• Title, Summary, Keyword: Mycelial viability

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Mycelial viability and cultivation characteristics of Hypsizygus marmoreus after long-term storage in different conditions (장기 저장 후 느티만가닥버섯(Hypsizygus marmoreus) 균주의 균사 생존력과 재배 특성에 대한 연구)

  • Lee, Hyae-Jung;Ro, Hyeon-Su;Kim, Jong-Guk;Lee, Chang-Yun
    • Journal of Mushroom
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    • v.12 no.1
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    • pp.29-34
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    • 2014
  • Mycelial viability and cultivation characteristics of strains of Hypsizygus marmoreus after long-term storage were investigated. The experimental conditions were the storage at $4^{\circ}C$ using a slant culture technique with or without mineral oil, and in liquid nitrogen tank in the presence of 10% glycerol or 10% glycerol with 5% trehalose. The myceila of four strains of H. marmoreus were thawed at 9, 21, 33, and 45 months after beginning of the storage, and then the growth of the mycelia was measured on a PDA plate with serial transfers to new plate for the recovery. The mycelial growth data after 45 months showed that the mycelia were mostly viable but not fully active particularly when they were stored in liquid nitrogen with 10% glycerol. The growth activity could be fully recovered after second transfer to new PDA plate. Cultivation of mushroom fruiting body using the recovered mycelia also demonstrated that the storage methods employed in this work were applicable for the long-term storage of H. marmoreus.

Preservation of Mushroom Cultures in Sterile Distilled Water (멸균 증류수를 사용한 담자균류 버섯 균주의 보존)

  • Lee, Dong-Hoon;Kim, Chang-Jin;Shin, Kwang-Soo
    • The Korean Journal of Mycology
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    • v.26 no.1
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    • pp.91-96
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    • 1998
  • Optimal conditions for the preservation of mushroom cultures in sterile distilled water were investigated. Mycelial disks from 12 mushroom cultures were preserved at room temperature, $5^{\circ}C,\;-20^{\circ}C$ and $-75^{\circ}C$ using sterile water and 6 cryoprotectants for 12 months. The maximal viability of mycelial disks was observed at room temperature and $5^{\circ}C$ with the value of 91.7%. At $-75^{\circ}C$, the viability was highest in polyethylene glycol (PEG) and lowest in 10% dimethyl sulfoxide (DMSO). The average mycelial growth was highest for disks preserved at $-75^{\circ}C$ in PEG. PEG was the most suitable for the preservation at $-75^{\circ}C$, although this chemical has rarely been used as a cryoprotectant in fungal culture preservation. The laccase isozyme patterns of the preserved isolate of H. ferrugineum were not changed during preservation at $5^{\circ}C$ in sterile distilled water as compared to that of slant culture.

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Effects of Preservation Period at Low Temperature on the Mycelial Growth and the Lignocellulolytic Enzyme Activities of Basidiomycetes (저온보존기간이 담자균류의 균사생장과 목질분해효소의 활성에 미치는 영향)

  • Jeong, Yeun Sug;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.322-327
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    • 2014
  • Subculture is the most common method for preservation fungi, but has a disadvantage of accumulation of spontaneous mutations during the repeated subculture. To reduce the subculture frequency, the effect of preservation period at $4^{\circ}C$ in a slant culture was examined on the mycelia growth and lignocellulolytic enzyme activities of various basidiomycetes. Mushrooms, including Stereum ostrea, Coprinellus micaeus, Trametes versicolor, Hypholoma fasciculare, Wolfiporia extensa, Pleurotus pulmonarius, Piptoporus betulinus and Ganoderma applanatum were not affected by the preservation period more than two years, indicating that they can be maintained by subculture every two years. Some other tested fungal strains showed a significant decrease in both viability and enzyme activity when they were maintained for two years, suggesting that they should be subcultured at least once in a year. A little correlation was found between the recovery of mycelial growth and extracellular enzyme activity. In conclusion, mycelial activity and enzyme activity according to storage period is expected to be a way of deciding on subculture times for fungal preservation.

Isolation and Characterization of Benomyl-Resistant Mutants in an Entomopathogenic Fungus, Metarhizium anisopliae

  • Kim Soon Kee;Shim Hee Jin;Roh Jong Yul;Jin Byung Rae;Boo Kyung Saeng;Je Yeon Ho
    • International Journal of Industrial Entomology
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    • v.10 no.2
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    • pp.119-123
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    • 2005
  • Benomyl-resistant mutants of entomopathogenic fungus, Metarhizium anisopliae were isolated and their physiological characteristics were investigated. These militants were obtained spontaneously or by UV irradiation in benomyl-treated media. Four spontaneous (S-2, S-11, S-18, S-19) and four UV-induced (UV-4, UV-5, UV-19, UV-24) mutants, which grow stably and normally were selected. No significant differences in conidia or hyphal shape, conidia viability, mycelial biomass, or virulent to the diamondback moth were observed between the wild type and their mutants. But differently from the mycelial growth of other benomyl-resistant mutants which was slower than that of the wild type on a modified Czapek-Dox, SDAY, $4\%$ chitin, or $1\%$ skim milk medium, that in the spontaneous mutants, S-18 and S-19, did not show any difference from the wild type. Especially, S-18 and S-19 grew well at benomyl concentrations up to 50 times or higher than that which inhibits wild type proliferation. These results suggested that S-18 and S-19 could potentially be used with the fungicide, benomyl.

