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Proteomic Analysis of Global Changes in Protein Expression During Exposure of Gamma Radiation in Bacillus sp. HKG 112 Isolated from Saline Soil

  • Gupta, Anil Kumar;Pathak, Rajiv;Singh, Bharat;Gautam, Hemlata;Kumar, Ram;Kumar, Raj;Arora, Rajesh;Gautam, Hemant K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.574-581
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    • 2011
  • A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

Keratinase Production by Recalcitrant Feather Degrading Pseudomonas Geniculata and Its Plant Growth Promoting Activity (난분해성 우모분해 Pseudomonas geniculata에 의한 케라틴 분해효소 생산 및 식물성장 촉진 활성)

  • Go, Tae-Hun;Lee, Sang-Mee;Lee, Na-Ri;Jeong, Seong-Yun;Hong, Chang-Oh;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.22 no.11
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    • pp.1457-1464
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    • 2013
  • We investigated the optimal conditions for keratinase production by feather-degrading Pseudomonas geniculata H10 using one variable at a time (OVT) method. The optimal medium composition and cultural condition for keratinase production were determined to be glucose 0.15% (w/v), beef extract 0.08% (w/v), $KH_2PO_4$ 0.12% (w/v), $K_2HPO_4$ 0.02% (w/v), NaCl 0.07% (w/v), $MgSO_4{\cdot}7H_2O$ 0.03%, $MgCl_2{\cdot}6H_2O$ 0.04% along with initial pH 10 at 200 rpm and $25^{\circ}C$, respectively. The production yield of keratinase was 31.6 U/ml in an optimal condition, showing 4.6-fold higher than that in basal medium. The strain H10 also showed plant growth promoting activities. This strain had ammonification activity and produced indoleacetic acid (IAA), siderophore and a variety of hydrolytic enzymes such as protease, lipase and chitinase. Therefore, this study showed that P. geniculata H10 could be not only used to upgrade the nutritional value of feather wastes but also useful in situ biodegradation of feather wastes. Moreover, it is also a potential candidate for the development of biofertilizing agent applicable to crop plant soil.

Induction of systemic resistance in Panax ginseng against Phytophthora cactorum by native Bacillus amyloliquefaciens HK34

  • Lee, Byung Dae;Dutta, Swarnalee;Ryu, Hojin;Yoo, Sung-Je;Suh, Dong-Sang;Park, Kyungseok
    • Journal of Ginseng Research
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    • v.39 no.3
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    • pp.213-220
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    • 2015
  • Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.

Phosphate Solubilization and Gene Expression of Phosphate-Solubilizing Bacterium Burkholderia multivorans WS-FJ9 under Different Levels of Soluble Phosphate

  • Zeng, Qingwei;Wu, Xiaoqin;Wang, Jiangchuan;Ding, Xiaolei
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.844-855
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    • 2017
  • Phosphate-solubilizing bacteria (PSB) have the ability to dissolve insoluble phosphate and enhance soil fertility. However, the growth and mineral phosphate solubilization of PSB could be affected by exogenous soluble phosphate and the mechanism has not been fully understood. In the present study, the growth and mineral phosphate-solubilizing characteristics of PSB strain Burkholderia multivorans WS-FJ9 were investigated at six levels of exogenous soluble phosphate (0, 0.5, 1, 5, 10, and 20 mM). The WS-FJ9 strain showed better growth at high levels of soluble phosphate. The phosphate-solubilizing activity of WS-FJ9 was reduced as the soluble phosphate concentration increased, as well as the production of pyruvic acid. Transcriptome profiling of WS-FJ9 at three levels of exogenous soluble phosphate (0, 5, and 20 mM) identified 446 differentially expressed genes, among which 44 genes were continuously up-regulated when soluble phosphate concentration was increased and 81 genes were continuously down-regulated. Some genes related to cell growth were continuously up-regulated, which would account for the better growth of WS-FJ9 at high levels of soluble phosphate. Genes involved in glucose metabolism, including glycerate kinase, 2-oxoglutarate dehydrogenase, and sugar ABC-type transporter, were continuously down-regulated, which indicates that metabolic channeling of glucose towards the phosphorylative pathway was negatively regulated by soluble phosphate. These findings represent an important first step in understanding the molecular mechanisms of soluble phosphate effects on the growth and mineral phosphate solubilization of PSB.

