• 제목/요약/키워드: Fungal isolation

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Isolation and Characterization of an Antifungal and Plant Growth-Promoting Microbe

  • Park, Se Won;Yang, Hee-Jong;Seo, Ji Won;Kim, Jinwon;Jeong, Su-ji;Ha, Gwangsu;Ryu, Myeong Seon;Yang, Hee Gun;Jeong, Do-Youn;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • 제49권4호
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    • pp.441-454
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    • 2021
  • Fungal diseases including anthracnose, stem rot, blight, wilting, and root rot of crops are caused by phytopathogens such as Colletotrichum species, Sclerotinia sclerotiorum, Phytophthora species, and Fusarium oxysporum and F. solani which threaten the production of chili pepper. In this study, to identify biological control agents (BCAs) of phytopathogenic fungi, potentially useful Bacillus species were isolated from the field soils. We screened out five Bacillus strains with antagonistic capacity that are efficiently inhibiting the growth of phytopathogenic fungi. Bacillus species were characterized by the production of extracellular enzymes, siderophores, and indole-3-acetic acid (IAA). Furthermore, the influence of bacterial strains on the plant growth promoting activity and seedling vigor index were assessed using Brassica juncea as a model plant. Inoculation with Bacillus subtilis SRCM 121379 significantly increased the length of B. juncea shoots and roots by 45.6% and 52.0%, respectively. Among the bacterial isolates, Bacillus subtilis SRCM 121379 showed the superior enzyme activities, antagonistic capacity and plant growth promoting effects. Based on the experimental results, Bacillus subtilis SRCM 121379 (GenBank accession no. NR027552) was finally selected as a BCA candidate.

Colletotrichum aenigma Associated with Apple Bitter Rot on Newly Bred cv. RubyS Apple

  • Lee, Seung-Yeol;Ten, Leonid N.;Ryu, Jung-Joo;Kang, In-Kyu;Jung, Hee-Young
    • Research in Plant Disease
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    • 제27권2호
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    • pp.70-75
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    • 2021
  • The abnormal brown sunken lesions were observed on cv. RubyS apple fruits in an orchard located in Gunwi, Gyeongbuk province, Korea. The primary observed symptoms such as small round sunken lesions and small black dots on the symptomatic area were different from the reported apple diseases. The affected apple fruits were sampled and subjected to isolation of the causal agent. Cultural and morphological characteristics of isolated fungal strain, designated KNUF-20GWA4, were similar to that of Colletotrichum spp. Based on multilocus sequence analyses using internal transcribed spacer regions and partial sequences of β-tubulin, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase, and actin genes, strain KNUF-20GWA4 showed 99.2-100% similarities with C. aenigma ICMP 18608 and the isolate clustered together with several other strains of this species in the phylogenetic tree. To our knowledge, this is the first report of bitter rot on apple fruits caused by C. aenigma.

Genetic localization of epicoccamide biosynthetic gene cluster in Epicoccum nigrum KACC 40642

  • Choi, Eun Ha;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • 제65권3호
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    • pp.159-166
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    • 2022
  • Epicoccum nigrum produces epipyrone A (orevactaene), a yellow polyketide pigment. Its biosynthetic gene cluster was previously characterized in E. nigrum KACC 40642. The YES liquid culture of this strain revealed high-level production of epicoccamide (EPC), with an identity that was determined using liquid chromatography-mass spectrometry analysis and molecular mass search using the SuperNatural database V2 webserver. The production of EPC was further confirmed by compound isolation and nuclear magnetic resonance spectroscopy. EPC is a highly reduced polyketide with tetramic acid and mannosyl moieties. The EPC structure guided us to localize the hypothetical EPC biosynthetic gene cluster (BGC) in E. nigrum ICMP 19927 genome sequence. The BGC contains genes encoding highly reducing (HR)-fungal polyketide synthase (fPKS)-nonribosomal peptide synthetase (NRPS), glycosyltransferase (GT), enoylreductase, cytochrome P450, and N-methyltrasnferase. Targeted inactivation of the HR-fPKS-NRPS and GT genes abolished EPC production, supporting the successful localization of EPC BGC. This study provides a platform to explore the hidden biological activities of EPC, a bolaamphiphilic compound.

