• Title/Summary/Keyword: Cultural mycelia

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Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

  • Alam, Nuhu;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Tae-Soo;Lee, U-Youn
    • Mycobiology
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    • v.38 no.1
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    • pp.17-25
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    • 2010
  • The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Production of HCN, Weed Control Substance, by Pseudomonas koreensis and its Plant Growth-Promoting and Termiticidal Activities (Pseudomonas koreensis에 의한 잡초제어활성물질인 HCN 생성과 이 균주의 식물성장 촉진 및 흰개미 살충 활성)

  • Yoo, Ji-Yeon;Jang, Eun-Jin;Park, Soo-Yeun;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.27 no.9
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    • pp.771-780
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    • 2018
  • To develope a microbial weed control agent, HCN-producing bacteria were isolated, and their characteristics were investigated. A selected strain of WA15 was identified as Pseudomonas koreensis by morphological, cultural, biochemical and 16S rRNA gene analyses. The conditions for HCN production was investigated by a One-Variable-at-a-Time (OVT) method. The optimal HCN production conditions were tryptone 1%, glycine 0.06%, NaCl 1%, and an initial pH and temperature of 5.0 and $30^{\circ}C$, respectively. The major component for HCN production was glycine. Under optimal conditions, HCN production was about 3 times higher than that of the basal medium. The WA15 strain had physiological activities, such as indoleacetic acid that was associated with the elongation of plant roots and siderophore and ammonification inhibiting fungal growth, and produced hydrolytic enzymes, such as cellulase, pectinase and lipase. The strain was able to inhibit the growth of phytopathogenic fungi, such as Rhizoctonia solani, Botrytis cinerea and Fusarium oxysporum, by the synergistic action of volatile HCN and diffusible antimicrobial compounds. A microscopic observation of R. solani that was teated with the WA15 strain showed morphological abnormalities of fungal mycelia, which could explain the role of the antimicrobial metabolites that were produced by the WA15 strain. The volatile HCN produced by the WA15 strain was also found to have insecticidal activity against termites. Our results indicate that Pseudomonas koreensis WA15 can be applied as a microbial agent for weed control and also as a termite repellent. Furthermore, it could be applied as a microbial termiticidal agent to replace synthetic insecticides.

Studies on Constituents of the Higher Fungi of Korea (LV) -The Antitumor Components and Culture of Lyophyllum decastes- (한국산(韓國産) 고등(高等) 균류(菌類)의 성분(成分) 및 배양(培養)에 관한 연구(硏究)(제55보)(第55報) -Lyophyllum decastes의 항암(抗癌) 성분(成分) 및 배양(培養)-)

  • Lee, Chong-Ock;Kim, Hyung-Soo;Choi, Eung-Chil;Kim, Byong-Kak
    • Korean Journal of Pharmacognosy
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    • v.17 no.1
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    • pp.23-34
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    • 1986
  • To find antitumor components of Lyophyllum decastes, the mycelia of L. decastes were cultured in artificial media and a new antitumor component which showed potent antitumor activity against sarcoma 180 implanted in mice, was isolated and named 'Lyophyllan A'. Lyophyllan A was composed of a polysaccharide moiety (86%) and a protein moiety (2%). The polysaccharide moiety was found to be a heteroglycan which consisted of glucose (48.1%), mannose (30.8%), galactose, xylose and fucose. The protein moiety contained 18 amino acids. Cultural characteristics of L. decastes were investigated by shake culture method. The best result was obtained when L. decastes was cultured in medium glucose 50g, peptone 10g, corn steep liquor 30ml, $KH_2PO_4$ 0.87g, $MgSO_4$ $7H_2O$ 0.5g, $CaCl_2$ 0.3g, $FeSO_4{\cdot}7H_2O$ 10mg, $MnCl_2{\cdot}4H_2O$ 7mg, $ZnSO_4{\cdot}7H_2O$ 4mg and $CuSO_4{\cdot}5H_2O$ 1mg per one liter at $26^{circ}C$, 180rpm, for 9 days.

