• Title/Summary/Keyword: ITS region of rDNA

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Characterization of a Novel Alga-Lytic Bacterium, Acidovorax temperans AK-05, Isolated from an Eutrophic Lake for Degradation of Anabaena cylindrica (부영양 호수에서 분리한 Acidovorax temperans AK-05의 Anabaena cylindrica 분해 특성)

  • Kim, Jeong-Dong;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.37 no.2 s.107
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    • pp.241-247
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    • 2004
  • Isolation and identification of alga-lytic bacteria were carried out. Fifteen isolates of alga-lytic bacteria were screened by the double layer method using A. cylindrica NIES-19 as a sole nutrient and four isolates among them were compared with their alga-lytic activity. The isolate AK-05 exhibiting the highest alga-lytic activity was identified as Acidovorax temperans base on its 16S rDNA sequence. The culture supernatant of the isolate AK-05 was reliable for the alga-lytic. Alga-lytic activity assays of culture supernatant revealed that the major substances for alga-lytic activity were non-proteins and heat stable. The highest alga-Iytic activity was practical under alkaline conditions and at 25${\sim}$$30^{\circ}C$. It is indicating an advantage for the application of water blooms by cyanobacteria in eutrophic lakes where the pH is generally in alkaline region.

New Species of the Genus Metschnikowia Isolated from Flowers in Indonesia, Metschnikowia cibodasensis sp. nov.

  • Sjamsuridzal, Wellyzar;Oetari, Ariyanti;Nakashima, Chiharu;Kanti, Atit;Saraswati, Rasti;Widyastuti, Yantyati;Ando, Katsuhiko
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.905-912
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    • 2013
  • A novel species, Metschnikowia cibodasensis, is proposed to accommodate eight strains (ID03-$0093^T$, ID03-0094, ID03-0095, ID03-0096, ID03-0097, ID03-0098, ID03-0099, and ID03-0109) isolated from flowers of Saurauia pendula, Berberis nepalensis, and Brunfelsia americana in Cibodas Botanical Garden, West Java, Indonesia. The type strain of M. cibodasensis is ID03-$0093^T$ (= NBRC $101693^T$ =UICC $Y-335^T$ = BTCC-$Y25^T$). The common features of M. cibodasensis are a spherical to ellipsoidopedunculate shaped ascus, which contains one or two needle-shaped ascospores, and lyse at maturity. Asci generally develop directly from vegetative cells but sometimes from chlamydospores. The neighbor-joining tree based on the D1/D2 domain of nuclear large subunit (nLSU) ribosomal DNA sequences strongly supports that M. cibodasensis (eight strains) and its closest teleomorphic species, M. reukaufii, are different species by a 100% bootstrap value. The type strain of M. cibodasensis, ID03-$0093^T$, differed from M. reukaufii NBRC $1679^T$ by six nt (five substitutions and one deletion) in their D1/D2 region of nLSU rDNA, and by 18 nt (five deletions, four insertions, and nine substitutions) in their internal transcribed spacer regions of rDNA, respectively. Four strains representative of M. cibodasensis (ID03-$0093^T$, ID03-0095, ID03-0096, and ID03-0099) showed a mol% G+C content of $44.05{\pm}0.25%$, whereas that of M. reukaufii NBRC $1679^T$ was 41.3%. The low value of DNA-DNA homology (5-16%) in four strains of M. cibodasensis and M. reukaufii NBRC $1679^T$ strongly supported that these strains represent a distinct species.

Plant growth-promoting activity and identification of endophytic fungi isolated from native plant in East coast (동해안 자생식물로부터 분리된 내생균류의 식물생장촉진활성 및 동정)

  • You, Young-Hyun;Jin, Yong Ju;Kang, Sang-Mo;Oh, Sejong;Lee, Myung-Chul;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.14-20
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    • 2015
  • Coastal plant species, Plantago camtschatica Cham. native to the coastal region of the East Sea were sampled and then morphologically different 20 endophytic fungal strains were purely isolated. Phylogenetic analysis of isolates was done by the Bayesian program based on sequenced internal transcribed spacer (ITS-rDNA) region. Culture filtrates of each of 20 isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity, respectively. As the results, E/PC/10/1 strain showed the highest plant growth-promoting activity among them. The culture filtrate of the strain E/PC/10/1 was revealed as containing gibberellins ($GA_1$, $GA_3$, $GA_4$) by using HPLC, and gas GC/MS with selected ion monitoring (SIM). Finally, this strain was identified as novel Penicillium spinulosum species that producing new GAs with microscopic observation and further molecular analysis with beta-tubulin gene sequence.

Two new species of Trichoderma isolated from commercially grown oyster mushroom, Pleurotus ostreatus (oral)

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.127.1-127
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    • 2003
  • We describe two new Trichoderma species associated with oyster mushroom in Korea. Trichoderma green mould has been one of the most serious diseases of oyster mushroom in Korea. Of these the predominant species are two unrecorded species. We designed as Trichoderma sp. Korean type 1 (Th K1) and Trichoderma sp. Korean type 2 (Th K2), respectively. Th K1 and Th K2 can be distinguished from previously reported Trichoderma species as well as each other in morphological characteristics including growth rate at 35$^{\circ}C$, colony morphology, conidia shape and branch pattern of phialides. Sequence of the ITS region of rDNA, the protein coding translation elongation factor gene(EF-1${\alpha}$), and RNA polymeraseII (RPB2) not only clearly separated Trichoderma sp. Korean types from their closely related T. harzianum biotype but also distinguished them from each other. Analyses of the EF-1${\alpha}$ and RPB2 sequences were found to be more useful for establishing systematic relationships among Trichoderma isolates than those of the ITS sequence. Based on the results of morphological and molecular characteristics. We propose the two Trichoderma sp. Korean types as the new species

