Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
권호 표지

Isolation and Culture of Meristotheca papulosa Protoplasts

갈래곰보, Meristotheca papulosa의 원형질체 분리와 배양

  • 정규화 (여수대학교 생물공학과) ;
  • 선상미 (여수대학교 생물공학과) ;
  • 조용철 (국립수산진흥원 남해수산연구소) ;
  • 공용근 (국립수산진흥원 남해수산연구소) ;
  • 윤장택 (국립수산진흥원 남해수산연구소)
  • Published : 1999.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Protoplasts were isolated from the vegetative thalli of Meristotheca papulosa using several commercial and crude enzymes. The suitable enzyme combination for the protoplast isolation was 4% abalone acetone powder, 4% Macerozyme R-10 and 4% Hemicellulase in the filtered seawater buffered with 50mM MES (pH 6.0) containing 0.6M mannitol and 0.5% potassium dextran sulfate. Yield of protoplast was $107.6{\times}10^4$ cells per gram of fresh thallus. Protoplasts were whitish ovoid in shape and ranged between $7{\mu} m$~ $24{\mu} m$ in diameter. Division of protoplasts was first observed 9 days after culture in $ASP_{12}$ medium, and the germination occurred within 25 days. The addition of Guillard's antibiotics in culture media was harmful to the regeneration of protoplasts.

갈래곰보의 원혈질체 분리와 배양 조건을 확립하여 생명공학기술에 의한 유전적 형질개량의 기초조건을 마련코자 하였다. Abalone acetone powder,소라의 내장조효소, Pseudomanas와 Vibrio의 조효소, Cellulas R-10, Macerozyme R-10, Hemicellulase, Pectinase, Driselase, Protease 등을 0.6M mannitol과 0.5% potassium dextran sulfate를 함유한 50mM MES 해수완충액(pH 6.0)에 단독 또는 조합하여 조제한 효소액을 갈래곰보의엽체에 처리하였을 때 Abalone acetone powder 4.0% + Macerozyme R-10 4.0% + Hemicellulase 4.0% 효소조합액의 처리로서 생체조직 1g당 $107.6{\times}10^4$개의 원혈질체를 분리시킬 수 있었다. 갓 분리한 원형질체는 투명한 타원형으로 $7{\mu} m$~ $24{\mu} m$의 범위에 분포하였다. 0.2M mannitol을 첨가한 $ASP_{12}$배지에 배양한 세포는 9일 후 분열되고 25일 후 발아하였다. $ASP_{12}$배지는 f/2배지보다 갈래 곰보의 원혈질체 배양에 효과적이었으며 Guillard 항생물질조합액의 처리는 분화에 장애가 되었다.

Keywords

References

  1. Fish. Sci. v.60 Effects of plant growth regulators, organic acids, and sugars on growth and chemical constituents in the tissue culture of sea lettuce Ulva pertusa Amano, H.;H. Noda
  2. Phycologia v.29 Preparation of alginate lyases from marine molluscs for protoplast isolation in brown algae Boyen, C.;B. Kloareg;M. Plone-Fuller;A. Gibor
  3. Bot. Mar. v.30 Protoplast morphogenesis of Porphyra leucosticta in culture Chen, L. C. M.
  4. Bot. Bull. Acad. Sin. v.35 Isolation and regeneration of protoplasts of Monostroma latissimum Wittrock (Monostromataceae, Chlorophyta) Chen, Y. C.;Y. M. Chiang
  5. Beihefte zur applications Hedwigia v.83 Genetic engineering in seaweeds: applications and current status Cheney, D. P.
  6. Int. Rev. Cytol. Suppl. v.16 Plant protoplast isolation and culture Evans, B. A.;J. E. Bravo
  7. Jpn. J. Phycol. v.35 Fusion of protoplasts from thalli of thw different color types in Porphya yezoensis Ueda and development of fusion products Fujita, V.;S. Migita
  8. Hydrobiologia v.204-205 Protoplast isolation and fusion inPorphyra (Bangiales, Rhodophyta) Fujita, Y.;M. Saito
  9. Kor. J. Phycol. v.8 Characterizaiton and genetic analysis of the violet type pigmentation mutant of Porphyra yezoensis Ueda (Bangiales, Rhodopyta) Niwa, K.;A. Miura;J. A. Shin;Y. Aruga
  10. Protoplasma v.97 Behaviour of cultured fusion products from Zygnema and Sprogyra protoplasts Ohiwa, T.
  11. Seaweed Cultivation for Renewable Resoources Tissue of seaweeds Polne-Fuller, M.;A. Gibor;K.T. Bird;P.H. Benson (eds)
  12. Plant Cell Culture Isolation, culture and genetic manipulation of plant protoplasts Power, J. B.;J. V. Chapman
  13. Culture and Collections of Algae, Proc. U.S.-Japan Conf. Hakone, Jap. Soc. Plant Physiol. Media and prospects for cultivation of marine algae Provasoli, L.;A. Watanabe;Hattori (eds)
  14. Bot. Mar. v.32 Protoplast isolation and regeneration of three species of Ulva in axenic culture Reddy, C. R. K.;S. Migita;Y. Fujita
  15. J. Appl. Phycol. v.3 Regeneration of plantlets ffrom Enteromorpha (Ulvales, Chlorophyta) protoplasts in axenic culture Reddy C. R. K.;Y. Fujita
  16. J. Appl. Phycol. v.4 Induction of fast-growing and morphologically different strains through intergeneric protoplast fusions of Ulva and Enteromorpha(Ulvales, Chlorphyta) Reddy, C. R. K.;M. Iima;Y. Fujita
  17. Beihefte zur Nova Hedwigia v.83 Protoplasts from seaweeds: production and fusion Saga, N.;M. Polen-Fuller;A. Gibor
  18. Nippon Suisan Gakkaishi v.59 Application of an alginate lyase from Alteromonas sp. for isolation fo protoplasts from a brown algae Laminaria japonica Sawabe, T.;Y. Ezura;T. Kimura
  19. Nippon Suisan Gakkaishi v.58 Alginate lyase form Vibrio alginolyticus ATCC 17749 Tseng, C.H.;K. Yamaguchi;M. Nishimura
  20. Algal Biotechnology The past, present, and future of tissue culture and biotechnology of seaweeds Van der Meer, J.;T. Stadler;J. Mollion;M.C. Verdus;Y. Karamanos;H. Morvan;D. Christiasn (eds)
  21. Adv. Agron. v.29 The progress, problems and prospects of plant protoplast reserch Vasil, I. K.
  22. Planta v.149 Isolation of acellfusion hormone from Griggithsia pacifica Kylin, a red alga Waaland, S. A.;B. A. Watson
  23. Planta v.181 Protoplasy isolation and regneration in the marine red alga Porphyra nereocystis Waaland, J. R.;L. G. Dickson;B.A. Watson
  24. 해양수산부 유용해조류(훗파래, 톳, 갈래곰부)양식개발시험 국립수산진흥원
  25. 藻類 v.22 トサカノリの胞子發生と生長 新村巖
  26. 藻類 v.22 トサカノリ生育の季節的消長と胞子放出期 新村巖
  27. トサカノリ(食用海藻の栽培) 喜田和四郞;谷口三津夫;三浦昭雄(編著)
  28. 硏究ジャ-ナル v.8 ノリのブロトプラストの作出と個體の再生 鬼頭均