• Title/Summary/Keyword: ITS rDNA sequences

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First Report of Allantophomopsiella pseudotsugae Isolated from Soil in Korea

  • Wajihi, Ally Hassan;Lee, Seung-Yeol;Das, Kallol;Eom, Ahn-Heum;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.29-34
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    • 2019
  • A fungal isolate designated 17E029 was isolated from a soil sample in Jeju, Korea. The strain was similar to other Allantophomopsiella species in its morphological characteristics such as grey mycelia, conidiophore, and conidia sizes. The isolate produced aerial mycelia, which appeared grey on the reverse side of the media surfaces and turned black on the front side of the colonies. The conidiophores emanating from the hyphae were hyaline, grey, aseptate, branched, and $6.7{\sim}9.2{\times}1.8{\sim}2.5{\mu}m$. Conidiogenous cells were ovoid to subcylindrical, discrete, guttulate, and hyaline. Conidia were hyaline, aseptate, smooth, guttulate, oval to subcylindrical, irregular in shape, and $6.0{\sim}7.8{\times}3.0{\sim}3.4{\mu}m$. The strain was confirmed based on phylogenetic analysis of the closest related organism, A. pseudotsugae CBS 288.37, using the partial 28S, internal transcribed spacer rDNA regions, and partial RNA polymerase II second largest subunit locus (RPB2) gene sequences along with its culture characteristics. Therefore, morphological observations and phylogenetic analysis revealed that strain 17E029 is similar to the previously identified A. pseudotsugae. Hence, this species was described as A. pseudotsugae strain 17E029, which is a new record in Korea.

First Report of Pectobacterium aroidearum Causing Soft Rot on Zamioculcas zamiifolia

  • Kyoung-Taek Park;Soo-Min Hong;Leonid N. Ten;Chang-Gi Back;Seung-Yeol Lee;In-Kyu Kang;Hee-Young Jung
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.445-451
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    • 2023
  • Zamioculcas zamiifolia is a popular indoor ornamental plant in Korea. In August 2021, a severe outbreak of soft rot disease affected Z. zamiifolia in Emseong, Chungcheongbuk-do, Korea. Infected plants displayed wilting, water-soaked lesions, stem collapse, and green-brown discoloration. The bacterial strain KNUB-05-21 was isolated from infected stems and identified as Pectobacterium aroidearum using 16S rRNA nucleotide sequencing and multilocus sequence analysis based on partial sequences of dnaX, leuS, and recA genes. Confirmation of its affiliation with P. aroidearum was also obtained through biochemical and morphological characterization. To confirm the pathogenicity of strain KNUB-05-21, its suspension was injected into Z. zamiifolia stems. Within a week, soft rot developed on the stems, exhibiting symptoms similar to those observed in field-infected plants. The reisolated strain was identical to those of P. aroidearum. Before this study, P. aroidearum was not reported as a causative pathogen of Z. zamiifolia soft rot in Korea.

Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.

Kitasatospora sp. MJM383 Strain Producing Two Antitumor Agents, Streptonigrin and Oxopropaline G

  • JIN YING-YU;YOON TAE-MI;KIM WON-KON;KIM KYOUNG-ROK;SONG JEA-KYOUNG;KIM JONG-GWAN;LIU JING;YANG YOUNG-YELL;KWON HYUNG-JIN;SUH JOO-WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1140-1145
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    • 2005
  • MJM383, a rare actinomycete sp. strain originated from Chinese soils, was isolated through an antimicrobial screening system. The analysis of 16S rDNA sequences and biochemical characterization determined the strain to belong to genus Kitasatospora. Both NMR and ESI mass data of its purified bioactive compounds revealed Kitasatospora sp. MJM383 to produce two antitumor agents, streptonigrin and oxopropaline G, which have been known to be produced from Streptomyces species. This is the first report to demonstrate the presence of antitumor agents produced by genus Kitasatospora.

Unrecorded Fungal Species Isolated from Greenhouses Used for Shiitake Cultivation in Korea (표고버섯 재배사에서 분리한 국내 미기록 진균 보고)

  • Ahn, Geum Ran;Kwon, Hyuk Woo;Ko, Han Kyu;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.8-15
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    • 2016
  • Fungal contamination is a detrimental factor affecting sawdust media-based shiitake cultivation in greenhouses. During fungal monitoring of greenhouses used for shiitake cultivation, eight fungal species were isolated and identified from indoor air and mushroom flies collected in the greenhouses. The current study reported five species as new in Korea, viz. Ascochyta hordei, Discosia artocreas, Mucor nidicola, Perenniporia medulla-panis, and Pseudozyma prolifica, and confirmed two species, Penicillium charlesii and Penicillium brevicompactum, which were previously recorded in Korea without molecular taxonomic validation. The morphological characteristics and phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer rDNA region or calmodulin gene were described for all identified species.

