• Title/Summary/Keyword: fungal mycelia

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Fungal Growth and Manganese Peroxidase Production in a Deep Tray Solid-State Bioreactor, and In Vitro Decolorization of Poly R-478 by MnP

  • Zhao, Xinshan;Huang, Xianjun;Yao, Juntao;Zhou, Yue;Jia, Rong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.803-813
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    • 2015
  • The growth of Irpex lacteus F17 and manganese peroxidase (MnP) production in a selfdesigned tray bioreactor, operating in solid-state conditions at a laboratory scale, were studied. The bioreactor was divided into three layers by three perforated trays. Agroindustrial residues were used both as the carrier of bound mycelia and as a nutrient medium for the growth of I. lacteus F17. The maximum biomass production in the bioreactor was detected at 60 h of fermentation, which was consistent with the CO2 releasing rate by the fungus. During the stationary phase of fungal growth, the maximum MnP activity was observed, reaching 950 U/l at 84 h. Scanning electron microscopy images clearly showed the growth situation of mycelia on the support matrix. Furthermore, the MnP produced by I. lacteus F17 in the bioreactor was isolated and purified, and the internal peptide sequences were also identified with mass spectrometry. The optimal activity of the enzyme was detected at pH 7 and 25℃, with a long half-life time of 9 days. In addition, the MnP exhibited significant stability within a broad pH range of 4-7 and at temperature up to 55℃. Besides this, the MnP showed the ability to decolorize the polymeric model dye Poly R-478 in vitro.

Histological Observation on the Symbiotic Relationships Between Gastrodia elata and Rhizomorph of Armillaria mellea (뽕나무버섯균 균사속과 천마의 공생관계에 대한 조직학적 관찰)

  • Kim, Yong-Kyoo;Km, Myung-Kon;Yoon, Sook;Hong, Jai-Sik
    • The Korean Journal of Mycology
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    • v.28 no.1
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    • pp.41-45
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    • 2000
  • The histological studies were conducted to investigate the symbiotic relationships between Gastrodia elata and Armillaria mellea by using light and electron microscopes. The fungus, A. mellea, penetrated into the cortex of G. elata, in which endomycorrhizal mycelia in the cortical cells appeared to be dissolved and digested, and seemed to be consequently used as nutritional sources for G. elata growth. Staining of infected tissues revealed that protein- and fat-like substances were localized in the cells. The nuclei of cells infected by the fungal mycelia were hypertrophied 1.5 to 2 times as those without the fungal infection.

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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.

Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione

  • Kim, Eunji;Lee, Hye Min;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.95-100
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    • 2017
  • Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.

Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Fusarium Fruit Rot of Posthavest Oriental Melon (Cucumis melo L. var. makuwa Mak.) Caused by Fusarium spp. (Fusarium spp.에 의한 수확 후 참외 열매썩음병)

  • Kim, Jin-Won;Kim, Hyun-Jin
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.260-267
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    • 2004
  • Fusarium spp. were isolated from the postharvest fruit rot of oriental melon fruits at commercial fruit markets in Korea during 2001 to 2003. The decayed fruits were covered with the fungal mycelia and eventually soft rotted. The disease started at the fruit stalk area, the calyx end of the fruit and skin of fruit. As the disease advanced, white to pinkish mycelia covered with the surface of decayed fruit. The cultural and morphological characteristic of Fusarium spp. were compared with descriptions of those reported previously, and identified as Fusarium equiseti, F. graminearum, F. moniliforme, F. proliferatum, F. sambucinum, and F. semitectum. Pathogenicity of the isolates was proved by artificial wound and unwound inoculation onto the healthy fruits. Two days after inoculation, aerial mycelia were noticed on the wound inocultion region of the fruit and developed soft rot symptoms. Although Fusarium spp. causing fruit rot disease in oriental melon have been reported in Korea, identification of the those species was not described. Therefore, this is the first report of Fusarium spp. causing postharvest fruit rot on oriental melon in Korea.

Isolation and Characterization of Airborne Mushroom Damaging Trichoderma spp. from Indoor Air of Cultivation Houses Used for Oak Wood Mushroom Production Using Sawdust Media

