• Title/Summary/Keyword: cultured mycelia

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Characteristics of Mycelial Growth and Enzyme Activities of Mattirolomyces terfezioides Collected from Robinia pseudoacacia Forest in Korea (국내 아까시나무 숲에서 수집한 감자덩이버섯(Mattirolomyces terfezioides)의 균사 생장 특성과 효소 활성)

  • Jeon, Sung-Min;Wang, Eun-Jin;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.165-173
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    • 2015
  • Mattirolomyces terfezioides is a type of sweet truffles that predominantly form ectomycorrhizae with Robinia pseudoacacia. It is also worthy of artificial cultivation. This is the first report on characteristics of mycelial growth and enzyme activities of M. terfezioides collected from R. pseudoacacia forest in Korea. M. terfezioides showed the highest mycelial growth when cultured on potato dextrose agar (PDA) at $30^{\circ}C$ or in modified Melin-Norkran's liquid medium (pH 8.0). The biomass of M. terfezioides was higher in liquid medium containing nitrate-nitrogen than ammonium-nitrogen by 1.8 fold. The mycelia of M. terfezioides showed both carboxymethylcellulase and laccase activities on solid media for enzyme screening.

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.

Availability of Sikhae Factory Wastewater as a Submerged Culture Medium for Lentinula edodes

  • Jung, Heon-Ho;Lee, Jae-Yun;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Lee, Sang-Joon;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.160-163
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    • 2001
  • Sikhae is a Korean traditional beverage of saccharified rice. Its factory waste(SFW) is usually thrown away instead of being used. We developed a cheap substrate of SFW for use in liquid spawn that is known for its higher fruit body yields than grain spawn in sawdust cultivation. Mycelia of Lentinula edodes ASI 3046, which is regarded as the most suitable strain for sawdust cultivation, were cultured on six kinds of previous known media and SFW. As the seven kinds of media were applied, a Sikhae Factory Waste(SFW) was most excellent in growth. The dried mycelial weight in SFW was almost four times as much as that in the other media. In the flask culture, optimum culture conditions for the mycelial growth were obtained after 13 days of cultivation at media volume of 100 ml, 100 rpm, initial pH 4.5, and $25^{\circ}C$. The best mycelial growth was observed when $MgSO_4{\cdot}7H_2O$ and D-sucrose were added as a supplement in SFW. SWM must be a remarkable medium for L. edodes because of its simple preparation and low cost.

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Effect of Growth and Culture Conditions of Paecilomyces japonica and Cordyceps militaris on the Formation of Bioactive Substance

  • Jeng, Yong-Young
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.138-139
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    • 2003
  • To investigated the effect of liquid culture conditions and nutrient sources on the formation with bioactive substance of Paeilomyces japonica and Cordyceps militaris cultivated in the country, the result are as follow; The growth temperature of two mycelia is 25$^{\circ}C$ and the proper temperature for cordycepin growth is around 20$^{\circ}C$. The formation amount of bioactive substance by nutrient sources reached its peak with using 2% glucose and 1% galactose in case of carbon sources and 0.4% inorganic compound in case of nitrogen sources. Also, the ratio of C/N was optimal with 3% glucose 1% peptone. For a natural medium, most grains were sufficient but the soybean oil was superb. The formation amount of protein-binding polysaccharide that are used for anticancer substance was in proportion to the growth rate of mycelium, had lots of aeration and showed a trend of increasing when the acidity lower. and the content of structural protein showed a trend of increasing when the acidity lower. However, the content of the structural hexosamin did not get a great the effect of culture conditions and nutrient sources. The constitution of monosaccharide that organizes a protein-binding polysaccharide greatly changed in proportion to carbon sources. When Paecilomyces japonica cultured in a silkworm larvae for 30 days, the content of cordycepin was 204.5 mg/100mL as a dry weight in the fruiting body, 41.8 mg/100mL in mycelium and larva, and the content for each bottle was average 29.5 mg/100mL. In case of Cordyceps militaris for 45days, the fruiting body was 563.5 mg/100mL, the larva and the mycelium was 86.1 mg/100mL, and the content for each bottle was average 65.0 mg/100mL.