Biodegradation of the Organic Solvents by an Isolated Pseudomonas sp. BCNU 154

  • Kim, Mi-Rim;Jeong, Mi-Yeon;Seong, Eun-Mi;Lee, Ho-Won;Lee, Yeong-Geun;Kim, Yong-Gyun;Park, Jeong-Uk;Ju, U-Hong
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.374-377
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    • 2000
  • A strain of Pseudomonas sp. BCNU 154 was isolated from contaminated soil with orangic solvents as the sole source of carbon and energy. It utilized an exceptionally wide aromatic substrates. The strain BCNU 154 was able to utilize toluene, p-xylene, ethylbenzene, cumene, as the only carbon and energy source. When toluene or p-xylene was used as the sole carbon and energy source, the compound was rapidly degraded with significant increase in biomass concentruction. The biodegradation of this compound was observed when ethylbenzene or cumene was supplied on the carbon source and energy source, which may be a candidate extremophilic bacterium for the bioremediation technology.

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Screening and Characterization of the High-Alcohol Producing Saccharornyces cerevisiae Dl (고농도 알콜발효효모 Saccharomyces cerevisiae D1의 분리 및 특성)

  • 양지영;박경호;백운화;유주현
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.511-516
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    • 1990
  • A high-alcohol producing yeast strain had sugar and alcohol tolerance was isolated from soil and identified as Saccharomyces cerevisiae Dl according to the Lodder's yeast taxonomic studies. On investigation of the characteristics of the strain, it could grow in 60% glucose, within 15% ethanol and in the YPD medium containing 2.0 mM copper. It had 39.1% the inhibition rate of fermentation efficiency and 8% viability after 2 days in the YPD medium containing 15% ethanol. Its optimum initial pH, growth temperature, initial glucose concentration for the production of alcohol showed pH 4.5, $30^{\circ}C$, and 30%, respectively. Saa:hwomyce8 mvisiae Dl produced 14.0% (wlv) alcohol when incubated at $30^{\circ}C$, with orbital shaking 150 rpm for 72 h in a medium (pH 4.5) containing 30% (wfv) glucose.

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Evaluation of induced systemic resistance agent, Bacillus subtilis strain BAC02-4 against Magnaporthe grisea in rice in field

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Han, Seong-Sook;Jung, Won-Kwon;Park, Jo-Im;Park, Sung-Tae;Kim, Soon-Chul
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.105.3-106
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    • 2003
  • Bacillus subtilis strain BAC02-4 was tested for its ability induced systemic resistance(ISR) in rice against Magnaporthe grisea We extend these studies to investigate the biological induction of systemic resistance in rice following treatment with the inducer isolate BAC02-4 and naturally infested with Pyricularia oryzae. We also determine levels of ISR activity during the period between disease development and the onset of systemic resistance. Comparition of lesion number according to applied concentration of BAC02-4 to 'Nagdongbyeo' when naturally infested with the conidia of P. grisea. Results from the blast nusery trial using the 'Nagdongbyeo' showed very low rice blast severity with the inducer concentration of 10$\^$8/ cfu level. Considering the low level of treatment and untreated control were observed to have developed typical susceptible lesion type. Highest protection against the rice blast pathogen when applied three times with 5 days interval as root drench at 5 to 6 leaf stage before pathogen challenge. But higher dose of bacterial inducer produced a little stunted plants with less number lesions and delayed disease development. Diseased leaf area of treated with suspension of the isolate which gave about 80% of control efficacy at 20 days later comparable to that in noninfested, inducer-free soil.