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection and Diagnosis of Epiphytic Didymella bryoniae in Cucurbits

  • Lee, Sun-Cheol;Han, Ki-Soo;Lee, Jung-Han;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.260-265
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    • 2003
  • Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

Cultural characteristics and antioxidant activity of wild-type collections of Hericium erinaceus (노루궁뎅이버섯 야생 수집균주의 배양적 특성과 항산화 활성)

  • Park, Soon-Ae;Kim, Sun-Joo;Kim, Ha-Lim;Kang, Hee-Wan
    • Journal of Mushroom
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    • v.16 no.1
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    • pp.9-15
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    • 2018
  • This study aimed to characterize the mycelial morphologies of Hericium erinaceus isolates, including 18 wild-type collections and cultivatable species 'Noru 1' and 'Noru 2', in Korea. The morphological characteristics were used to classify the species based on aerial or branched mycelia and their brownish or whitish colors when grown on potato dextrose agar. Of the wild-type collections, the isolates KFRI 509, KFRI 1091, KFRI 1093, and KFRI 1623 showed faster mycelial growth than the cultivars 'Noru 1' and 'Noru 2'. Moreover, 60% ethanol extracts of the fruiting body of the mushroom showed the highest phenol content and DPPH and ABTS radical scavenging activity. The DPPH and ABTS radical scavenging activities of KFRI 507, KFRI 508, KFRI 842, and KFRI 1623 were 10-60% higher than those of 'Noru 1' and 'Noru 2', depending on the extract concentrations. Thus, results suggest that these wild-type collections could be useful for breeding genetic sources or processed food materials with high antioxidant activity.

A New Record of Epicoccum draconis Isolated from the Soil in Korea

  • Ayim, Benjamin Yaw;Das, Kallol;Cho, Young-Je;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.48 no.1
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    • pp.39-45
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    • 2020
  • A fungal isolate US-18-11 was isolated from the soil in Uiseong, Korea. The mycelium growth measured after 7 days of incubation at 22℃ on malt extract agar (MEA) and oatmeal agar (OA) media was 42-43 mm and 41-44 mm in diameter, respectively. The fungal colony formed white to dull green aerial mycelia that were floccose with regular margins and olivaceous black with leaden gray patches on the reverse side. The conidia were hyaline to brown in color, ellipsoidal to ovoid, guttulate, abundant, globose, solitary, or confluent measuring 3.2-7.2×1.1-2.3 ㎛. A BLAST search of the large subunit (LSU), internal transcribed spacer (ITS) region, second largest subunit of DNA-directed RNA polymerase II (RPB2) and β-tubulin (TUB2) gene sequences revealed that the isolate US-18-11 has similarities of 99, 100, 97, and 99% with those of Epicoccum draconis CBS 186.83, respectively. A neighbor-joining phylogenetic tree constructed based on the concatenated dataset of above-mentioned sequences showed that isolate US-18-11 clustered with Epicoccum draconis CBS 186.83 in the same clade. Based on the results of morphological, cultural, and phylogenetic analysis, the isolate US-18-11 was identical to the previously described E. draconis CBS 186.83. To our knowledge, this is the first report of E. draconis in Korea.

Weeding Efficacy of Melanized Formula with Epicoccosorus nematosporus on Eleocharis kuroguwai in the Field

  • Hong, Yeon-Kyu;Cho, Jae-Min;Uhm, Jae-Youl;Hyun, Jong-Nae;Lee, Bong-Choon;Song, Seok-Bo;Lee, Dong-Chang
    • The Plant Pathology Journal
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    • v.19 no.2
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    • pp.92-96
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    • 2003
  • The study was conducted to determine the cultural conditions and the effect of inert fillers for melanization and sporulation abilities of sodium alginate pellets, and the weeding efficacy of the formula in the field. Melanin production of E. nematosporus was affected by striking frequency. Percentage of melanized beads was increased to 80.6% at higher rpm up to 180. The melanized pellets produced more conidia with abundant mucilage than unmelanized pellets. Shaker culture of Epicoccosorus nematosporus with sodium alginate yielded a total of 55 mg per 100 pellets. Percentage of melanized pellets was highest with 81.0% and 83.3% of melanization, when wheat bran and rice polish were amended and produced the conidia with 65.4 and 68.4 mg per 100 pellets, respectively. When 1 L of conidial suspension of 6.0$\times$$10^5$ conidia per ml was applied on 30-day-old plants in a plot, 74.5% of the plants were killed within 20 days, whereas, its melanized sodium alginate pellets killed 57.8% of the plants in the same period. The number of tuber formation of Eleocharis kuroguwai in the untreated control plots was 128.5 per plot, but those of the plots treated with conidial suspension and melanized pellets were 22.1 and 39.7, respectively, at the end of the season. Results of this study showed that melanization of mycelia-mixed sodium alginate are an important sporulation factor in E. namatosporus as a mycoherbicide.

