• Title/Summary/Keyword: ITS rDNA sequences

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Comparative Molecular Phylogenetic Relationships in Different Strains of Pleurotus spp. (느타리속 버섯 계통의 분자생물학적 유연관계의 비교연구)

  • Cho, Hae-Jin;Lee, Jae-Seong;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.112-119
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    • 2010
  • Pleurotus spp. have been used for edible and medicinal purposes in Asian countries for a long time. The fruiting bodies of the Pleurotus ostreatus, Pleurotus citrinopileatus and Pleurotus salmoneostramineus contained many physiologically beneficial substances for human health. Therefore, it is necessary to study the genetic diversity of Pleurotus mushroom cultivars commercially cultivated in Korea. Eleven strains of Pleurotus spp. were collected from different geographical regions in South-East Asia and ITS regions of rDNA and RAPD of genomic DNA were analyzed. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 167 to 254 bp and 156 to 213 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into 4 clusters. Eleven Pleurotus species were also analyzed by RAPD with 20 arbitrary primers. Ten of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.1 to 2.0kb. The results revealed that genetic diversity of selected strains of P. ostreatus, P. citrinopileatus and P. salmoneostramineus is low.

Isolation of Streptomyces sp. YU100 Producing Extracellular Phospholipase D

  • Lim, Si-Kyu;Choi, Jae-Woong;Lee, Eun-Tag;Khang, Yong-Ho;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.71-76
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    • 2002
  • Soil samples were screened for actinomycete strains capable of producing phospholipase D, and a strain, Streptomyces sp. YU100, showing a high transphosphatidylation activity was isolated. This strain secreted phospholipase D in a culture broth after 12 h of cultivation, and its productivity continued to increase for 36 h of fermentation. In addition, its transphosphatidylation rate of phosphatidylcholine to phosphatidylserine was almost $68\%$ within 1 h. The morphological and chemotaxonomical characteristics showed that this strain could be classified as a number of the Streptomycetaceae family, particularly due to the spiral form of its spore chain consisting of 60-70 smooth spores $(0.75{\times}1.0{\mu}m$) on an aerial mycelium, FA-2c type of fatty acid profile in the cell wall, and LL-DAP component in the cell wall peptidoglycan. A phylogenetic analysis of the 16S rDNA provided a clue that the strain YU100 was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyes sp. ASB27, S. peucetius JCM9920, and S. griseus ATCC10137. A dendrogram based on the 16S rDNA sequences also showed a phylogenetic relationship between the strain YU100 and these strains. However, the strain YU100 has not yet been assigned to a particular species, because of absence of any other classified species with a high matching score.

Usability of DNA Sequence Data: from Taxonomy over Barcoding to Field Detection. A Case Study of Oomycete Pathogens

  • Choi, Young-Joon;Thines, Marco
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.41-41
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    • 2015
  • Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

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The Determination of the Partial 28S Ribosomal DNA Sequences and Rapid Detection of Phellinus linteus and Related species

  • Park, Hyung-Sik;Kim, Gi-Young;Nam, Byung-Hyouk;Lee, Sang-Joon;Lee, Jae-Dong
    • Mycobiology
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    • v.30 no.2
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    • pp.82-87
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    • 2002
  • Species of Phellinus were known to harmful fungi causing white pocket rot and severe plant disease such as canker or heartrot in living trees in the West, but some species have been used to traditional medicines in the Orient for a long time. In this study the partial D1-D2 nucleotide sequences of 28S ribosomal DNA from 13 Phellinus strains were determined and compared with the sequences of 21 strains obtained from GenBank database. According to the neighbor-joining(NJ) method comparing the sequence data the phylogenetic tree was constructed. The phylogenetic tree displayed the presence of four groups. Group I includes P. ferreus, P. gilvus and P. johnsonianus, Group II contains P. laevigatus, P. conchatus and P. tremulae, Group III possesses P. linteus, P. weirianus, P. baumii, P. rhabarbarinus and P. igniarius, and Group IV comprises P. pini, P. chrysoloma. P. linteus and P. baumii, which were used mainly in traditional medicine, belong to the same group, but exactly speaking both were split into two different subgroups. To detect P. linteus only, we developed the PCR primer, D12HR. The primer showed the specific amplification of P linteus, which is permitted to medicinal mushroom in the East. The results make a potential to be incorporated in a PCR identification system that could be used for the rapid identification of this species from its related species, P. linteus especially.

Cloning and Expression of Alginate Lyase from a Marine Bacterium, Streptomyces sp. M3 (해양미생물 Streptomyces sp. M3로부터 alginate lyase의 클로닝 및 발현)

  • Kim, Hee-Sook
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1522-1528
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    • 2009
  • A marine bacterium was isolated from brown seaweeds for its ability to degrade alginate. Analysis of 16S ribosomal DNA sequence revealed that the strain belongs to Streptomyces like strain ALG-5 which was reported previously. New alginate lyase gene of Streptomyces sp. M3 was cloned by using PCR with the specific primers designed from homologous nucleotide sequences. The consensus sequences of N-terminal YXRSELREM and C-terminal YFKAGXYXQ were conserved in the M3 alginate lyase amino acid sequences. The homology model for the M3 alginate lyase showed a characteristic structure of $\beta$-jelly roll fold main domain like alyPG from Corynebacterium sp. ALY-1. The homogenate of the recombinant E. coli with the alginate lyase gene showed more degrading activity for polyguluronate block than polymannuronate block. The results from the multiple alignments and the homology modeling elucidated in the M3 alginate lyase can be classified into family PL-7.