Effect of Temperature on Sclerotia Formation and Viability of Sclerotinia sclerotiorum Causing Sclerotiorum Rot of Cryptotaenia japonica (파드득나물 균핵병균(Sclerotinia sclerotiorum)의 균핵 헝성 및 생존에 미치는 온도의 영향)

  • Chang, Seog-Won;Lee, Han-Bum;Kim, Sung-Kee
    • Research in Plant Disease
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    • v.9 no.1
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    • pp.47-51
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    • 2003
  • Sclerotinia rot of C. japonica caused by Sclerotinia sclerotiorum was observed throught the growing season in the vinylhouse at the Namyangiu, Yangju and Yangpyung areas. The disease was commonly occurred at late autumn after transplanting and continuously increased until spring in next year, The disease was especially severe at the Yangpyung area among three areas. Three isolates of S. sclerotiorum obtained from C. japonica in different geographic areas were evaluated for ability of sclerotia formation and viability under different temperature. Temperature for mycelial growth ranged from 5 to 3$0^{\circ}C$, with optimum temperature at 15~3$0^{\circ}C$. Sclerotia were fewer formed at low temperature, but their size was larger than that at high temperature. Small size of sclerotia was more viable than those of large size at high temperature. Based on the differences in viability of sclerotia, the relative ranks of sclerotia were in order ~2 mm > 3~4 mm > 5 mm~.

Comparison of anti-inflammatory, antioxidant and anti-allergic effects of Ganoderma species mycelial extracts (영지버섯 균주별 균사체 추출물의 항염, 항산화 및 항알러지 효능 비교 분석)

  • Park, Young-Jin;Nam, Jae-Young;Yoon, Dae-Eun;Kwon, O-Chul;Kim, Hong-Il;Yoo, Young-Bok;Kong, Won-Sik;Lee, Chang-Soo
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.111-115
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    • 2013
  • This study wascarried out to compare the medicinal effects of various Ganoderma species mycelial extracts. Among 6 Ganoderma species mycelial extracts by using 100% MeOH, G. species ASI-7150 showed the highest antioxidant effect. In nitric oxide (NO) production and ${\beta}$-hexosaminidase release inhibition assay, the treatment of G. lucidum ATCC64251 (Taiwan) mycelia extracts most effectively inhibited NO production in LPS-stimulated RAW264.7 cells and ${\beta}$-hexosaminidase release. In addition, the treatment of all 6 Ganoderma species mycelial extracts were not affect on RAW264.7 cell viability. Although this preliminary research has thrown up many questions in need of further investigation, it will serve as a base for further studies of medicinal effects of various Ganoderma species.

Effects of Ionizing Radiation on Postharvest Fungal Pathogens

  • Jeong, Rae-Dong;Shin, Eun-Jung;Chu, Eun-Hee;Park, Hae-Jun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.176-180
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    • 2015
  • Postharvest diseases cause losses in a wide variety of crops around the world. Irradiation, a useful nonchemical approach, has been used as an alternative treatment for fungicide to control plant fungal pathogens. For a preliminary study, ionizing radiations (gamma, X-ray, or e-beam irradiation) were evaluated for their antifungal activity against Botrytis cinerea, Penicillium expansum, and Rhizopus stolonifer through mycelial growth, spore germination, and morphological analysis under various conditions. Different fungi exhibited different radiosensitivity. The inhibition of fungal growth showed in a dose-dependent manner. Three fungal pathogens have greater sensitivity to the e-beam treatment compared to gamma or X-ray irradiations. The inactivation of individual fungal-viability to different irradiations can be considered between 3-4 kGy for B. cinerea and 1-2 kGy for P. expansum and R. stolonifer based on the radiosensitive and radio-resistant species, respectively. These preliminary data will provide critical information to control postharvest diseases through radiation.

Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Effect of Ganoderma lucidum Solid-state Fermented Salix gracilistyla Extract on Type I Procollagen Biosynthesis in HDFn Cells (불로초 균사배양 갯버들 추출물이 인간 피부 섬유아세포의 제1형 프로콜라겐 생성에 미치는 영향)

  • Jeong, Yong-Un;Park, Young-Jin
    • The Korean Journal of Mycology
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    • v.47 no.2
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    • pp.153-163
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    • 2019
  • This study was carried out to investigate the feasibility of Salix gracilistyla production for cosmetic use through mycelial fermentation. The efficacy of this method was confirmed by fermentation using the mycelia of Ganoderma lucidum (a representative medicinal mushroom). Total polyphenol and flavonoid content and DPPH radical scavenging activity of S. gracilistyla extract (SGE) were found to be higher than those of G. lucidum fermented S. gracilistyla extract (GLSGE). GLSGE had relatively lower collagenase activity than SGE. However, GLSGE increased HDFn cell viability more potently than SGE, and increased the biosynthesis of type I procollagen. Thus, GLSGE could be used as an anti-aging cosmetic active ingredient. These results indicate that extract fermentation using G. lucidum mycelia can effectively enhance some beneficial effects of functional materials.