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A Study on the Relationship between Void Ratio and Permeability by Constant Strain Rate Consolidation Test (일정변형률 압밀시험을 이용한 간극비-투수계수의 관계 연구)

  • Joo, Jong-Jin;Lim, Hyung-Duk;Lee, Woo-Jin;Kim, Dae-Kyu;Kim, Nak-Kyung;Kim, Hyung-Joo
    • Journal of the Korean GEO-environmental Society
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    • v.3 no.3
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    • pp.15-25
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    • 2002
  • The permeability coefficient is one of the fundamental engineering properties of soft clays. Consolidation process as well as migration of pollutants in soil are affected the permeability coefficient, which generally decreases with the reduction in void ratio during consolidation. After Kozeny(1927) and Carman(1956), many researchers have proposed the relationships between void ratio and permeability in such forms as; (1) log e - log k(1+e), (2) e - log k, or (3) log e - log k. Constant rate of strain (CRS) tests was performed with undisturbed samples obtained at Kunsan and Kimhae deposits, which are representative Korean marine clay. From the results of the tests, the relationships were found valid for Kunsan and Kimhae clays. The experimental correlation $C_k=0.5e_o$ was satisfied with Kimhae clay but not with Kunsan clay.

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The Effects of Sample Disturbance on Consolidation Properties of Yang-San Clay (시료교란이 양산 점토의 압밀특성에 미치는 영향)

  • 박정규;임형덕;이우진
    • Proceedings of the Korean Geotechical Society Conference
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    • 1999.10a
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    • pp.95-102
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    • 1999
  • It is important to estimate the magnitude of settlement and the time of primary consolidation in geotechnical engineering projects. For these purposes, site explorations and laboratory tests are usually performed. However, the mechanical properties determined from laboratory tests on the samples obtained by different sampling techniques show different trends. In this study, three types of consolidation test were carried out on the soil samples obtained by three different sampling techniques (76mm tube, 76mm piston, and block samples), to verify the effect of sample disturbance on consolidation properties. It was found that sampling methods have influence on the $\varepsilon$ - log $\sigma$'$_{v}$ relationship. While insignificant difference of compression indices from the samples obtained by different sampling techniques was observed in compression region, these values showed a different trend in precompression region. The values of $c_{v}$ and k of block samples had a tendency to be larger than those of piston and tube samples. At consolidation pressure larger than $\sigma$'$_{p}$, however, these properties became similar regardless of sampling methods. The block and piston samples gave slightly higher values of $C_{a/}$ $C_{c}$ than tube samples. In the results of I $L_{EOP}$ and CRS test, it was observed that the values of $\sigma$'$_{p measured}$/$\sigma$'$_{p best estimated}$ of Yang-San clay decreases when strain becomes larger than 1.0% and that precompression strain of block samples is in the range of 1.5~2.0% while those of piston and tube samples are 1.75~3.75%. It was also shown that the values of $\sigma$'$_{p}$ of block samples exceed those of piston and tube samples by about 6~10%.6~10%.%.%.%.%.

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The Effects of Sample Disturbance on Undrained Properties of Yangsan Clay (양산점토의 비배수 특성에 대한 시료교란의 효과)

  • 김길수;임형덕;이우진
    • Proceedings of the Korean Geotechical Society Conference
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    • 2000.03b
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    • pp.639-646
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    • 2000
  • It is important to estimate the mechanical properties of clay since it is directly related to the design and the construction of geotechnical structures. Site exploration, which is composed of boring, sampling, in-situ, or laboratory tests, is preformed to estimate the mechanical properties. However, mechanical properties of clay measured from laboratory test may be different from in-situ properties due to disturbances occurred during sampling, transportation, storage, and trimming. In this study, the degree of disturbance according to sampling method was estimated with the test results of CK/sub o/U triaxial compression test on Yangsan clay. The soil samples were obtained by three types of sampling method, j.e., 76mm-tube sampler, 76mm-piston sampler, and block sampler. In order to evaluate the quality of samples, volumetric strain, undrained shear strength, secant Young's modulus, and pore pressure coefficient at peak measured from each sample were compared with one another. From the test results, it was observed that mechanical properties of the block and piston samples were more reliable than those of tube samples. But it was observed that the water content of piston was similar to that of tube samples at given depths while the water content of block samples was 14.3∼15.8% smaller than that of piston and tube samples. In addition to the evaluation of the quality of samples, relationship between c/sub u// σ/sub vc/'and OCR was established from the results of the CK/sub o/U triaxial compression tests, which were carried out using SHANSEP method. And also undrained shear strength was analyzed using the in-situ test data such as Cone Penetration Test(CPT), Dilatometer Test(DMT), and Field Vane Test(FVT) and was compared with that evaluated from CK/sub o/U triaxial compression test.

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