• Title/Summary/Keyword: and BIOLOG

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Sigma S Involved in Bacterial Survival of Ralstonia pseudosolanacearum (Ralstonia pseudosolanacearum 생존에 관여하는 Sigma S 역할)

  • Hye Kyung Choi;Eun Jeong Jo;Jee Eun Heo;Hyun Gi Kong;Seon-Woo Lee
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.148-156
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    • 2024
  • Ralstonia pseudosolanacearum, a plant pathogenic bacterium that can survive for a long time in soil and water, causes lethal wilt in the Solanaceae family. Sigma S is a part of the RNA polymerase complex, which regulates gene expression during bacterial stress response or stationary phase. In this study, we investigated the role of sigma S in R. pseudosolanacearum under stress conditions using a rpoS-defective mutant strain of R. pseudosolanacearum and its wild-type strain. The phenotypes of rpoS-defective mutant were complemented by introducing the original rpoS gene. There were no differences observed in bacterial growth rate and exopolysaccharide production between the wild-type strain and the rpoS mutant. However, the wild-type strain responded more sensitively to nutrient deficiency compared to the mutant strain. Under the nutrient deficiency, the rpoS mutant maintained a high bacterial viability for a longer period, while the viability of the wild-type strain declined rapidly. Furthermore, a significant difference in pH was observed between the culture supernatant of the wild-type strain and the mutant strain. The pH of the culture supernatant for the wild-type strain decreased rapidly during bacterial growth, leading to medium acidification. The rapid decline in the wild-type strain's viability may be associated with medium acidification and bacterial sensitivity to acidity during transition to the stationary phase. Interestingly, the rpoS mutant strain cannot utilize acetic acid, D-alanine, D-trehalose, and L-histidine. These results suggest that sigma S of R. pseudosolanacearum regulates the production or utilization of organic acids and controls cell death during stationary phase under nutrient deficiency.

Cloning and Expression of an Insecticidal Crystal Protein CryIIA Gene from Bacillus thuringiensis subsp. kurstaki HD-1 (Bacillus thuringiensis subsp. kurstaki HD-1 CryIIA의 내독소 단백질 유전자의 클로닝 및 발현)

  • 김호산;김상현;제연호;유용만;서숙재;강석권;조용섭
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.300-306
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    • 1993
  • The CryIIA gene encoding the insecticidal crystal protein of Bacillus thuringiens!s subsp. kurstalri HD-l has been cloned in Escherichia col!, and its nucleotide sequences were determined completely. 5kb Hindlli fragment harboring CryIIA gene was screened in the large ca. 225kb plasmid DNA by southern blot. HindlIT digested 5kb fragment was ligated into pUC19 and transformed in E. coli. The 4kb BamHI-HindlIT fragment containing the CryIIA gene was subcloned and named pSKIIA. DNA sequence analysis demonstrates that pSKIIA is the gene of an operon which is comprised of Lhree open reading frames (designated orn, orf2 and or£3). The CrylIA gene is composed of 3,952bp-long BamHI-Hindill DNA restriction fragment. The orf3 code for a polypeptide of 633 amino acid residues. The protoxin protein has a predicted molecular weight of 70,780. The E. coli derived protoxin gene product is biologICally active against three species of Lepidopteran (Plu.lelia maculipennis, He/iolhis assulta, Spodoptera litura) and a species of Dip Leran( Culex pipines) larvae in bioassay.

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A Multi-microbial Biofungicide for the Biological Control against Several Important Plant Pathogenic Fungi (진균성 식물병해 방제를 위한 항생물질 생산 길항미생물의 복합제제화)

  • Jung, Hee-Kyoung;Ryoo, Jae-Cheon;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.40-47
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    • 2005
  • In order to develop a multi-microbial biofungicide against several important plant pathogenic fungi, strains were isolated from the phtophthora blight suppressive red-pepper field soil of Gyeongsangbuk-do, Korea. Strains AY1, AY6, AB1, BB2 and F4, which had strong antagonistic ability against Phytophthota capsici and Fusarium oxysporum, were selected for their involvement with strains of biocontrol fungicide. There were no antagonism among the selected strains and were compatible for making the biofungicide. Their antagonistic mechanisms, except for strain BB2, were an antibiosis by the production of antibiotic, while BB2 produced not only an antibiotic but also cellulase as an antagonistic mechanism against blight causing P. capsici. They were identified as Halobacterium sp. AB1, Xenorhadus sp. AY1, Bacillus sp. AY6, Bacillus sp. BB2, Zymomonas sp. F4 by various cultural, biochemical test and $Biolog^{TM}$ System 4.0. The highest levels of antifungal antibiotic could be produced after 48 hrs of incubation under the optimal medium which were 0.1% galactose, 0.1% $NaNO_2$, 5 mM $Na_2{\cdot}HPO_4$ (pH 5.5). The cultured multi-microbial biofungicide showed strong biocontrol activity against bacterial wilt disease and fusarium wilt disease in cucumber and tomato fields.

