• Title/Summary/Keyword: Sequence of 16S rDNA.

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Induced of Systemic Resistance against Gray Leaf Spot in Pepper by Enterobacter Species Isolated from Family Gramineae Plants in Dok-do (독도의 벼과식물로부터 분리된 Enterobacter spp.에 의한 고추의 흰별무늬병에 대한 전신유도저항성)

  • Son, Jin-Soo;Sumayo, Marilyn;Kang, Hyun-Uk;Kim, Byung-Soo;Kwon, Duck-Kee;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.135-143
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    • 2012
  • This study's aim is to isolate and characterize plant growth promoting Enterobacter species for the biological control of gray leaf spot in pepper. Screening was carried out from the rhizosphere of Agropyron tsukushiensi var. transiens (Hack.) Ohwi in Dok-do. Rhizobacterial isolates were partially identified by 16S rDNA sequencing and Enterobacter species were tested for plant growth promoting capabilities and the induction of systemic resistance in pepper against gray leaf spot caused by Stemphylium solani. Isolates were tested for production of indole-acetic acid and siderophore, and for phosphate solubilization. The application of isolates was effective in controlling gray leaf spot in pepper with E. asburiae (KNUC5007) and E. cancerogenes (KNUC5008 and KNUC5010) having the highest efficacy in reducing gray leaf spot severity. This is the first report of the biological control of gray leaf spot in pepper using rhizobacteria and it is hoped that this study will increase the utilization of Enterobacter species as plant growth promoters and biocontrol agents.

Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Gene Cloning and Expression of Trehalose Synthase from Thermus thermophilus HJ6 (Thermus thermophilus HJ6 유래 내열성 Trehalose Synthase의 유전자 클로닝 및 발현)

  • Kim, Hyun-Jung;Kim, Han-Woo;Jeon, Sung-Jong
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.182-188
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    • 2008
  • A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type ($2-4{\mu}m$), about $0.4{\mu}m$ in diameter. The pH and temperature for optimal growth were 6.5 and $80^{\circ}C$, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be $80^{\circ}C$ and 7.5, respectively. The half-life of heat inactivation was about 40 min at $90^{\circ}C$. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.

First Report of Pseudomonas viridiflava Causing Leaf Spot of Cucumber in Korea (Pseudomonas viridiflava에 의한 오이 점무늬병의 발생 보고)

  • Seo, Yunhee;Park, Mi-Jeong;Back, Chang-Gi;Park, Jong-Han
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.328-331
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    • 2018
  • A severe disease with leaf spots and necrotic symptoms was observed in cucumber (Cucumis sativus L.) seedlings in April 2018 at a nursery in Kimjae, Korea (35o 47'09.8"N 127o 2'24.3"E). The infected plants initially showed spots on water-soaked cotyledons which, at later stages, enlarged and spread to the leaves, which the lesions becoming dry and chlorotic. The symptomatic samples were collected from cucumber and the isolates were cultured on LB agar. The representative bacterial strain selected for identification showed fluorescent on King's medium B, was potato rot-positive, levan and arginine dihydrolase-negative, oxidase-negative and tobacco hypersensitivity-positive in LOPAT group 2 as determined by LOPAT tests. A pathogenicity test was carried out on a 3-week-old cucumber. After 3 days of inoculation, leaf spots and necrotic symptoms appeared on the cucumber, similar to the originally infected plants. The infecting bacterial strain was identified as Pseudomonas viridiflava, by 16S rDNA sequence analysis. This is the first report of leaf spot diseases on cucumber caused by P. viridiflava.

Prodigiosin Production From Serratia sp. PDGS120915 Isolated From Daeyeon Stream Water in Busan (하천에서 분리한 Serratia sp. PDGS120915의 프로디지오신 생산)

  • Keunho Ji;Young Tae Kim
    • Journal of Life Science
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    • v.34 no.6
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    • pp.377-384
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    • 2024
  • Prodigiosin is a red pigment characterized by a common pyrrolylpyrromethane skeleton. It is produced by Serratia marcescens, Vibrio psychroerythrus, Hahella chejuensis, etc. Prodigiosin has been reported to possess anticancer, immunosuppressant, antifungal antimalarial, and algicidal activities. However, despite prodigiosin's diverse range of activities, its production rate is significantly low and biosynthesis conditions are difficult. Consequently, the selling price is high, and its usability is limited. This study aimed to increase the efficiency of prodigiosin production according to the culture conditions of Serratia. In this study, a bacterial strain PDGS120915 producing prodigiosin was isolated from lightly contaminated stream water in Busan and identified as a strain of Serratia sp. based on 16S rDNA gene sequence analysis and physiological characteristics. The reddish pigment from PDGS120915 was directly extracted using acidified ethanol, and a characterization analysis confirmed that it was a prodigiosin compound. The optimal conditions for pigment production were 25℃, pH 7, and 0% NaCl concentration for a duration of 14 hr. Furthermore, by treating carbon and nitrogen sources, such as fructose and beef extract, respectively, prodigiosin production increased approximately six-fold and four-fold. Among the minerals tested, 0.1% KCl was found to be the most effective for prodigiosin production. Moreover, casein was identified as the most suitable source for prodigiosin production.

