• Title/Summary/Keyword: Isolation resistance

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

Changes in Sensitivity Levels of Botrytis spp. Population Isolated from Lily to Fungicides and Control under Field Condition (나리에서 분리한 잎마름병균의 살균제에 대한 감수성 변화와 포장 방제)

  • Hahm, Soosang;Kyeong, Kicheon;Kim, Byungryun;Han, Kwangseop;Choi, Jongjin;Nam, Yunkyu;Yu, Seunghun
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.7-11
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    • 2013
  • Forty eight isolates of Botrytis elliptica and 23 isolates of B. cinerea from several locations in Korea were tested for resistance to fungicides used in the farmer's fields. Isolation frequency of B. elliptica having $EC_{50}$ (effective concentration of 50%) value $500-1000{\mu}g/ml$ to benomyl and mancozeb appeared highly, suggesting that the two fungicides are not effective in controlling leaf blight of lily in the field. The isolates were tested for resistance to fungicides procymidone and iprodione which were most commonly used in the farmer's fields. The rates of $EC_{50}$ value $5-50{\mu}g/ml$ to procymidome and iprodione were 93.7% and 100%, respectively, and those of $0-0.1{\mu}g/ml$ to diethofencarb+carbendazim and fludioxonil were 98.0% and 93.8%, respectively. In the rain-protected cultivation, control of leaf blight of lily was the most effective when iprodine, diethofencarb+carbendazim, and fludioxonil were sprayed alternately four times during the growing season.

Isolation and Identification of a New Gene Related to Salt Tolerance in Chinese Cabbage (배추에서 신규 염 저항성 관련 유전자 분리 및 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.748-755
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    • 2013
  • This study was conducted to find a salt tolerance gene in Brassica rapa. In order to meet this objective, we analyzed data from a KBGP-24K oligo chip [BrEMD (Brassica rapa EST and microarray database)] of the B. rapa ssp. pekinensis 'Chiifu' under salt stress (250 mM NaCl). From the B. rapa KBGP-24K microarray chip analysis, 202 salt-responsive unigenes were primarily selected under salt stress. Of these, a gene with unknown function but known full-length sequence was chosen to closely investigate the gene function. The selected gene was named BrSSR (B. rapa salt stress resistance). BrSSR contains a 285 bp open reading frame encoding a putative 94-amino acid protein, and a DUF581 domain. The pSL94 vector was designed to over-express BrSSR, and was used to transform tobacco plants for salt tolerance analysis. T1 transgenic tobacco plants that over-expressed BrSSR were selected by PCR and DNA blot analyses. Quantitative real-time RT PCR revealed that the expression of BrSSR in transgenic tobacco plants increased by approximately 3.8-fold. Similar results were obtained by RNA blot analysis. Phenotypic characteristics analysis showed that transgenic tobacco plants with over-expressed BrSSR were more salt-tolerant than the wild type control under 250 mM NaCl for 5 days. Based on these results, we hypothesized that the over-expression of BrSSR may be closely related to the enhancement of salt tolerance.

Interaction between the Rice Pathogens, Fusarium graminearum and Burkholderia glumae

  • Lee, Jungkwan;Jung, Boknam;Park, Jungwook;Kim, Sungyoung;Youn, Kihun;Seo, Young-Su
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.13-13
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    • 2014
  • Species belonging to the genus Fusarium are widely distributed and cause diseases in many plants. Isolation of fungal strains from air or cereals is necessary for disease forecasting, disease diagnosis, and population genetics [1]. Previously we showed that Fusarium species are resistant to toxoflavin produced by the bacterial rice pathogen Burkholderia glumae while other fungal genera are sensitive to the toxin, resulting in the development of a selective medium for Fusarium species using toxoflavin [2]. In this study, we have tried to elucidate the resistant mechanism of F. graminearum against toxoflavin and interaction between the two pathogens in nature. To test whether B. glumae affects the development of F. graminearum, the wild-type F. graminearum strains were incubated with either the bacterial strain or supernatant of the bacterial culture. Both conditions increased the conidial production five times more than when the fungus was incubated alone. While co-incubation resulted in dramatic increase of conidial production, conidia germination delayed by either the bacterial strain or supernatant. These results suggest that certain factors produced by B. glumae induce conidial production and delay conidial germination in F. graminearum. To identify genes related to toxoflavin resistance in F. graminearum, we screened the transcriptional factor mutant library previously generated in F. graminearum [3] and identified one mutant that is sensitive to toxoflavin. We analyzed transcriptomes of the wild-type strain and the mutant strain under either absence or presence of toxoflavin through RNAseq. Expression level of total genes of 13,820 was measured by reads per kilobase per million mapped reads (RPKM). Under the criteria with more than two-fold changes, 1,440 genes were upregulated and 1,267 genes were down-regulated in wild-type strain than mutant strain in response to toxoflavin treatment. A comparison of gene expression profiling between the wild type and mutant through gene ontology analysis showed that genes related to metabolic process and oxidation-reduction process were highly enriched in the mutant strain. The data analyses will focus on elucidating the resistance mechanism of F. graminearum against toxoflavin and the interaction between the two pathogens in rice. Further evolutionary history will be traced through figuring out the gene function in populations and in other filamentous fungi.