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Erysiphe lonicerigena sp. nov., a Powdery Mildew Species Found on Lonicera harae

  • In-Young Choi;Lamiya Abasova;Joon-Ho Choi;Jung-Hee Park;Hyeon-Dong Shin
    • Mycobiology
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    • v.51 no.2
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    • pp.67-71
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    • 2023
  • A powdery mildew (Erysiphaceae) has been continuously collected on the leaves of Lonicera harae in the southern part of the Korean Peninsula, where this shrub is indigenous. Microscopic examination of the asexual morphs revealed that the current collections are differentiated from the all known Erysiphe species on Lonicera spp. by its longer conidiophores and longer conidia. Although the morphology of the chasmothecia is reminiscent of Erysiphe ehrenbergii and E. lonicerae, the specimens on L. harae differ from them in having smaller ascospores. A phylogenetic tree generated from a combined dataset of the internal transcribed spacer region and 28S rDNA gene sequences demonstrates that sequences obtained from three powdery mildew collections on L. harae clustered together as an independent species clade with high bootstrap values distant from other Erysiphe species on Lonicera, representing a species of its own. Based on morphological differences and molecular-phylogenetic results, the powdery mildew on L. harae is proposed as a new species, Erysiphe lonicerigena, and the holomorph of the fungus is described and illustrated in this study.

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

  • Kim Cuc Thi Nguyen;Phuc Hung Truong;Cuong Tu Ho;Cong Tuan Le;Khoa Dang Tran;Tien Long Nguyen;Manh Tuan Nguyen;Phu Van Nguyen
    • Mycobiology
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    • v.51 no.6
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    • pp.379-387
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    • 2023
  • In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1T could highly tolerate Cu2+ up to 10 mM, and the isolates could also grow in a wide range of pH (3-7), and temperature (10-40 ℃). Dried biomass of GL1 was able to remove Cu2+ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 ℃, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

Site-directed Mutagenesis Analysis Elucidates the Role of 223/227 Arginine in 23S rRNA Methylation, Which Is in 'Target Adenine Binding Loop' Region of ErmSF (위치 지정 치환 변이를 이용한 ErmSF의 '타깃 Adenine Binding Loop'을 형성하는 부위에 존재하는 223/227 Arginine 잔기의 23S rRNA Methylation 활성에서의 역할 규명)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.79-86
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    • 2012
  • ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.

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.

Genetic Diversity of Endophytic Fungi Isolated from the Roots of Halophytes Naturally Growing in Suncheon Bay (순천만에 자생하는 염생식물의 뿌리로부터 분리된 내생진균의 유전적 다양성)

  • Seo, Yeong-Gyo;Kim, Mi-Ae;You, Young-Hyun;Yoon, Hyeok-Jun;Woo, Ju-Ri;Lee, Gyeong-Min;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.7-10
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    • 2012
  • Endophytic fungi were isolated from the roots of halophytes, Suaeda japonica and Carex scabrifolia in the Suncheonbay. The ITS region in rDNA of 15 endophytic fungal strains were amplified using PCR with universal primers ITS1 and ITS4, and those amplified fragments were sequenced. Based on ITS sequence, five fungal genera were identified in S. japonica and seven fungal genera were identified in C. scabrifolia. The Shannon's diversity index (H') of endophytic fungi isolated from S. japonica and C. scabrifolia was 1.561 and 1.889, respectively. In phylogenetic analysis, it was shown that Ascomycota and Pezizomycotina was widely distributed both in S. japonica and C. scabrifolia. Also, Sordariomycetes, Dothideomycetes and Eurotiomycetes were shown to be distributed in these halophytes used in this experiment.

DNA Polymorphism of Insulin-like Growth Factor-binding Protein-3 Gene and Its Association with Cashmere Traits in Cashmere Goats

  • Liu, Haiying;Liu, Chao;Yang, Guiqin;Li, Hui;Dai, Jin;Cong, Yuyan;Li, Xuejian
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1515-1520
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    • 2012
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is important for regulation of growth and development in mammals. The present investigation was carried out to study DNA polymorphism by PCR-RFLP of IGFBP-3 gene and its effect on fibre traits of Chinese Inner Mongolian cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Four hundred and forty-four animals were used to detect polymorphisms in the hircine IGFBP-3 gene. A 316-bp fragment of the IGFBP-3 gene in exon 2 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in the populations. The frequency of AA, AB and BB genotypes was 0.58, 0.33 and 0.09 respectively. The allelic frequency of the A and B allele was 0.75 and 0.25 respectively. Nucleotide sequencing revealed a C>G transition in the exon 2 region of the IGFBP-3 gene resulting in R158G change which caused the polymorphism. Least squares analysis revealed a significant effect of genotypes on cashmere weight (p<0.0001), cashmere fibre length (p<0.001) and hair length (p<0.05) of the animals. The effect of genotypes on cashmere fibre diameter was not statistically significant (p>0.05). The animals of AB and BB genotypes showed higher cashmere weight, cashmere fibre length and hair length than the animals possessing AA genotype. These results suggested that polymorphisms in the hircine IGFBP-3 gene might be a potential molecular marker for cashmere weight in cashmere goats.