• 제목/요약/키워드: rDNA ITS sequence

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Isolation of Pseudoalteromonas sp. HJ 47 from Deep Sea Water of East Sea and Characterization of its Extracellular Protease (동해 심층수로부터 Pseudoalteromonas sp. HJ 47의 분리 및 체외단백질분해효소 특성)

  • Cha, In-Tae;Lim, Hayung-Joon;Roh, Dong-Hyun
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.272-278
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    • 2007
  • Proteases are enzymes that break peptide bonds between amino acids of other proteins and occupy a crucial position with respect to their applications in both physiological and commercial fields. In order to screen new source of protease, bacteria producing extracellular proteases at low temperature were isolated from deep sea water of East Sea, Korea. A bacterium showing the best growth rate and production of an extracellular protease at low temperature was designated HJ 47. The DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology led to the placement of this organism in the genus Pseudoalteromonas. Although maximal growth was observed at $37^{\circ}C$, enzyme production per culture time was maximum at $20^{\circ}C$. At this temperature, extracellluar protease production was detected from the end of the exponential phage to stationary phase, and maximal at 15 hours after initial production. The optimum temperature and pH of the protease were found to be $35^{\circ}C$ and 8.

Antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi (김치 분리 유산균의 치즈 곰팡이 항진균 활성)

  • Choi, Ha Nuel;Oh, Hyun Hee;Yang, Hee Sun;Huh, Chang Ki;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Jeong, Eun Jeong;Jung, Hoo Kil
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.727-734
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    • 2013
  • The antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi was investigated. Eight fungi were isolated from cheese in the cheese ripening room. Two of them were identified as Penicillium and Cladosporium via ITS-5.8S rDNA analysis. Twenty-two species of lactic acid bacteria with antifungal activity were isolated from kimchi. Two of them were identified as Lactobacillus and Pediococcus via 16S rRNA sequence analysis. Of the 22 lactic acid bacteria species, six were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024 and P. pentosaceus ALJ026) due to their higher activity against the eight fungi isolated from cheese in the cheese ripening room; and among the six species, the P. pentosaceus ALJ015 and P. pentosaceus ALJ024 isolates from the Jeonju area kimchi and the L. sakei subsp. ALI033 isolate from the Iimsil area kimchi had higher antifungal activity than the other lactic acid bacteria. The minimum inhibitory concentration (MIC) of L. sakei subsp. ALI033 against the eight fungi isolated from cheese in the cheese ripening room was $62.5{\mu}g/mL$.

Isolation and Characterization of Yam-Putrefactive Psychrotrophic Bacteria from Rotted Yam (생마 저온부패 원인세균의 분리 및 부패균의 특성)

  • Ryu Hee-Young;Kim Young-Sook;Park Sang-Jo;Lee Bong-Ho;Kwon Soon-Tae;Sohn Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.109-114
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    • 2006
  • Yam has been recognized as healthy food due to its various biological activities, such as anti-obesity, antimicrobial, anticancer and immuno-stimulation activities, and its consumption has been increased during last decades. In this study, to investigate low-temperature, long-term storage of yam and to develop processed yam products, yam-putrefactive psychrotrophic bacteria were isolated from rotted yam and identified based on BBL identification system, fatty acid analysis in cell membrane and 16S rDNA sequence analysis. The putrefaction activity of isolated thirteen bacteria was evaluated using yam-slices (NaOCl-treated, autoclaved yam and without treatment), and YAM-10 and YAM-12 were identified as major psychrotrophic putrefactive bacteria. Both YAM-10 (Pseudomonas cepacia) and YAM-12 (Pseudomonas rhodesiae) bacteria grew well at 4$\sim$12$^{\circ}C$ and showed strong activity of polymer degrading enzymes, especially amylase, carboxy methyl cellulase and xylanase, at 20$^{\circ}C$. But they failed to grow at acidic pH (<5) or alkaline pH (>10). Our results suggested that the control of psychrotrophic Pseudomonas sp. by pH change and inhibition of polymer degrading enzymes, such as amy-lase, are necessary to long-term storage of yam.