PCR Detection of Terephthalic Acid Degrading Comamonas testosteroni in Soil (PCR을 이용한 토양 중 Terephthalic Acid 분해 Comamonas testosteroni의 검출)

  • 이종훈;강동주;홍연표
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.177-181
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    • 2003
  • Eleven bacterial strains which are able to utilize terephthalic acid as a carbon and an energy source for growth were isolated from the soil of 7 water quality evaluation points in Kyonggi area of Korea. Phthalic acid isomer degrading activity of the isolates from the 4 contaminated points was higher than those from the 3 clean points. Among 11 isolates, 4 isolates which have high terephthalic acid degrading activity and degrade two phthalic acid isomers were identified by partal 16S rDNA sequence determination. One of them was identified as Pseudomonas putida, and the others as Comamonas testosteroni. Thus a large number of phthalic acid isomer degrading bacteria in domestic soil were inferred as C. testosteroni. On the basis of these results, the PCR detection of C. testosteroni in soil was applied to monitor soil contamination by phthalic acid isomers. The DNA of C. test-osteroni extracted from 4 g soil was directly detected by PCR with C. testosteroni specific primer pair. The amount of PCR products was different according to sampling sites and more PCR products were obtained from contaminated sites than those from clean sites (Gulpo-chun>Anyang-chun>Hwangguji-chun>Shin-chun>Huk-chun>Pukhan-river>Kapyeong-chun). This result was coincided with that of the viable cell counts for terephthalic acid degrading bacteria.

Diversity and physiological properties of soil actinobacteria in Ulleung Island (울릉도 유래 토양 방선균의 다양성과 생리활성)

  • Yun, Bo-Ram;Roh, Su Gwon;Kim, Seung Bum
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.242-250
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    • 2017
  • Actinobacteria tolerating extreme conditions can be a rich source of bioactive compounds and enzymes. In this study filamentous actinobacteria were isolated from soils of Ulleung Island, and their physiological properties were examined. Soil samples were collected, serially diluted and spread on various agar media. The average viable counts of total bacteria were $1.28{\times}10^7CFU/g$ for soil sample 1 (ULS1) and $2.05{\times}10^7CFU/g$ for soil sample 2 (ULS2). As a result, 34 strains of actinobacteria were isolated and assigned to the genera Streptomyces (16 strains), Isoptericola (5 strains), Rhodococcus (4 strains), Agromyces (3 strains), Micrococcus (2 strains), Arthrobacter (1 strain), Williamsia (1 strain), Microbacterium (1 strain), and Oerskovia (1 strain) based on 16S rRNA gene sequence analysis. Enzyme activity and plant growth promoting potential were tested for representative isolates. Multiple strains of Streptomyces degraded starch, casein and Tween 80. As for plant growth promoting potential, strains of Oerskovia, Williamsia, Isoptericola, and Streptomyces solubilized phosphate, and those of Agromyces, Oerskovia, Micrococcus, Rhodococcus, Streptomyces, and Isoptericola produced 3-indole-acetic acid (IAA), respectively. Selected strains of Streptomyces exhibited strong antagonistic activity against Staphylococcus aureus and Bacillus subtilis as well as Candida albicans. This study confirms that actinobacteria from Ulleung Island can be a good source of novel bioactive compounds.