First Report of Bacterial Root Rot Caused by Serratia plymuthica on Panax ginseng (Serratia plymuthica에 의한 인삼 세균뿌리썩음병 발생)

  • Jung, Won Kwon;Kim, Young Soo;Choi, Jin Kook;Kim, Seung-Han;Jang, Myeong-Hwan;Kwon, Tae Lyong;Jeon, Yongho
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.332-338
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    • 2018
  • In August of 2011, a wilting disease of ginseng was observed at Bongwha, Gyeongbuk province, Korea. Affected plants initially show withering symptoms on leaves of ginseng. As the disease progresses, withering leaves spread downward, eventually encompassing the whole plant. Leaves lose vigor but remain pale green. Symptoms of roots were brown, and soft rots characterized by moist and watery decay of the whole ginseng root, which initiated as small brown, water-soaked lesions of hairy roots and enlarged to the entire roots. The causal organism isolated from the infected roots was identified as Serratia plymuthica based on its physiological and biochemical characteristics, by cellular fatty acid composition (GC-FAME), the utilization of carbon sources (BioLog System), and 16S rRNA sequence of the isolated bacterium were 99% homologous to those of Serratia plymuthica strains. Artificial inoculation of the bacterium produced the same brown or soft rot symptoms on the ginseng roots, from which the same bacterium was isolated. This is the first report of bacterial root rot caused by the Serratia plymuthica in ginseng in Korea. Serratia plymuthica has been used as antagonistic microorganism for biological control on several crop plants. But it was proved pathogen of ginseng at humid condition in this study.

Effect of Water Activity and Temperature on Growth, Germination, Sporulation, and Utilization of Carbon Source of Penicillium oxalicum (PENOX) as a Biocontrol Agent(BCA) for control of Clover(Trifolium repens L.) (토끼풀(Trifolium repens L.) 방제용 생물제제 Penicillium oxalicum (PENOX)의 발아, 생장, 포자생성 및 탄소원이용에 미치는 수분활성 및 온도의 영향)

  • Lee, Hyang-Burm;Kim, Chang-Jin
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.68-74
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    • 2000
  • Penicillium oxalicum (PENOX) has shown the potential as a biocontrol agent(5CA) for control of a weed, clover(Trifolium repens L.) in grass plots. The bioherbicidal activity may be due to germinative and growth capacities and substrate availability of the agent over a range of environmental factors. The influences of different water activities($0.94{\sim}0.995\;a_w$) and temperatures($18{\sim}30^{\circ}C$) on mycelial growth, conidial germination, sporulation oil 2% MEA(malt extract agar) adjusted to different water activities with glycerol, and carbon source utilization using BIOLOG GN MicroPlate were determined in vitro. Decreases in $a_w$ on MEA caused a reduction in mycelial growth and conidial germination depending on temperature. The mycelial growth of PENOX was greatest at $30^{\circ}C/0.995\;a_w$. At some lowered water activity($0.97\;a_w$), the growth was similar between 25 and $30^{\circ}C$, and considerably decreased at lowered temperature($20^{\circ}C$). The germination rate was also greatest at $30^{\circ}C/0.995\;a_w$. Lag phase times for PENOX at $18^{\circ}C$ on MEA were >6hrs at tile whole $a_w$ level tested, and at 18 and $25^{\circ}C$ they were >18hrs and >12hrs at $0.94\;a_w$, respectively. However, its sporulation was some better at $0.97\;a_w$ than $0.995\;a_w$ or $0.94\;a_w$, and better at $20^{\circ}C$ than $30^{\circ}C$. In contrast, the number of carbon sources(niche size) utilized by PENOX varied with $a_w$ and temperature. Under some water stress condition($0.95\;a_w$), the agent utilized smaller number of carbon sources than $0.995\;a_w$ depending on temperature. The niche size at 0.995 and $0.95\;a_w$ were highest at $25^{\circ}C$, and showed 86 and 65, respectively. At $30^{\circ}C$, the niche size at 0.995 and $0.95\;a_w$ showed 84 and 50, respectively. There was no carbon source utilized by PENOX at $0.90\;a_w$ regardless of temperature. These information of tile fungal ecophysiology will be useful for the effective development of BCA.

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Assessment of Soil Properties and Growth of Organically Cultivated Cucumber (Cucumis sativus L.) with Applications of Livestock Manure Compost and Fish Meal Liquid Fertilizer (가축분 퇴비와 어분 액비 시용이 유기농 오이 생육 및 토양환경에 미치는 영향)

