Almeida José M, Fidalgo Fernanda, Confraria Ana, Santos Arlete, Pires Helena, Santos Isabel
Institute for Molecular and Cellular Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal.
Funct Plant Biol. 2005 Sep;32(8):707-720. doi: 10.1071/FP04235.
The effect of hydrogen peroxide (HO) on catalase (CAT) isoform activities and amounts and on mRNA levels was studied in leaves from potato plants untreated and treated with homobrassinolide (HBR). Northern blot analysis revealed that 100 mm HO supplied through the leaf petiole for 4 h did not induce CAT expression. In contrast, CAT1 and CAT2 responded differently to longer treatment, as CAT2 transcript levels increased markedly whereas CAT1 transcript levels remained unchanged. Western blot analysis showed disparity between the level of CAT1 transcript and CAT1 amount, which actually decreased after 28 h. CAT2 amount correlated well with transcript accumulation and CAT2 activity as visualised by zymogram analysis. HO modified the relative importance of CAT isoforms. After 4 h, CAT1 was prevalent in untreated and HO-treated leaves. After 28 h, CAT2 was prevalent in HO-treated leaves; therefore, the quantified increase in total CAT activity in these leaves was due to the rise in CAT2. HBR pre-treatment increased CAT2 basal level not changing the pattern of CAT responses to HO, only lowering its amplitude. Even so, ultrastructural studies showed that HBR significantly reduced HO negative effects on cellular sub-structures, allowing better recovery of affected structures and reducing the macroscopic injury symptoms on leaves, thus data point to a HBR protective role.
研究了过氧化氢(HO)对未经处理和用高油菜素内酯(HBR)处理的马铃薯植株叶片中过氧化氢酶(CAT)同工型活性、含量及mRNA水平的影响。Northern印迹分析表明,通过叶柄供应100 mM HO 4小时未诱导CAT表达。相反,CAT1和CAT2对较长时间处理的反应不同,因为CAT2转录水平显著增加而CAT1转录水平保持不变。蛋白质免疫印迹分析显示CAT1转录水平与CAT1含量之间存在差异,实际上在28小时后CAT1含量下降。通过酶谱分析可见,CAT2含量与转录物积累及CAT2活性密切相关。HO改变了CAT同工型的相对重要性。4小时后,CAT1在未处理和HO处理的叶片中占主导。28小时后,CAT2在HO处理的叶片中占主导;因此,这些叶片中总CAT活性的定量增加是由于CAT2的增加。HBR预处理增加了CAT2基础水平,并未改变CAT对HO的反应模式,只是降低了其幅度。即便如此,超微结构研究表明,HBR显著降低了HO对细胞亚结构的负面影响,使受影响的结构能更好地恢复,并减轻叶片上的宏观损伤症状,因此数据表明HBR具有保护作用。