Isolation and Characterization of Diesel Oil Degrading Bacterium, Pseudomonas sp. GENECO 1 Isolated from Oil Contaminated Soil (유류 오염 토양으로부터 분리한 디젤 분해 세균 Pseudomonas sp. GENECO 1의 분리 및 특성 규명)

  • 이종광;김무훈;박형수
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.102-107
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    • 2003
  • With the enrichment culture technique, bacterial strains which degrade diesel oil were isolated from soil contaminated with diesel oil. One of the isolates named GENECO 1 showed the highest activity for emulsification of diesel oil as well as the highest growth rate. This strain, GENECO 1, was identified as a Pseudomonas sp. based on its biochemical, physiological characteristics and 16S rDNA sequences. The optimal cultural conditions for cell growth and oil emulsifying activity of its culture were as follow; $30^{\circ}C$ for temperature, 7.0 for pH. Diesel oil degradation was analysed by the gas chromatography. More than 95% of 1% treated diesel oil were converted into a form no longer extractable by mixed organic solvents after 96 hours incubation.

Bacterial Canker of Japanese Apricot (Prunus mume) Caused by Pseudomonas syringae pv. morsprunorum (Pseudomonas syringae pv. morsprunorum에 의한 매실의 세균성궤양성)

  • Kim Doo Young;Han Hyo Shim;Koh Young Jin;Jung Jae Sung
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.135-139
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    • 2005
  • Bacterial canker of Japanese apricot (Prunus mume Sieb. et Zucc.) was found in all orchards located at southern area of Korea. Typical symptoms were characterized by dark spots formed on fruits, brown lesions on leaves, and bacterial exudate oozed out of the cracked bark of diseased tree. Thirty-eight isolates from 16 different areas were identified on the basis of biochemical and physiological characteristics (LOPAT and GATTa test) and also on the basis of 165 rDNA and ITS sequences. Pathogenicity tests confirmed that bacterial canker of Japanese apricot in Korea is caused by Pseudomonas syringae pv. morsprunorum.

Diversity of Foliar Endophytic Fungi Isolated from Lindera obtusiloba in Korea (생강나무(Lindera obtusiloba)의 잎에서 분리한 내생균의 다양성)

  • Kim, Chang-Kyun;Eo, Ju-Kyeong;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.136-140
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    • 2012
  • Leaves of Lindera obtusiloba were collected from four sites in Gangwon Province, Korea. Endophytic fungi were isolated from the leaves and identified using ITS sequences of rDNA. Total twelve species belonging of endophytic fungi were identified; Alternaria alternata, Annulohypoxylon annulatum, Creosphaeria sassafras, Diaporthe eres, Discosia sp., Epicoccum nigrum, Glomerella acutata, Glomerella cingulata, Paraconiothyrium brasiliense, Pestalotiopsis neglecta, Phomopsis amygdali, Xylaria sp. The endophytic fungus, Phomopsis amygdali, was the most dominant species isolated from L. obtusiloba in this study and the fungal diversities varied in the different sites.

Leavening Ability of the Isolate Saccharomyces cerevisiae MF10003 in Bakery Dough (분리 효모 Saccharomyces cerevisiae MF10003의 빵반죽 발효 팽창력)

  • Oh, Jung-Suk;Min, Eung-Ki;Ahn, Chang-Hyun;Han, Yeong-Hwan
    • Journal of Life Science
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    • v.23 no.2
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    • pp.222-227
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    • 2013
  • An effective leavening yeast was isolated from raisin broth. The isolate was identified as Saccharomyces cerevisiae by comparing the homology of 18S rDNA ITS sequences and named as S. cerevisiae MF10003. S. cerevisiae MF10003 showed a 1.9-fold and 3.1-fold increase in $CO_2$ production and leavening ability, respectively, compared with the wild yeast S. ellipsoideus KCTC7243, and the dough had a rich and volatile flavor. When glucose, sucrose, fructose, and maltose were added to the culture broth as a carbon and energy source, $CO_2$ was produced in 4 hr.

Identification and Genotypic Characterization of Potentially Pathogenic Acanthamoeba Isolated from Tap Water in Wuxi, China

  • Wang, Meixu;Sun, Guangxu;Sun, Yangkai;You, Xiaomin;Li, Xiaoxue;Cheng, Yang;Xuan, Yinghua
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.615-618
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    • 2018
  • Members of genus Acanthamoeba are widely distributed in the environment. Some are pathogenic and cause keratitis and fatal granulomatous amoebic encephalitis. In this study, we isolated an Acanthamoeba CJW/W1 strain from tap water in Wuxi, Jiangsu Province, China. Its 18S rDNA was sequenced and a phylogenetic tree was constructed. The isolated cysts belonged to morphologic group II. Comparison of 18S rDNA sequences of CJW/W1 strain and other isolates showed high similarity (99.7%) to a clinical isolate Asp, KA/E28. A phylogeny analysis confirmed this isolate belonged to the pathogenic genotype T4, the most common strain associated with Acanthamoeba-related diseases. This is the first report of an Acanthamoeba strain isolated from tap water in Wuxi, China. Acanthamoeba could be a public health threat to the contact lens wearers and, therefore, its prevalence should be monitored.