Effect of salt on ROS homeostasis, lipid peroxidation and antioxidant mechanisms in Pinus pinaster suspension cells
Effect of salt on ROS homeostasis, lipid peroxidation and antioxidant mechanisms in Pinus pinaster suspension cells
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DOI：
10.1051/forest/2008093
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发表时间：
2009-03-01
影响因子：
3
通讯作者：
Tavares, Rui M.
中科院分区：
文献类型：
--
作者：
Azevedo, Herlander;Amorim-Silva, Vitor;Tavares, Rui M.
在松属植物中，缺乏关于暴露于盐期间氧化防御机制有效性的信息。使用海松 (Pinus pinaster Ait.) 悬浮细胞作为模型系统，研究盐胁迫对 ROS 稳态的影响。将细胞维持在基于 MS 的培养基中，暴露于盐（50、100 和 150 mM NaCl），并分析生物量产生，证明生长能力下降。使用 100 mM NaCl 会施加严重的盐胁迫，但不会影响细胞活力，被选择用于盐休克处理的悬浮细胞的 ROS 稳态的后续研究。在盐暴露的第二天，总 ROS 水平明显增加，但超氧离子瞬时爆发立即引人注目。此外，脂质过氧化物的形成似乎与超氧离子分解相关。凝胶内超氧化物歧化酶活性证明 FeSOD 同二聚体在施加盐胁迫的 12-48 小时之间活性急剧增加。随后，从 cDNA 文库中分离出 P. pinaster Fe-Sod1 和 csApx1 基因，表达在 12-24 小时内增加。结果表明，尽管诱导了抗氧化系统，但严重的盐处理仍会在 P. pinaster 细胞中产生氧化应激，并表明 ROS 可能参与盐应激信号传导。
In the Pinus genus, information on the effectiveness of oxidative defence mechanisms during exposure to salt is lacking. The effect of salt stress imposition on ROS homeostasis was investigated using maritime pine (Pinus pinaster Ait.) suspension cells as a model system.Cells were maintained in MS-based medium, exposed to salt (50, 100 and 150 mM NaCl) and analysed for biomass production, evidencing a decreasing growth capacity. Use of 100 mM NaCl imposed severe salt stress without affecting cell viability, being chosen for subsequent studies on the ROS homeostasis of salt shock-treated suspension cells.Increased total ROS levels were evident on the second day of salt exposure, but a superoxide ion transient burst was immediately noticeable. Additionally, lipid peroxide formation seemed to correlate with superoxide ion breakdown. In-gel superoxide dismutase activity evidenced a FeSOD homodimer with strongly increasing activity between hours 12-48 of salt stress imposition. Subsequently, P. pinaster Fe-Sod1 and csApx1 genes were isolated from a cDNA library and expression was shown to increase within 12-24 h.Results show that severe salt treatment generates oxidative stress in P. pinaster cells despite the induction of antioxidant systems, and suggest a putative involvement of ROS in salt stress signalling.