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Cultural Characteristics of Ectomycorrhizal Mushrooms

  • Jeon, Sung-Min;Ka, Kang-Hyeon
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.16-16
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    • 2015
  • Ectomycorrhizal (ECM) mushrooms play a major role in plant growth promotion through symbiotic association with roots of forest trees. They also provide an economically important food resource to us and therefore they have been studied for their artificial cultivation for decades in Korea. We have secured bio-resources of ECM mushrooms from Korean forests and performed their physiological studies. To investigate the cultural characteristics, the fungi were cultured under different conditions (medium, temperature, pH of the medium, inorganic nitrogen source). More than 90% of total 160 strains grew on three solid media (potato dextrose agar, PDA; sabouraud dextrose agar, SDA; modified Melin-Norkrans medium, MMN). The rate of mycelial growth on malt extract agar (MEA) was lower than those of three media (PDA, SDA, MMN). None of the Tricholomataceae strains grew on MEA. Many strains of ECM mushrooms were able to grow at the temperature range of $15{\sim}25^{\circ}C$ on PDA, while they showed poor growth at $10^{\circ}C$ or $30^{\circ}C$. In particular, the growth rates of both Gomphaceae and Tricholomataceae were significantly lower at $10^{\circ}C$ than at $30^{\circ}C$. The optimal pH of many strains was pH 5.0 when they cultured in potato dextrose broth (PDB). Fifty-seven percent of tested strains grew well on medium containing ammonium source than nitrate source. Many strains of Tricholomataceae showed a notable growth on ammonium medium than nitrate medium. Twenty-three percent of strains preferred nitrate source than ammonium source for their mycelial growth. The production and activity of two enzymes (cellulase and laccase) by ECM fungi were also assayed on the enzyme screening media containing CMC or ABTS. Each strains exhibited different levels of enzymatic activities as well as enzyme production. The number of laccase-producing strains was less than that of cellulase-producing strains. We found that 77% of tested strains produced both cellulase and laccase, whereas 2% of strains did not produce any enzymes. The morphological characteristics of mycelial colony were also examined on four different solid media. Yellow was a dominant color in mycelial colony and followed by white and brown on all culture media. ECM mushrooms formed mycelial colonies with a single or multiple colors within a culture medium depending on the strains and culture media. The most common shape of mycelial colony was a circular form on all media tested. Other families except for Amanitaceae formed an irregular colony on MMN than PDA. All strains of Tricholomataceae did not form a filamentous colony on all media. The pigmentation of culture media by mycelial colonies was observed in more than 50% of strains tested on both PDA and SDA. The degree of pigmentation on PDA or SDA was higher than MMN and brown color was dominant than yellow color. The production of exudates from mycelial colony was higher on PDA than MMN. Brown exudates were mainly produced by many strains on PDA or SDA, whereas transparent exudates were mainly produced by strains on MMN. We observed the mycelial colonies with a single or multiple textures in just one culture plate. Wrinkled or uneven colony surfaces were remarkably observed in many strains on PDA or SDA, while an even colony surface was observed in many strains on MMN. Sixty percent of Tricholomaceae strains formed wrinkled surface on PDA. However, they did not form any wrinkle on MMN plate. Cottony texture was observed in mycelia colonies of many strains. Velvety texture was often observed in the mycelial colonies on SDA than PDA and accounted for 60% of Suillaceae strains on SDA.

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Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology (반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화)

  • Yang, Hee-Jong;Park, Chang-Su;Yang, Ho-Yeon;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn;Kang, Dae-Ook;Moon, Ja-Young;Choi, Nack-Shick
    • Journal of Life Science
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    • v.25 no.6
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    • pp.680-685
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    • 2015
  • Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R2) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

Selection and Mechanisms of Indigenous Antagonistic Microorganisms against Sheath Rot and Dry Rot Disease of Garlic (마늘 잎집썩음병과 마른썩음병을 길항하는 토착길항미생물의 선발 및 기작)

  • Jeong, Hee-Young;Lim, Jong-Hui;Kim, Byung-Keuk;Lee, Jung-Jong;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.295-301
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    • 2010
  • Sheath rot and dry rot disease caused by Pseudomonas marginalis and Fusarium oxysporum were serious problems in garlic farmland. In this study, total of 160 indigenous antagonistic bacteria were isolated from 16 farmlands in Yeongcheon, Korea. Among these, 15 strains were able to inhibited P. marginalis and F. oxysporum. The 16s rDNA genes of the selected 15 strains were amplified and sequenced. The strains has strong antagonistic ability against garlic pathogens was achieved Bacillus subtilis YC82, B. vallismortis YC84, B. amyloliquefaciens YC240. The selected 3 strains tested for investigation of antifungal mechanisms further analyses; 3 strains of these validated for production of siderophore, ${\beta}$-glucanase and chitinase using CAS (chrome azurol S) blue agar, CMC-congo red agar and DNS method. The 3 strains were able to utilized insoluble phosphate as dertermined by vanado-molybdate method. The 3 strains verified for production of auxin and gibberellic acid using Salkowski test and holdbrook test. Also, 3 strains showed stimulation germination, stem growth promoting activity on the in vivo test. The 3 strains were able to effectively suppress P. marginalis and F. oxysporum causing sheath rot and dry rot diseases on the in vivo pot test.

Study on Improvement of Mechanical Property, Oxidation and Erosion Resistance of SiC Matrix Ceramic Composites Reinforced by Hybrid Fabric Composed of SiC and Carbon Fiber (탄화규소섬유와 탄소섬유 하이브리드 직물을 강화재로 한 SiC 매트릭스 세라믹복합재의 기계적물성, 산화 및 삭마 저항성 개선 연구)

  • Yoon, Byungil;Kim, Myeongju;Kim, Jaesung;Kwon, Hyangjoo;Youn, Sungtae;Kim, Jungil
    • Composites Research
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    • v.32 no.3
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    • pp.148-157
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    • 2019
  • In this study, $C_f/SiC$, $SiC_f/SiC$ and $C_f-SiC_f/SiC$ ceramic composites reinforcing carbon fiber, SiC fiber and hybrid fiber were fabricated by hybrid TGCVI and PIP process. After the thermal shock cycle, 3-point bending and Oxy-Acetylene torch test, their mechanical behavior, oxidation and erosion resistance were evaluated. The $C_f/SiC$ composite showed a decrease in mechanical property along with increasing temperature, a pseudo-ductile fracture mode and a large quantity of erosion. The $SiC_f/SiC$ composite exhibited stronger mechanical property and lower erosion rate compared to the $C_f/SiC$, but brittle fracture mode. On the other hand, hybrid type of $C_f-SiC_f/SiC$ composite gave the best mechanical property, more ductile failure mode than the $SiC_f/SiC$, and lower erosion rate than the $C_f/SiC$. During the Oxy-Acetylene torch test, the $SiO_2$ formed by reaction of the SiC matrix with oxygen prevented further oxidation or erosion of the fibers for $C_f-SiC_f/SiC$ and $SiC_f/SiC$ composites particularly. In conclusion, if a hybrid composite with low porosity is prepared, this material is expected to have high applicability as a high temperature thermo-structural composite under high temperature oxidation atmosphere by improving low mechanical property due to the oxidation of $C_f/SiC$ and brittle fracture mode of $SiC_f/SiC$ composite.

Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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