  • Kim, Jun Young;Kwon, Hyuk Woo;Lee, Dong Hyeung;Ko, Han Kyu;Kim, Seong Hwan
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.674-683
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    • 2019
  • Some species of the Trichoderma genus are reported as the major problem in oak wood mushroom production in Korea. In spite of economic loss by the fungi, scientific information on airborne Trichoderma species is not much available. To generate information for disease management development we analyzed airborne Trichoderma. A total of 1,063 fungal isolates were purely obtained from indoor air sampling of cultivation houses used for oak wood mushroom using sawdust media. Among the obtained isolates, 248 isolates were identified as Trichoderma fungi including T. harzianum, T. atroviride, T. citrinoviride, and T. pseudokoningii, by morphological and molecular analysis. T. harzianum was dominant among the four identified species. All the four Trichoderma species grew fast on solid nutrient media tested (potato dextrose agar [PDA], malt extract agar [MEA], Czapek's Dox + yeast extract agar [CYA] and cornmeal dextrose agar). Compact mycelia growth and mass spore production were better on PDA and CYA. In addition, T. harzianum and T. citrinoviride formed greenish and yellowish mycelium and spores on PDA and CYA. Greenish and yellowish pigment was saturated into PDA only by T. pseudokoningii. These four Trichoderma species could produce extracellular enzymes of sawdust substrate degradation such as β-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease. Their mycelia inhibited the growth of oak wood mushroom mycelia of two tested cultivars on dual culture assay. Among of eleven antifungal agents tested, benomyl was the best to inhibit the growth of the four Trichoderma species. Our results demonstrate that the airborne Trichoderma fungi need to be properly managed in the cultivation houses for safe mushroom production.

Proliferation of Tricholoma matsutake Mycelial Mats in Pine Forest Using Mass Liquid Inoculum

  • Lee, Won-Ho;Han, Sang-Kuk;Kim, Beom-Seok;Shrestha, Bhushan;Lee, Soo-Yong;Ko, Cheol-Soon;Sung, Gi-Ho;Sung, Jae-Mo
    • Mycobiology
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    • v.35 no.2
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    • pp.54-61
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    • 2007
  • Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

Antifungal Properties of Rhizopus oligosporus Against Apple Anthracnose Fungi

  • Bajpai, Vivek K.;Choi, Seak-Won;Kang, Sun-Chul
    • Korean Journal of Environmental Agriculture
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    • v.29 no.1
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    • pp.86-91
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    • 2010
  • This study was carried out to assess the antifungal potential of R. oligosporus and its ethyl acetate (EtOAc) extract against the fungal pathogens causing anthracnose disease in apple fruits using disc diffusion, antagonistic effect and morphological abnormalities in fungal mycelia. The percentage of inhibition of antifungal effect of the ethyl acetate extract (5 ${\mu}l$ $disc^{-1}$) of the R. oligosporus against C. acutatum KACC 40848, C. gloeosporioides KACC 40897, C. higginsianum KACC 40806, C. orbiculare KACC 40808, C. coccodes KACC 40008, C. musae KACC 40947, C. boninense KACC 40893, C. liliacearum KACC 40981, C. caudatum KACC 41028 and Colletotrichum sp. KACC 40811 was found to be 44.4, 35.5, 40, 31.1, 33.3, 37.7, 40, 51.1, 28.8 and 28.8%, respectively. Also the fungus R. oligosporus showed potential antagonistic effect of antifungal activity against the tested pathogens of Colletotrichum spp. Further, R. oligosporus had a potential detrimental effect on the morphology of the tested fungi of Colletotrichum spp. such as wrinkle abnormalities, abnormal cell formation, lysis of mycelium, empty cell formation, distorted cell formation and breakage of the mycelium. These findings strongly support the role of R. oligosporus to serve as a potential antifungal agent to control plant pathogenic fungi causing anthracnose disease in apple fruits.

Survey of Fungicide Resistance for Chemical Control of Botrytis cinerea on Paprika

  • Yoon, Cheol-Soo;Ju, Eun-Hee;Yeoung, Young-Rog;Kim, Byung-Sup
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.447-452
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    • 2008
  • Four hundred and sixty six isolates of B. cinerea were obtained from infected leaves, stems and fruits of paprika grown in greenhouses or plastic film houses in Gangwon province, Korea, between August and November in 2006 and 2007. These isolates were classified into five representative phenotypes of resistant (R) and sensitive (S) reactions as SSR, SRR, RSS, RRS and RSR according to the responses of isolates against benzimidazole, dicarboximide and N-phenyl-carbamate fungicide in order. The percentage of five phenotypes were 51.3, 2.4, 35.6, 8.1 and 2.6%, respectively. The SSR phenotype (51.3%) was the most common. Among the nineteen fungicide products evaluated to compare their efficacy against gray mold pathogen on the paprika fruit inoculated with fungal mycelia, the mixture of diethofencarb and carbendazim was the most effective followed by iprodione, boscalid, the mixture of iprodione and thiophanate-methyl, fludioxonil, polyoxin-B, fluazinam, the mixture of tebuconazole and tolyfluanid and procymidone; while in the assay methods inoculated with fungal spores, the mixture of tebuconazole and tolyfluanid was the most effective in controlling gray mold followed by boscalid, fludioxonil, the mixture of diethofencarb and carbendazim and the mixture of pyrimethanil and chlorothalonil.