• 한국미생물·생명공학회지
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• 제38권3호
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• pp.322-328
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• 2010

발효홍어(홍어회)에 존재하는 박테리아 집단의 다양성을 확인하기 위해, 박테리아의 16S rDNA 절편들을 증폭하고 클로닝하여 라이브러리를 구축하였다. 삽입 서열의 염기서열을 결정한 후, BLAST 분석에 의해 미생물 동정을 하였다. 동일한 삽입서열 빈도를 계산하였을 때, 발효홍어(홍어회)에는 uncultured bacterium clone 054E11.b가 57.1% 나타남으로써 우점균으로 존재하고 있다고 추정하였다. 또한 발효홍어(홍어회) 현탁액을 한천배지에 도말하여 형성된 집락을 콜로니 PCR을 수행하였을 때, Pseudomonas sp. KC-EPS13 등 12 종의 박테리아가 동정되었다. 배양 의존적 방법과 배양 비의존적 방법으로 홍어회를 분석하였을 때, Psychrobacter sp. J466만이 두 가지 방법에서 모두 검출되었고 나머지 박테리아들의 균총은 상이하였다.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.66-78
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• 2019

In this study, strain KNU17Pc1 was tested for its antifungal activity against Rhizoctonia solani AG-1(IA), which causes banded leaf and sheath blight (BLSB) of maize. KNU17Pc1 was tested further for its broad-spectrum antifungal activity and in vitro plant growth promoting (PGP) traits. In addition, the in vivo effects of KNU17Pc1 on reduction of BLSB severity and seedling growth promotion of two maize cultivars under greenhouse conditions were investigated. On the basis of multilocus sequence analysis (MLSA), KNU17Pc1 was confirmed as P. chlororaphis subsp. aurantiaca. The study revealed that KNU17Pc1 had strong in vitro antifungal activity and was effective toward all in vitro PGP traits except phosphate solubilization. In this study, for the first time, a strain of P. chlororaphis against Colletotrichum dematium, Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. melonis, Fusarium subglutinans and Stemphylium lycopersici has been reported. Further biochemical studies showed that KNU17Pc1 was able to produce both types of phenazine derivatives, PCA and 2-OH-PCA. In addition, solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis identified 13 volatile organic compounds (VOCs) in the TSB culture of KNU17Pc1, 1-undecene being the most abundant volatile. Moreover, for the first time, Octamethylcyclotetrasiloxan (D4), dimethyl disulfide, 2-methyl-1,3-butadiene and 1-undecene were detected in P. chlororaphis. Furthermore, this study reported for the first time the effectiveness of P. chlororaphis to control BLSB of maize. Hence, further studies are necessary to test the effectiveness of KNU17Pc1 under different environmental conditions so that it can be exploited further for biocontrol and plant growth promotion.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.79-90
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• 2019

Lichens are generally known as self-sufficient, symbiotic life-forms between fungi and algae/cyanobacteria, and they also provide shelter for a wide range of beneficial bacteria. Currently, bacterial-derived biodegradable polyhydroxyalkanoate (PHA) is grabbing the attention of many researchers as a promising alternative to non-degradable plastics. This study was conducted to develop a new method of PHA production using unexplored lichen-associated bacteria, which can simultaneously degrade two ubiquitous industrial toxins, anthracene and naphthalene. Here, 49 lichen-associated bacteria were isolated and tested for PHA synthesis. During the GC-MS analysis, a potential strain of EL19 was found to be a 3-hydroxyhexanoate (3-HHx) accumulator and identified as Pseudomonas sp. based on the 16S rRNA sequencing. GC analysis revealed that EL19 was capable of accumulating 30.62% and 19.63% of 3-HHx from naphthalene and anthracene, respectively, resulting in significant degradation of 98% and 96% of naphthalene and anthracene, respectively, within seven days. Moreover, the highly expressed phaC gene verified the genetic basis of $PHA_{mcl}$ production under nitrogen starvation conditions. Thus, this study strongly supports the hypothesis that lichen-associated bacteria can detoxify naphthalene and anthracene, store energy for extreme conditions, and probably help the associated lichen to live in extreme conditions. So far, this is the first investigation of lichen-associated bacteria that might utilize harmful toxins as feasible supplements and convert anthracene and naphthalene into eco-friendly 3-HHx. Implementation of the developed method would reduce the production cost of $PHA_{mcl}$ while removing harmful waste products from the environment.

