Yannarelli Gustavo G, Noriega Guillermo O, Batlle Alcira, Tomaro Maria L
Departamento de Química Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, Buenos Aires, 1113, Argentina.
Planta. 2006 Oct;224(5):1154-62. doi: 10.1007/s00425-006-0297-x. Epub 2006 May 16.
Ultraviolet-B (UV-B) radiation has a negative impact on plant cells, and leads to the generation of reactive oxygen species (ROS). Heme oxygenase (HO, EC 1.14.99.3) plays a protective role against oxidative stress in mammals, but little is known about this issue in plants. Here, we report for the first time the response of HO in leaves of soybean (Glycine max L.) plants subjected to UV-B radiation. Under 7.5 and 15 kJ m(-2 )UV-B doses, HO, catalase (CAT, EC 1.11.1.6) and ascorbate peroxidase (APX, EC 1.11.1.11) activities were increased and the production of thiobarbituric acid reactive substances (TBARS) regain control values after 4 h of plant recuperation. Treatment with 30 kJ m(-2) UV-B provoked a decrease in these antioxidant enzyme activities. Immunoblot analysis showed a 4.3 and 3.7-fold increase in HO-1 protein expression after irradiation with 7.5 and 15 kJ m(-2), respectively. HO-1 transcript levels were enhanced (up to 77%) at these doses, as assessed by semi-quantitative RT-PCR. These data demonstrated that increased HO activity was associated with augmented protein expression and transcript levels. Plants pre-treated with the antioxidant ascorbic acid did not show the UV-B-induced up-regulation of HO-1 mRNA, but hydrogen peroxide treatment could mimic this reaction. Our results indicate that HO is up-regulated in a dose-depending manner as a mechanism of cell protection against oxidative damage and that such response occurred as a consequence of HO-1 mRNA enhancement involving ROS.
紫外线B(UV-B)辐射对植物细胞有负面影响,并导致活性氧(ROS)的产生。血红素加氧酶(HO,EC 1.14.99.3)在哺乳动物中对氧化应激起保护作用,但在植物中对此问题了解甚少。在此,我们首次报道了大豆(Glycine max L.)植株叶片中HO对UV-B辐射的响应。在7.5和15 kJ m(-2)的UV-B剂量下,HO、过氧化氢酶(CAT,EC 1.11.1.6)和抗坏血酸过氧化物酶(APX,EC 1.11.1.11)的活性增加,并且在植株恢复4小时后,硫代巴比妥酸反应性物质(TBARS)的产生恢复到对照值。用30 kJ m(-2)的UV-B处理导致这些抗氧化酶活性下降。免疫印迹分析表明,在分别用7.5和15 kJ m(-2)照射后,HO-1蛋白表达分别增加了4.3倍和3.7倍。通过半定量RT-PCR评估,在这些剂量下HO-1转录水平增强(高达77%)。这些数据表明,HO活性的增加与蛋白质表达和转录水平的提高相关。用抗氧化剂抗坏血酸预处理的植株未显示出UV-B诱导的HO-1 mRNA上调,但过氧化氢处理可模拟此反应。我们的结果表明,HO以剂量依赖的方式上调,作为细胞保护免受氧化损伤的一种机制,并且这种反应是由于涉及ROS的HO-1 mRNA增强而发生的。