Morphological and Molecular Characterization of Alternaria Isolates from Solanaceous Crops (가지과 작물에서 분리한 Alternaria 속 균의 형태적, 분자생물학적 특징)

  • Yu, Seung-Hun;Cho, Hye-Sun;Kim, Byung-Ryun;Park, Myung-Soo
    • The Korean Journal of Mycology
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    • v.31 no.2
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    • pp.103-113
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    • 2003
  • Twenty five isolates of Alternaria were obtained from various solanaceous crops in Korea. For all isolates, morphological characteristics of the conidia were determined and compared with those of representative isolates of A. solani and A. tomatophila. A selection of the isolates and the representative Alternaria isolates were evaluated for Pathogenicity to potato, tomato, egg plant and red pepper. Molecular characteristics of 17 isolates of Alternaria inculding the representative isolates were determined using sequence analysis of IRS rDNA and histone H3 gene, and URP-PCR analysis. Based on morphological characteristics, the isolates from the solanaceous crops were grouped as identical or very similar to either A. tomatophila (ATO), A. solani (ASO), and unidentified Alternaria sp. (ASP). Isolates of ASO were moderately pathogenic to all the solanaceous crops tested, but ATO isolates were highly pathogenic to tomato and the ASP isolate was pathogenic only to potato. Among the molecular markers used in this study, the URP-PCR analysis was found to be appropriate for taxonomic resolution of these species. Based on the conidial morphology, pathogenicity test and molecular characteristics, A. tomatophila (early blight of tomato) could be distinguished from A. solani (early blight of potato), and the Alternaria sp. (ASP) from potato, which was closely related to ASO in conidial morphology, was considered as a new species.

Identification and Characterization of Entomopathogenic Fungi Isolated from Bemisia tabaci in Korea (담배가루이(Bemisia tabaci )에서 분리한 곤충병원성진균의 동정 및 병원성 검정)

  • Park, Hyun-Rho;Ryu, Young-Hyun;Yeon, Il-Kyen;Nam, Sung-Hee;Kim, Dong-Geun;Han, Myung-Sae
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.27-34
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    • 2014
  • Entomopathogenic fungi were isolated from Bemisia tabaci in an Oriental melon field, and their growth characteristics, factors related to a natural outbreak, and infectivity against Bemisia tabaci, Tetranychus urticae, and Frankliniella intonsa were investigated. The isolates had erect conidiophores bearing whorls of 4-6 phialides with a swollen base where cylindrical conidia of $3.0-3.4{\mu}m$ were attached. The isolates were identified as Isaria fumosorosea on the basis of morphological characteristics and an ITS sequence with 99% similarity. I. fumosorosea IFs-08 grew well on Sabouraud dextrose agar+yeast extract medium(3.2 mm/day/$24^{\circ}C$); it grew better at $35^{\circ}C$ than at $15^{\circ}C$. The isolates of I. fumosorosea-IFs were highly infective and killed 93.9-96.7% B. tabaci, 84.9-92.0% T. urticae, and 81.5-84.4% F. intonsa in bioassay, whereas three isolates (Isaria tenuipes, Isaria farinosa, and Isaria fumosorosea) from KACC showed a low infectivity of 10-20%. To the best of our knowledge, this is the first report of I. fumosorosea isolated from B. tabaci in Korea.

In vivo Antifungal Activity of Pyrrolnitrin Isolated from Burkholderia capacia EB215 with Antagonistic Activity Towards Colletotrichum Species (탄저병균에 대하여 길항작용을 보이는 Burkholderia cepacia EB215로부터 분리한 Pyrrolnitrin의 항균활성)

  • Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.31-38
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    • 2004
  • An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

