• Title/Summary/Keyword: 16S-rRNA

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Identification of Endophytic Bacteria in Panax ginseng Seeds and Their Potential for Plant Growth Promotion (인삼종자로부터 분리된 내생균의 동정과 식물생장 촉진 관련 활성의 평가)

  • Um, Yurry;Kim, Bo Ra;Jeong, Jin Ju;Chung, Chan Moon;Lee, Yi
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.4
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    • pp.306-312
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    • 2014
  • Endophytes are microorganisms that live in the internal tissues of plants without harming the host plants. In this symbiotic relationship, the host plants provide nutrients and shelter to the endophytes, in turn, endophytes can promote the growth of host plants and act as a biological control agents against plant pathogens. Plant-microbe interactions like this are noted for natural methods for sustainable agriculture and environmental conservation. However, in spite of the infinite potential, there are only a few reports on the endophytes present in ginseng. In this study, we isolated and identified the endophytes from Panax ginseng seeds and evaluated the biological activities (IAA production ability, nitrogen fixation ability, phosphate solubilization capacity, siderophore production ability, and antifungal activities) of the endophyte isolates. Eight different endophytes were identified by 16S rRNA sequencing. Most of the endophytes have antibiotic and plant growth promoting (PGP) activities. Particularly, PgSEB5-37E have the highest antibiotic activity, both PgSEB5-37B and PgSEB5-37H have high PGP traits such as an abilities to produce IAA, solubilize phosphate and fix nitrogen. These results indicated that the endophytes from P. ginseng seeds may have applicable value to many industries. In order to use the isolated endophytes, quantitative analysis and field tests are needed to be performed.

Characterization of an Alkaline Protease from an Alkalophilic Bacillus pseudofirmus HS-54 (호알칼리성 Bacillus pseudofirmus HS-54가 생산하는 알칼리성 Protease의 특성)

  • Bang, Seong-Ho;Jeong, In-Sil
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.194-199
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    • 2011
  • An alkalophilic bacterium producing alkaline protease was isolated from waste water and solar saltern sample and identified as Bacillus pseudofirmus HS-54 based on morphological, biochemical characteristics as well as 16S-rRNA gene sequencing. The HS-54 protease was purified to homogeneity using ammonium sulfate precipitation, DEAE cellulose column chromatography, and sephadex G-100 gel filtration with a 4.0 purification fold. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 27 kDa. The optimal pH and temperature for the purified protease activity were 10.0 and $50^{\circ}C$, respectively. The purified enzyme was relatively stable at the pH range of 6.0-11.0 and at the temperature below $50^{\circ}C$. This enzyme was activated by $Ca^{2+}$ and $Mg^{2+}$ and inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Al^{3+}$, $Ag^{2+}$. And this enzyme was strongly inhibited by PMSF, suggesting that it belongs to the serine protease superfamily.

Effect of button mushroom compost on mobilization of heavy metals by sunflower

  • Kyeong, Ki-Cheon;Kim, Yong-Gyun;Lee, Chan-Jung;Lee, Byung-Eui;Lee, Heon-Hak;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.163-170
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    • 2014
  • The potential ability of Button mushroom compost (BMC) to solubilize heavy metals was estimated with metal contaminated soils collected from abandoned mines of Boryeong area in South Korea. The bacterial strains in BMC were isolated for investigating the mobilization of metals in soil or plant by the strains and identified according to 16S rRNA gene sequence analysis. When metal solubilization potential of BMC was assessed in a batch experiment, the BMC was found to be capable of solubilizing metals in the presence of metals (Co, Pb and Zn) and the results showed that inoculation of BMC could increase the concentrations of water soluble Co, Pb and Cd by 35, 25 and 45% respectively, than those of non-inoculated soils. BMC-assisted growth promotion and metal uptake in sunflower (Helianthus annuus) was also evaluated in a pot experiment. In comparison with non-inoculated seedlings, the inoculation led to increase the growth of H. annuus by 27, 25 and 28% respectively in Co, Pb and Zn contaminated soils. Moreover, enhanced accumulation of Co, Pb and Zn in the shoot and root systems was observed in inoculated plants, where metal translocation from root to the above-ground tissues was also found to be enhanced by the BMC. The apparent results suggested that the BMC could effectively be employed in enhancing phytoextraction of Co, Pb and Zn from contaminated soils.

