• Title/Summary/Keyword: Ralstonia sp.

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Enhancement of Tomato Tolerance to Biotic and Abiotic Stresses by Variovorax sp. PMC12 (Variovorax sp. PMC12 균주에 의한 토마토의 생물학 및 비생물학적 스트레스 저항성 증진)

  • Kim, Hyeon Su;Lee, Shin Ae;Kim, Yiseul;Sang, Mee kyung;Song, Jaekyeong;Chae, Jong-Chan;Weon, Hang-Yeon
    • Research in Plant Disease
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    • v.24 no.3
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    • pp.221-232
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    • 2018
  • Rhizobacteria play important roles in plant growth and health enhancement and render them resistant to not only biotic stresses but also abiotic stresses, such as low/high temperature, drought, and salinity. This study aimed to select plant growth promoting rhizobacteria (PGPR) with the capability to mitigate biotic and abiotic stress effects on tomato plants. We isolated a novel PGPR strain, Variovorax sp. PMC12 from tomato rhizosphere. An in vitro assay indicated that strain PMC12 produced ammonia, indole-3-acetic acid (IAA), siderophore, and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, which are well-known traits of PGPR. The aboveground fresh weight was significantly higher in tomato plants treated with strain PMC12 than in non-treated tomato plants under various abiotic stress conditions including salinity, low temperature, and drought. Furthermore, strain PMC12 also enhanced the resistance to bacterial wilt disease caused by Ralstonia solanacearum. Taken together, these results indicated that strain PMC12 is a promising biocontrol agent and a biostimulant to reduce the susceptibility of plants to both abiotic and biotic stresses.

Polyhydroxyalkanoate (PHA) Production Using Waste Vegetable Oil by Pseudomonas sp. Strain DR2

  • Song, Jin-Hwan;Jeon, Che-Ok;Choi, Mun-Hwan;Yoon, Sung-Chul;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1408-1415
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    • 2008
  • To produce polyhydroxyalkanoate (PHA) from inexpensive substrates by bacteria, vegetable-oil-degrading bacteria were isolated from a rice field using enrichment cultivation. The isolated Pseudomonas sp. strain DR2 showed clear orange or red spots of accumulated PHA granules when grown on phosphate and nitrogen limited medium containing vegetable oil as the sole carbon source and stained with Nile blue A. Up to 37.34% (w/w) of intracellular PHA was produced from corn oil, which consisted of three major 3-hydroxyalkanoates; octanoic (C8:0, 37.75% of the total 3-hydroxyalkanoate content of PHA), decanoic (C10:0, 36.74%), and dodecanoic (C12:0, 11.36%). Pseudomonas sp. strain DR2 accumulated up to 23.52% (w/w) of $PHA_{MCL}$ from waste vegetable oil. The proportion of 3-hydroxyalkanoate of the waste vegetable-oil-derived PHA [hexanoic (5.86%), octanoic (45.67%), decanoic (34.88%), tetradecanoic (8.35%), and hexadecanoic (5.24%)] showed a composition ratio different from that of the corn-oil-derived PHA. Strain DR2 used three major fatty acids in the same ratio, and linoleic acid was the major source of PHA production. Interestingly, the production of PHA in Pseudomonas sp. strain DR2 could not occur in either acetate- or butyrate-amended media. Pseudomonas sp. strain DR2 accumulated a greater amount of PHA than other well-studied strains (Chromobacterium violaceum and Ralstonia eutropha H16) when grown on vegetable oil. The data showed that Pseudomonas sp. strain DR2 was capable of producing PHA from waste vegetable oil.

MaoC Mediated Biosynthesis of Medium-chain-length Polyhydroxyalkanoates in Recombinant Escherichia coli from Fatty Acid (재조합 대장균에서 MaoC를 이용한 지방산으로부터의 중간사슬길이 폴리하이드록시알칸산 생산 연구)

  • Park, Si Jae;Lee, Seung Hwan;Oh, Young Hoon;Lee, Sang Yup
    • KSBB Journal
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    • v.29 no.4
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    • pp.244-249
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    • 2014
  • Biosynthesis pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHA) from fatty acid ${\beta}$-oxidation pathway was constructed in recombinant Escherichia coli by introducing the Pseudomonas sp. 61-3 PHA synthase gene (phaC2) and the maoC genes from Pseudomonas putida, Sinorhizobium meliloti, and Ralstonia eutropha. The metabolic link between fatty acid ${\beta}$-oxidation pathway and PHA biosynthesis pathway was constructed by MaoC, which is homologous to P. aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1). When the E. coli W3110 strains expressing the phaC2 gene and one of the maoC genes from P. putida, Sinorhizobium meliloti, and Ralstonia eutropha were cultured in LB medium containing 2 g/L of sodium decanoate as a carbon source, MCL-PHA that mainly consists of 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD), was produced. The monomer composition of PHA and PHA contents varied depending on MaoC employed for the production of PHA. The highest PHA content of 18.7 wt% was achieved in recombinant E. coli W3110 expressing the phaC2 gene and the P. putida maoC gene. These results suggest that MCL-PHA biosynthesis pathway can be constructed in recombinant E. coli strains from the b-oxidation pathway by employing MaoC able to supply (R)-3-hydroxyacyl-CoA, the substrate of PHA synthase.