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Isolation and Identification of Pseudomonas aeruginosa in Natural Environments by International Organization for Standardization ISO/NP 16266 (국제표준화기구 ISO/NP 16266 방법을 이용한 환경 중 Pseudomonas aeruginosa의 분리 및 동정)

  • Lee, Siwon;Kim, Ji Hye;Lee, Bo-Ram;Joo, Youn-Lee;Choe, Byeol;Park, Su Jeong;Chung, Hyen-Mi;Jheong, Weon Hwa
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.384-386
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    • 2014
  • Pseudomonas aeruginosa is an opportunistic pathogen that inhabits various natural and artificial environments, such as pathogenesis, water, soil and air. They can cause serious problems, such as pathogenic infection. In this study, 220 colonies were isolated from water and soil environment that assumed to be P. aeruginosa using a membrane filter method based on International Organization for Standardization (ISO/NP 16266). Identification of the isolates was determined by physiobiochemical characteristics using newly modified ISO method which includes the resistance to 1,10 phenanthroline test. Only one of 220 presumed P. aeruginosa strains isolated from effluence water using a drain swab was determined as P. aeruginosa-positive by the ISO/NP 16266 method. Subsequently, the resistance to 1,10 phenanthroline test, which was newly proposed by ISO in 2014 and applied in this study, was considered as more precise and improvable method for identification of P. aeruginosa.

Isolation and Degradation Activity of a TBTCl (Tributyltin Chloride) Resistant Bacteriain Gwangyang Bay (광양만에서 TBTCl (Tributyltin Chloride) 내성세균의 분리 및 분해활성)

  • Jeong, Seong-Yun;Son, Hong-Joo;Jeoung, Nam-Ho
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.424-431
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    • 2011
  • BACKGROUND: Tributyltin chloride is among the most toxic compounds known for aquatic ecosystems. Microorganisms are responsible for removal of TBTCl. Nevertheless, only a limited number of marine bacteria were investigated for biodegradation of TBTCl in Korea. METHODS AND RESULTS: The number of TBTCl resistant bacteria ranged from $2.5{\times}10^3$ to $3.8{\times}10^3$ cfu/mL in the seawater, and ranged from $3.2{\times}10^5$ to $9.1{\times}10^5$ cfu/g in the surface sediment, respectively. The morphological, physiological, and biochemical characteristics of TBTCl resistant bacteria were investigated by API 20NE and other tests. The most abundant species of TBTCl resistant bacteria were Vibrio spp. (19.2%), Bacillus spp. (16.2%), Aeromonas spp. (15.2%), and Pseudomonas spp. (13.1%), etc. Eleven TBTCl resistant isolates also had a resistance to heavy metals (Cd, Cu, Hg, and Zn). Among them, isolate T7 showing the strong TBTCl-resistance was selected. This isolate was identified as the genus Pantoea by 16S rRNA gene sequencing and designated as Pantoea sp. T7. In addition, this bacterium was cultivated up to the growth of 50.7% after 60 hrs at TBTCl concentration of $500{\mu}M$. TBTCl-degrading activity of Pantoea sp. T7 was measured by GC-FPD analysis. As a result of biological TBTCl-degradation at TBTCl concentration of $100{\mu}M$, TBTCl-removal efficiency of Pantoeasp. T7 was 62.7% after 40 hrs. CONCLUSION(S): These results suggest that Pantoea sp. T7 is potentially useful for the bioremediation of TBT contamination.

Prevalence and Antibiotic Resistance Patterns in Listeria Monocytogenes Isolated from Food (식품에서 분리된 리스테리아 모노사이토젠스의 분포 및 항생제 내성)

  • Jeong, Hyo-Won;Park, Sang-Hun;Lee, Jib-Ho;Kim, Soo-Jin;Ryu, Seung-Hee;Song, Mi-Ok;Park, Sun-Hee;Jo, Jeong-Yun;Park, Geon-Yong;Choi, Sung-Min
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.26-30
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    • 2014
  • Listeria monocytogenes continues to be a important food safety concern. The aims of the present study were to investigate the prevalence and antimicrobial susceptibility of L. monocytogenes. A total of 1,042 samples was collected from 2010 to 2011 in Seoul and twelve L. monocytogenes were isolated. Isolation rate was Gimbap (0.8%), Beef (yukhoe) (2.6%), Pork (cooked) (5.9%), Fish(cooked) (6.3%), and Salmon (11.1%), respectively. In this study, most of the isolates were susceptible to antibiotics. The most common resistance was cefotetan on 11 isolates, followed by cefotaxime on 7 isolates, cepefime on 6 isolates and tetracyclin on 3 isolates.

