• Title/Summary/Keyword: Fungal rDNA

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Bioprospecting Endophytic Fungi and Their Metabolites from Medicinal Tree Aegle marmelos in Western Ghats, India

  • Mani, Vellingiri Manon;Soundari, Arockiamjeyasundar Parimala Gnana;Karthiyaini, Damodharan;Preethi, Kathirvel
    • Mycobiology
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    • v.43 no.3
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    • pp.303-310
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    • 2015
  • The increasing emergence of lead drugs for the resistance produced by the pathogenic strains and arrival of new diseases have initiated the need for searching novel metabolites with best anticancer and antimicrobial properties than the existing one. With this view, the investigation was conducted for the isolation, identification, and biological evaluation of potential endophytic fungi of Aegle marmelos, a medicinal tree used for more than three decades, for curing various disorders. A total of 169 endophytic fungal strains obtained from sampling and among those 67 were pigmented strains. Upon antagonistic screening, five endophytic fungal strains exhibited antagonistic potentiality by inhibiting the pathogens. These five potent strains were characterized at molecular level by sequencing the amplified internal transcribed spacer (ITS) 1 and ITS 4 regions of rDNA and they were grouped under order Pleosporales, Eurotiales, and Capnodiales. The metabolites from the respective strains were produced in fungal culturing media and extracted using polar solvents. Further, the extracts of five endophytes manifested antimicrobial activity against tested clinical pathogens and Alternaria alternata (FC39BY), Al. citrimacularis (FC8ABr), and Curvularia australiensis (FC2AP) exhibited significant antimicrobial profile against 9 of 12 tested pathogens, showing broad spectrum activity. The antioxidant levels of all the five endophytes revealed the highest activity at least concentrations, and major activity was unveiled by the members of order Pleosporales FC2AP and FC8ABr. This research explains the value of endophytic fungal extracts and its significance of antimicrobial and antioxidant properties.

Isolation and Identification of Fungi from a Meju Contaminated with Aflatoxins

  • Jung, Yu Jung;Chung, Soo Hyun;Lee, Hyo Ku;Chun, Hyang Sook;Hong, Seung Beom
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1740-1748
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    • 2012
  • A home-made meju sample contaminated naturally with aflatoxins was used for isolation of fungal strains. Overall, 230 fungal isolates were obtained on dichloran rosebengal chloramphenicol (DRBC) and dichloran 18% glycerol (DG18) agar plates. Morphological characteristics and molecular analysis of a partial ${\beta}$-tubulin gene and the internal transcribed spacer (ITS) of rDNA were used for the identification of the isolates. The fungal isolates were divided into 7 genera: Aspergillus, Eurotium, Penicillium, Eupenicillium, Mucor, Lichtheimia, and Curvularia. Three strains from 56 isolates of the A. oryzae/flavus group were found to be aflatoxigenic A. flavus, by the presence of the aflatoxin biosynthesis genes and confirmatory aflatoxin production by high-performance liquid chromatography (HPLC). The predominant isolate from DRBC plates was A. oryzae (42 strains, 36.2%), whereas that from DG18 was A. candidus (61 strains, 53.5%). Out of the 230 isolates, the most common species was A. candidus (34.3%) followed by A. oryzae (22.2%), Mucor circinelloides (13.0%), P. polonicum (10.0%), A. tubingensis (4.8%), and L. ramosa (3.5%). A. flavus and E. chevalieri presented occurrence levels of 2.2%, respectively. The remaining isolates of A. unguis, P. oxalicum, Eupenicillium cinnamopurpureum, A. acidus, E. rubrum, P. chrysogenum, M. racemosus, and C. inaequalis had lower occurrence levels of < 2.0%.

Identification of fungal species in oak mushroom cultivation houses during the rainy season (장마철 표고(Lentinula edodes) 재배사내 발생 유해 진균류의 동정)

  • Jeong, Sang-Wook;Jang, Eun-Gyeong;Choi, Sun-Gyu;Lee, Won-Ho;Ban, Seung-Eon
    • Journal of Mushroom
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    • v.19 no.1
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    • pp.66-70
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    • 2021
  • We monitored the occurrence of fungal species on bed-logs inoculated with oak mushroom (Lentinula edodes) and the environmental conditions of temperature and humidity in the cultivation houses during monsoon. Six fungal species, viz., Cladosporium sp., C. cladosporioides, C. anthropophilum, Pleosporales sp., Trichoderma harzianum, and Acremonium sp., wereidentified from the cultivation houses located in Jangheung, Jeonnam province. This identification was confirmed by performing nucleotide sequence analysis of the internal transcribed spacer and 28S rDNA regions. Our study presents significant findings that can help in preventing fungal damage induced by inappropriate temperature and humidity in oak mushroom cultivation houses.

Characteristics and Antimicrobial Effects of Novel Burkholderia cepacia No. 15-2 Isolated from Compost (퇴비로부터 분리된 Burkholderia cepacia No.15-2의 특성과 항균 효과)

  • Yun, Soon-Il
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.421-428
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    • 2003
  • To develop the functional-compost containing antifungal substance by using antagonistic microorganisms, Spinacia oleracea L and Rhizoctonia solani Kuhn O-28 were used as a model plant and phytopathogen, respectively. Total 80 strains were isolated from the compost of various waste foods mixture processed for a year. Among them, No.15-2 strain was selected due to its highest antifungal activity against R. solani Kuhn O-28 and was identified phyno- and phylogenotypically as Burkholderia cepacia genomovar V. which is rare probability in pathogen, by 16S rDNA sequencing and specific primer pair PCR method. B. cepacia No.15-2 preferentially dominated during the compost and its cell numbers were maintained almost $${\times}$10^{13}$ cuf/g for 15 days. The morbidity caused by R. solani Kuhn O-28 in S. oleracea L cultivation was reduced to 40% by addition of B. cepacia No.15-2. In conclusion, the antifungal compost using B. cepacia No.15-2 could be applied to biocontrol of various crops blights caused by fungal pathogen.

Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba

  • Yang, Hye-Won;Lee, Yu-Ran;Inoue, Noboru;Jha, Bijay Kumar;Sylvatrie Danne, Dinzouna-Boutamba;Kim, Hong-Kyun;Lee, Junhun;Goo, Youn-Kyoung;Kong, Hyun-Hee;Chung, Dong-Il;Hong, Yeonchul
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.269-277
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    • 2013
  • Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

Characteristics of an Entomopathogenic Fungus Infecting Corythucha ciliata (Hemiptera: Tingidae) (버즘나무방패벌레 기생성 곰팡이의 특성 구명)

  • Koo, Chang-Duck;Lee, Seung-Kyu;Kim, Seong Hwan
    • Journal of Korean Society of Forest Science
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    • v.96 no.1
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    • pp.58-64
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    • 2007
  • Overwintering adults of sycamore lace bug (Corythucha ciliata) infected by an unidentified pathogenic fungus were found on the stems of street trees of sycamore in Cheongju city. The objective of this study was to describe this entomopathogenic fungus infecting overwintering sycamore lace bug adults. This unidentified fungus colonized the insect adult body and formed white colony with subglobose clusters of conidiocarps. The size of conidiocarps was 300 to $400{\mu}m$ and each conidium was 15 to $20{\mu}m$. The conidiospore was globus and 2.5 to $3.0{\mu}m$ in diameter, and the hyphae were 1 to $5{\mu}m$ thick. This fungus was successfully isolated and cultivated on potato dextrose agar medium (PDA). The fungal colony was white and then became light yellow. When conidia from this pure culture were inoculated into the overwintering adults, the fungus formed conidiocarps with the same morphology on the insect body and the lethal rate by the fungus was $88{\pm}16%$. This fungus has over 99% homology with Cordyceps bassiana (imperfect fungal name is Beauveria bassiana) in ITS-5.8s rDNA base sequence. The fungal ecology and the infection process of the fungus into its host need to be clarified before using this fungus as a biological control agent.

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.

Molecular Diversity of Fungal Endophytes Isolated from Garcinia mangostana and Garcinia parvifolia

  • Sim, Jiun-Horng;Khoo, Chai-Hoon;Lee, Learn-Han;Cheah, Yoke-Kqueen
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.651-658
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    • 2010
  • Garcinia is commonly found in Malaysia, but limited information is available regarding endophytic fungi associated with this plant. In this study, 24 endophytic fungi were successfully recovered from different parts of two Garcinia species. Characterization of endophytic fungi was performed based on the conserved internal transcribed spacer (ITS) region sequence analysis and the antimicrobial properties. Results revealed that fruits of the plant appeared to be the highest inhabitation site (38%) as compared with others. Glomerella sp., Guignardia sp., and Phomopsis sp. appeared to be the predominant endophytic fungi group in Garcinia mangostana and Garcinia parvifolia. Phylogenetic relationships of the isolated endophytic fungi were estimated from the sequences of the ITS region. On the other hand, antibacterial screening showed 11 of the isolates possessed positive response towards pathogenic and nonpathogenic bacteria. However, there was no direct association between certain antibacterial properties with the specific genus observed.

A New Record of Nannizziopsis mirabilis from Field Soil in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Sang Woo;Lee, Hyun Goo;Ju, Han Jun;Gwon, Byeong Heon;Kosol, San;Bazie, Setu;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.249-254
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    • 2018
  • Nannizziopsis mirabilis (KNU17-67) was isolated from a field soil sample collected from Jejudo in 2017. This fungal isolate was identified and described by its morphological characters and the internal transcribed spacer rDNA gene sequencing. Potato dextrose agar medium was used to study its macro and micromorphology. In addition, a molecular phylogenetic tree was constructed for its precise confirmation as N. mirabilis. This fungal isolate has not officially been reported previously in Korea.

Unidentified Mycosis of Kelp Saccharina japonica Gametophytes (다시마(Saccharina japonica) 배우체의 미동정 진균증)

  • Jeong, Ha-Na;Oh, Myung-Joo;Choi, Sung-Je;Seo, Jung-Soo;Park, Myoung-Ae;Kim, Wi-Sik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.2
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    • pp.219-221
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    • 2017
  • In 2015, white cottony tufts were observed on gametophytes of the kelp Saccharina japonica. Wet mount and histopathology examination revealed numerous fungal hyphae and mycelium around the gametophytes. The gametophytes surrounded by fungal hyphae were generally round and empty. A specific 610-bp fragment of the internal transcribed spacer (ITS)-5.8S rDNA-ITS gene of fungi was amplified by polymerase chain reaction and the nucleotide sequence showed 100% identity with those of Acremonium sclerotigenum, Acremonium sp. and Ascomycota sp. When fungus-infected gametophytes were mixed with healthy gametophytes, a high transmission rate (100%) resulted. This is the first report of mycosis of gametophytes in Korea.