• 한국동물위생학회지
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• 제38권2호
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• pp.101-106
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• 2015

Murine mycoplasmosis, caused by Mycoplasma (M.) pulmonis, is a prominent disease in rodent animals. The aim of this study was to develop a sensitive and specific PCR assay to detect M. pulmonis in animals and to assess the suitability of this assay for the detection of mycoplasmal infection in rats experimentally infected with M. pulmonis. A new PCR assay using the M. pulmonis-specific primer pairs MPul-F and MPul-R was developed. The primers and probe for the assay were designed from regions in the 16S rRNA gene that are unique to M. pulmonis. The novel PCR assay was very specific and sensitive for M. pulmonis, detecting the equivalent of 5 pg of target template DNA. It detected only M. pulmonis and no other Mycoplasma species or other bacterial species. The newly developed PCR assay also effectively detected M. pulmonis infection in rats. These results suggest that this PCR assay using M. pulmonis-specific primer pairs of MPul-F and MPul-R will be useful and effective for monitoring M. pulmonis infection in animals.

• Animal Systematics, Evolution and Diversity
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• 제36권3호
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• pp.268-273
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• 2020

A spionid polychaete, Boccardiella hamata (Webster, 1879) has been found from mud in crevices between the shells of oysters and adherent substrates in South Korea. The sequences of mitochondrial DNA (mtDNA) cytochrome c oxidase subunit 1 (CO1), 16S ribosomal DNA (16S), and the nuclear 18S ribosomal DNA (18S) from Korean individuals of Boccardiella hamata were determined in the present study. The molecular analysis based on the 18S rRNA gene sequences showed clear separation among the spionid polychaete species, and the sequences of Korean and Japanese individuals are completely identical. The morphological diagnosis and photographs of B. hamata are also provided.

• The Plant Pathology Journal
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• 제40권2호
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• pp.171-191
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• 2024

Identification of Helicotylenchus species is very challenging due to phenotypic plasticity and existence of cryptic species complexes. Recently, the use of rDNA barcodes has proven to be useful for identification of Helicotylenchus. Molecular markers are a quick diagnostic tool and are crucial for discriminating related species and resolving cryptic species complexes within this speciose genus. However, DNA barcoding is not an error-free approach. The public databases appear to be marred by incorrect sequences, arising from sequencing errors, mislabeling, and misidentifications. Herein, we provide a comprehensive analysis of the newly obtained, and published DNA sequences of Helicotylenchus, revealing the potential faults in the available DNA barcodes. A total of 97 sequences (25 nearly full-length 18S-rRNA, 12 partial 28S-rRNA, 16 partial internal transcribed spacer [ITS]-rRNA, and 44 partial cytochrome c oxidase subunit I [COI] gene sequences) were newly obtained in the present study. Phylogenetic relationships between species are given as inferred from the analyses of 103 sequences of 18S-rRNA, 469 sequences of 28S-rRNA, 183 sequences of ITS-rRNA, and 63 sequences of COI. Remarks on suggested corrections of published accessions in GenBank database are given. Additionally, COI gene sequences of H. dihystera, H. asiaticus and the contentious H. microlobus are provided herein for the first time. Similar to rDNA gene analyses, the COI sequences support the genetic distinctness and validity of H. microlobus. DNA barcodes from type material are needed for resolving the taxonomic status of the unresolved taxonomic groups within the genus.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1571-1576
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• 2010

Two bacterial strains designated as CT2 and CT5 were isolated from highly alkaline cement samples using the enrichment culture technique. On the basis of various physiological tests and 16S rRNA sequence analysis, the bacteria were identified as Bacillus species. The urease production was 575.87 U/ml and 670.71 U/ml for CT2 and CT5, respectively. Calcite constituted 27.6% and 31% of the total weight of sand samples plugged by CT2 and CT5, respectively. Scanning electron micrography analysis revealed the direct involvement of these isolates in calcite precipitation. This is the first report of the isolation and identification of Bacillus species from cement. Based on the ability of these bacteria to tolerate the extreme environment of cement, they have potential to be used in remediating the cracks and fissures in various building or concrete structures.

• 생명과학회지
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• 제28권12호
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• pp.1477-1488
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• 2018

