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用1.0mmol/L FeSO_4/0.5mmol/L H_2O_2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(ΔΨ_m)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(phosphatidylserine,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。
Protoplasts of tobacco (Nicotiana tabacum L., cultivar BY-2) were treated with 1.0 mmol / L FeSO 4 / 0.5 mmol / L H 2 O 2 and found that hydroxyl radical induced the apoptosis of tobacco protoplasts. Specific manifestations of nuclear shrinkage, DNA Ladder, TUNEL-positive and other typical characteristics of apoptosis. Mitochondria play a very important role in animal apoptosis, in which the change of membrane potential (ΔΨ_m) and the opening of the permeable pore (PTP) located on the mitochondrial membrane are related to the release of Cyt c. In addition, in animal apoptotic cells, phosphatidylserine (PS) turns outwards from the inside of the cell membrane. In order to determine the change of membrane potential during plant cell apoptosis and the degree of eversion of PS, we used flow cytometry. The results showed that the mitochondrial membrane potential of tobacco protoplasts decreased gradually with the prolongation of treatment time. The results showed that the apoptosis system consisting of hydroxyl radical and tobacco protoplast is a reliable combination of apoptosis and can be used to further study the mechanism of apoptosis in plant cells.