Light and Electron Microscopy Studies Elucidating Mechanisms of Tomato Leaf Infection by Pseudocercospora fuligena

  • Zelalem Mersha;Girma Birru;Bernhard Hau
    • The Plant Pathology Journal
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    • 제39권2호
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    • pp.181-190
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    • 2023
  • The fungal pathogen Pseudocercospora fuligena, known to affect tomatoes in the tropics and subtropics, has been reported from temperate climates including the United States and Turkey in recent years. In this study, an isolate from fresh tomatoes and the disease it causes were characterized and infection mechanisms investigated. Macroscopically, both sides of tomato leaves show indistinct effuse patches but prolific production of fuliginous lesions is conspicuous on the abaxial side first but also on the adaxial side later on as infection progressed. Microscopically, fascicles of conidiophores (11-128 ㎛ × 3.5-9 ㎛) arising from stromata and conidia with up to 12 septations were observed. Molecular characterization of the isolate revealed high homology (99.8%) to other P. fuligena isolated from tomatoes in Turkey. Out of the 10 media tested, P. fuligena grew significantly well and sporulated better on unsealed tomato oatmeal agar and carrot leaf decoction agar, both supplemented with CaCO3. Direct transfer of conidia from profusely sporulating lesions was the easiest and quickest method of isolation for in-vitro studies. Light and scanning electron microscopy on cleared and intact tomato leaves further confirmed stomatal penetration and egress as well as prevalence of primary and secondary infection hyphae. In situ, blocked stomatal aperture areas of 154, 401, and 2,043 ㎛2 were recorded at 7, 12, and 17 days after inoculation, respectively. With the recent expanded horizon of the pathosystem and its consequential impact, such studies will be useful for a proper diagnosis, identification and management of the disease on tomato worldwide.

지류 및 섬유질 문화재의 미생물에 관한 연구 (紙類 및 纖維質 文化財의 微生物에 關한 硏究) ­경남지방(慶南地方)을 중심(中心)으로-

  • Min, Gyeong-Hui;An, Hui-Gyun
    • Korean Journal of Heritage: History & Science
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    • 제14권
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    • pp.225-250
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    • 1981
  • The investigation of air fungal population in the storages to keep papers and textiles that are designated as important folk life materials or treasures was carried out from Dec. 17 to. 23, 1980. Isolation media was used for malt extract agar with chloramphnicol to prevent bacterial contamination. Isolation and identification of air fungi from the four preserved rooms were Cladosporium cladosporioides, Alternaria chlamydospora, Aspergillus fumigatus, A. versicolor, Eurotium chevalieri, Penicillium charlesii var. rapidum, P. oxalicum. P. viridicatum, Trichoderma viride, Acremomium sp., Mucor sp. and Yeast. It was found that nine species in eight genera was isolated. Among them, underscribed species in Korea was two species ; Eurotium chevalieri and Penicillium visidicatum. The fungal population of four storages was showed to be dominant species such as Cladosporium cladosporioides and the order was Acremonium, Penicillium, Aspergillus, Trichoderma, Alternaria and Eurotium. Eurotium chevalieri was ascomycetous fungi including distinctive ascospores in cleistothecia, the filamentous fungi was directly isolated from the papers and cellulose materials showing to be fourteen species in eight genera. The most species of the fungi isolated was also Cladosporium cladosporioides and the other fungi were found as Acremonium, Penicillium, Aspergillus and Trichoderma. It was confirmed that underscribed fungi were two species ; Mucor racemosus and Penicillium spinulosvm. The effect of four antifungal agents, benzoic acid, sorbic acid, dehydroacetic acid and thymol was also examined on eight species of Aspergillus, Penicillium, Cladosporium. and Tricoderma. this results were shown that more than 0.5% concentration of thymol inhibited the grow of all fungalspecies and other three chemicals appeared various inhibition zones of fungal growth depending in their different concentrations.

