The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Suitable Conditions for Mycelial Culture of Tremella fuciformis

흰목이 균사체 배양 적합 조건 설정

  • Lee, Eun-ji (Mushroom Research Division, National Institute of Horticultural & Herbal Science, RDA) ;
  • Park, Hye-Sung (Mushroom Research Division, National Institute of Horticultural & Herbal Science, RDA) ;
  • Lee, Chan-Jung (Mushroom Research Division, National Institute of Horticultural & Herbal Science, RDA) ;
  • Kong, Won-Sik (Mushroom Research Division, National Institute of Horticultural & Herbal Science, RDA) ;
  • Koo, Chang-Duck (Department of Forestry Science, Chungbuk National University)
  • 이은지 (국립원예특작과학원 버섯과) ;
  • 박혜성 (국립원예특작과학원 버섯과) ;
  • 이찬중 (국립원예특작과학원 버섯과) ;
  • 공원식 (국립원예특작과학원 버섯과) ;
  • 구창덕 (충북대학교 산림학과)
  • Received : 2019.01.30
  • Accepted : 2019.03.08
  • Published : 2019.03.01
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Abstract

This study was conducted to investigate the optimum culture conditions, for mass production of Tremella fuciformis in M9 basic medium. The strain KMCC04674, used in this study was identified T. fuciformis by internal transcribed spacer (ITS). To define the optimum conditions for the mass production of T. fuciformis, we investigated the effects of different culture conditions and various nutrient sources on the fungal growth. The optimum initial pH and temperature for the fungal growth were 5.0 and $25^{\circ}C$, respectively. The optimal composition of the growth medium was 4.0% mannitol, 3.0% $NH_4H_2PO_4$, 1.0% malt extract, 1.0% glutamic acid, 5mM $CaCl_2$, and 0.5% glucanic acid.

본 연구에서는 흰목이 균사배양을 위한 최적조건을 분석하였다. 흰목이균주(KMCC04674) 균사체의 대량 배양에 효율적인 영양원 선발을 위하여 M9 기본배지에탄소원 fructose 등 21종, 무기질소원 $(NH_4)_2C_2O_4$, 등 7종, 유기질소원 peptone 등 7종, 아미노산 asparagine 등 14종, 그리고 유기산 acetic acid 등 8종을 각각 1%씩 그리고, 무기염류 KCl 등 13종을 1 mM 농도로 첨가하여 각각에 대한 균사체의 생육을 조사하였다. 또한 선발된 영양성분들에 대한 최적 농도를 알기 위하여 0.1%에서 4.0%까지 배지에 첨가하고 배양 후 흰목이 균사체생육정도를 조사하였다. 그결과, 균사체의 대량배양을 위한 생육최적조건은 온도 $25^{\circ}C$, pH5, 탄소원 4.0% mannitol, 유기질소원 1.0% malt extract, 무기질소원 3.0% $NH_4H_2PO_4$, 아미노산 1.0% glutamic acid, 무기염류는 5mM $CaCl_2$ 그리고유기산 0.5% gluconic acid였다.

Keywords

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Fig. 1. Phylogenetic relationships of T. fuciformis based on internal transcribed spacer(ITS) sequences. Numerical values on branches are the bootstrap values as percentage of bootstrap replication from 1000 replicate analysis. Bar = 0.02 genetic distance between samples.

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Fig. 2. Effect of temperature (n = 3; error bar = one standard deviation) and pH (n = 9; error bar = one standard deviation) on the growth of T. fuciformis.

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Fig. 3. Effect of carbon sources for the growth of T. fuciformis in basal medium. 1: Fructose, 2: Galactose, 3: Saccharose, 4: Soluble starch, 5: Inositol, 6: Glycerol, 7: Xylose, 8: Dextrose, 9: Lactose, 10: Dextrin, 11: Na-CMC, 12: Adonitol. 13: Mannitol, 14: Mannose, 15: Maltose, 16: Raffinose, 17: Cellobiose, 18: Ethanol, 19: Salicin, 20: Glucose, 21: Arabinose (n = 21; error bar = one standard deviation).

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Fig. 4. Effect of mannitol concentration by T. fuciformis in basal medium (n = 14; error bar = one standard deviation).

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Fig. 5. Effect of inorganic nitrogen sources for the growth of T. fuciformis in basal medium. 1: (NH4)2C2O4, 2: NH4H2PO4, 3: (NH4)2HPO4, 4: NH4NO3, 5: NaNO3, 6: (NH4)2SO4, 7: (NH4)2C4H4O6 (n = 7; error bar = one standard deviation).

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Fig. 6. Effect of NH4H2PO4 concentration by T. fuciformis in basal medium (n = 8; error bar = onestandard deviation).

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Fig. 7. Effect of organic nitrogen sources for the growth of T. fuciformis in basal medium. 1: Peptone, 2: Soytone 3: Urea, 4: Casamino acid 5: Tryptone, 6: Yeast extract, 7: Malt extract (n = 7; error bar = one standard deviation).

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Fig. 8. Effect of Malt extract concentration by T. fuciformis in basal medium (n = 8; error bar = one standard deviation).

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Fig. 9. Effect of amino acid sources for the growth of T. fuciformis in basal medium. 1: Alanine 2:Tyrosine, 3: Asparagine, 4: Methionine, 5: Proline, 6: Leucine, 7: Glutamine, 8: Valine, 9: Aspartic acid, 10: Glutamic acid, 11: Arginine, 12: Histidine, 13: Cysteine, 14: Threonine (n = 14; error bar = one standard deviation)

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Fig. 10. Effect of Glutamic acid concentration by T. fuciformis in basal medium (n = 8; error bar = one standard deviation).

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Fig. 11. Effect of inorganic salts for the growth T. fuciformis in basal medium. 1: KCl, 2: BaCl2, 3: CaCl2, 4: CoCl2, 5: Li2SO4, 6: MnSO4 7: MgSO4, 8: ZnSO4, 9: FeSO4, 10: AgNO3, 11: Na2MoO4, 12: KH2PO4, 13: FeCl3 (n = 13; error bar = one standard deviation).

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Fig. 12. Effect of CaCl2 concentration by T. fuciformis in basal medium (n = 10; error bar = one standard deviation).

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Fig. 13. Effect of organic acid for the growth T. fuciformis in basal medium. 1: Acetic acid, 2: Citric acid,3: Gluconic acid, 4: Glutamic acid, 5: Lactic acid, 6: Maleic acid, 7: Propionic acid, 8: Succinic acid (n =8; error bar = one standard deviation).

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Fig. 14. Effect of gluconic acid concentration by T. fuciformis in basal medium (n = 8; error bar = one standard deviation).

Table1. Tremella fuciformis used in this study

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Table 2. Culture condition of T. fuciformis for the maximal production of inhibition substance

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