Kobayashi Yuriko, Hoekenga Owen A, Itoh Hirotaka, Nakashima Midori, Saito Shoichiro, Shaff Jon E, Maron Lyza G, Piñeros Miguel A, Kochian Leon V, Koyama Hiroyuki
Laboratory of Plant Cell Technology, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido Gifu 501-1193, Japan.
Plant Physiol. 2007 Nov;145(3):843-52. doi: 10.1104/pp.107.102335. Epub 2007 Sep 20.
Malate transporters play a critical role in aluminum (Al) tolerance responses for some plant species, such as Arabidopsis (Arabidopsis thaliana). Here, we further characterize AtALMT1, an Arabidopsis aluminum-activated malate transporter, to clarify its specific role in malate release and Al stress responses. Malate excretion from the roots of accession Columbia was sharply induced by Al, which is concomitant with the induction of AtALMT1 gene expression. The malate release was specific for Al among rhizotoxic stressors, namely cadmium, copper, erbium, lanthanum, sodium, and low pH, which accounts for the specific sensitivity of a null mutant to Al stress. Al-specific malate excretion can be explained by a combined regulation of AtALMT1 expression and activation of AtALMT1 protein, which is specific for Al. Although low pH treatment slightly induced gene expression, other treatments did not. In addition, malate excretion in Al-activated seedlings was rapidly stopped by removing Al from the solution. Other rhizotoxic stressors were not effective in maintaining malate release. Protein kinase and phosphatase inhibitor studies indicated that reversible phosphorylation was important for the transcriptional and posttranslational regulation of AtALMT1. AtALMT1 promoter-beta-glucuronidase fusion lines revealed that AtALMT1 has restricted expression within the root, such that unnecessary carbon loss is likely minimized. Lastly, a natural nonsense mutation allele of AtALMT1 was identified from the Al-hypersensitive natural accession Warschau-1.
苹果酸转运蛋白在某些植物物种(如拟南芥)对铝(Al)的耐受性反应中起着关键作用。在这里,我们进一步表征了拟南芥铝激活苹果酸转运蛋白AtALMT1,以阐明其在苹果酸释放和铝胁迫反应中的具体作用。哥伦比亚生态型根中的苹果酸分泌受到铝的强烈诱导,这与AtALMT1基因表达的诱导同时发生。在镉、铜、铒、镧、钠和低pH等根毒性胁迫因子中,苹果酸释放对铝具有特异性,这解释了一个无效突变体对铝胁迫的特异性敏感性。铝特异性苹果酸分泌可以通过AtALMT1表达的联合调控和AtALMT1蛋白的激活来解释,AtALMT1蛋白对铝具有特异性。虽然低pH处理轻微诱导了基因表达,但其他处理没有。此外,通过从溶液中去除铝,铝激活幼苗中的苹果酸分泌迅速停止。其他根毒性胁迫因子在维持苹果酸释放方面无效。蛋白激酶和磷酸酶抑制剂研究表明,可逆磷酸化对AtALMT1的转录和翻译后调控很重要。AtALMT1启动子-β-葡萄糖醛酸酶融合系表明,AtALMT1在根内的表达受到限制,从而可能最大限度地减少不必要的碳损失。最后,从对铝敏感的自然生态型Warschau-1中鉴定出AtALMT1的一个自然无义突变等位基因。