• Title/Summary/Keyword: rhizobium

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PQQ-Dependent Organic Acid Production and Effect on Common Bean Growth by Rhizobium tropici CIAT 899

  • Cho, Young-Shin;Park, Ro-Dong;Kim, Yong-Woong;Hwangbo, Hoon;Jung, Woo-Jin;Suh, Jang-Sun;Koo, Bon-Sung;Krishnan, Hari-B.;Kim, Kil-Yong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.955-959
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    • 2003
  • Rhizobium tropici CIAT 899 is capable of synthesizing inactive apo-glucose dehydrogenase (GDH). To become an active holo enzyme, the GDH requires a cofactor, PQQ. When R. tropici CIAT 899 was grown in a broth culture medium containing hydroxyapatite and pyrrolo quinoline quinone (PQQ), pH decreased while the concentration of soluble P increased. The solubilization of hydroxyapatite was associated with the production of gluconic acid and 2-ketogluconic acids. The organic acid production and P solubilization were greatly enhanced when the bacterium was grown with air supply. Effect of R. tropici CIAT 899 with (CI+PQQ) and without PQQ (CI) on the common bean growth was examined. Shoot and root weight, and N and P contents in CI+PQQ treatment, were significantly higher than those in control and CI treatment. Nodule weight and acetylene reducing activities were also significantly higher in CI+PQQ treatment than in other treatments.

Investigation of Possible Gene Transfer to Soil Microorganisms for Environmental Risk Assessment of Genetically Modified Organisms

  • Kim, Young-Tae;Park, Byoung-Keun;Hwang, Eui-Il;Yim, Nam-Hui;Kim, Na-Rae;Kang, Tae-Hoon;Lee, Sang-Han;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.498-502
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    • 2004
  • The current study was conducted to monitor the possibility of the gene transfer among soil bacteria, including the effect of drift due to rain and surface water, in relation to the release of genetically modified organisms into the environment. Four types of bacteria, each with a distinct antibiotic marker, kanamycin-resistant P. fluorescens, rifampicin-resistant P. putida, chloramphenicol-resistant B. subtilis, and spectinomycin-resistant B. subtilis, were plated using a small-scale soil-core device designed to track drifting microorganisms. After three weeks of culture in the device, no Pseudomonas colonies resistant to both kanamycin and rifampicin were found. Likewise, no Bacillus colonies resistant to both chloramphenicol and spectinomycin were found. The gene transfer from glyphosate-tolerant soybeans to soil bacteria, including Rhizobium spp. as a symbiotic bacteria, was examined by hybridization using the DNA extracted from soil taken from pots, in which glyphosate-tolerant soybeans had been growing for 6 months. The results showed that 35S, T-nos, and EPSPS were observed in the positive control, but not in the DNA extracted from the soilborne microorganisms. In addition, no transgenes, such as the 35S promoter, T-nos, and EPSPS introduced into the GMO soybeans were detected in soilborne bacteria, Rhizobium leguminosarum, thereby strongly rejecting the possibility of gene transfer from the GMO soybeans to the bacterium.

Dissimilatory Nitrate Reduction Characteristics of Indigenous Soybean Rhizobia Distributed in Korea Soils (한국에 분포되어 있는 토착대두 근류균의 질산 환원 특성)

  • Choi, Young-Ju;Choi, Yong-Lark;Yun, Han-Dae;Ryu, Jin-Chang;Lee, Sang-Kyu;Cho, Moo-Je
    • Applied Biological Chemistry
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    • v.29 no.2
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    • pp.175-181
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    • 1986
  • Eightyseven strains of indigenous Rhizobia were isolated from the nodule of soybean cultivar, Danyup, cultivated in four different soils sampled from continuously soybean cultivated and newly reclaimed fields. The strains were grouped into Bradyrhizobium japanicum (slow-grower:55 strains) and Rhizobium fredii (fast-grower: 32 strains). The both groups could be divided into two sub-groups according to the denitrification characteristics, that is, denitrifying fast-grower (F-I), nitrate respiring fast-grower (F-II), denitrifying slow-grower (S-I). and nitrate respiring slow-grower (S-II). Among the 87 isolates, F-I, F-II, S-I and S-II sub-groups were 10, 22, 48 and 7 strains, respectively. The one-and two-dimensional polyacrylamide gel electrophoretic pattern of the four sub-groups were compared and discernible difference was observed between fast and slow-grower, but the difference was not discernible between subgroups within the same growth rate group.