Suppression of Bacterial Wilt with Fuorescent Pseudomonads, TS3-7 strain (Fluorescent siderophore 생산균주, TS3-7에 의한 풋마름병 발병 억제)

  • Kim, Ji-Tae;Cho, Hong-Bum;Kim, Shin-Duk
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.296-300
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    • 2005
  • Among the root colonizing and plant growth promoting bacteria isolated from the bacterial wilt suppressive soil, five strains were detected to produce siderophores by CAS agar assay. The most effective isolate, TS3-7 strain induced significant suppression of bacterial wilt disease in tomato and pepper plants. Seed treatment followed by soil drench application with this strain resulted in over 80% reduction of bacterial wilt disease compared with the control. Significant disease suppression by TS3-7 strain was related to the production of siderophore. Besides iron competition, induction of resistance of the host plant with siderophore was suggested to be another mode of action that suppress bacterial wilt, based on the lack of direct antibiosis against pathogen in vitro. According to Bergey's Manual of Systemic Bacteriology and 16S rDNA sequence data, TS3-7 stain was identified as Pseudomonas sp. TS3-7.

Biological Control of Soilborne Diseases on Tomato, Potato and Black Pepper by Selected PGPR in the Greenhouse and Field in Vietnam

  • Thanh, D.T.;Tarn, L.T.T.;Hanh, N.T.;Tuyen, N.H.;Srinivasan, Bharathkumar;Lee, Sang-Yeob;Park, Kyung-Seok
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.263-269
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    • 2009
  • Bacterial wilt, Fusarium wilt and Foot rot caused by Ralstonia solanacearum, Fusarium oxysporum, and Phytophthora capsici respectively, continue to be severe problems to tomato, potato and black pepper growers in Vietnam. Three bio-products, Bacillus vallismortis EXTN-1 (EXTN-1), Bacillus sp. and Paenibacillus sp. (ESSC) and Bacillus substilis (MFMF) were examined in greenhouse bioassay for the ability to reduce bacterial wilt, fusarium wilt and foot rot disease severity. While these bio-products significantly reduced disease severities, EXTN-1 was the most effective, providing a mean level of disease reduction 80.0 to 90.0% against bacterial wilt, fusarium wilt and foot rot diseases under greenhouse conditions. ESSC and MFMF also significantly reduced fusarium wilt, bacterial wilt and foot rot severity under greenhouse conditions. Bio-product, EXTN-1 with the greatest efficacy under greenhouse condition was tested for the ability to reduce bacterial wilt, fusarium wilt and foot rot under field condition at Song Phuong and Thuong Tin locations in Ha Tay province, Vietnam. Under field condition, EXTN-1 provided a mean level of disease reduction more than 45.0% against all three diseases compared to water treated control. Besides, EXTN-1 treatment increased the yield in tomato fruits 17.3% than water treated control plants.

Earthworm Enhanced Bioaugmentation of PCB Contaminated Soil

  • Crowley, David E.;Luepromchai, Ekawan;Singer, Andrew S.;Yang, Chang Sool
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.100-107
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    • 2000
  • In a recently developed strategy for in-situ treatment of polychlorinated biphenyls (PCB), bioaugmentation was used in conjunction with a surfactant, sorbitan trioleate, as a carbon source for the degrader bacteria, along with the monoterpene, carvone, and salicylic acid as inducing substrates. Two bacteria were used for soil inoculants, including Arthrobacter sp. st. B1B and Ralstonia eutrophus H850. This methodology achieved 60% degradation of PCBs in Aroclor 1242 after 18 weeks in soils receiving 34 repeated applications of the degrader bacteria. However, an obvious limitation was the requirement for soil mixing after every soil inoculation. In the research reported here, bioaugmentation and biostimulation treatment strategies were modified by using the earthworm, Pheretima hawayana, as a vector for dispersal and mixing of surface-applied PCB-degrading bacteria and soil chemical amendments. Changes in microbial biomass and microbial community structure due to earthworm effects were examined using DNA extraction and PCR-DGGE of 16S rDNA. Results showed that earthworms effectively promoted biodegradation of PCBs in bioaugmented soils to the same extent previously achieved using physical soil mixing, and had a lesser, but significant effect in promoting PCB biodegradation in biostimulated soils treated with carvone and salicylic acid. The effects of earthworms were speculated to involve many interacting factors including increased bacterial transport to lower soil depths, improved soil aeration, and enhanced microbial activity and diversity.