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Effect of Kp, an Antitumor Protein-Polysaccharide from Mycelial Culture of Phellinus Linteus on the Humoral lammune Response of Tumor-Bearing ICR Mice to Sheep Red Blood Cells

  • Chung, Kyeong-Soo;Kim, Shin-Sook;Kim, Hee-Soo;Kim, Kye-Yang;Han, Man-Woo;Kim, Kil-Hyun
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.336-338
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    • 1993
  • The immunomodulating activity of Kp, an antitumor protein-polysacchanide preparation from the shake-cultured mycelia of Phellinus linteus, was investigated in ICR mice subcutaneously implanted wit $1\times10^6$ cells of sarcoma 180. The mice were intraperitoneally administered with Kp at a does of 100 mg/kg once daily for five consecutive days starting from 24 hrs after the tumor implantation. Ten days after the last injection, the mice were immunized with $1\times10^7$ or $4\times10^8$ sheep red blood cells (SRBC) and five days later, the antibody-forming immune response were assessed by direct hemolytic plaque assay. To an immunization does of $1\times10^7$ SRBC, the Kp-treated mice elicied a successful humoral immune response despite the turmor-burden and produced $259\times10^3$ plaque-forming cells (PFC)/spleen, while the corresponding tumor-bearing control mice showed virtually no reponse $(2.0\times10^3$ PFC/spleen) (the stimulation index=129.5). However, to an immunization dose of $4\times10^8$ SRBC, both of the control mice and Kp-treated mice showed almost the same level of strong humoral immune response. From these data it is clear that Kp effectively restores the humoral immune response of the turmor-bearing ICR mice.

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Factors Affecting Spore Formation of Carrot Leaf Blight Caused by Alternaria dauci In Vitro (배지에서 당근검은잎마름병균의 포자 형성에 영향을 미치는 요인)

  • Sena Yoon;Jiyoung Min;Heung Tae Kim
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.251-257
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    • 2023
  • In order to examine the pathogenicity of Alternaria dauci, the causal agent of carrot leaf blight, it is necessary to standardize sporulation conditions to ensure the optimal quantity and quality of spore inoculum. Therefore, in this study, the effects of the growth medium, aeration treatment, and UV treatment with 12-hr photoperiod on the sporulation of A. dauci KACC42997 were investigated. A. dauci KACC42997 was pre-cultured for 7 days in a potato dextrose agar medium at 25℃ in the dark condition. When the pre-culture, after removing aerial mycelia, was re-incubated for 5 days, with simultaneous aeration treatment and 12-hr cycle UV treatment at 20℃, the largest number of spores was produced. One hundred seventy isolates of A. dauci were isolated from major carrot growing regions such as Pyeongchang, Gumi and Jeju and tested for sporulation under the conditions established in this study. Except for 20 strains, all strains produced spores. Statistically significant differences in the extent of sporulation were found among local populations of A. dauci isolates, but no difference was observed in their pathogenicity on carrots.

Transcriptome Analysis of Antrodia cinnamomea Mycelia from Different Wood Substrates

  • Jiao-Jiao Chen;Zhang Zhang;Yi Wang;Xiao-Long Yuan;Juan Wang;Yu-Ming Yang;Yuan Zheng
    • Mycobiology
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    • v.51 no.1
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    • pp.49-59
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    • 2023
  • Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

Changes in Polysaccharides Content and Cell Morphology of Fomitopsis pinicola Mycelium during Submerged Culture (소나무잔나비버섯(Fomitopsis pinicola) 균사체 배양에 따른 함량 및 세포의 형태학적 변화)

