Roth Udo, von Roepenack-Lahaye Edda, Clemens Stephan
Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany.
J Exp Bot. 2006;57(15):4003-13. doi: 10.1093/jxb/erl170. Epub 2006 Oct 30.
Cadmium is a major environmental pollutant that enters human food via accumulation in crop plants. Responses of plants to cadmium exposure--which directly influence accumulation rates--are not well understood. In general, little is known about stress-elicited changes in plants at the proteome level. Alterations in the root proteome of hydroponically grown Arabidopsis thaliana plants treated with 10 microM Cd(2+) for 24 h are reported here. These conditions trigger the synthesis of phytochelatins (PCs), glutathione-derived metal-binding peptides, shown here as PC2 accumulation. Two-dimensional gel electrophoresis using different pH gradients in the first dimension detected on average approximately 1100 spots per gel type. Forty-one spots indicated significant changes in protein abundance upon Cd(2+) treatment. Seventeen proteins found in 25 spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Selected results were independently confirmed by western analysis and selective enrichment of a protein family (glutathione S-transferases) through affinity chromatography. Most of the identified proteins belong to four different classes: metabolic enzymes such as ATP sulphurylase, glycine hydroxymethyltransferase, and trehalose-6-phosphate phosphatase; glutathione S-transferases; latex allergen-like proteins; and unknown proteins. These results represent a basis for reverse genetics studies to better understand plant responses to toxic metal exposure and to the generation of internal sinks for reduced sulphur.
镉是一种主要的环境污染物,可通过在农作物中积累进入人类食物。植物对镉暴露的反应(直接影响积累速率)尚未得到充分了解。一般来说,对于胁迫引发的植物蛋白质组水平的变化知之甚少。本文报道了水培生长的拟南芥植株在10微摩尔镉离子处理24小时后根蛋白质组的变化。这些条件会触发植物螯合肽(PCs)的合成,PCs是谷胱甘肽衍生的金属结合肽,此处表现为PC2积累。在第一维使用不同pH梯度的双向凝胶电泳平均每类凝胶检测到约1100个斑点。41个斑点表明镉离子处理后蛋白质丰度有显著变化。通过基质辅助激光解吸/电离飞行时间质谱法鉴定了在25个斑点中发现的17种蛋白质。选定的结果通过蛋白质免疫印迹分析以及通过亲和色谱法对一个蛋白质家族(谷胱甘肽S-转移酶)进行选择性富集得到独立验证。大多数鉴定出的蛋白质属于四个不同类别:代谢酶,如ATP硫酸化酶、甘氨酸羟甲基转移酶和海藻糖-6-磷酸磷酸酶;谷胱甘肽S-转移酶;乳胶过敏原样蛋白;以及未知蛋白。这些结果为反向遗传学研究奠定了基础,以更好地理解植物对有毒金属暴露的反应以及对还原态硫内部汇的产生。