• Title/Summary/Keyword: Internal transcribed spacer region

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Genetic Diversity of Culturable Endophytic Fungi Isolated from Halophytes Naturally Growing in Muan Salt Marsh (무안 염습지에 자생하는 염생식물(halophyte)의 뿌리로부터 분리된 내생진균(endophytic fungi)의 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Seo, Yeong-Gyo;Kim, Mi-Ae;Kang, Myung-Suk;Kim, Chang-Mu;Ha, Sang-Chul;Cho, Ga-Youn;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.7
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    • pp.970-980
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    • 2012
  • Native halophytes, such as Suaeda maritima, Limonium tetragonum, S. japonica, Zoysia sinica, and Phragmites australis were collected from the Muan salt marsh. Ninety endophytic fungi were isolated from the roots of the collected halophytes. Molecular insights inferred by internal transcribed spacer containing ITS1, 5.8s, and the ITS2 region showed that all the fungal strains belong to ten orders, i.e., Capnodiales (4.44%), Cystofilobasidiales (1.11%), Dothideales (3.33%), Eurotiales (53.33%), Glomerellales (3.33%), Hypocreales (8.89%), Mucorales (1.11%), Pleosporales (15.56%), Sordariales (1.11%), and Trichosphaeriales (1.11%). The rest (6.67%) of all fungal isolates were not identified. Ninety fungal strains were confirmed at the genus level, containing Acremonium, Alternaria, Aspergillus, Aureobasidium, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Didymella, Dothideomycete, Emericellopsis, Epicoccum, Eupenicillium, Fusarium, Gibberella, Gongronella, Macrophoma, Microsphaeropsis, Nigrospora, Paecilomyces, Paraconiothyrium, Penicillium, Phaeomyces, Phoma, Pleosporales, Purpureocillium, and Talaromyces. Of all the endophytic fungi identified from the various halophytes, Aspergillus and Penicillium of Eurotiales had the highest abundance.

Characterisation of fungal contamination sources for use in quality management of cheese production farms in Korea

  • Kandasamy, Sujatha;Park, Won Seo;Yoo, Jayeon;Yun, Jeonghee;Kang, Han Byul;Seol, Kuk-Hwan;Oh, Mi-Hwa;Ham, Jun Sang
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.1002-1011
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    • 2020
  • Objective: This study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea. Methods: Floor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region. Results: The fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillus in a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms. Conclusion: The environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

The Possible Mechanisms Involved in Citrinin Elimination by Cryptococcus podzolicus Y3 and the Effects of Extrinsic Factors on the Degradation of Citrinin

  • Zhang, Xiaoyun;Lin, Zhen;Apaliya, Maurice Tibiru;Gu, Xiangyu;Zheng, Xiangfeng;Zhao, Lina;Abdelhai, Mandour Haydar;Zhang, Hongyin;Hu, Weicheng
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2119-2128
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    • 2017
  • Citrinin (CIT) is a toxic secondary metabolite produced by fungi belonging to the Penicillium, Aspergillus, and Monascus spp. This toxin has been detected in many agricultural products. In this study, a strain Y3 with the ability to eliminate CIT was screened and identified as Cryptococcus podzolicus, based on the sequence analysis of the internal transcribed spacer region. Neither uptake of CIT by cells nor adsorption by cell wall was involved in CIT elimination by Cryptococcus podzolicus Y3. The extracellular metabolites of Cryptococcus podzolicus Y3 stimulated by CIT or not showed no degradation for CIT. It indicated that CIT elimination was attributed to the degradation of intracellular enzyme(s). The degradation of CIT by C. podzolicus Y3 was dependent on the type of media, yeast concentration, temperature, pH, and initial concentration of CIT. Most of the CIT was degraded by C. podzolicus Y3 in NYDB medium at 42 h but not in PDB medium. The degradation rate of CIT was the highest (94%) when the concentration of C. podzolicus Y3 was $1{\times}10^8cells/ml$. The quantity of CIT degradation was highest at $28^{\circ}C$, and there was no degradation observed at 3$5^{\circ}C$. The study also showed that acidic condition (pH 4.0) was the most favorable for CIT degradation by C. podzolicus Y3. The degradation rate of CIT increased to 98% as the concentration of CIT was increased to $20{\mu}g/ml$. The toxicity of CIT degradation product(s) toward HEK293 was much lower than that of CIT.