  • Jung Yoo-Kyung;Shin Kyung-Ok;Park Hong-Duok;Kim Soon-Dong
    • Food Science and Preservation
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    • v.13 no.3
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    • pp.397-403
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    • 2006
  • This experiment was conducted to examine the changes in polysaccharide concentration and morphological variation of Fomitopsis pinicola mycelium during submerged-culture in the citrus peel medium (CP). On the 12 days culture, the yields of mycelium and alcohol insoluble substance were 40.21%(w/v) and 6.94%(w/w), respectively, which were much higher than 11.29%(w/v, wet basis) and 3.17%(w/w, wet basis) obtained from YM medium. A large amount of acid soluble polysaccharides was derived from YM medium while a larger amount of alkali soluble polysaccharide was produced from CP medium. Yields of the mycelium were higher when cultured in CP medium However, there was no significant difference in formation of membranous vesicle between mycelia cultured in CP medium and YM medium. It was also observed that the formation of vacuole was closely related to the activation of the multivescular body known as cytolysome. As a result activation of mycelium and cell wall biosynthesis were more accelerated in CP medium.

Protoplast Formation, Regeneration and Reversion in Pleurotus ostreatus and P. sajor-caju (느타리버섯과 여름느타리버섯의 원형질체(原形質體) 나출(裸出)과 재생(再生))

  • Go, Seung-Joo;Shin, Gwan-Chull;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.169-177
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    • 1985
  • The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.

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Mycelial Culture of Lentinus edodes Alleviates Rat Liver Toxicity Induced by Carbon Tetrachloride and Ethanol (표고버섯균사체의 사염화탄소 및 알콜로 처리된 흰쥐 간기능 보호 효과)

  • Ha, Yeong-L.;Kim, Young-S.;Ahn, Chae-R.;Kweon, Jung-M.;Park, Cherl-W.;Ha, Young-K.;Kim, Jeong-O.
    • Journal of Life Science
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    • v.20 no.1
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    • pp.133-141
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    • 2010
  • The protective effect of a mixed powder from solid-cultured and liquid-cultured Lentinus edodes mycelia (2:1, w/w) (designate LED) on the carbon tetrachloride ($CCl_4$)- and ethanol-induced hepatotoxicity of male Sprague-Dawley (SD) rat was investigated. In the $CCl_4$-induced rat hepatotoxicity experiment, rats of 4 groups (6 rats/group) were administere with Normal (0.2 ml distilled water), Control (0.2 ml distilled water), LED (LED 200 mg/kg BW + 0.2 ml distilled water), and Silymarin (200 mg/kg BW + 0.2 ml distilled water), p.o., daily for 2 weeks. Afterwards, all groups except for the Normal group were subjected to abdominal injection with $CCl_4$ ($CCl_4$ : corn oil, 1:1 v/v; 0.5 ml/kg BW). For the ethanol- induced rat hepatotoxicity experiment, rats were divided into 5 groups (5 rats/group): Normal; Pair-fed control (PFC); Control (ethanol); LED (ethanol + LED 200 mg/kg BW); and Silymarin (ethanol + silymarin 200 mg/kg BW). Rats of the Normal and PFC groups were fed a basal liquid diet, and rats of the Control, LED, and Silymarin groups were fed a liquid ethanol diet containing LED or Silymarin. Eight weeks later, blood and liver samples were collected to analyze biomarkers. In $CCl_4$-induced SD rats, LED elevated hepatic superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH peroxidase) activities and thiobarbituric reactive substances (TBARS) were reduced, resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic dehydrogenase (LDH) activities in plasma. Similar results of these enzymes and biochemical markers in both liver tissues and plasma were seen in ethanol-induced hepatotoxicity of SD rats. In addition, elevated alcohol dehydrogenase (ADH) activity and reduced expression of cytochrome p450 mixed monooxygenase enzyme (CYP2E1) were seen in liver tissues from ethanol-treated rats by LED treatment. These effects of LED were similar to those of Silymarin. In in vitro experiments, LED showed antioxidant activity in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) system and mouse liver mitochondria system induced by NADPH/$Fe^{2+}$ and cumine hydroperoxide (CuOOH). These results indicate that LED protected SD rat hepatotoxicity, induced by $CCl_4$ and ethanol, through its antioxidative activity and might be useful as a material for protection from hepatoxicity in humans.