• Title/Summary/Keyword: Agar media

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Studies on Protoplast Isolation and Regeneration of Lyophyllum ulmarium (느티만가닥 버섯의 원형질체 분리와 재생에 관한 연구)

  • 최혜진;김병각;현진원
    • Journal of Life Science
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    • v.13 no.2
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    • pp.143-149
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    • 2003
  • This experiment was undertaken to investigate proper conditions for protoplast isolation and regeneration from the mycelia of Lyophyllum ulmnrium. Protoplast isolation and regeneration are influenced by a variety of factors such as enzyme, osmotic stabilizer, reaction time and age of mycelia. A combination of Novozyme 234 (10mg/ml) and cellulase Onozuka R-10 (10 mg/ml) with 0.6 M $MgSO_4$ was most effective for isolation of the protoplasts. The optimum reaction time of the mycelia with the lytic enzymes was 3.5~4 hours at $28^{\circ}C$ in shaking condition at 120 strokes per min. High yield of the protoplasts were obtained from its 4~5 days old mycelia on complete agar media. Its protoplasts were regenerated to normal hyphae. Regeneration media with 0.6 M sucrose were proper for regeneration of the protoplasts. Their regeneration frequency on complete agar media was 2.3~2.7%.

Development of an improved selective media for differentiation of emetic and diarrheal type Bacillus cereus

  • Hong, Yong-Gun;Lee, Jin-Joo;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.815-818
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    • 2021
  • The objective of this study was to develop a differential medium with improved selectivity for the isolation of Bacillus cereus. Mannitol egg yolk polymyxin medium supplemented with D-galactose allowed the differentiation of diarrheal- and emetic-type B. cereus through pH monitoring. The pH of the medium decreased significantly when incubating the emetic-type B. cereus, whereas the pH change was not significant when incubating the diarrheal-type. The addition of pH indicators, such as methyl red and phenol red, to the medium allowed visual differentiation between diarrheal- and emetic-type B. cereus. A solid agar medium was also developed by optimizing the concentrations of medium components such as monosaccharides, agar, egg yolk enrichment, pH indicators, and antibiotics. This study indicates the possibility of applying selective media for the differentiation of diarrheal- and emetic-type B. cereus.

Studies on Distribution and Utilization of Cordyceps militaris and C. nutans (동충하초속균의 분포 및 Cordyceps militaris와 C. nutans의 이용에 관한 연구)

  • Sung, Jae-Mo;Kim, Chun-Hwan;Yang, Kun-Joo;Lee, Hyun-Kyung;Kim, Yang-Sup
    • The Korean Journal of Mycology
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    • v.21 no.2
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    • pp.94-105
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    • 1993
  • The genus Cordyceps known as an insect parasite forms a sclerotium in insect bodies and then produces perithecia on the single or multiple stromata produced from sclerotium. Collected Cordyceps were identified into 5 species: Cordyceps militaris, C. nutans, Cordyceps sphecocephala, Isaria japonica, and Torrubiella sp. The fruit bodies of Cordyceps in petri-dish cover were fixed by tape and put the lid on water agar plates to isolate these collected Cordyceps. The germinated spores were transferred from water agar to Potato dextrose agar(PDA) after six hours. Mycelial growth of C. nutans and C. militaris was the most successful on Hamada media and was also good on Complete media and PDA. Mannose as a carbon source was good for two species and Glutamic acid as a nitrogen source was satisfactory to C. militaris and Asparagine gave a good result to C. nutans. C. militaris and C. nutans showed similar mycelial growth rate on the media that contained thiamine-HCI, biotine or nicotinic acid as a vitamine. When conidia of C. nutans were inoculated to insects, mortality was high in Artogeia napi L, Hemiptera, Plutella xylostella and 50% in Orthoptera, 12% in Acantholyda posticalise M, but not Agelastica coerulea B. in Aphididae, C. nutans was collected from only Hemiptera in nature, but killing effect on other insects was proved. Mycelial growth and fruit-body formation were good on the media that consist of rice powder 5g, wheat flour 5g, water 100ml, but formed fruit-body was not complete stromata but a mass of conidia according to results of observing microscope.