The Significance of Cancer Stem Cells in Canine Mammary Gland Tumors (개 유선종양 내 종양줄기세포의 중요성)

  • Park, Seo-Young;Baek, Yeong-Bin;Park, Sang-Ik;Lee, Chang-Min;Kim, Sung-Hak
    • Journal of Life Science
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    • v.31 no.2
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    • pp.248-255
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    • 2021
  • Mammary gland tumors are one of the most common cancers in female dogs, and there are various types of cells depending on the tumor type. Complex carcinoma consists of a combination of luminal epithelial and myoepithelial cells with intra-tumoral heterogeneity. However, the origins of these tumor cells and their effects on the malignancies of tumors have not been identified. Recently, it has been reported that cancer stem cells, identified in several types of human tumors, are involved in tumor heterogeneity and may also contribute to malignancies such as tumor recurrence and metastasis. Interestingly, cancer stem cells share several abilities of self-renewal and cell differentiation into multiple types of cancer cells, but they have abnormal genetic mutation and signal transduction pathways to regulate the maintenance of stem cell characters. Moreover, it is known that these cell populations contribute to cell metastasis as well as cell resistance against chemo- and radio-therapeutics that promote tumor recurrence. The existence of cancer stem cells might explain the intra-tumoral heterogeneity and cancer aggressiveness during tumorigenesis in canine mammary gland tumors. This review summarizes the characteristics and types of canine mammary gland tumors, the definition of tumor stem cells, methods of isolation, and clinical significance.

Isolation, Identification, and Probiotic Properties of Lactobacillus reuteri HY701 from Human Feces

  • Kim, Jun-Tae;Jung, Hwang-Young;Lee, Na-Kyoung;Rhim, Seong-Lyul;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.677-682
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    • 2006
  • Strain HY701 was isolated from human feces for probiotic use by selecting highly resistant isolates to artificial gastric acid and bile acid. Strain HY701 was identified as Lactobacillus reuteri using 16S rDNA sequencing, and tentatively named L. reuteri HY701. The resistance of L. reuteri HY701 to artificial gastric acid (PH 2.5) was high with a survival rate of over 90%. L. reuteri HY701 also showed high tolerance to artificial bile acid after incubation in artificial gastric acid. Using the API ZYM test kit, the carcinogenic enzymes (${\beta}$-glucuronidase and (${\beta}$-glucosidase were not detected with L. reuteri HY70l, while the beneficial enzyme (${\beta}$-galactosidase was weakly detected. L. reuteri HY701 was sensitive to $100\;{\mu}g/mL$ nisin, $20\;{\mu}g/mL$ roxithromycin, $15\;{\mu}g/mL$ erythromycin, but resistant to $20\;{\mu}g/mL$ streptomycin, $10\;{\mu}g/mL$ tetracycline, $20\;{\mu}g/mL$ ciprofloxacin, $20\;{\mu}g/mL$ nystatin, $20\;{\mu}g/mL$ gentamycin, $10\;{\mu}g/mL$ doxycycline, $10\;{\mu}g/mL$ chloramphenicol, and $20\;{\mu}g/mL$ ampicillin. L. reuteri HY701 was shown to possess bactericidal activity as it inhibited the growth of Listeria monocytogenes ATCC 19111 and Escherichia coli JM109 completely within 24 hr of incubation. These results indicate that L. reuteri HY701 could be used as a probiotic strain.

Isolation and Identification of Antifungal Compounds from $Bacillus$ $subtilis$ C9 Inhibiting the Growth of Plant Pathogenic Fungi

  • Islam, Md. Rezuanul;Jeong, Yong-Tae;Lee, Yong-Se;Song, Chi-Hyun
    • Mycobiology
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    • v.40 no.1
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    • pp.59-65
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    • 2012
  • Antagonistic microorganisms against $Rhizoctonia$ $solani$ were isolated and their antifungal activities were investigated. Two hundred sixteen bacterial isolates were isolated from various soil samples and 19 isolates were found to antagonize the selected plant pathogenic fungi with varying degrees. Among them, isolate C9 was selected as an antagonistic microorganism with potential for use in further studies. Treatment with the selected isolate C9 resulted in significantly reduced incidence of stem-segment colonization by $R.$ $solani$ AG2-2(IV) in Zoysia grass and enhanced growth of grass. Through its biochemical, physiological, and 16S rDNA characteristics, the selected bacterium was identified as $Bacillus$ $subtilis$ subsp. $subtilis$. Mannitol (1%) and soytone (1%) were found to be the best carbon and nitrogen sources, respectively, for use in antibiotic production. An antibiotic compound, designated as DG4, was separated and purified from ethyl acetate extract of the culture broth of isolate C9. On the basis of spectral data, including proton nuclear magneric resonance ($^1H$ NMR), carbon nuclear magneric resonance ($^{13}C$ NMR), and mass analyses, its chemical structure was established as a stereoisomer of acetylbutanediol. Application of the ethyl acetate extract of isolate C9 to several plant pathogens resulted in dose-dependent inhibition. Treatment with the purified compound (an isomer of acetylbuanediol) resulted in significantly inhibited growth of tested pathogens. The cell free culture supernatant of isolate C9 showed a chitinase effect on chitin medium. Results from the present study demonstrated the significant potential of the purified compound from isolate C9 for use as a biocontrol agent as well as a plant growth promoter with the ability to trigger induced systemic resistance of plants.