• 식물병연구
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• 제17권1호
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• pp.38-43
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• 2011

채소류가 소비되기까지 약 10~30%에 이르는 양이 부패에 의해 버려지고 있다. 채소류는 수확 후 수많은 세균이나 곰팡이와 같은 다양한 미생물들에 의해 부패가 이루어진다. 채소류의 부패와 세균과의 관계를 알아보고자 세균의 다양성에 관한 조사를 하였다. 신선한 상추와 깻잎, 치커리에서의 총 호기성 균 수는 각각 $2.6{\sim}2.7{\times}10^6$, $4.6{\times}10^5$, $1.2{\times}10^6\;CFU/g$ of fresh weight이었으며, Pseudomonas spp., Alysiella spp., Burkholderia spp.와 그 외의 18개의 다양한 속들이 확인되었다. 신선한 채소류를 $28^{\circ}C$에서 일주일 동안 배양하였을 때 세균 다양성에 변화가 생겼다. 총 호기성 균의 수는 상추와 깻잎, 치커리에 대하여 각각 $1.1{\sim}4.6{\times}10^8$, $4.9{\times}10^7$, $7.6{\times}10^8\;CFU/g$ of fresh weight로 나타났으며, 이는 약 $10^2$배 정도 증가한 수치이다. 하지만 세균 다양성은 단순해져서 보다 적은 수의 세균이 분리, 동정되었다. Pseudomonas spp.가 대부분을 차지하였으며(~48%), Arthrobacter sp., Bacillus sp.가 그 뒤를 이었다. 동정된 세균 각각의 부패능을 검정하고자 무균배양한 상추에 접종하였으며, 그 결과 Pseudomonas fluorescence와 Pantoea agglomerans가 상당한 부패를 야기하였다.

• 한국어병학회지
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• 제10권2호
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• pp.165-176
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• 1997

양식넙치의 병원바이러스인 HRV 및 FBV, 연어과 유래 바이러스이면서 해산어에 병원성을 나타내는 RVS를 대상으로 넙치사육해수, 2차증류수 및 Hanks' BSS중에서의 바이러스 감염가의 변화를 관찰하고 사육해수중에 존재하는 미생물과의 상호작용을 검토하였다. HRV, FBV, RVS의 감염가는 2차증류수 및 Hanks' BSS중에서 21일간 안정적이었으나, 사육해수 중에서 HRV는 $0^{\circ}C$를 제외한 5, 10, 15, $20^{\circ}C$의 실험구에서 바이러스 현탁후 7일째 부터 감염가의 저하를 확인할 수 있었고, RVS에서도 감염가의 하락이 확인되었다. HRV 및 RVS를 고압멸균, 여과처리 및 무처리 사육해수에 현탁시켜 본 결과 무처리 사육해수에서 바이러스감염가의 하락을 나타내는데 비하여 고압멸균 및 여과 처리한 사육해수중의 바이러스 감염가는 비교적 안정적이었다. 사육해수에 1% MEM10을 첨가하여 $15^{\circ}C$에서 7일간 배양한 후 여과 처리하여 얻은 여과액에 HRV를 현탁시켜 감염가의 변화를 관찰한 결과 1일에서 3일째에 감염가가 검출한계 이하로 하락하였으며, 해수배양액내의 세균 수는 $8.1{\times}10^8$ cells/ml이었고, Pseudomonas, Achromobacter, Flavobacterium 및 Vibrio속 세균이 분리되었다. 분리된 Pseudomonas 및 Vibrio속의 대표주에서 항바이러스작용 물질의 생산이 확인되었다.

• 상하수도학회지
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• 제13권4호
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• pp.45-53
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• 1999

The microbial organisms named "Pseudomonas sp. LK-14" were isolated from farm land and shallow river sediment, activated, augmented and identified; which were using 2,4-D (2,4-Dichlorophenoxyacetic acid) as a sole carbon source and energy source. 2,4-D removal efficiency of LK-14 with 2,4-D sole carbon source (reactor S) were higher than that of Activated Sludge with 2,4-D sole carbon source (reactor A). Dynamic bioligical reaction kinetic parameters (sole carbon source was 2,4-D) obtained from batch reactor experiments were ${\mu}_{max}$ $0.105hr^{-1}$, $K_{s,24D}$ 15.64mg/L, $K_{i,24D}$ $1.94h^{r-1}$, $Y_{24D}$ 0.39 for LK-14 and ${\mu}_{max}$ $0.008hr^{-1}$, $K_{s,24D}$ 26.95mg/L, $K_{i,24D}$ $1.75hr^{-1}$, $Y_{24D}$ 0.10 for Activated Sludge. Using these parameters, we could predict the behaviors of 2,4-D substrate utilized by LK-14 and Activated Sludge in batch reactors. The kinetic parameters are enable to predict the 2,4-D substrate and microbial population behavior entering into wastewater treatment plants by using unsteady states dynamic simulation modeling technique.