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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.

Differential Growth Response of $A_1\;and\;A_2$ Mating Types of Phytophthora infestans on Rye A and V-8 Juice Agar Media Supplemented with Rhizome Powder of Cyperus rotundus

  • Singh, U.P.;Sarma, B.K.;Nishimura, Ruo;Kobayashi, Kiroku;Ogoshi, Akira;Zinkernagel, Volker;Schlenzig, Alexendra;Schober-Butin, Barbel;Aust, H.J.
    • Mycobiology
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    • v.29 no.3
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    • pp.164-169
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    • 2001
  • A new medium for studies of diversity among populations of $A_1\;and\;A_2$ mating types of Phytophthora infestans has been evolved. The rye A agar and V-8 juice agar media on which P. infestans grows well have been amended with rhizome powder of Cyperus rotundus. A total of 259 isolates of $A_1\;and\;A_2$ mating types representing Japan, Korea, India, Taiwan, Indonesia, Thailand, China, Nepal, U.K and Medico were screened for their growth response on these two media. Most of the A1 isolates did not grow well on them except Thailand while growth of $A_2$ mating types differed as some grew on it whereas others did not. It is quite likely that the populations of $A_2$ mating types that did not grow well on rhizome-amended medium are of different clonal lineage. This suggests that this medium can be used for the study of diversification among the isolates of the same or both the mating types as well as to detect the newly introduced genetically different isolates of P. infestans in a locality where it was not reported earlier.

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Three New Records of Ascomycetes Isolates from Field Soils in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Hyun Seung;Bazie, Setu;Lee, Hyun Gu;Lee, Hyang Burm;Lee, Youn Su
    • Mycobiology
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    • v.45 no.4
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    • pp.327-337
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    • 2017
  • Three new records of Ascomycota species (Chaetomium acropullum, Phialemonium globosum, Phialemonium atrogriseum) from field soils in Korea are presented in this study. These newly discovered fungal isolates were isolated from field soils from various places across Gyeongnam, Korea in 2016. All the isolates were identified and described based on morphological characteristics, and rDNA internal transcribed spacer and ${\beta}$-tubulin gene sequence data. Morphological features of these fungal species were studied on different agar media: potato dextrose agar, oatmeal agar, malt extract agar, Czapek yeast extract agar, and yeast extract sucrose agar. Full description and illustrations of their morphological characters are provided. These fungal species have not officially been previously reported in Korea.

Studies on the microbiological assay method for tabtoxin produced in pseudomonas syringae pv. tabaci (Pseudomonas syringae pv. tabaci가 생산하는 tabtoxin의 미생물학적 검색방법에 관한 연구)

  • 백형석;구재관;전홍기
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.310-315
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    • 1989
  • Tabtoxin produced in Pseudomonas syringae pv. tabace irreversibly inhibits its known physiological target, glutamine synthetase so that causes wildfire disease on leaves of host plant. In this study, we examined a rapid and sensitive microbiological method for tabtoxin assay in several media. In minimal A agar medium nd minimal glucose agar medium, growth inhibition zone of Agrobacterium tumefaciens was larger than that of other indicator strain. However, mostly, growth inhibition zone of indicator strains on the minimal glucose agar medium was smaller than that of on the miniaml A agar medium. In complex agar medium, growth inhibithiton zone was not observed in all the tested indicator strains. Pseudomonas syringae pv. tabaci produced more tabtoxin according to the incubation time. When glutamine was added to the minimal glucose agar medium, growth inhkbition zone of Agrobacterium tumefaciens was reduced.

