烟草悬浮细胞在镉胁迫下微丝形态与黏弹性变化的实时监测
Real-time Monitoring of Microfilament Morphology and Viscoelasticity of Tobacco Suspension Cells under Cadmium Stress
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摘要: 本文利用增强型绿色荧光蛋白标记微丝(Lifeact-EGFP)的烟草为供试材料，在LumascopeTM720全自动活细胞成像系统下观察了10-4~10-8 mol·L-1镉离子胁迫下及空白对照的Lifeact-EGFP烟草悬浮细胞的微丝聚合-解聚情况的实时变化。使用石英晶体微天平技术监测了10-4~10-8 mol·L-1镉离子胁迫下及空白对照的烟草悬浮细胞黏弹性实时变化。进而测定了10-4~10-8 mol·L-1镉离子胁迫下及空白对照的烟草悬浮细胞的过氧化氢酶活力变化情况。研究发现，在镉逆境胁迫早期，烟草悬浮细胞的微丝结构、细胞黏弹性指数变化和逆境相关酶活力变化存在一定程度的关联。本研究尝试从细胞层面探索镉毒害对烟草细胞生理特征的影响，为镉胁迫下植物细胞生理生化研究提供了新证据。Abstract: In this paper, using tobacco with enhanced green fluorescent protein labeled microfilament (Lifeact-EGFP) as the test material, real-time changes in the polymerization and de-polymerization of microfilaments of Lifeact-EGFP tobacco suspension cells in the presence and absence of cadmium stress (10-4~10-8 mol·L-1) were observed under the LumascopeTM 720 automatic live cell imaging system. Quartz crystal microbalance (QCM) technology was used to monitor the real-time changes of viscoelasticity of tobacco suspension cells under the stress of 10-4~10-8 mol·L-1 cadmium ion and in the absence of cadmium ion. Moreover, the changes in catalase activity of tobacco suspension cells under the stress of 10-4 to 10-8 mol·L-1 cadmium ion and the blank control without cadmium ion were measured. It was found that changes in the microfilament structure, cell viscoelasticity index and cadmium stress related enzyme activity of tobacco suspension cells were correlated to some extent in the early stage of cadmium stress. This study attempts to explore the effects of cadmium toxicity on the physiological characteristics of tobacco cells from the cellular level, and provides new evidence for the physiological and biochemical research of plant cells under cadmium stress.
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Key words:
- cadmium stress /
- tobacco suspension cells /
- cytoskeleton /
- microfilament /
- cell viscoelastic index /
- CAT /
- dynamic monitoring
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