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Binding between Lipopolysaccharide of Rhizobia and Lectins from Soybean (대두 근류균의 리포 다당과 Lectin의 결합성)

  • Kang, Sang-Jae;Kim, Jin-Ho;Park, Woo-Churl
    • Current Research on Agriculture and Life Sciences
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    • v.15
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    • pp.25-32
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    • 1997
  • This study was carried out to research the biological characteristics among rhizobia and soybean seed and root lectins, and to obtain a basic imformation of host specificity in biological nitrogen symbiosis system. The results obtained were as follows: Purified seed lectin from soybean varieties of paldal, backwoon and hwangkeum formed immunoprecipitin lines with standard soybean seed lectin and the root lectins from soybean seedlings immunoelectrophoretically. Soybean seed and root lectins interacted with Rhizobium japonicum and Bradyrhizobium japonicum, but didn't interacted with Rhizobium. viceae, whereas pea lectin conjugated with R. viceae, but didn't bind with R. japonicum and B. japonicum. Lipopolysaccharides of B. japonicum and R. viceae were fractionated into LPS I and LPS II on the sephadox G-50. Lipopolysaccharides from B. japonicum showed rhe binding acitivity both with soybean seed lectin and root lectin, but those from R. viceae didn't show it with soybean seed and root lectins.

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Isolation of an Indigenous Imidacloprid-Degrading Bacterium and Imidacloprid Bioremediation Under Simulated In Situ and Ex Situ Conditions

  • Hu, Guiping;Zhao, Yan;Liu, Bo;Song, Fengqing;You, Minsheng
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1617-1626
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    • 2013
  • The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

Identification and Expression Profiles of Six Transcripts Encoding Carboxylesterase Protein in Vitis flexuosa Infected with Pathogens

  • Islam, Md. Zaherul;Yun, Hae Keun
    • The Plant Pathology Journal
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    • v.32 no.4
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    • pp.347-356
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    • 2016
  • Plants protect themselves from pathogen attacks via several mechanisms, including hypersensitive cell death. Recognition of pathogen attack by the plant resistance gene triggers expression of carboxylesterase genes associated with hypersensitive response. We identified six transcripts of carboxylesterase genes, Vitis flexuosa carboxylesterase 5585 (VfCXE5585), Vf-CXE12827, VfCXE13132, VfCXE17159, VfCXE18231, and VfCXE47674, which showed different expression patterns upon transcriptome analysis of V. flexuosa inoculated with Elsinoe ampelina. The lengths of genes ranged from 1,098 to 1,629 bp, and their encoded proteins consisted of 309 to 335 amino acids. The predicted amino acid sequences showed hydrolase like domains in all six transcripts and contained two conserved motifs, GXSXG of serine hydrolase characteristics and HGGGF related to the carboxylesterase family. The deduced amino acid sequence also contained a potential catalytic triad consisted of serine, aspartic acid and histidine. Of the six transcripts, Vf-CXE12827 showed upregulated expression against E. ampelina at all time points. Three genes (VfCXE5585, VfCXE12827, and VfCXE13132) showed upregulation, while others (VfCXE17159, VfCXE18231, and VfCXE47674) were down regulated in grapevines infected with Botrytis cinerea. All transcripts showed upregulated expression against Rhizobium vitis at early and later time points except VfCXE12827, and were downregulated for up to 48 hours post inoculation (hpi) after upregulation at 1 hpi in response to R. vitis infection. All tested genes showed high and differential expression in response to pathogens, indicating that they all may play a role in defense pathways during pathogen infection in grapevines.

Effect of Herbicides on the Survival of Soybean Nodule Bacteria(Rhizobium japonicum) in vitro. (제초제의 처리가 콩 근류균의 생존율에 미치는 영향)

  • Jeong-Hae Oh
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.1
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    • pp.86-91
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    • 1989
  • Since most chemicals effect to the non-target organisms, present study was conducted to determine the effects of some herbicides to soybean nodule bacteria(Rhizobium japonicum) in vitro. Selective upland herbicides Alachlor, Linuron, Simazine and nonselective Paraquat were medicated to the YEMA media and survival rate of the bacteria in the medium was measured a week after incubation. Survival of soybean nodule bacteria in the media medicated with recommended concentration 400 ppm of Alachlor and Linuron were decreased significantly by 27.4 % and 57.8 %, respectively. While little effect was observed in Simazine, a marked reduction of survival was observed in 200 ppm of Paraquat. Effects of Alachlor was different with isolates of the nodule bacteria. Isolate I -122 was relatively resistant and I -145 was more sensitive. than other isolates tested. Intermediate K-5 was the most resistant at the recommended dose and the survival dose was drastically reduced with increment of concentration. Nodule formation of the soybean plants was different with the varieties and isolates. The most nodulation was observed in the resistant isolates I -122 and K-S, for which the nodulation might be related with the resistance to the environmental stress factors.