Morphology, Phylogeny and Ecology of Hyphomycetes Hyperparasitic to Rusts

  • Park, Mi-Jeong;Park, Jong-Han;Hong, Seung-Beom;Shin, Hyeon-Dong
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.55-55
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    • 2015
  • Rust is one of the most destructive diseases on economically important plants such as agricultural and horticultural crops, as well as forest trees [1]. Chemical treatment is the most effective means to control rust, but use of the chemical fungicides involves inevitable risks to human health and environment [2]. Unfortunately, biocontrol is currently impracticable for rust disease management [3]. It is necessary to exploit biocontrol agents to help prevent rust diseases. As a fundamental research for future development of biocontrol agents for rusts, biodiversity of hyperparasites occurring on rust fungi was investigated. During 2006-2010, 197 fungal isolates of the rust hyperparasites were collected and isolated from various combinations of mycohosts and plant hosts in many regions of Korea. Based on morphological and molecular data, they were identified as 8 genera and 12 species. Besides, phylogenetic relationships between the hyperparasites and related taxa were inferred. A total of 114 isolates of Pseudovirgaria were obtained from rust pustules of Phragmidium spp. and Pucciniastrum agrimoniae infecting rosaceous plants. Phylogenetic analysis using multigene sequences revealed a high level of genetic variability among many isolates of Pseudovirgaria and close correlation between the isolates and mycohosts. Only two species of Pseudovirgaria, P. hyperparasitica and P. grisea are often difficult to distinguish by their morphological similarity, but on the molecular basis they were clearly differentiated from each other. There had been no previous record of P. grisea outside Europe, but the present study has proved its presence in Korea. Among six distinct groups (five of P. hyperparasitica and one of P. grisea) within the Pseudovirgaria isolates, each lineage of P. hyperparasitica was closely associated with specific mycohosts and thus might have cospeciated with their mycohosts, which probably led to coevolution. Although P. grisea possesses a host preference for Phragmidium species occurring on Rubus, it was not specific for a mycohost. P. grisea seems to evolve in the direction of having a broad mycohost range. Seventeen isolates of Verticillium-like fungi were isolated from rust sori. Based on morphological data and DNA sequence analysis, the isolates were identified as three Lecanicillium species, viz. L. attenuatum, Lecanicillium sp. 1, Lecanicillium sp. 2, and V. epiphytum. The unidenified two species of Lecanicillium appear to be previously unknown taxa. Sixty-six isolates of miscellaneous hyphomycetes belonging to 6 species of 5 genera were obtained from pustules of rust fungi. On the basis of morphological and molecular analyses, the miscellaneous hyphomycetes growing on rusts were identified as Acrodontium crateriforme, Cladophialophora pucciniophila, Cladosporium cladosporioides, Phacellium vossianum, Ramularia coleosporii, and R. uredinicola.

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Stem Rot on Ligularia fischeri Caused by Sclerotium rolfsii in Korea (Sclerotium rolfsii에 의한 곰취 흰비단병)

  • Moon, Youn-Gi;Kim, Se-Won;Choi, Jun-Keun;Kwon, Soon-Bae;Shim, Hong-Sik;Ju, Ho-Jong;Choi, In-Young
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.36-39
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    • 2015
  • In June 2012 and 2013, a destructive stem rot symptoms of Ligularia fischeri occurred sporadically in Hoengseong-gun and Pyeongchang-gun Gangwon-do, Korea. The typical symptom included water-soaking on the main stem, rotting, wilting and blighting, which eventually leads to death of the plant. White mycelial mats were spread over lesions and brown sclerotia were formed on stems and near soil surface. The sclerotia were white to brown, spherical or irregular, 1-3 mm in size on potato dextrose agar (PDA), The optimum temperature range of hyphal growth was $25-30^{\circ}C$ and the hyphal diameter was $4-10{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. The resulting sequence of 695 bp was deposited in GenBank. A BLAST search revealed that sequences of the this isolates showed >99% identity with those of Sclerotium rolfsii. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers ITS rDNA, the fungi were identified as S. rolfsii. A pathogenicity test was carried out to fulfill Koch's postulates. To our knowledge, this is the first report of S. rolfsii on Ligularia fischeri in Korea.

Induced Systemic Resistance in plants by Bacillus sp. Isolated from Dok-do Islands (독도 자생식물 번행초로부터 분리한 바실러스 속 식물생장촉진근권 세균에 의한 식물병 저항성 유도)

  • Kim, Seung-Kun;Son, Jin-Soo;Kwon, Duck-Kee;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.596-602
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    • 2019
  • In September 2017, the rhizospheric soil of Tetragonia tetragonoides (Pall.) Kuntze was further sampled. One hundred and thirty eight species of microorganisms were isolated from the soil. Indole-3-acetic acid (IAA) production, siderophore production, and phosphate degradation were examined in order to confirm bacterial growth from isolated microorganisms. As a result, most strains were able to produce auxins or siderophores and to solubilize phosphate. In addition, 138 isolated strains were treated with tobacco extract and conferred pathogen resistance to host plants upon treatment. As a result, 35 strains that were able to reduce pathophysiology by more the 60% were selected. Among them, 6 strains with high induced systemic resistance (ISR) activity were found. All of these strains belong to the genus Bacillus according to the 16S rDNA sequence analysis. Bacillus aryabhattai KUDC6619 showed outstanding effects with reduced infection in tobacco and pepper plants. Probably, these Bacillus species play a beneficial role by association with T. tetragonoides for its survival in the harsh conditions found on the island of Dokdo.

The Isolation and Culture Characterization of a Lipolytic Enzyme Producing Strain from Meju (메주로부터 지질분해 효소 생산 균주의 분리 및 배양학적 특성)

  • Yun, Hye-Ju;Lee, You-Jung;Yeo, Soo-Hwan;Choi, Hye-Sun;Park, Hye-Young;Park, Heui-Dong;Baek, Seong-Yeol
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.98-103
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    • 2012
  • For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.