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Cultural Characteristics of Ophiocordyceps heteropoda Collected from Korea

  • Sung, Gi-Ho;Shrestha, Bhushan;Han, Sang-Kuk;Sung, Jae-Mo
    • Mycobiology
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    • v.39 no.1
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    • pp.1-6
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    • 2011
  • Isolates of Ophiocordyceps heteropoda (Kobayasi) collected from Mt. Halla on Jeju-do, Korea were tested for mycelial growth on different agar media and in the presence of different carbon and nitrogen sources. Similarly, isolates were also incubated at different temperatures as well as under continuous light and dark conditions. Growth was better on Hamada agar, basal medium, and malt-yeast agar, but poor on Czapek-Dox agar. Different carbon sources such as dextrin, saccharose, starch, lactose, maltose, fructose, and dextrose resulted in better growth. Complex organic nitrogen sources such as yeast extract and peptone revealed the most effective growth. Mycelial growth was best at $25^{\circ}C$. The growth rate was faster in the dark than the light, but mycelial density was less compact in the dark.

Cultural Characteristics and Ascospore Density in Soil of Monosporascus Cannonballus on Cucurbitaceae Plants (박과류 검은점뿌리썩음병균의 배양적 특성 및 토양내 자낭포자 밀도)

  • 허노열;류경열;이용범
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.16-19
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    • 2001
  • The cultural caracteristics of Monosporascus cannonballus causing root rot of Cucurbitaceae plants were examined in vitro, and population density of the fungal ascospores were measured and compared aoming diferent host species and soil depths. Potato-dextrose agar(PDA) was the most appropriate medium for the mycelial growth and perithecial formation among the 5 media tested. Corn-meal agar(CMA), oat-meal agar (OMA) and V-8 juice agar were moderate media for the mycelial growth and perithecial formation, whereas water agar(WA) was poor medium. Perithecia were not formed on WA. Optimum temperature for the formation of perithecia was about 25 to 30$^{\circ}C$. distribution of ascospores in the infested fields was variable amount the curcubitaceae plants and within the same plant species, ranging from 1.7 to 14.6 ascospores in 20 g of soil, but no ascospore was detected in the uninfested field soils. Ascospores were distributed more at 20cm of soil depth than at 10cm or 30cm of soil depth.

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Isolation of Nematophagous Fungi against Root-knot Nematode and Their Growth in Vitro (뿌리혹 선충에 대한 기생 천적 진균 분리 및 이들의 생장에 미치는 환경조사)

  • 정미정;김희규
    • Korean journal of applied entomology
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    • v.27 no.3
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    • pp.149-158
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    • 1988
  • Nematophagous fungi were successfully isolated by baited plating, centrifugation technique of soil, and direct isolation from naturally ingested nematodes. Predominant seven fungi isolated were identified as Artheobotrys arthroboteyides, A.conoides, A. oligospora, Dactylella lobata, Fusatium oxysporum, Monacrosporium ellopsoporum and Harposporium anguillu-lae. Of these, six fungi were tested for cultural characteristics except. H, anguillulae, extre-mely fastidious fungus in artificial media. Among 14 media tested in this experiment, Corn-meal Agar (CMA) and Oatmeal Agar (OMA) were the most suitable media for growing all six nematophagous fungi. Weakly saprophytic M. ellipsospoyum also grew vigoroualy on these two media. The radial growth, dry weight and sporulation of the fungi tested were quite diverse depending on the culture media. D. lobata revealed good growth and abundantly sporulated on Glucoes Peptone Agar (GPA). Although over-all growth of F, oxysporum was not satisfactory on Sucrose Nitrate Agar (SNA), the sporulation was best on this medium. Optimum conditious for mycelial growth and sporulation of nematophagous fungi ranged pH 5-8 and 20-$30^{\circ}C$ on SNA. D. lobata and F, oxysporum grew vigorously and most profusely sporulated on all media tested. They turned out an most promising biocontrol agents for their aggressive growth and sporulation over the ranges of temperature and pH ranges.

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