한국균학회지 (The Korean Journal of Mycology)

  • 제47권1호
  • /
  • Pages.1-12
  • /
  • 2019
  • /
  • 0253-651X(pISSN)
  • /
  • 2383-5249(eISSN)
한국균학회 (The Korean Society of Mycology)
권호 표지
DOI QR코드

DOI QR Code

흰목이 균사체 배양 적합 조건 설정

Suitable Conditions for Mycelial Culture of Tremella fuciformis

  • 이은지 (국립원예특작과학원 버섯과) ;
  • 박혜성 (국립원예특작과학원 버섯과) ;
  • 이찬중 (국립원예특작과학원 버섯과) ;
  • 공원식 (국립원예특작과학원 버섯과) ;
  • 구창덕 (충북대학교 산림학과)
  • 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)
  • 투고 : 2019.01.30
  • 심사 : 2019.03.08
  • 발행 : 2019.03.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

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.

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.

키워드

GNHHDL_2019_v47n1_1_f0001.png 이미지

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.

GNHHDL_2019_v47n1_1_f0002.png 이미지

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.

GNHHDL_2019_v47n1_1_f0003.png 이미지

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).

GNHHDL_2019_v47n1_1_f0004.png 이미지

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

GNHHDL_2019_v47n1_1_f0005.png 이미지

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).

GNHHDL_2019_v47n1_1_f0006.png 이미지

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

GNHHDL_2019_v47n1_1_f0007.png 이미지

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).

GNHHDL_2019_v47n1_1_f0008.png 이미지

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

GNHHDL_2019_v47n1_1_f0009.png 이미지

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)

GNHHDL_2019_v47n1_1_f0010.png 이미지

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

GNHHDL_2019_v47n1_1_f0011.png 이미지

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).

GNHHDL_2019_v47n1_1_f0012.png 이미지

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

GNHHDL_2019_v47n1_1_f0013.png 이미지

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).

GNHHDL_2019_v47n1_1_f0014.png 이미지

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

GNHHDL_2019_v47n1_1_t0001.png 이미지

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

GNHHDL_2019_v47n1_1_t0002.png 이미지

참고문헌

  1. Chang HY. Artificial cultivation of Tremella fuciformis Berk. using associated fungus, Hypoxylon sp. [dissertation]. Chuncheon, Korea: Kangwon National University; 1997.
  2. Cheng HH, How WC, Lu ML. Interactions of lipid metabolism and intestinal physiology with Tremella fuciformis Berk. edible mushroom in rats fed a high cholesterol diet with or without Nebacitin. J Agric Food Chem 2003;50:7438-43. https://doi.org/10.1021/jf020648q
  3. Choi SW, Chang HY, Yoon JW, Lee C. Optimization of artificial cultivation of Tremella fuciformis in closed culture bottle. J Mushrooms 2008;6:20-6.
  4. Huang NL. Cultivation of Tremella fuciformis in Fujian, China. Mushroom Newsl Trop 1982;2:2-5.
  5. Huang NL. Cultivation of Tremella fuciformis in China. 1st ed. Beijing: China Agricultural Publishing Company; 2000.
  6. Jukes TH, Cantor CR. Evolution of protein molecules, Munro HN, editor. Mammalian protein metabolism. New York: Academic Press; 1969. p. 21-132.
  7. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018;35:1547-9. https://doi.org/10.1093/molbev/msy096
  8. Kwon SY, Whang K, Lee SP. Anti-inflammatory effects and GABA production of old antler and Auricularia auricula-judae extract fermented by Lactobacillus plantarum. Korean J Food Preserv 2017;24: 274-81. https://doi.org/10.11002/kjfp.2017.24.2.274
  9. Lee CJ, Moon JW, Yoo YM, Han JY, Cheong JC, Kong WS. Optimum cultivation conditions for mass production of antagonistic bacterium Pseudomonas azotoformans HC5 effective in antagonistic of brown blotch disease caused by Pseudomonas tolaasii . J Mushrooms 2015;13:97-102. https://doi.org/10.14480/JM.2015.13.2.97
  10. Luo XC. Review of studies on Tremella fuciformis in China. Mycosystema 2013;32:14-9.
  11. Peng WH, Wang Y, Huang ZQ, Gan BC. The Present situation of Tremella fuciformis research and problems existed in China. Acta Edulis Fungi 2005;12
  12. Cheung PCK. The hypocholesterolemic effect of two edible mushrooms: Auricularia auricular (tree ear) and Tremella fuciformis (white jelly leaf) in hypocholesterolemic rats. Nutr Res 1996;16:1721-5 https://doi.org/10.1016/0271-5317(96)00191-1
  13. Ryu YH, Yoon YS, Jo WS, Park SD, Choi BS, Kim JK. Effect of liquid spawn on Flammulina veluipes cultivation. J Mycol 1998;26:20-4.
  14. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406-25.
  15. Stanier, RY, Palleroni, NJ, Doudoroff M. The aerobic pseudomonads: a taxonomic study. Microbiology 1966;43:159-271.
  16. Thompson JD, Higgins DG, Gibson TJ. Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighing position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994;11:4673-80. https://doi.org/10.1093/nar/22.22.4673
  17. White TJ, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, editors. PCR protocols: a guide to methods and applications. San Diego: Academic Press; 1990. p. 315-22.