본 연구는 제주 연안에서 채집한 오분자기(Sulculus diversicolor supertexta)를 구성하는 미생물 군집의 다양성을 알아보기 위하여 근육, 장, 생식소 각 부위별로 조사하였다. 배지로 1차 순수 분리한 결과 근육은 MA, 장 1% BHIA, 생식소 1% TSA에서 각각 최대 군락 계수가 나타났다. 16S rRNA sequence로 표준 균주와 비교 유사도 분석 결과 총 190개의 순수 colony가 분리되었다. NBLAST program 분석 결과 크게 5문 25과 39속 71종으로 나타났다. 표준 균주와 상동성은 91-100%를 나타냈다. 오분자기 내 미생물 군집은 크게 Probacteria (Gamma-proteobacteria, Alpha-proteobacteria) 48%, Actinobacteria 32.5%, Firmicutes 16.9%, Bacteroide 1.3%, Deinococcus-thermus 1.3%로 나타났다. 근육, 장, 생식소 모든 부위에서 Moraxellaceae과 Psychrobacter cibarius가 우점하였다. 근육, 장, 생식소 모든 부위에서 Alteromonadaceae, Enterobacteriaceae, Pasturellaceae, Moraxellaceae, Rhodobacteraceae, Geminicoccaceae, Dietziaceae, Intrasporangiaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Streptomycetaceae, Aerococcaceae, Bacillaceae, Paenibacillaceae, Planococcaceae, Staphylcoccaceae가 공통적으로 분리되었으며, 장에서 Flavobacteriaceae, Corynebacteriaceae, Yesiniaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae가 추가 분리되었다. 분리 균주로부터 인체 유해 질병 세균에 대한 항균활성 모니터링 결과 Sterptomyces albus (96%)가 4균주 모두 항균활성을 보였고 Agrococcus baldri (99%), Psychrobacter nivimaris (99%)가 E. coli, E. aerogens에 대한 항균활성을 나타냈다. 그 외 상동성이 낮은 일부 균주가 분리되어 신균주 실험을 비롯한 항균활성물질 정제 등 추가 실험이 필요한 것으로 사료된다. 본 실험은 오분자기 미생물 군집의 다양성과 유전학적 자원을 확보하는데 의의를 두며, 응용 미생물의 개발 가능성에 있어 기초 자료를 제공하고자 하였다.

• Journal of Microbiology
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• 제44권2호
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• pp.155-161
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• 2006

Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.301-310
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• 2012

Prosopis juliflora and Parthenium hysterophorus are the two arid, exotic weeds of India that are characterized by distinct, profuse growth even in nutritionally poor soils and environmentally stressed conditions. Owing to the exceptional growth nature of these two plants, they are believed to harbor some novel bacterial communities with wide adaptability in their rhizosphere. Hence, in the present study, the bacterial communities associated with the rhizosphere of Prosopis and Parthenium were characterized by clonal 16S rRNA gene sequence analysis. The culturable microbial counts in the rhizosphere of these two plants were higher than bulk soils, possibly influenced by the root exudates of these two plants. The phylogenetic analysis of V1_V2 domains of the 16S rRNA gene indicated a wider range of bacterial communities present in the rhizosphere of these two plants than in bulk soils and the predominant genera included Acidobacteria, Gammaproteobacteria, and Bacteriodetes in the rhizosphere of Prosopis, and Acidobacteria, Betaproteobacteria, and Nitrospirae in the Parthenium rhizosphere. The diversity of bacterial communities was more pronounced in the Parthenium rhizosphere than in the Prosopis rhizosphere. This culture-independent bacterial analysis offered extensive possibilities of unraveling novel microbes in the rhizospheres of Prosopis and Parthenium with genes for diverse functions, which could be exploited for nutrient transformation and stress tolerance in cultivated crops.

• 생명과학회지
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• 제28권8호
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• pp.955-961
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• 2018

불가사리를 이용하여 다양한 생리활성에 대한 연구가 많이 진행되었지만, 다른 천연물 연구에 비해 불가사리로 부터 분리한 미생물에 대한 연구는 부족하다. 이에 본 연구에서는 제주도에서 채집한 극피동물인 빨강불가사리(Certonardoa semiregularis)의 내장으로부터 Marine Agar와 R2A를 이용하여 총 103개의 균주를 분리하여 세균군집에 대해 조사하였다. 분리된 균주들은 16S rRNA유전자를 이용하여 염기서열을 분석 및 동정하였다. 그 결과 주요 계통군은 Proteobacteria (Alpha-proteobacteria 24%, Beta-proteobacteria 4%, Gamma-proteobacteria 65%) 93%, Bacteroidetes 4%, Actinobacteria 2%, Firmicutes 1%로 4개의 문이 확인되었고, 7개의 강(Actinobacteria, Flavobacteria, Bacilli, Sphingobacteria, Alpha-proteobacteria, Beta-proteobacteria, Gamma-proteobacteria), 15개의 목, 19개의 과, 24속이 관찰되었다. 또한 계통 분석 결과 2개의 균주(Lysobacter sp., Pedobacter sp.)가 각각 97.55%, 97.58%로 상동성이 98% 이하로 나타나 새로운 속 또는 종으로 분류될 가능성이 있다고 여겨지며, 표준 균주와 함께 신종 후보 균주에 대한 생화학적, 형태학적 등의 추가적인 신종실험을 향후 진행해야 할 것으로 사료 된다.

• The Plant Pathology Journal
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• 제17권6호
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• pp.361.4-361
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• 2001

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.205-211
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• 2004

A 16S rDNA clone library was generated to investigate the bacterial diversity in tidal flat sediment in Ganghwa Island, Republic of Korea. A total of 103 clones were sequenced and analyzed by comprehensive phylogenetic analyses. No clones were identical to any of known 16S rRNA sequences in public databases. Sequenced clones fell into thirteen lineages of the domain Bacteria: the alpha, beta, gamma, delta, and epsilon Proteobacteria, Actinobacteria, CFB group, Chloroflexi, Acidobacteria, Planctomycetes, Verrucomicrobia, and known uncultured candidate divisions (OP11, BRC1, KSB1, and WS1). Two clones were not associated with any known bacterial divisions. The majority of clones belonged to the gamma and delta Proteobacteria (46.7%). Clones of Actinobacteria were distantly related to known taxa. It is evident from 16S rDNA-based community analysis that the bacterial community in tidal flat sediment is remarkably diverse and unique among other marine environments examined so far.