyces sp. A020645 의 대량 균체생산 및 항더뎅이병 활성증진을 위한 고체배지 조성에 관한 연구)

  • Lee, Hyang-Burm;Roh, Hyo-Young;Park, Dong-Jin;Lee, So-Keum;Ko, Young-wan;Koh, Jeong-Sam;Kim, Chang-Jin
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.66-71
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    • 2005
  • The effect of medium components such as wheat bran, rice bran, oat meal, and soybean meal as basic ingredients and KH2PO4, glucose, and molasses as additives on mass production and anti-potato common scab activ ity of a streptomycete A020645 strain as a biocontrol agent (BCA) candidate was investigated. Of basicingredients, oat meal was the best one for mass poduction and enhancement of anti-potato common scabactivity. The biomass production of the active strain was more enhanced when 0.1-0.01.% glucose or molassesas additive were added into the basic medium. These information may have important implications in applying for effective formulation of BCA.

Isolation and Characterization of Indole-3-acetic acid- and 1-aminocylopropane-1-carboxylyic Acid Deaminase-producing Bacteria Related to Environmental Stress (환경스트레스와 관련된 indole-3-acetic acid 및 1-aminocylopropane-1-carboxylyic acid deaminase 활성을 갖는 박테리아의 분리와 특성 연구)

  • Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.390-400
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    • 2019
  • In this study, strains isolated from soil samples collected from Busan, Changwon, and Jeju Island were examined to verify their abilities of phosphate solubilization and nitrogen fixation, production of indole-3-acetic acid (IAA), siderophore, and 1-aminocylopropane-1-carboxylyic acid (ACC) deaminase in order to select strains that promote plant growth and play a role in biocontrol of pests or pathogens. According to the results of this study, most of the isolated strains were found to have ability of phosphate solubilization, nitrogen fixation, IAA production, siderophore production, and production of ACC deaminase. These isolated strains might help plant growth by directly improving absorption of nutrients essential for phosphate solubilization and nitrogen fixation. In addition, they can promote plant growth and control resistance to plant diseases through extracellular enzyme activity and antifungal activity. In addition, most of the selected strains were found to survive in various environmental conditions such as temperature, salinity, and pH. Therefore, Pseudomonas plecoglossicida ANG14, Pseudarthrobacter equi ANG28, Beijerinckia fluminensis ANG34, and Acinetobacter calcoaceticus ANG35 were finally selected through a comparative advantage analysis to suggest their potential as novel biological agents. Further studies are necessary in order to prove their efficacy as novel biological agents through formulation and optimization of effective microorganisms, their preservation period, and crop cultivation tests.