Plant Growth Promotion and Gibberellin A3 Production by Aspergillus flavus Y2H001 (Aspergillus flavus Y2H001의 식물생육촉진과 Gibberellin A3의 생산)

  • You, Young-Hyun;Park, Jong Myong;Kang, Sang-Mo;Park, Jong-Han;Lee, In-Jung;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.200-205
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    • 2015
  • Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new $GA_3$-producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.

Yeasts Diversity of Wild Flowers in Mountains of Korea and Their Physiological Functionalities (한국의 야생화에서 분리한 효모의 종 다양성과 생리기능성)

  • Hyun, Se-Hee;Han, Sang-Min;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.137-141
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    • 2015
  • This study focused on isolation of wild yeasts from natural flowers and elucidation of yeast diversity. Wild yeasts were isolated from various flowers collected from mountains on the islands including Jejudo, Ulleungdo, Yokjido, and Seonyudo as well as inlands including Gyejoksan, Oseosan, Beakamsan, and Deogyusan in Korea. Isolated yeasts were identified by comparison of nucleotide sequences for polymerrase chain reaction-amplified D1/D2 region of 26S rDNA or internal transcribed spacer 1 and 2 including 5.8S rDNA using BLAST. 289 strains belonging to 134 yeast species were isolated. Cryptococcus genus strains were the most frequently isolated species among the identified yeasts. Metschnikowia reukaufii was also frequently isolated. Twenty three species including Cryptococcus aureus were overlapped between those of mountains on islands and inland. Physiological functionalities such as antioxidant activity, xanthine oxidase inhibitory activity, and tyrosinase inhibitory activity for the 289 identified yeast strains were investigated using their supernatant and cell-free extracts. The supernatants of Candida sp. 78-J-2 and Metschnikowia reukaufii SY44-6 showed antioxidant activity of 22.5%, and anti-gout xanthine oxidase inhibitory activity of 49.6%, respectively.

Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Morphology and molecular characterization of the epiphytic dinoflagellate Amphidinium massartii, isolated from the temperate waters off Jeju Island, Korea

  • Lee, Kyung Ha;Jeong, Hae Jin;Park, Kila;Kang, Nam Seon;Yoo, Yeong Du;Lee, Moo Joon;Lee, Jin-Woo;Lee, Soojin;Kim, Taekyung;Kim, Hyung Seop;Noh, Jae Hoon
    • ALGAE
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    • v.28 no.3
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    • pp.213-231
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    • 2013
  • Amphidinium massartii Biecheler is an epiphytic and toxic dinoflagellate. Prior to the present study, A. massartii has been reported in the waters off the Mediterranean, Australian, USA, and Canadian coasts. We isolated Amphidinium cells from the coastal waters of Jeju Island, Korea and their morphology and rDNA sequences indicated that they were A. massartii. Herein, we report for the first time the occurrence of A. massartii in the waters of the temperate region in the northwestern Pacific Ocean. The large subunit (LSU) rDNA sequences of the Korean strains were 0.7% different from those of an Australian strain of A. massartii CS-259, the closest species, but were 4.1-5.8% different from those of the other Australian strains and the USA strains of A. massartii and from those of Amphidinium sp. HG115 that was isolated from subtropical Okinawan waters. In phylogenetic trees based on LSU, internal transcribed spacer, small subunit rDNA, and cytochrome b sequences, the Korean strains belonged to the A. massartii clade, which was clearly divergent from the A. carterae clade. The morphology of the Korean A. massartii strains was similar to that of the originally described French strain and recently described Australian strain. However, we report for the first time here that scales were observed on the surface of the flagella. In conclusion, the Korean A. massartii strains have unique rDNA sequences, even though they have a very similar morphology to that of previously reported strains. This report extends the known range of this dinoflagellate to the temperate waters of the northwestern Pacific Ocean.

Analysis of Genomic Diversity of Endophytic Fungal Strains Isolated from the Roots of Suaeda japonica and S. maritima for the Restoration of Ecosystems in Buan Salt Marsh (부안갯벌 생태계 복원을 위한 칠면초와 해홍나물의 내생진균류에 대한 유전학적 다양성 분석)