Morphological and Physiological Characteristics of Amanita hemibapha subsp. hemibapha (Berk. & Broome) Sacc. (달걀버섯의 형태 및 생리적 특성)

  • Sou, Hong-Duck;Hur, Tae-Chul;Joo, Sung-Hyun
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.41-48
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    • 2009
  • This study was conducted to investigate optimum cultural conditions for Amanita hemibapha isolated in Korea and its hyphae characteristics. Micrographs shows the presence of clamp connection. A. hemibapha grows as mycelial form(M-phase) 2-4 ${\mu}m$ and yeast-like form(Y-phase) 7-8 ${\mu}m$. The fungal spores were broadly elliptical and papillate, 8-11 ${\times}$ 6-9 ${\mu}m$ in size. The nucleotide sequence analysis of the ITS of nuclear ribosomal DNA from sporocarps and in-vitro-grown mycelium supported the fungal species is Amanita hemibapha. A. hemibapha showed sequence similarity in the ITS rDNA with A. caesarea(97.5) and A. jacksonii(98.5%) which are morphologically similar species to A. hemibapha. The optimal pH and temperature for mycelial growth of A. hemibapha were pH 6.0 and $28^{\circ}C$, respectively. The fungal species showed best growth in SYP and GYS medium. A. hemibapha grew well with mannitol and glucose as carbon sources and peptone as a nitrogen source.

Phylogenetic Placement and Morphological Characterization of Sclerotium rolfsii (Teleomorph: Athelia rolfsii) Associated with Blight Disease of Ipomoea batatas in Korea

  • Paul, Narayan Chandra;Hwang, Eom-Ji;Nam, Sang-Sik;Lee, Hyeong-Un;Lee, Joon-Seol;Yu, Gyeong-Dan;Kang, Yong-Gu;Lee, Kyeong-Bo;Go, San;Yang, Jung-Wook
    • Mycobiology
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    • v.45 no.3
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    • pp.129-138
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    • 2017
  • In this study, we aimed to characterize fungal samples from necrotic lesions on collar regions observed in different sweetpotato growing regions during 2015 and 2016 in Korea. Sclerotia appeared on the root zone soil surface, and white dense mycelia were observed. At the later stages of infection, mother roots quickly rotted, and large areas of the plants were destroyed. The disease occurrence was monitored at 45 and 84 farms, and 11.8% and 6.8% of the land areas were found to be infected in 2015 and 2016, respectively. Fungi were isolated from disease samples, and 36 strains were preserved. Based on the cultural and morphological characteristics of colonies, the isolates resembled the reference strain of Sclerotium rolfsii. Representative strains were identified as S. rolfsii (teleomorph: Athelia rolfsii) based on phylogenetic analysis of the internal transcribed spacer and large subunit genes along with morphological observations. To test the pathogenicity, sweetpotato storage roots were inoculated with different S. rolfsii strains. 'Yulmi' variety displayed the highest disease incidence, whereas 'Pungwonmi' resulted in the least. These findings suggested that morphological characteristics and molecular phylogenetic analysis were useful for identification of S. rolfsii.

Leaf Blight of Kudzu (Pueraria lobata) Caused by Rhizoctonia solani (Rhizoctonia solani 에 의한 칡 잎마름병)

  • Kim, Hyung-Moo;Song, Wan-Yeob;Seo, Byung-Soo;Ko, Jeong-Ae
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.77-79
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    • 2005
  • Leaf blight of kudzu was found in Jeonbuk province in 2002. Water-soaked lesion appeared on leaves, and young stems and gray green blight sypmtom expanded from margin to main vine of leaves. As the disease became severe, blighted leaves and shoots turned dark brown and then collapsed. The causal pathgen showed initially white aerial mycelium and turned brown to gray. Mycelia were branched at 90o angles and multinucleate in one cell. It formed sclerotia on PDA. Slerotia were irregular, globose and 0.5~3.0 mm in diameter. The causal fungus of leaf blight was identified as Rhizoctonia solani Kuhn based on the cultural and morphological characteristics. This is the first report on leaf blight of kudzu caused by R. solani in Korea.