  • An, Nan-Hee;Cho, Jung-Rai;Lee, Sang-min;Nam, Hong-Sik
    • Journal of the Korea Organic Resources Recycling Association
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    • v.26 no.3
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    • pp.23-31
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    • 2018
  • This study was carried out to investigate the effects of livestock manure compost and fish-meal liquid fertilizer on the growth of cucumber and the soil properties for the stable production of organic cucumber. Cucumber was transplanted in greenhouse on the $6^{th}$ of April in 2017, and this experiment contained five treatments: livestock manure compost 100% (LC 100%), livestock manure compost 50% + fish-meal liquid fertilizer 50% (LC50 + LF50), livestock manure compost 50% (LC50), chemical fertilizer (NPK), and no fertilizer (NF). As a result, it was shown that soil chemical properties of LC50 + LF50 plot is not different from that of LC100 plot except for the EC content, but soil chemical properties of LC50 + LF50 plot is statistically significantly different from that of NPK plot except for pH. As a result of evaluating the functional diversity of soil microbial communities using Biolog system, the substrate richness (S) and the diversity index (H) were the highest in LC50 + LF50 plot. As a result of comparing the cucumber growth and yield, it was found that there was no statistically significant difference between the plant height and the fresh weight of LC100, LC50 + LF50, and NPK plot, but the plant height and the fresh weight of LC100, LC50 + LF50, and NPK plot were different from that of LC50 and NF plot. The yield of cucumber was the highest in NPK plot r(7,397 kg/10a), but there was no statistically significant difference in the yield of cucumber between NPK plot and LC100, LC50 + LF50 plot. The above-described results suggested that the livestock manure compost and fish meal liquid fertilizer can be used for organic cucumber production under greenhouse condition.

Effect of Fish Meal Liquid Fertilizer Application on Soil Characteristics and Growth of Cucumber(Cucumis sativus L.) for Organic Culture (유기농 오이재배를 위한 어분액비 공급이 토양특성 및 오이 수량에 미치는 영향)

  • An, Nan-Hee;Cho, Jung-Rai;Gu, Ja-Sun;Kim, Young-ki;Han, Eun-Jung
    • Journal of the Korea Organic Resources Recycling Association
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    • v.25 no.3
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    • pp.13-21
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    • 2017
  • This study was carried out to evaluate the application effects of fish meal liquid fertilizer on soil characteristics and growth of cucumber for organic cultivation. Cucumber in greenhouse was transplanted on March $31^{th}$ in 2016, and the experimental treatments involve six treatments: No fertilizer, 0, 25, 50, and 100 mg/L N application by fish meal liquid fertilizer and chemical fertilizer. In the results of soil chemical property, application of 100 mg/L of fish meal liquid fertilizer showed a significant differences in pH, K, and Mg contents. The soil microbial community varied in relation to the fish meal liquid fertilizer treatments. Microbial biomass was lower in the chemical fertilizer than in the liquid fertilizer treatment. Result of principal component analysis obtained from Ecoplate showed that fish meal liquid fertilizer treatments, no liquid fertilizer, chemical fertilizer, and no fertilizer were divided into distinct groups, with the no fertilizer treatment located furthest from the other treatments. There were no significant differences in plant height of cucumber between the fish meal liquid fertilizer treatments and chemical fertilizer treatments. Also, the cucumber yield did not vary significantly between the concentrations of liquid fertilizers, and there were also no significant differences in the yield among the fish meal liquid and chemical fertilizer treatments. In conclusion, it is suggested that the application of fish meal liquid fertilizer can be used as a additional fertilizer for cucumber production with organic culture in greenhouse.

In vivo Antifungal Activity of Pyrrolnitrin Isolated from Burkholderia capacia EB215 with Antagonistic Activity Towards Colletotrichum Species (탄저병균에 대하여 길항작용을 보이는 Burkholderia cepacia EB215로부터 분리한 Pyrrolnitrin의 항균활성)

  • Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.31-38
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    • 2004
  • An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

Biological Control of Garlic Blue Mold using Pantoea agglomerans S59-4 (Pantoea agglomerans S59-4를 이용한 마늘 푸른곰팡이병의 생물학적 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Jee, Hyung-Jin;Park, Jong-Ho;Han, Eun-Jung;Park, Kyung-Seok;Lee, Sang-Yeob;Lee, Seong-Don
    • The Korean Journal of Pesticide Science
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    • v.14 no.2
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    • pp.148-156
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    • 2010
  • S59-4 isolate was evaluated as a potential biocontrol agent using in vivo wounded garlic bulb assay. When the spore suspension ($10^5$ spores/$m\ell$) of Penicillium hirsutum was co-inoculated with cell suspension of S59-4 isolate on wounded garlics, the isolate showed high suppressive effect to disease development. The isolate was identified as Pantoea agglomerans S59-4(Pa59-4) through Biolog system. Furthermore, soaking garlic bulbs in the suspension of Pa59-4 significantly reduced garlic decay caused by P. hirsutum. The optimal concentration of Pa59-4 for controlling garlic blue mold was $10^7\sim10^8$ cfu/$m\ell$. And suppressive effect of Pa59-4 on garlic storage decay reduced as inoculation concentration of Penicillium hirsutum increased. In addition in order to investigate population dynamics of Pa59-4 on application site of garlic cloves, two antibiotic markers, pimaricin and vancomycin were selected. Bacterial density of Pa59-4 on the wounded garlic cloves increased continuously both under room temperature condition and low temperature condition until 30days after application of Pa59-4, meanwhile that of Pa59-4 on intact garlic cloves increased until 15days after application of Pa59-4 and thereafter decreased continuously. Two culture media for mass-production of Pa59-4, LB medium and TSB medium, were selected. By-product of bio-fungicide formulated by mixing white carbon and bacterial suspension of Pa59-4 suppressed by 40 to 50% garlic blue mold. Above results suggest that Pa59-4 be a promising control agent against garlic blue mold.