Talarczyk Andrzej, Krzymowska Magdalena, Borucki Wojciech, Hennig Jacek
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.
Plant Physiol. 2002 Jul;129(3):1032-44. doi: 10.1104/pp.010960.
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 (H(2)O(2)) in the TMV infected tissue. Presence of H(2)O(2) 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 H(2)O(2) 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.
研究了过氧化氢酶活性升高的烟草(Nicotiana tabacum L. cv Xanthi-nc)植株在感染烟草花叶病毒(TMV)后的反应。这些植株含有受花椰菜花叶病毒35S启动子控制的酵母(Saccharomyces cerevisiae)过氧化物酶体过氧化氢酶基因CTA1。在正常生长条件下,转基因株系的体外总过氧化氢酶活性比未转化的对照植株高2至4倍。使用免疫细胞化学分析确定了CTA1蛋白的细胞定位。金颗粒主要在过氧化物酶体内被检测到,而在其他细胞区室或细胞间隙中未检测到明显的标记。从生长速率、总体外观、碳水化合物含量和干重方面评估了转基因植株的生理状态。与非转基因烟草植株相比,未记录到显著差异。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(H(2)O(2))。在非转基因植株中,可在TMV感染引起的坏死病斑周围检测到H(2)O(2),但在CTA1转基因植株中检测到的程度要小得多。此外,转基因植株受感染叶片上坏死病斑的尺寸明显更大。水杨酸产生的变化并未反映H(2)O(2)分布和病斑形成的变化。与局部反应不同,CTA1转基因植株和对照植株上部未接种叶片的系统反应相似。这表明细胞过氧化氢酶活性的增加影响对TMV感染的局部反应而非系统反应。