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Genetic and Phenotypic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Sugarcane Plants Growing in Pakistan

  • Mehnaz, Samina;Baig, Deeba N.;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1614-1623
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    • 2010
  • Bacteria were isolated from roots of sugarcane varieties grown in the fields of Punjab. They were identified by using API20E/NE bacterial identification kits and from sequences of 16S rRNA and amplicons of the cpn60 gene. The majority of bacteria were found to belong to the genera of Enterobacter, Pseudomonas, and Klebsiella, but members of genera Azospirillum, Rhizobium, Rahnella, Delftia, Caulobacter, Pannonibacter, Xanthomonas, and Stenotrophomonas were also found. The community, however, was dominated by members of the Pseudomonadaceae and Enterobacteriaceae, as representatives of these genera were found in samples from every variety and location examined. All isolates were tested for the presence of five enzymes and seven factors known to be associated with plant growth promotion. Ten isolates showed lipase activity and eight were positive for protease activity. Cellulase, chitinase, and pectinase were not detected in any strain. Nine strains showed nitrogen fixing ability (acetylene reduction assay) and 26 were capable of solubilizing phosphate. In the presence of 100 mg/l tryptophan, all strains except one produced indole acetic acid in the growth medium. All isolates were positive for ACC deaminase activity. Six strains produced homoserine lactones and three produced HCN and hexamate type siderophores. One isolate was capable of inhibiting the growth of 24 pathogenic fungal strains of Colletotrichum, Fusarium, Pythium, and Rhizoctonia spp. In tests of their abilities to grow under a range of temperature, pH, and NaCl concentrations, all isolates grew well on plates with 3% NaCl and most of them grew well at 4 to $41^{\circ}C$ and at pH 11.

Selection of R. japonicum Strains for Developing Soybean Inoculant and Plasmid Characterization (대두접종제(大豆接種劑) 개발(開發)을 위한 우수근류균(優秀根瘤菌)의 선발(選拔) 및 plasmid 특성(特性))

  • Kim, Chang-Jin;Kim, Sung-Hoon;Rhee, Yoon;Yoo, Ick-Dong;Mheen, Tae-Ick
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.149-155
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    • 1985
  • 590 strains of Rhizobia were isolated from root nodules of the legumes collected at 223 sites in Korea. According to their host specificities they were classified into R. japonicum(218 strains), R. phaseoli(101 strains), R. trifolii(97 strains), R. meliloti(4 strains), R. leguminosarium(1 strain), Rhizobium species(101 strains), and unidentified species(159 strains). 3 potent strains R-138, R-168, and R-214 of R. japonicum have been selected based on the infectivity to soybean cultivar and effeciency of nitrogen fixation. It was observed that the fast-growing strains of R. japonicum contained 1 to 4 plasmids of M.W. of 35-300 Md. However, plasmids were hardly detected for the slow-growing strains.

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Kinetics of Malonyl-CoA Synthetase from Rhizobium trifolii and Evidences for Malonyl-AMP Formation as a Reaction Intermediate

  • Kang, Sang-Won;Hong, Sung-Yu;Ryoo, Hyung-Don;Rhyu, Gyung-Ihm;Kim Yu-Sam
    • Bulletin of the Korean Chemical Society
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    • v.15 no.5
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    • pp.394-399
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    • 1994
  • The catalytic mechanism of malonyl-CoA synthetase from Rhizobium trifolii was investigated by the steady state kinetics and intermediate identification. Initial velocity studies and the product inhibition studies with AMP and PPi strongly suggested ordered Bi Uni Uni Bi Ping-Pong Ter Ter system as the most probable steady state kinetic mechanism of malonyl-CoA synthetase. Michaelis constants were $0.17{\pm}0.04 {\mu}M,\;0.24{\pm}0.18 {\mu}M\;and\;0.045{\pm}0.26 {\mu}$M for ATP, malonate and CoA, respectively. The TLC analysis of the $^{32}P-labelled$ products in reaction mixture containing $[{\gamma}-^{32}P]$ ATP in the absence of CoA showed that PPi was produced after the sequential addition of ATP and malonate. Formation of malonyl-AMP, suggested as an intermediate in the kinetically deduced mechanism, was confirmed by the analysis of $^{31}P-NMR$ spectra of AMP product isolated from the $^{18}O$ transfer experiment using $[^{18}O]$malonate. Two resonances were observed, corresponding to AMP labelled with zero and one atom of $^{18}O$, indicating that one atom of $^{18}O$ transferred from $[^{18}O]$malonate to AMP through the formation of malonyl-AMP. Formation of malonyl-AMP was also confirmed through the TLC analysis of reaction mixture containing $[{\alpha}-^{32}P]$ATP. These results strongly support the ordered Bi Uni Uni Bi Ping-Pong Ter Ter mechanism deduced from the initial velocity and product inhibition studies.