애기장대에서 액포막 존재 Ca2+-ATPase11 (ACA11) 형질전환체의 세포사멸 표현형 분석
Cell death phenotype of vacuole Ca2+-ATPase11 (ACA11) transgenic plant in Arabidopsis
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작물 스트레스 내성 연구의 궁극적인 과제는 가진 형질을 극대 화하여 생산량을 증대하고 외부의 환경적인 요소로부터 피해를 최 소화하는 것이다. 따라서 작물의 다양한 외부 환경적 스트레스에 대응하기 위한 연구들이 진행되고 있다. 특히 세포 내 신호전달 과 정의 이차매개체인 칼슘에 대한 다양한 연구들이 진행되고 있지만 아직까지 많은 부분이 밝혀져 있지 않다. 본 연구는 모델식물인 애 기장대를 이용하여 세포 내 칼슘의 주요 저장소인 액포로 칼슘을 수송하는 역할을 수행하는 Ca2+-ATPase의 형질전환 식물을 이용하 여 세포 내 칼슘의 신호전달과 식물 생물학적 기능을 알아보았다. ACA11-GFP 유전자가 형질전환된 식물에서 흥미롭게도 ACA11 유전자가 발현 침묵됨으로써 세포 내 칼슘농도 항상성 조절과 신 호전달 과정에 문제가 발생하고 세포질 내 활성산소가 증가되어 결국 형질전환체의 잎에서 HR과 같은 세포사멸을 유발한다는 것 을 제시하였다.
Abstract
Calcium ion (Ca2+) is thought to play the important role as a second messenger for signal transduction that results in various physiological responses to cope with developmental programs and environmental changes in plant. In plant cells, the central vacuole functions as a major calcium store, which is important for both signal transduction and preventing cytotoxicity. Although there is evidence for the biochemical characterizations of a calmodulin-regulated Ca2+-ATPase (ACA11) localized to vacuole membrane, the biological function to ACA11 in plant has not been verified. In this study, we show that the cell death as the hypersensitive response (HR) in mature leaves is induced in transgenic plant of a vacuole ACA-type Ca2+-ATPase, ACA11. Evidence that cell death phenotype is the result of ACA11 gene silencing is provided by Western blot assay using membrane fraction proteins extracted from transgenic plant. The 3, 3’-diaminobenzidine (DAB) staining study provides that the cell death is caused by the increase of reactive oxygen species (ROS) in mature leaves of transgenic plants. Key words: Ca2+-ATPase, vacuole, hypersensitive response, Arabidopsis
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