Organic acid production and phosphate solubilization by Enterobacter intermedium 60-2G (Enterobacter intermedium 60- 2G의 유기산 생성과 불용성인의 가용화)

  • Kim, Kil-Yong;Hwangbo, Hoon;Kim, Yong-Woong;Kim, Hyo-Jeong;Park, Keun-Hyung;Kim, Young-Cheol;Seong, Ki-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.1
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    • pp.59-67
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    • 2002
  • A phosphate solubilizing bacterium. strain 60-2G, possessing a strong ability to solubilize insoluble phosphate was isolated from the rhizosphere of grass. On the basis of GC-FAME profile, carbon utilization pattern, and the DNA sequence of a conserved partial 16S rRNA gene, the 60-2G was identified as Enterobacter intermedium. The analysis by HPLC revealed that the strain 60-2G produced mainly gluconic and 2-ketogluconic acids with small amounts of lactic acid in broth culture medium containing hydroxyapatite. During the incubation period of the strain 60-2G in broth culture, pH of the medium decreased upto 3.8 while the soluble phosphate concentration increased. The reversed correlation between pH and soluble phosphate concentration indicated that the solubility of P was due to the produced organic acids. The sequence homology of the deduced amino acids suggested that E. intermedium 60-2G synthesized PQQ which is essential for the oxidation of glucose by glucose dehydrogenase.

Antibacterial Activity of Bacteria Isolated from Rocks on the Seashore (갯바위에서 분리한 미생물의 항균활성 분석)

  • Park, In-Suk;Oh, Ryunkyoung;Lee, Min Jeong;Moon, Ji Young;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Woo-Jin;An, Cheul Min;Kim, Dong-Gyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.6
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    • pp.904-912
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    • 2015
  • There is a great deal of research interest regarding substitutes for antibiotics because of various obstacles to the efficacy and use of antibiotics. We isolated and analyzed diversity of microbiota which exhibited antibacterial activity against 23 pathogenic bacteria, to develop alternative agent of antibiotics. By investigating the microbiota from rocks on the seashore, we characterized and obtained various antibacterial material-producing bacteria. Thirty-one isolates belong to four genera and seven species, according to 16S rDNA sequence analysis, showed antibacterial activities against 23 pathogenic bacteria. The Identity of 16S rDNA sequences indicated three species of Bacillus, one species of Paenibacillus, one species of Pseudomonas and two species of Enterobacter. Two isolates were similar to Bacillus aerophilus, four isolates were similar to Bacillus pumilus, seven isolates were similar to Bacillus safensis, 15 isolates were similar to Paenibacillus polymyxa, respectively. In addition, one isolate was similar with Pseudomonas poae, one isolate was similar to Enterobacter asburiae, and one isolate was similar to Enterobacter ludwigii, respectively. Variations of antibacterial activity and level among the same species were indicated the diverse strains of isolates. Vibrio vulnificus showed the highest degree of growth inhibition by 29 isolates. Further studies regarding antibacterial materials and bacteria suggest that development of probiotic strains or alternative agents to antibiotics.

Molecular and Ecological Analyses of Microbial Community Structures in Biofilms of a Full-Scale Aerated Up-Flow Biobead Process

  • Ju, Dong-Hun;Choi, Min-Kyung;Ahn, Jae-Hyung;Kim, Mi-Hwa;Cho, Jae-Chang;Kim, Tae-Sung;Kim, Tae-San;Seong, Chi-Nam;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.253-261
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    • 2007
  • Molecular and cultivation techniques were used to characterize the bacterial communities of biobead reactor biofilms in a sewage treatment plant to which an Aerated Up-Flow Biobead process was applied. With this biobead process, the monthly average values of various chemical parameters in the effluent were generally kept under the regulation limits of the effluent quality of the sewage treatment plant during the operation period. Most probable number (MPN) analysis revealed that the population of denitrifying bacteria was abundant in the biobead #1 reactor, denitrifying and nitrifying bacteria coexisted in the biobead #2 reactor, and nitrifying bacteria prevailed over denitrifying bacteria in the biobead #3 reactor. The results of the MPN test suggested that the biobead #2 reactor was a transition zone leading to acclimated nitrifying biofilms in the biobead #3 reactor. Phylogenetic analysis of 16S rDNA sequences cloned from biofilms showed that the biobead #1 reactor, which received a high organic loading rate, had much diverse microorganisms, whereas the biobead #2 and #3 reactors were dominated by the members of Proteobacteria. DGGE analysis with the ammonia monooxygenase (amoA) gene supported the observation from the MPN test that the biofilms of September were fully developed and specialized for nitrification in the biobead reactor #3. All of the DNA sequences of the amoA DGGE bands were very similar to the sequence of the amoA gene of Nitrosomonas species, the presence of which is typical in the biological aerated filters. The results of this study showed that organic and inorganic nutrients were efficiently removed by both denitrifying microbial populations in the anaerobic tank and heterotrophic and nitrifying bacterial biofilms well-formed in the three functional biobead reactors in the Aerated Up-Flow Biobead process.