  • You, Young-Hyun;Yoon, Hyeokjun;Seo, Yeonggyo;Kim, Miae;Shin, Jae-Ho;Lee, In-Jung;Choo, Yeon-Sik;Kim, Jong-Guk
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.287-295
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    • 2012
  • Eighty-four endophytic fungal strains were isolated and identified from the roots of halophytes collected in Buan salt marsh. All halophyte samples, such as Suaeda japonica, and Suaeda maritima were isolated from Buan salt marsh. All endophytic fungi isolated were analyzed by sequences of internal transcribed spacer (ITS) containing ITS1, 5.8s and ITS2 region. All endophytic fungi expressed that fungal strains belong to eight orders; Pleosporales (45%), Eurotiales (27%), Incertae sedis (11%), Dothideales (6%), Capnodiales (5%), Hypocreales (5%), and Agaricales (1%). All endophytic fungi were confirmed at the genus level of Ascomycota and Basidiomycota, containing Alternaria, Ascomycota, Aspergillus, Aureobasidium, Cladosporium, Eupenicillium, Fusarium, Gibberella, Hypocrea, Lewia, Macrophoma, Penicillium, Peyronellaea, Phoma, Pleospora, Pleosporales, Pseudeurotium, Schizophyllum, and Talaromyces. Alternaria (21%) and Penicillium (13%) were the dominant endophytic fungal strains. In this study, endophytic fungal strains analyzed from S. japonica and S. maritime, Alternaria (21%), and Penicillium (13%) of Pleosporales and Eurotiales in halophytes were very abundant.

Diversity of Endophytic Fungal Strains from Jeju Aquatic Plants (제주 수생식물에서 분리한 내생균류의 다양성)

  • Oh, Yoosun;Mun, Hye Yeon;Goh, Jaeduk;Chung, Namil
    • Journal of Life Science
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    • v.27 no.6
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    • pp.661-672
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    • 2017
  • Endophytic fungi are present in host plants and contribute to resistance to biotic and abiotic stress. Aquatic plants are living in aquatic environment such as saltwater or freshwater and exposed more water stress than other land plants. In this study, we investigated 4 wetlands in Jeju and collected 11 aquatic plants. Exogenous microbes were removed by preprocessing of plants and endophytic fungal strains were isolated from the plants. We isolated 126 fungal strains from Namsaengi-pond, 22 fungal strains from Sujangdong-marsh, 44 fungal strains from Yongsu-reservoir and 32 fungal strains from Gangjeongcheon. The fungal strains were identified using internal transcribed spacer (ITS) region and analyzed the phylogeny and diversity. Endophytic fungi isolated from plants of Namsaengi-pond were classified to 30 genera, 19 families, 12 orders, 7 classes and 4 phyla. Endophytic fungi of Sujangdong-marsh were classified to 11 genera, 11 families, 6 orders, 5 classes and 4 phyla. Endophytic fungi of Yongsu-reservoir were classified to 13 genera, 12 families, 7 orders, 5 classes and 4 phyla. Endophytic fungi isolated from Gangjeongcheon were classified to 9 genera, 7 families, 5 orders, 2 classes and 1 phyla. Overall, they were divided 40 genera and Alternaria, Colletotrichum and Fusarium were isolated from 4 sites in common. By investigating the endophytic fungi in aquatic plants, it is for baseline data that determination of diversity and the ecological distribution of endophytic fungi.

Optimal Culture Conditions for Penicillium rubefaciens NNIBRFG5039 Possessing Antimicrobial Activity (항균활성 보유 Penicillium rubefaciens NNIBRFG5039의 최적배양 조건)

  • Hwang, Hye Jin;Mun, Hye Yeon;Hwang, Buyng Su;Nam, Young Ho;Chung, Eu Jin
    • The Korean Journal of Mycology
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    • v.48 no.1
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    • pp.15-27
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    • 2020
  • In screening for antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) KCCM 40510 and Bacillus cereus KCTC 3624, NNIBRFG5039 was isolated from the air in Sangju-si, Gyeongsangbuk-do. Based on a high sequence similarity of the internal transcribed spacer (ITS) region, NNIBRFG5039 was determined to be closely related to Penicillium rubefaciens CBS 139145. The optimal media, initial pH, and temperature for mycelial growth and antimicrobial activity of P. rubefaciens NNIBRFG5039 were determined as follows: potato dextrose broth (PDB), pH 6.5, and 30℃, respectively. Under the optimal culture conditions, maximum mycelial growth (12.4 g L-1) and antibacterial activity (7.5 mm zone of inhibition against MRSA KCCM 40510, and 5.0 mm zone of inhibition against B. cereus KCTC 3624) were observed in a 5 L stirred-tank fermenter. We also isolated the antimicrobial compound from an ethyl acetate fraction, and its chemical structure was identified as (S)-6-hydroxymellein (1) by ESI-MS, 1H-NMR, and 13C-NMR. Consequently, the extract from P. rubefaciens NNIBRFG5039 may be used in functional materials for antimicrobial-related applications.