A Report of Five Unrecorded Fungal Species of Korea (국내 미기록 진균 5종 보고)

  • Ahn, Geum Ran;Kim, Bo Young;Lee, Geun Sick;Hyun, Min Woo;Lee, Chan Jung;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.240-246
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    • 2016
  • During a survey of the activities of fungi in steamed sweet potato, stored garlic, agricultural by-products for mushroom cultivation media, and pinewood chips from a pinewood nematode-infected tree, numerous fungal samples were isolated and identified. This study identified five species that have not been previously reported in Korea, namely Geomyces pannorum, Neopestalotiopsis javaensis, Penicillium allii, Penicillium chermesinum, and Ophiognomonia setacea. For all identified species, the cultural features of colonies formed on growth media, their morphological characteristics observed by a light microscope, and their molecular phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer rDNA region or calmodulin gene were described.

Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae;Kim, Hyung-Kwoun;Park, Sun-Yang;Koo, Bon-Tag;Oh, Tae-Kwang;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.85-91
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    • 2001
  • To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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One unusual species, Coilia sp. (Engraulidae, Pisces) from the Yellow Sea

  • Kwun, Hyuck-Joon;Kim, Yeong-Hye;Kim, Jong-Bin;Jeong, Choong-Hoon;Kim, Jin-Koo
    • Animal cells and systems
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    • v.14 no.2
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    • pp.137-145
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    • 2010
  • Four specimens of unknown Coilia sp. were collected for the first time from the Yellow Sea in 2008 and compared with Coilia mystus and Coilia nasus. Coilia sp. showed similar morphology to C. mystus and C. nasus, but differed in that its tail was considerably shorter. We conducted an analysis of the morphological and genetic characteristics in an effort to clarify the taxonomic position of Coilia sp. In counts and measurements, Coilia sp. were well distinguished from C. nasus by the number of scutes (42-44 in Coilia sp. vs. 40-45 in C. mystus vs. 45-55 in C. nasus), ratio of dorsal base length to head length (43.4-47.6 vs. 37.9-47.6 vs. 33.0-41.0), and eye length to head length (19.2-20.8 vs. 17.0-22.4 vs. 13.8-18.2). In caudal skeleton of Coilia sp., urostyle, hypural and epural bones were not observed; instead of them, caudal fin rays were supported by the last vertebra, neural and haemal spines' extension. The molecular phylogenetic relationship was analyzed using 414 base-pair 12S rRNA mitochondrial DNA sequences. The Kimura-2-parameter distance between Coilia sp. and C. mystus was 0.3%, but was 1.3% between Coilia sp. and C. nasus. Both the neighbor-joining tree and maximum-likelihood tree showed that Coilia sp. are closely clustered with C. mystus. Therefore, our results suggest that the Coilia sp. may be a deformed fish of C. mystus.

Pucciniastrum hydrangeae-petiolaris, a Newly Found Rust Fungus on Hydrangea petiolaris in Korea (국내 미기록 등수국 녹병균 Pucciniastrum hydrangeae-petiolaris)

  • Lee, Jae Sung;Choi, Young-Joon;Choi, Byoung-Ki;Jung, Bok-Nam;Park, Ji-Hyun;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.119-125
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    • 2021
  • Hydrangea petiolaris, belonging to the family Hydrangeaceae, is a vine plant distributed in Ulleung, Jeju, and other southern islands of Korea. In October 2017, a rust fungus was discovered on H. petiolaris in Jeju Island, Korea. To identify the rust fungus, we performed a morphological examination and molecular phylogenetic analysis of the internal transcribed spacer and 28S large subunit rDNA sequences. As a result, the fungus was identified as Pucciniastrum hydrangeae-petiolaris, consistent with previous reports from Japan and Russia, but showed a significant phylogenetic distance from Pucciniastrum hydrangeae reported on Hydrangeae spp. in North America. This is the first record of P. hydrangeae-petiolaris on H. petiolaris in Korea.

Rust Fungus Pucciniastrum styracinum on Styrax japonicus in Korea Confirmed by Morphological and Molecular Data (형태 및 분자적 특징에 의한 때죽나무 녹병균(Pucciniastrum styracinum)의 재확인)

  • Park, Ji-Hyun;Jung, Bok-Nam;Lee, Jae Sung;Choi, Young-Joon;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.531-537
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    • 2021
  • Styrax japonicus is a deciduous shrub or a small tree belonging to the family Styracaceae. Since 1990, a rust fungus has continuously been found on S. japonicus leaves across the country. The morphological characteristics of the rust fungus were most consistent with those of Pucciniastrum styracinum reported on Styrax obassia in Japan. The species identification was confirmed by molecular phylogenetic analysis of the internal transcribed spacer and large subunit rDNA sequences. The current study provides mycological information with morphological measurements and molecular phylogenetic data of P. styracinum found on S. japonicus in Korea. Therefore, this is the first confirmed report on the presence of P. styracinum on S. japonicus in Korea.