• Journal of Microbiology and Biotechnology
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• 제18권9호
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• pp.1492-1495
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• 2008

Successful control of Xanthomonas oryzae pv. oryzicola, the causal agent of bacterial leaf streak, requires a specific and reliable diagnostic tool. A pathovar-specific PCR assay was developed for the rapid and accurate detection ofthe plant pathogenic bacterium Xanthomonas oryzae pv. oryzicola in diseased plant. Based on differences in a membrane fusion protein gene of Xanthomonas oryzae pv. oryzicola and other microorganisms, which was generated from NCBI (http://www.ncbi.nlm.nih.gov/) and CMR (http://cmr.tigr.org/) BLAST searches, one pair of pathovar-specific primers, XOCMF/XOCMR, was synthesized. Primers XOCMF and XOCMR from a membrane fusion protein gene were used to amplity a 488-bp DNA fragment. The PCR product was only produced from 4 isolates of Xanthomonas oryzae pv. oryzicola among 37 isolates of other pathovars and species of Xanthomonas, Pectobacterium, Pseudomonas, Burkholderia, Escherichia coli, and Fusarium oxysporum f.sp. dianthi. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1765-1771
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• 2007

A polymerase chain reaction (PCR)-based method was developed to detect the DNA of Ralstonia solanacearum, the causal agent of bacterial wilt in various crop plants. One pair of primers (RALSF and RALSR), designed using cytochrome c1 signal peptide sequences specific to R. solanacearum, produced a PCR product of 932 bp from 13 isolates of R. solanacearum from several countries. The primer specificity was then tested using DNA from 21 isolates of Ralstonia, Pseudomonas, Burkholderia, Xanthomonas, and Fusarium oxysporum f. sp. dianthi. The specificity of the cytochrome c1 signal peptide sequences in R. solanacearum was further confirmed by a DNA-dot blot analysis. Moreover, the primer pair was able to detect the pathogen in artificially inoculated soil and tomato plants. Therefore, the present results indicate that the primer pair can be effectively used for the detection of R. solanacearum in soil and host plants.

• Journal of Microbiology
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• 제38권1호
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• pp.8-10
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• 2000

In order to rapidly identify and differentiate Salmonella typhimurium from the intestinal gram-negative bacteria, randomly amplified polymorphic DNA (RAPD) fingerprinting of Salmonella typhimurium was carried out using random primers designated OPA-13 (5'-CAGCACCCAC-3'), OPB-10 (5'-CGTCTGGGAC-3'), OPB-18 (5'-CCACAGCAGT-3'), and OPJ-10 (5'-AAGCCCGAGG-3'), and its patterns compared with 6 representive intestinal, gram-negative bacterial strains, Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, Escherichia coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., which are often found in foods. S. typhimurium had unique and distinct fingerprinting patterns. RAPD fingerprinting is thus concluded to be a rapid and sensitive method for the identification of S. typhimurium compared to conventional culturing procedures or immunoassays.

• Journal of Microbiology and Biotechnology
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• 제20권5호
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• pp.853-861
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• 2010

High substrate costs decrease the profitability of polyhydroxyalkanoates (PHAs) production, and thus low-cost carbon substrates coming from agricultural and industrial residuals are tested for the production of these biopolymers. Among them, crude glycerol, formed as a by-product during biodiesel production, seems to be the most promising source of carbon. The object of this study was to characterize the mixed population responsible for the conversion of crude glycerol into PHAs by cultivation-dependent and -independent methods. Enrichment of the microbial community was monitored by applying the Ribosomal Intergenic Spacer Analysis (RISA), and the identification of community members was based on 16S rRNA gene sequencing of cultivable species. Molecular analysis revealed that mixed populations consisted of microorganisms affiliated with four bacterial lineages: ${\alpha}$, ${\gamma}$-Proteobacteria, Actinobacteria, and Bacteroides. Among these, three Pseudomonas strains and Rhodobacter sp. possessed genes coding for polyhydroxyalkanoates synthase. Comparative analysis revealed that most of the microorganisms detected by direct molecular analysis were obtained by the traditional culturing method.