CAJ | 학술논문

用1.0 mmol/L FeSO4/0.5 mmol/L H202处理烟草(Nicotiana tabacum L.cultivar BY 2)原生质体,发现羟自由基能够诱导烟草原生质体的凋亡.具体表现为细胞核皱缩、DNA Ladder、TUNEL阳性反应等典型的凋亡特征.在动物细胞凋亡过程中,线粒体起着非常重要的作用,其中膜电位(△ψm)的变化以及由其引起的位于线粒体膜上的通透性孔(PTP)的开放与Cyt c的释放有关.另外,在动物凋亡细胞中,磷脂酰丝氨酸(phosphatidyl serine,PS)会从细胞膜内侧向外翻转.为了判断植物细胞凋亡过程中膜电位的变化情况以及PS的外翻程度,我们采用了流式细胞法.结果表明,随着处理时间的延长,烟草原生质体线粒体的膜电位逐渐降低;膜内PS大量外翻.说明由羟自由基和烟草原生质体组成的凋亡体系是一种可靠的凋亡组合,可以用来对植物细胞凋亡机理做进一步研究.
용1.0 mmol/L FeSO4/0.5 mmol/L H202처리연초(Nicotiana tabacum L.cultivar BY 2)원생질체,발현간자유기능구유도연초원생질체적조망.구체표현위세포핵추축、DNA Ladder、TUNEL양성반응등전형적조망특정.재동물세포조망과정중,선립체기착비상중요적작용,기중막전위(△ψm)적변화이급유기인기적위우선립체막상적통투성공(PTP)적개방여Cyt c적석방유관.령외,재동물조망세포중,린지선사안산(phosphatidyl serine,PS)회종세포막내측향외번전.위료판단식물세포조망과정중막전위적변화정황이급PS적외번정도,아문채용료류식세포법.결과표명,수착처리시간적연장,연초원생질체선립체적막전위축점강저;막내PS대량외번.설명유간자유기화연초원생질체조성적조망체계시일충가고적조망조합,가이용래대식물세포조망궤리주진일보연구.
Protoplasts prepared from tobacco (Nicotiana tabacum L., cultivar BY-2) suspension cellshave similar morphological characteristics to those in animal cells. The hallmarks of apoptosis such ascondensation and peripheral distribution of nuclei, TUNEL positive reaction, and DNA ladders were ob-served when tobacco protoplasts were treated with the hydroxyl radical generating system (1.0 mmol/LFeSO4/0.5 mmol/L H2O2). In animals, the loss of transmembrane potential (△ψm ) and the exposure ofphospholipid phosphatidylserine (PS) are believed to be the main apoptosis events. To test whether thesesignificant processes take place in plants, flow cytometry was used to detect annexin V binding andchanges in △ψm. Results showed that the PS turned out from inner membrane and △ψm graduallydecreased during the apoptosis. All these apoptotic characteristics proved that hydroxyl radicals cancause typical programmed cell death (PCD) in tobacco protoplasts and this design can be served as aneffective experiment system to explore the mechanism of plant apoptosis.