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Effect of Yeast CTA1 Gene Expression on Response of Tobacco Plants to Tobacco Mosaic Virus Infection

机译:酵母CTA1基因表达对烟草植株对烟草花叶病毒感染应答的影响

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摘要

The response of tobacco (Nicotiana tabacum L. cv Xanthi-nc) plants with elevated catalase activity was studied after infection by tobacco mosaic virus (TMV). These plants contain the yeast (Saccharomyces cerevisiae) peroxisomal catalase gene CTA1 under the control of the cauliflower mosaic virus 35S promoter. The transgenic lines exhibited 2- to 4-fold higher total in vitro catalase activity than untransformed control plants under normal growth conditions. Cellular localization of the CTA1 protein was established using immunocytochemical analysis. Gold particles were detected mainly inside peroxisomes, whereas no significant labeling was detected in other cellular compartments or in the intercellular space. The physiological state of the transgenic plants was evaluated in respect to growth rate, general appearance, carbohydrate content, and dry weight. No significant differences were recorded in comparison with non-transgenic tobacco plants. The 3,3′-diaminobenzidine-stain method was applied to visualize hydrogen peroxide (H2O2) in the TMV infected tissue. Presence of H2O2 could be detected around necrotic lesions caused by TMV infection in non-transgenic plants but to a much lesser extent in the CTA1 transgenic plants. In addition, the size of necrotic lesions was significantly bigger in the infected leaves of the transgenic plants. Changes in the distribution of H2O2 and in lesion formation were not reflected by changes in salicylic acid production. In contrast to the local response, the systemic response in upper noninoculated leaves of both CTA1 transgenic and control plants was similar. This suggests that increased cellular catalase activity influences local but not systemic response to TMV infection.
机译:烟草花叶病毒(TMV)感染后,研究过氧化氢酶活性升高的烟草(Nicotiana tabacum L. cv Xanthi-nc)植物的响应。这些植物在花椰菜花叶病毒35S启动子的控制下含有酵母(Saccharomyces cerevisiae)过氧化物酶体过氧化氢酶基因CTA1。在正常生长条件下,转基因品系的总体外过氧化氢酶活性比未转化的对照植物高2-4倍。使用免疫细胞化学分析确定CTA1蛋白的细胞定位。主要在过氧化物酶体内部检测到金颗粒,而在其他细胞室或细胞间空间中未检测到明显的标记。关于生长速率,总体外观,碳水化合物含量和干重,评估了转基因植物的生理状态。与非转基因烟草植物相比,没有明显差异。应用3,3'-二氨基联苯胺染色法观察TMV感染组织中的过氧化氢(H2O2)。在非转基因植物中,TMV感染引起的坏死病灶周围可以检测到H2O2的存在,而在CTA1转基因植物中,H2O2的存在程度要小得多。另外,在转基因植物的感染叶片中,坏死病灶的大小明显更大。水杨酸产量的变化并未反映出H2O2分布和病变形成的变化。与局部反应相反,在CTA1转基因植物和对照植物的上部未接种叶片中的系统反应相似。这表明增加的细胞过氧化氢酶活性影响对TMV感染的局部反应,但不影响全身反应。

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