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Monitoring of Geothermal Systems Wells and Surrounding Area using Molecular Biological Methods for Microbial Species (분자생물학적 방법을 이용한 지열시스템 관정 및 주변지역 미생물종 모니터링)

  • Ahn, Chang-Min;Han, Ji-Sun;Kim, Chang-Gyun;Park, Yu-Chul;Mok, Jong-Koo;Jang, Bum-Ju
    • Journal of Soil and Groundwater Environment
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    • v.17 no.6
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    • pp.23-32
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    • 2012
  • This study was conducted to monitor microbial species dynamics within the aquifer due to long term operation of geothermal heat pump system. The species were identified by molecular biological methods of 16S rDNA. Groundwater sample was collected from both open (S region) and closed geothermal recovery system (J region) along with the control. J measured and control as well as S measured found Ralstonia pickettii as dominant species at year 2010. In contrast, Rhodoferax ferrireducens was dominantly observed for the control of S. In 2011, Sediminibacterium sp. was universely identified as the dominant species regardless of the monitoring places and type of sample, i.e., measured or control. The difference in the dynamics between the measured and the control was not critically observed, but annual variation was more strikingly found. It reveals that possible environmental changes (e.g. ORP and DO) due to the operation of geothermal heat recovery system in aquifer could be more exceedingly preceded to differentiate annual variation of microbial species rather than positional differences.

Effects of Long-Term Fertilizer Practices on Rhizosphere Soil Autotrophic CO2-Fixing Bacteria under Double Rice Ecosystem in Southern China

  • Tang, Haiming;Wen, Li;Shi, Lihong;Li, Chao;Cheng, Kaikai;Li, Weiyan;Xiao, Xiaoping
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1292-1298
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    • 2022
  • Soil autotrophic bacterial communities play a significant role in the soil carbon (C) cycle in paddy fields, but little is known about how rhizosphere soil microorganisms respond to different long-term (35 years) fertilization practices under double rice cropping ecosystems in southern China. Here, we investigated the variation characteristics of rhizosphere soil RubisCO gene cbbL in the double rice ecosystems of in southern China where such fertilization practices are used. For this experiment we set up the following fertilizer regime: without any fertilizer input as a control (CK), inorganic fertilizer (MF), straw returning (RF), and organic and inorganic fertilizer (OM). We found that abundances of cbbL, 16S rRNA genes and RubisCO activity in rhizosphere soil with OM, RF and MF treatments were significantly higher than that of CK treatment. The abundances of cbbL and 16S rRNA genes in rhizosphere soil with OM treatment were 5.46 and 3.64 times higher than that of CK treatment, respectively. Rhizosphere soil RubisCO activity with OM and RF treatments increased by 50.56 and 45.22%, compared to CK treatment. Shannon and Chao1 indices for rhizosphere soil cbbL libraries with RF and OM treatments increased by 44.28, 28.56, 29.60, and 23.13% compared to CK treatment. Rhizosphere soil cbbL sequences with MF, RF and OM treatments mainly belonged to Variovorax paradoxus, uncultured proteobacterium, Ralstonia pickettii, Thermononospora curvata, and Azoarcus sp.KH33C. Meanwhile, cbbL-carrying bacterial composition was obviously influenced by soil bulk density, rhizosphere soil dissolved organic C, soil organic C, and microbial biomass C contents. Fertilizer practices were the principal factor influencing rhizosphere soil cbbL-carrying bacterial communities. These results showed that rhizosphere soil autotrophic bacterial communities were significantly changed under conditions of different long-term fertilization practices Therefore, increasing rhizosphere soil autotrophic bacteria community with crop residue and organic manure practices was found to be beneficial for management of double rice ecosystems in southern China.

Comparative Genomic Analysis and BTEX Degradation Pathways of a Thermotolerant Cupriavidus cauae PHS1

  • Chandran Sathesh-Prabu;Jihoon Woo;Yuchan Kim;Suk Min Kim;Sun Bok Lee;Che Ok Jeon;Donghyuk Kim;Sung Kuk Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.875-885
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    • 2023
  • Volatile organic compounds such as benzene, toluene, ethylbenzene, and isomers of xylenes (BTEX) constitute a group of monoaromatic compounds that are found in petroleum and have been classified as priority pollutants. In this study, based on its newly sequenced genome, we reclassified the previously identified BTEX-degrading thermotolerant strain Ralstonia sp. PHS1 as Cupriavidus cauae PHS1. Also presented are the complete genome sequence of C. cauae PHS1, its annotation, species delineation, and a comparative analysis of the BTEX-degrading gene cluster. Moreover, we cloned and characterized the BTEX-degrading pathway genes in C. cauae PHS1, the BTEX-degrading gene cluster of which consists of two monooxygenases and meta-cleavage genes. A genome-wide investigation of the PHS1 coding sequence and the experimentally confirmed regioselectivity of the toluene monooxygenases and catechol 2,3-dioxygenase allowed us to reconstruct the BTEX degradation pathway. The degradation of BTEX begins with aromatic ring hydroxylation, followed by ring cleavage, and eventually enters the core carbon metabolism. The information provided here on the genome and BTEX-degrading pathway of the thermotolerant strain C. cauae PHS1 could be useful in constructing an efficient production host.

Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli from Sucrose (재조합 대장균에서 수크로즈로부터의 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Oh, Young Hoon;Kang, Kyoung-Hee;Shin, Jihoon;Song, Bong Keun;Lee, Seung Hwan;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.29 no.6
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    • pp.443-447
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    • 2014
  • Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.