Institute of Plant Nutrition, Leibniz University Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany.
Ann Bot. 2010 Jun;105(7):1119-28. doi: 10.1093/aob/mcq049. Epub 2010 Mar 17.
Aluminium (Al) resistance in common bean is known to be due to exudation of citrate from the root after a lag phase, indicating the induction of gene transcription and protein synthesis. The aims of this study were to identify Al-induced differentially expressed genes and to analyse the expression of candidate genes conferring Al resistance in bean.
The suppression subtractive hybridization (SSH) method was used to identify differentially expressed genes in an Al-resistant bean genotype ('Quimbaya') during the induction period. Using quantitative real-time PCR the expression patterns of selected genes were compared between an Al-resistant and an Al-sensitive genotype ('VAX 1') treated with Al for up to 24 h.
Short-term Al treatment resulted in up-regulation of stress-induced genes and down-regulation of genes involved in metabolism. However, the expressions of genes encoding enzymes involved in citrate metabolism were not significantly affected by Al. Al treatment dramatically increased the expression of common bean expressed sequence tags belonging to the citrate transporter gene family MATE (multidrug and toxin extrusion family protein) in both the Al-resistant and -sensitive genotype in close agreement with Al-induced citrate exudation.
The expression of a citrate transporter MATE gene is crucial for citrate exudation in common bean. However, although the expression of the citrate transporter is a prerequisite for citrate exudation, genotypic Al resistance in common bean particularly depends on the capacity to sustain the synthesis of citrate for maintaining the cytosolic citrate pool that enables exudation.
已知普通豆科植物对铝具有抗性,这种抗性是由于根在滞后阶段后分泌柠檬酸,表明基因转录和蛋白质合成的诱导。本研究的目的是鉴定铝诱导的差异表达基因,并分析赋予豆科植物铝抗性的候选基因的表达。
采用抑制性消减杂交(SSH)方法鉴定在铝诱导期耐铝基因型(“Quimbaya”)中差异表达的基因。使用定量实时 PCR 比较用铝处理长达 24 小时的耐铝和敏感基因型(“VAX 1”)中选定基因的表达模式。
短期铝处理导致应激诱导基因的上调和代谢相关基因的下调。然而,柠檬酸代谢相关酶编码基因的表达不受铝的显著影响。铝处理显著增加了耐铝和敏感基因型中柠檬酸转运体基因家族 MATE(多药和毒素外排家族蛋白)的普通豆科表达序列标签的表达,与铝诱导的柠檬酸分泌密切一致。
柠檬酸转运体 MATE 基因的表达对于普通豆科植物的柠檬酸分泌至关重要。然而,尽管柠檬酸转运体的表达是柠檬酸分泌的前提条件,但普通豆科植物的基因型耐铝特别取决于维持细胞溶质柠檬酸池所需的柠檬酸合成能力,这使得柠檬酸能够分泌。