Superoxide Dismutase Activity in Suspension Cultured Cells of Tomato (Lycopersicon esculentum Mill)

토마토(Lycopersicon esculentum Mill) 현탁배양세포에서 Superoxide Dismutase 활성

  • 유순희 (한국과학기술연구원 생명공학연구소 식물생화학 Research Unit, 충남대학교 생물학과) ;
  • 허경혜 (한국과학기술연구원 생명공학연구소 식물생화학 Research Unit) ;
  • 권석윤 (한국과학기술연구원 생명공학연구소 식물생화학 Research Unit) ;
  • 이행순 (한국과학기술연구원 생명공학연구소 식물생화학 Research Unit) ;
  • 방재욱 (충남대학교 생물학과) ;
  • 곽상수 (한국과학기술연구원 생명공학연구소 식물생화학 Research Unit)
  • Published : 1997.01.01

Abstract

We investigated changes in the superoxide dismutase (SOD) activity and SOD isoenzyme pattern in suspension cultures of tomato (Lycopersicon esculentum), which were compared with those of intact tomato plants. two grams (fr wt) of cells subcultured at 15-day intervals were inoculated into 50 mL MS medium containing l mg/L 2,4-D and 30 g/L sucrose in a 300 mL flask and maintained at $25^{\circ}C$ in the dark (100 rpm). The cell growth reached a maximum at 20 days after subculture (DAS), followed by a rapid decrease with further cultures. The cell colour changed from white to black from 23 DAS. The intracellular SOD activity (units/g cell dry wt) was significantly increased from 23 DAS and reached a maximum at 28 DAS (52,400 units), followed by a decrease with further cultures, whereas the extracellular SOD activity showed a maximum at 25 DAS (27,800 units/50 mL medium). The total SOD activity per flask showed a maximum at 25 DAS (35,700 units), in which the extracellular SOD activity occupied about 75%. The tomato cultured cells had four SOD isoenzymes and their patterns were well correlated with SOD activity without a qualitative change during the cell cultures. The intact tomato plants had an additional CuZnSOD isoenzyme, showing the different isoenzyme patterns from cultured cells.

Superoxide dismutase (SOD) 고생산세포주로 선발된 토마토(Lycopersicun esculentum) 배양세포를 사용하여 현탁배양에 따른 SOD 활성과 isoenzyme변화를 조사하고 토마토 식물체의 것과 비교하였다. 현탁배양은 세포생중량 2 g을 1 mg/L 2,4-D, 30 g/L sucrose를 함유한 MS 배지 50 mL과 함께 mL flask에서 $25^{\circ}C$암상태로 배양(100 rpm)하였다. 세포생장은 계대배양후 20일에 최고점에 도달한 후, 급격히 감소하며 배양 후 23일부터 세포가 검게 변하였다. 세포 단위무게당 SOD활성(unit/g dry cell wt)은 배양 후 23일부터 증가하여 28일째에 최고활성(52,400 unit)을 나타낸 후 급격히 감소하였다. 세포 밖으로 분비되는 extracellular SOD활성은 배양 후 25일에 최고치(27,800 unit/so mL medium)를 나타낸 후 감소하였다. Flask 전체의 SOD활성은 배양 후 25일에 최대치(35,700 unit)를 나타내었으며 extracellular SOD 활성이 약 75%을 차지하였다. 토마토 배양세포에는 4개의 SOD isoenzyme이 존재하며, isoenzyme의 패턴변화는 세포생장에 따른 효소활성의 변화와 일치하였다. 토마토 식물체는 배양세포에 없는 CuZnSOD가 존재하며 배양세포와 식물체 조직사이에는 서로 다른 isoenzyme 패턴이 존재함을 알 수 있었다.

Keywords

References

  1. Plant Physiol v.107 Dissection of oxidative stress tolerance using transgenic plants Allen RD
  2. Antioxidants in Higher Plants Alscher RG;Hess JL
  3. Anal Biochem v.44 Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels Beauchamp C;Fidovich I
  4. Crit Rev Plant Sci v.13 Superoxide dismutase in plants Bowler C
  5. Annu. Rev. Plant Physiol. Plant Mol. Biol v.43 Superoxide dismutase and stress tolerance Bowler C;Van Montagu M;Inze D
  6. Anal Biochem v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford MM
  7. Biosci Biotech Biochem v.59 Formation of phenolic compounds from the callus of bangah (Isodon japonicus Hara) Choi KS;Lee JG;Lee HY
  8. Plant Cell Physiol v.25 Paraquat resistant tobacco calluses with enhanced superoxide dismutase activity Furusawa I;Tanaka K;Thanutong P;Mizuguchi A;Yazaki M;Asada K
  9. Molecular and General Genetics Molecular cloning and characterization of anioric and neutral peroxidase cDNAs from sweet potato suspension-cultured cells and their differential expression in response to stress Huh, GH;Lee SJ;Bae YS;Liu JR;Kwak SS
  10. Korean J Plant Tissue Culture v.23 Comparison of catalase and other antioxidant enzyme activities in various plant cell lines Jang MS;Huh GH;Kim SW;Park IH;Liu JR;Kwak SS
  11. Molecular Aspects of Enzyme Catalysis Superoxide dismutase Kanematsu S;Asada K;T Fukui(ed.);K Soda(ed.)
  12. Korean J Bioch v.27 Selection of plant cell lines for high yields of peroxidase Kim SK;Kwak SS;Jung KH;Min SR;Park IH;Liu JR
  13. Phytochmistry v.39 Acidic peroxidase from suspension cultures of sweet potato Kwak SS;Kim SK;Lee MS;Jung KH;Park IH;Liu JR
  14. Phytochmistry v.43 Enhancement of peroxidase activity by stress-related chemicals in suspension cultures of sweet potato Kwak SS;Kim SK;Park IH;Liu JR
  15. Biochim Biophys Acta v.874 Isolation and characterization of cytosolic and chloroplastic isozymes of Cu, Zn superoxide dismutase from tomato leaves and their relationships to other Cu, Zn superoxide dismutases Kwiatowski J;Kaniuga Z
  16. Eur J Biochem v.146 Isolation and characterization of an iron-containing superoxide dismutase from tomato leaves Lycopersicun esculentum Kwiatowsk J;Safianowska A;Kaniuga Z
  17. J. Biol Chem v.244 Superoxide dismutase. An enzymic function for erythrocuprein (Hemocuprein) McCord JM;Fridovich I
  18. Physiol Plant v.15 A revised medium for rapid growth and bioassay with tobacco tissue cultures Murashige T;Skoog F
  19. SOD and Active Oxygen Modulators: pharmacology and clinical trials Oyanagui Y
  20. Plant Molecular Biology v.17 The tomato Cu, Zn superoxide dismutase genes are developmentally regulated and respond to light and stress Perl-Treves R;Galun E
  21. Arch Biochem Biophys v.225 Positive correlation between superoxide dismutase and resistance to paraquat toxicity in the green alga Chlorella sorokiniana Rabinowitch HD;Clare DA;Crapo JD;Fridovich I
  22. Plant Physiol v.101 Oxygen stress and superoxide dismutase Scandalios JG
  23. Plant Physiol v.103 Overexpression of superoxide dismutase protects plants from oxidative stress Sen Gupta A;Webb RP;Holaday AS;Allen RD
  24. Korean J Pant Tissue Culture v.23 Selection and isenzyme analysis of plant cell lines for high yields of superexide dismutase You SH;Kim SW;Kim SH;Liu JR;Kwak SS