Abstract

The aim of the study was to study the state and reaction of cytoskeletal elements, microtubules and actin filaments, in root cells of Samsun tobacco plants and its transgenic line expressing the FeSOD1 gene from Arabidopsis thaliana with the pea rbcS leader sequence for the localization of the gene product in chloroplasts encoding Fe-dependent superoxide dismutase, constitutively inducing intracellular oxidative stress, by increasing the H₂O₂ pool for a long-term effect of moderate concentrations of NaCl and Na₂SO₄. The main hypothesis was to identify the positive protective effect of controlled constant oxidative stress on the stability of the most sensitive system that provides growth by division and growth by extension (the tubulin cytoskeleton) and effective intracellular transport and structural stability (the actin filament system). Localization of the microtubule cytoskeleton and actin filaments using antibodies to tubulin clone DM1α and actin clone 10-B3 by transmission electron microscopy and immunocytologically, detected by treatment with the second antibodies conjugated with Alexa‑488, made it possible to establish signs of reorganization and disassembly of the actin filament network under the action of NaCl and Na₂SO₄ as in control and in transgenic plants. At the same time, in transgenic plants, differences can be noted even without exposure, which indicates the effectiveness of this method for stimulating a protective response. These data suggest that the state of the system of the tubulin cytoskeleton and actin filaments may be an indicator of the resistance of FeSOD1 transgenic plants to salinity. A relationship has also been established between the reorganization of the cytoskeleton and vacuolization, especially with Na₂SO₄.

中文翻译：

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盐度下Samsun原品种与FeSOD1过表达转基因株系烟草根细胞骨架重组的差异

摘要

本研究的目的是研究萨姆松烟草植物及其转基因系的根细胞中细胞骨架元件、微管和肌动蛋白丝的状态和反应，该转基因系表达来自拟南芥的 FeSOD1 基因，并带有豌豆rbcS前导序列，以定位叶绿体中编码 Fe 依赖性超氧化物歧化酶的基因产物，通过增加 H ₂ O _{2库来持续诱导细胞内氧化应激，以实现中等浓度的 NaCl 和 Na 2} SO ₄的长期作用。主要假设是确定受控恒定氧化应激对最敏感系统稳定性的积极保护作用，该系统提供分裂生长和延伸生长（微管蛋白细胞骨架）以及有效的细胞内运输和结构稳定性（肌动蛋白丝系统） 。使用微管蛋白克隆 DM1α 和肌动蛋白克隆 10-B3 的抗体，通过透射电子显微镜和免疫细胞学方法定位微管细胞骨架和肌动蛋白丝，并通过与 Alexa-488 缀合的第二抗体处理进行检测，从而可以建立重组和分解的迹象对照和转基因植物中肌动蛋白丝网络在 NaCl 和 Na ₂ SO _{4作用下的变化。}同时，在转基因植物中，即使没有暴露，也可以注意到差异，这表明该方法对于刺激保护反应的有效性。这些数据表明微管蛋白细胞骨架和肌动蛋白丝系统的状态可能是FeSOD1转基因植物耐盐性的指标。细胞骨架的重组和空泡化之间也建立了关系，特别是在Na ₂ SO ₄的情况下。