A Bacterial Endophyte, Pseudomonas brassicacearum YC5480, Isolated from the Root of Artemisia sp. Producing Antifungal and Phytotoxic Compounds

  • Chung, Bok-Sil;Aslam, Zubair;Kim, Seon-Won;Kim, Geun-Gon;Kang, Hye-Sook;Ahn, Jong-Woong;Chung, Young-Ryun
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.461-468
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    • 2008
  • An endophytic bacterial strain YC5480 producing antifungal and phytotoxic compounds simultaneously was isolated from the surface sterilized root of Artemisia sp. collected at Jinju area, Korea. The bacterial strain was identified as a species of Pseudomonas brassicacearum based on its 16S rRNA gene sequence analysis and physiological and biochemical characteristics. The seed germination and growth of monocot and dicot plants were inhibited by culture filtrate (1/10-strength Tryptic Soy Broth) of the strain. The germination rate of radish seeds in the culture filtrate differed in various culture media. Only 20% of radish seeds germinated in the culture media of 1/2 TSB for 5 days incubation. Mycelial growth of fungal pathogens, Colletotrichum gloeosporioides, Fusarium oxysporum and Phytophthora capsici was also inhibited by the culture filtrate of the strain YC5480. An antifungal compound, KS-1 with slight inhibitory activity of radish seed germination at 1,000 ppm and a seed germination inhibitory compound, KS-2 without suppression of fungal growth were produced simultaneously in TSB. The compounds KS-1 and KS-2 were identified to be 2,4-diacetylphloroglucinol (DAPG) and 2,4,6-trihydroxyacetophenone (THA), respectively.

Microbial Community Dynamics in Batch High-Solid Anaerobic Digestion of Food Waste Under Mesophilic Conditions

  • Yi, Jing;Dong, Bin;Xue, Yonggang;Li, Ning;Gao, Peng;Zhao, Yuxin;Dai, Lingling;Dai, Xiaohu
    • Journal of Microbiology and Biotechnology
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    • v.24 no.2
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    • pp.270-279
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    • 2014
  • Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g $VS_{removal}$. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

Phylogenetics, Safety and In Vitro Functional Properties of Bacillus Species Isolated from Iru, a Nigerian Fermented Condiment

  • Adewumi, Gbenga Adedeji;Grover, Sunita;Isanbor, Chukwuemeka;Oguntoyinbo, Folarin Anthony
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.498-508
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    • 2019
  • Bacillus species were isolated from iru, a traditional fermented condiment in Nigeria. Polyphasic approach was used to evaluate the phylogenetic relationship and strain sub-type of the isolated species. Additionally, the phylogenetic profiles of the species isolated from iru were compared with those of bacilli isolated from different continents. The phylogenetic diversity analysis was performed using the combination of 16S rRNA gene sequencing, ITS-PCR, ITS-PCR-RFLP, and M13 RAPD-PCR. The analysis revealed that Bacillus subtilis U170B and B. subtilis U146A isolated from iru were the closest relatives of strains belonging to the phylogeny of B. subtilis sensu stricto and were related to other bacilli isolated from different continents that had functional benefits. The two isolated species exhibited resistance to acidic pH (pH 2.0). The survival rates of B. subtilis U170B, B. subtilis U146A, and B. clausii UBBC-07 (commercial probiotic strain) cultured at pH 2.0 for 3 h were 33.45, 12.44, and 9.53%, respectively. The strains were highly tolerant to bile salts [0.3% (w/v)]. B. subtilis U170B exhibited the highest cell viability (43.45%) when cultured for 3 h in the presence of bile salts, followed by B. subtilis U146A (25%) and B. clausii UBBC-07 (18.94%). B. subtilis U170B and B. subtilis U146A did not exhibit haemolytic activity and were susceptible to different antibiotics. Additionally, these two strains exhibited weak antagonistic activity against B. cereus. The diverse wild strains of B. subtilis can be used as a safe multifunctional starter culture for the industrial production of condiments with health benefits.

