Departamento de Biologia Vegetal, Instituto de Biologia, CP 6109, Universidade Estadual de Campinas, Campinas, SP, 13083-970, Brazil.
Biometals. 2011 Apr;24(2):225-37. doi: 10.1007/s10534-010-9388-z. Epub 2010 Nov 3.
Iron (Fe) is an essential nutrient for plants, but it can generate oxidative stress at high concentrations. In this study, Coffea arabica L. cell suspension cultures were exposed to excess Fe (60 and 240 μM) to investigate changes in the gene expression of ferritin and antioxidant enzymes. Iron content accumulated during cell growth, and Western blot analysis showed an increase of ferritin in cells treated with Fe. The expression of two ferritin genes retrieved from the Brazilian coffee EST database was studied. CaFER1, but not CaFER2, transcripts were induced by Fe exposure. Phylogenetic analysis revealed that CaFER1 is not similar to CaFER2 or to any ferritin that has been characterised in detail. The increase in ferritin gene expression was accompanied by an increase in the activity of antioxidant enzymes. Superoxide dismutase, guaiacol peroxidase, catalase, and glutathione reductase activities increased in cells grown in the presence of excess Fe, especially at 60 μM, while the activity of glutathione S-transferase decreased. These data suggest that Fe induces oxidative stress in coffee cell suspension cultures and that ferritin participates in the antioxidant system to protect cells against oxidative damage. Thus, cellular Fe concentrations must be finely regulated to avoid cellular damage most likely caused by increased oxidative stress induced by Fe. However, transcriptional analyses indicate that ferritin genes are differentially controlled, as only CaFER1 expression was responsive to Fe treatment.
铁(Fe)是植物必需的营养物质,但在高浓度下会产生氧化应激。本研究中,采用过量铁(60 和 240 μM)处理咖啡悬浮细胞培养物,以研究铁处理对铁蛋白和抗氧化酶基因表达的变化。细胞生长过程中铁含量增加,Western blot 分析显示铁处理细胞中铁蛋白增加。从巴西咖啡 EST 数据库中检索到的两个铁蛋白基因的表达进行了研究。CaFER1 而非 CaFER2 的转录物被铁暴露诱导。系统发生分析表明,CaFER1 与 CaFER2 或任何详细表征的铁蛋白均不相似。铁蛋白基因表达增加伴随着抗氧化酶活性的增加。超氧化物歧化酶、愈创木酚过氧化物酶、过氧化氢酶和谷胱甘肽还原酶的活性在含有过量铁的细胞中增加,特别是在 60 μM 时,而谷胱甘肽 S-转移酶的活性降低。这些数据表明,铁诱导咖啡悬浮细胞培养物的氧化应激,铁蛋白参与抗氧化系统以保护细胞免受氧化损伤。因此,细胞内铁浓度必须精细调节,以避免可能由铁诱导的氧化应激增加引起的细胞损伤。然而,转录分析表明铁蛋白基因受到不同的调控,因为只有 CaFER1 的表达对铁处理有反应。