Antifungal Activity of Bacterial Strains isolated from Tidal Mudflat and Salted Seafood (traditional Jeotgal) Against Six Major Plant Pathogens (갯벌 및 젓갈에서 분리한 세균의 작물 주요 병원균에 대한 항균활성 효과 검정)

  • Kim, Tack-Soo;Lee, Ga-Hyung;Kim, Gyun-Jang;Lee, Se-Won;Park, Kyung-Seok;Park, Jin-Woo
    • The Korean Journal of Pesticide Science
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    • 제14권4호
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    • pp.421-426
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    • 2010
  • Many bacterial strains inhabit strong saline condition, such as tidal mudflat and salted seafoods, were identified and reported for the proposed protease activities and salt resistance; however antifungal activities against plant fungal pathogen have not well been studied until now. In this study, primary screening was performed for the isolation of promising strains against major plant pathogens like Sclerotinia sclerotiorum, Fusarium oxysporum, Phytophthora capsici, Botrytis cineria, Collectotrichum acutatum and Pythium ultimum. Totally 423 bacterial strain were isolated from laboratory media which was based on different morphological characteristics and all the strains were dual cultured against major fungal pathogens on PDA, finally 40 strains were selected as antifungal bacterial strain and identified by fatty acid phylogenic difference analysis from MIDI shorlock gas chromatography system. As a result, antifungal strains from tidal mudflat were 10 species of 6 genus. Paenibacillus macerans was dominant species; 5 strains among the 17 isolates from tidal mudflat. Antifungal strains from salted seafoods were 7 species of 3 genus and Bacillus atrophaeus was dominant species; 12 strains among the 23 isolates from salted fishes.

Isolation and Morphological Characterization of Ttichoderma harzianum SJG-99721, a Powerful Biocontrol Agent (길항작용을 나타내는 Trichoderma harzianum SJG-99721의 분리 및 형태학적 특징)

  • 이호용;민봉희
    • Korean Journal of Environmental Biology
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    • 제20권2호
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    • pp.130-135
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    • 2002
  • Species of Genus Trichoderma are commercially applied as biological control agents against fungal Pathogens. A powerful biocontrol agent, Trichoderma sp. SJG-99721 was isolated from 305 isolates by morphological characters, chitinase activities and antifungal activities against Phytophthora capsiei. The isolate was identified as Trichoderma harzianum from various features such as growth rate at $27^\circ{C}$, significant growth ratio of $27^\circ{C}$ to $17^\circ{C}$, amount of aerial mycelium, types of branching: system, and disposition patterns of phialide and phialospore. Trichoderma harzianum SJG-99721 have been shown to act as a powerful biological agent against fungal phytopathogens; Botrytis cinerea, Rhizoctonia solani, Phytophthora cryptogea, Phytophthora capsiei, Sclerotinia sclerotiorum, Mycoshaerella melonis, Alternaria sotani, Fusarium oxysporum, Collectotrichum gloesporioodes, Alternaria alternata, Phythium ultimum, Phytophthora drechsleri, Pyricularia grisea.

Production of Fungal Lipids (Part 2) Isolation of Starch Utilizing Mold and Its Optimum Compositions of Growth Media (곰팡이 유지 생산에 관한 연구 (제 2 보) 전분 이용성 곰팡이의 분리 및 배지조성에 관하여)

  • 신동화;김창식
    • Microbiology and Biotechnology Letters
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    • 제10권1호
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    • pp.15-26
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    • 1982
  • A potential fungal lipid producer from starch, which was identified as Muror plumbeus, was isolated from natural sources and its optimum cultivation condition for lipid production was investigated. The Mucor plumbeus FRI 0007 showed the highest felt weight and lipid content which were 2.09 $\pm$ 0.24g per 50$m\ell$ of medium and 37.43% on dry weight basis respectively after 20 days incubation on the medium containing 21% of starch as a carbon source. The urea was the best nitrogen source as compared with sodium nitrate, potassium nitrate, magnesium nitrate, ammonium nitrate and ammonium acetate and its optimum concentration was 2.14g/$\ell$, showing 2.39 $\pm$ 0.07 g felt/50$m\ell$ of medium and 50.73% lipid content on dry weight basis after 25 days incubation. Besides the starch as a carbon source and urea as a nitrogen source, the Mucor plumbeus FRI 0007 utilized ZnSO$_4$, MgSO$_4$, NaH$_2$PO$_4$, $K_2$SO$_4$and FeCl$_3$as mineral sources. However, it did not require ail the above 5 minerals in group in-dispensably for its growth and lipid accumulation. The lipid and economic coefficient of Mucor plumbeus FRI 0007 grown on the medium containing 0.44g $K_2$SO$_4$or 5.00g MgSO$_4$/$\ell$solely were 14.96 and 15.37 and 31.12 and 26.10 which was higher than those on the medium containing the above 5 minerals.