A Sporolactobacillus-, Clostridium-, and Paenibacillus- Dominant Microbial Consortium Improved Anaerobic RDX Detoxification by Starch Addition

  • Khan, Muhammad Imran;Yoo, Keunje;Kim, Seonghoon;Cheema, Sardar Alam;Bashir, Safdar;Park, Joonhong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.839-847
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    • 2020
  • In the present study, an anaerobic microbial consortium for the degradation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) was selectively enriched with the co-addition of RDX and starch under nitrogen-deficient conditions. Microbial growth and anaerobic RDX biodegradation were effectively enhanced by the co-addition of RDX and starch, which resulted in increased RDX biotransformation to nitroso derivatives at a greater specific degradation rate than those for previously reported anaerobic RDX-degrading bacteria (isolates). The accumulation of the most toxic RDX degradation intermediate (MNX [hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine]) was significantly reduced by starch addition, suggesting improved RDX detoxification by the co-addition of RDX and starch. The subsequent MiSeq sequencing that targeted the bacterial 16S rRNA gene revealed that the Sporolactobacillus, Clostridium, and Paenibacillus populations were involved in the enhanced anaerobic RDX degradation. These results suggest that these three bacterial populations are important for anaerobic RDX degradation and detoxification. The findings from this work imply that the Sporolactobacillus, Clostridium, and Paenibacillus dominant microbial consortium may be valuable for the development of bioremediation resources for RDX-contaminated environments.

Relationship Between Genome Similarity and DNA-DNA Hybridization Among Closely Related Bacteria

  • Kang, Cheol-Hee;Nam, Young-Do;Chung, Won-Hyong;Quan, Zhe-Xue;Park, Yong-Ha;Park, Soo-Je;Desmone, Racheal;Wan, Xiu-Feng;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.945-951
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    • 2007
  • DNA-DNA hybridization has been established as an important technology in bacterial species taxonomy and phylogenetic analysis. In this study, we analyzed how the efficiency with which the genomic DNA from one species hybridizes to the genomic DNA of another species (DNA-DNA hybridization) in microarray analysis relates to the similarity between two genomes. We found that the predicted DNA-DNA hybridization based on genome sequence similarity correlated well with the experimentally determined microarray hybridization. Between closely related strains, significant numbers of highly divergent genes (>55% identity) and/or the accumulation of mismatches between conserved genes lowered the DNA-DNA hybridization signal, and this reduced the hybridization signals to below 70% for even bacterial strains with over 97% 16S rRNA gene identity. In addition, our results also suggest that a DNA-DNA hybridization signal intensity of over 40% indicates that two genomes at least shared 30% conserved genes (>60% gene identity). This study may expand our knowledge of DNA-DNA hybridization based on genomic sequence similarity comparison and further provide insights for bacterial phylogeny analyses.

Genetic and Phenotypic Diversity of Parathion-Degrading Bacteria Isolated from Rice Paddy Soils

  • Choi, Min-Kyeong;Kim, Kyung-Duk;Ahn, Kyong-Mok;Shin, Dong-Hyun;Hwang, Jae-Hong;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1679-1687
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    • 2009
  • Three parathion-degrading bacteria and eight pairs of bacteria showing syntrophic metabolism of parathion were isolated from rice field soils, and their genetic and phenotypic characteristics were investigated. The three isolates and eight syntrophic pairs were able to utilize parathion as a sole source of carbon and energy, producing p-nitrophenol as the intermediate metabolite during the complete degradation of parathion. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Burkholderia, Arthrobacter, Pseudomonas, Variovorax, and Ensifer. The chromosomal DNA patterns of the isolates obtained by polymerasechain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences were distinct from one another. Ten of the isolates had plasmids. All of the isolates and syntrophic pairs were able to degrade parathion-related compounds such as EPN, p-nitrophenol, fenitrothion, and methyl parathion. When analyzed with PCR amplification and dot-blotting hybridization using various primers targeted for the organophosphorus pesticide hydrolase genes of previously reported isolates, most of the isolates did not show positive signals, suggesting that their parathion hydrolase genes had no significant sequence homology with those of the previously reported organosphophate pesticide-degrading isolates.

Characterization of a Phenazine and Hexanoyl Homoserine Lactone Producing Pseudomonas aurantiaca Strain PB-St2, Isolated from Sugarcane Stem

  • Mehnaz, Samina;Baig, Deeba Noreen;Jamil, Farrukh;Weselowski, Brian;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1688-1694
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    • 2009
  • A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.