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Response of Monodehydroascorbate Reductase in Lettuce Leaves Subjected to Low Temperature Stress

저온 처리한 상추 잎에서 monodehydroascorbate 환원효소의 반응

  • Kang, Sang-Jae (School of Applied Ecological Resources, Kyungpook National University)
  • 강상재 (경북대학교 생태자원응용학부)
  • Received : 2010.11.05
  • Accepted : 2011.03.16
  • Published : 2011.03.30

Abstract

The relationship between MDHA reductase activity and ascorbate, dehydroascorbate, and hydrogen peroxide content was evaluated, and this experiment was conducted to determine the change of MDHA reductase activity and the level of steady-state mRNA abundance of MDHA reductase in lettuce leaves subjected to low temperature stress. MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ for 24 hr increased, followed by a steady decrease during the duration of recovery to $20^{\circ}C$ for 48 hr. The content of ascorbate slowly increased during low temperature treatment, followed by a rapid increase during the duration of recovery to $20^{\circ}C$ for 48 hr, while dehydroascorbate content rapidly decreased. The relationship between MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ and ascorbate content correlated positively ($R^2$=0.9240, 0.9108, respectively), but MDHA reductase activity of chloroplastic and cytosolic fraction and dehydroascorbate were reversely correlated ($R^2$=0.8638, 0.8980, respectively). Hydrogen peroxide content and MDHA reductase activity of chloroplastic and cytosolic fraction in lettuce leaves subjected to $4^{\circ}C$ correlated positively ($R^2$=0.9443, 0.9647, respectively). Northern blot analysis showed that the level of mRNA transcript of MDHA reductase was similar to total activity of MDHA reductase, and also that the level of mRNA of MDHA reductase after recovery to $20^{\circ}C$ for 24 hr decreased.

식물의 저온 적응 메카니즘에서 아스코브산과 관련된 효소 중 MDHA 환원효소의 활성도와 과산화수소, 아스코브산의 함량, mRNA의 발현수준과의 연관성을 연구한 결과는 다음과 같다. MDHA 환원효소의 활성도 변화는 저온에 노출되는 시간이 길어질수록 증가하였으며 6시간 이후에 엽록체분획과 세포질분획에서 급격하게 증가하는 경향을 보였으나 실온으로 회복시켰을 때 효소의 활성도가 상대적으로 감소하는 경향을 보였다. 저온에 노출된 동안 아스코브산의 함량은 비교적 일정한 경향을 보이다가 실온으로 회복시키면 그 이후에는 급격하게 증가하는 경향을 보였다. 반면 저온에 노출되는 동안 급격히 dehydroascorbate 함량이 감소하였다가 실온으로 회복되면 약간 증가하는 경향을 보였다. 아스코브산의 함량과 엽록체분획과 세포질분획의 MDHA 환원효소의 활성도와의 상관관계는 각각 정의 상관($R^2$=0.9240, 0.9108)을 나타내었으나 디하이드로아스코브산의 함량과 MDHA 환원효소의 활성도 사이에는 각각 부의 상관($R^2$=0.8638, 0.8980)을 나타내었다. MDHA 환원효소 활성도와 과산화수소의 함량과의 상관관계를 과산화수소의 생성량이 증가하면 MDHA 환원효소의 활성도가 증가하는 정의 상관($R^2$=0.9443, 0.9647)을 나타내었다. 저온스트레스 처리 시간이 증가할수록 MDHA 환원효소의 mRNA의 발현 수준과 총 MDHA 환원효소의 활성도가 증가하는 경향을 나타내었다.

Keywords

References

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