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Microfungal flora of Tricholoma matsutake producing and nonproducing sites in the forest of Pinus densiflora (적송 (Pinus densiflora) 림내 송이(Tricholoma matsutake) 발생지와 미발생지의 토양 균류의 수직 분포)

  • Song, Hyun-Soon;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • 제19권2호
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    • pp.109-119
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    • 1991
  • The vertical distribution of the fungal population for the soil samples from two sites of producing and nonproducing of Tricholoma matsutake, song-yi mushroom, were examined at Yang­yang and Myung-joo, Gangweon province. By the dilution plate method, a total number of propagu­les of fungi per gram of soil was observed to be low at the song-yi producing sites but high at the song-yi nonproducing sites under the communities of Pinus densiflora. The tendency of the number of fungal propagules were decreased with the increasing vertical depth. In the incuhation method at $42^{\circ}C$, six genera and nineteen species of the fungi were isolated from two sites; Aspergillus fumigatus, Acremonium sp., Talaromyces stipitatus, Penicillium lilacinum, P. oxalicum and Westerdykella multispora. The most dominant species by this method was A. fumigatus. From heat treatment method at $70^{\circ}C$, seven genera and nineteen species were isolated; Aspergillus fumigatus, Alternaria alternata, Neurospora sitophila and Mucor sp.. In the ethanol treatment method, one genera and one species was isolated Mortierella sp.. From the three isolation methods, it was found that the total number of the soil fungi and the frequency of species appeared were the highest at the soil of upper layer whereas the lowest at the soils of lower layer in its vertical distribution.

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Interaction between the Rice Pathogens, Fusarium graminearum and Burkholderia glumae

  • Lee, Jungkwan;Jung, Boknam;Park, Jungwook;Kim, Sungyoung;Youn, Kihun;Seo, Young-Su
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2014년도 추계학술대회 및 정기총회
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    • pp.13-13
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    • 2014
  • Species belonging to the genus Fusarium are widely distributed and cause diseases in many plants. Isolation of fungal strains from air or cereals is necessary for disease forecasting, disease diagnosis, and population genetics [1]. Previously we showed that Fusarium species are resistant to toxoflavin produced by the bacterial rice pathogen Burkholderia glumae while other fungal genera are sensitive to the toxin, resulting in the development of a selective medium for Fusarium species using toxoflavin [2]. In this study, we have tried to elucidate the resistant mechanism of F. graminearum against toxoflavin and interaction between the two pathogens in nature. To test whether B. glumae affects the development of F. graminearum, the wild-type F. graminearum strains were incubated with either the bacterial strain or supernatant of the bacterial culture. Both conditions increased the conidial production five times more than when the fungus was incubated alone. While co-incubation resulted in dramatic increase of conidial production, conidia germination delayed by either the bacterial strain or supernatant. These results suggest that certain factors produced by B. glumae induce conidial production and delay conidial germination in F. graminearum. To identify genes related to toxoflavin resistance in F. graminearum, we screened the transcriptional factor mutant library previously generated in F. graminearum [3] and identified one mutant that is sensitive to toxoflavin. We analyzed transcriptomes of the wild-type strain and the mutant strain under either absence or presence of toxoflavin through RNAseq. Expression level of total genes of 13,820 was measured by reads per kilobase per million mapped reads (RPKM). Under the criteria with more than two-fold changes, 1,440 genes were upregulated and 1,267 genes were down-regulated in wild-type strain than mutant strain in response to toxoflavin treatment. A comparison of gene expression profiling between the wild type and mutant through gene ontology analysis showed that genes related to metabolic process and oxidation-reduction process were highly enriched in the mutant strain. The data analyses will focus on elucidating the resistance mechanism of F. graminearum against toxoflavin and the interaction between the two pathogens in rice. Further evolutionary history will be traced through figuring out the gene function in populations and in other filamentous fungi.

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