Protoplasts Isolation and Reversion of Fomitella fraxinea

장수버섯(Fomitella fraxinea)의 원형질체 분리 및 재생

  • Kim, Kyung-Soo (Division of Applied Microbiology, National Institute of Agricultural Science and Technology, RDA) ;
  • You, Chang-Hyun (Division of Applied Microbiology, National Institute of Agricultural Science and Technology, RDA) ;
  • Kong, Won-Sik (Division of Applied Microbiology, National Institute of Agricultural Science and Technology, RDA) ;
  • Kim, Young-Ho (Division of Applied Microbiology, National Institute of Agricultural Science and Technology, RDA) ;
  • Cha, Dong-Yeul (Division of Applied Microbiology, National Institute of Agricultural Science and Technology, RDA)
  • 김경수 (농업과학기술원 응용미생물과) ;
  • 유창현 (농업과학기술원 응용미생물과) ;
  • 공원식 (농업과학기술원 응용미생물과) ;
  • 김영호 (농업과학기술원 응용미생물과) ;
  • 차동열 (농업과학기술원 응용미생물과)
  • Published : 1998.06.30

Abstract

Factors affecting protoplasts isolation and regeneration of Fomitella fraxinea were investigated. Lytic enzyme mixture of Novozym 234, Cellulase onozuka R-10 and ${\beta}-Glucuronidase$ was found to be the best for the protoplasts isolation. Osmotic stabilizer of 0.6 M sucrose was observed as the best for protoplasts isolation. The highest number of protoplasts was obtained from the F. fraxinea mycelium with lytic enzyme mixture and osmotic stabilizer that had been cultured for 3 hours. The highest regeneration rate of 0.02 % was achieved when the 0.6 M sorbitol was employed as osmotic stabilizer.

장수버섯(Fomitella fraxinea)의 원형질체 분리와 재생에 영향을 미치는 요인을 조사하고자 본 연구를 수행하였다. 세포벽 분해 효소 종류별 원형질체 분리 시험을 수행한 결과 장수버섯 ASI 17001은 Novozym 234+Cellulase onozuka R-10+${\beta}-Glucuronidase$를 혼합하여 처리하였을 때 원형질체 분리율이 $8.9{\times}10^4/ml$로 가장 높았다. 원형질체 분리시 사용하는 삼투압 조절제로는 0.6 M sucrose가 가장 양호하였으며, 처리시간은 3시간이 가장 양호하였다. 원형질체 재생율은 0.6 M sorbitol에서 0.02%로 다른 조절제에 비하여 양호한 결과를 나타내었으나 KCl는 전혀 재생이 되지 않았다.

Keywords

References

  1. 표고버섯과 구름버섯 이목간의 원형질체 융합 및 핵 전이에 관한 연구 김채균
  2. 영지와 구름버섯 이속간의 우너형질체 융합 및 핵 전이에 관한 연구 박설희
  3. 체세포 융합에 의한 약용버섯 우량 품종 육성연구 유창현(등)
  4. きのこの化學, 生化學 水野卓;川合正允
  5. Agric. Biol. Chem. v.46 no.7 Regeneration and fusion of mycelial protoplasts of Tricholoma matsutake Abe, M.;Umetsu, M.;Nakai, T.;Sasage, D.
  6. Arch. Pharm. Res. v.10 no.3 Studies on protoplast formation and regeneration of Ganoderma lucidum Choi, S.H.;Kim, B.K.;Kim, H.W.;Kwak, J.H.;Choi, E.C.;Kim, Y.C.;Yoo, Y.B.;Park, Y.H.
  7. Journal of General Microbiology v.73 Release of protoplasts from Schizophyllum commune by a lytic enzyme preparation from Trichoderma viride De Vris, O.M.H.;Wessels, J.G.H.
  8. J. Gen. Microbiol. v.44 Regeneration of mycelial protoplast of Fusarium culmorum Garcia, A.;Lopez-Belmonte, I.F.;Villanueva, J.R.
  9. Appl. Environ. Microbiol. v.46 no.1 Formation fusion and regeneration of protop lasts from wild-type and auxotrophic strains of the white rot basidiomycetes Phanerochaete chrysosporium Gold, M.H.;Cheng, T.M.;Alic, M.
  10. Studies on the strain improvement by mycelial fusion and nuclear transfer of Ganoderma lucidum, and its pharmacological characteristics Jeong, H.
  11. Kor. J. Mycol. v.14 no.3 Protoplast regeneration and reversion in Pleurotus cornucopiae Lee, Y.H.;You, C.H.;Cha, D.Y.;Yoo, Y.B.;Min, K.H.
  12. Trans. Br. Mycol. Sec. v.65 no.1 Production of Coprinus protoplasts by use of chitinase or helicase Moore, D.
  13. Kor. J. Appl. Microbiol. Bioeng. v.15 no.4 Protoplast formation and reversion of Ganoderma applanatum Park, Y.D.;Yoo, Y.B.;Park, Y.H.;Chang, M.W.;Lee, J.S.
  14. Ann. Reu. Microbiol. v.33 Fungal protoplasts: isolation, reversion and fusion Peberdy, J.F.
  15. Mycol. Res. v.93 Fungi without coats-pro-toplasts as tools for mycological research Peberdu, J.F.
  16. Res. Rep. Agri. Sci. Tech. Chungnam Natl. Univ. v.13 no.2 Some factors affecting the protoplast formation and regeneration from the mycelium of Ganoderma lucidum (Fr) Karsten Shin, G.C.;Yeo, U.Y.;Yoo, Y.B.;Park, Y.H.
  17. Biol. Rundsch Uber entstehung und reversion enzymatisch erzeugter protoplasten von Polysticus versicolor Strunk, C.
  18. Kor. J. Mycol. v.16 no.1 Protoplast isolation and reversion from Ganoderma lucidum and Ganoderma sp Um, S.D.;Chae, Y.A.;Yoo, Y.B.;You, C.H.;Cha, D.Y.
  19. Eup. J. Appl. Microbiol. Biotech. v.17 Preparation and regeneration of mycelial protoplasts of Collybia velutipes and Pleurotus ostreatus Yamada, D.;Magae, Y.;Kashiwagi, Y.;Kakimoto, Y.;Sasagi, T.
  20. Kor. J. Mycol. v.13 no.1 Studies on protoplast isolation from edible fungi Yoo, Y.B.;Peberdy, J.F.;You, C.H.
  21. Kor. J. Mycol. v.13 no.2 Studies on protoplast regeneration and reversion of Pleurotus ostreatus and Pleurotus florida Yoo, Y.B.;Peberdy, J.F.;Cha, D.Y.
  22. Kor. J. Mycol. v.16 no.4 Studies on protoplast formation and reversion of Pleurotus sapidus Kalhbr You, C.H.;Yoo, Y.B.;Park, Y.H.