a Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere , Kyoto University , Kyoto , Japan.
b Department of Molecular Pharmacology, Graduate School of Pharmaceutical Sciences , Kyoto University , Kyoto , Japan.
Int J Phytoremediation. 2018;20(14):1427-1437. doi: 10.1080/15226514.2018.1501333. Epub 2019 Jan 17.
Large quantities of Fe and Cd accumulate in the leaves of the metal-accumulating leguminous plant, Crotalaria juncea. A member of the metal transporter NRAMP family was cloned from C. juncea. The amino acid sequence of this clone, designated CjNRAMP1, was similar to the sequence of Arabidopsis AtNRAMP1, which is involved in Fe and Cd transport. Organ-specific analysis showed that CjNRAMP1 mRNA was expressed mainly in the leaves of C. juncea plants, as well as in stems and roots. Use of green fluorescent protein fused to CjNRAMP1 suggested its localization to the plasma membranes of plant cells. Complementation experiments using yeast strains with impaired metal transport systems showed that CjNRAMP1 transported both Fe and Cd in an inward direction within the cells. Transgenic Arabidopsis plants overexpressing CjNRAMP1 showed high tolerance to Cd, with Cd translocation from roots to leaves being substantially greater in transgenic than in wild-type plants. Overexpression of CjNRAMP1 resulted in a greater accumulation of Fe in shoots and roots, suggesting that CjNRAMP1 recognizes Fe and Cd as substrates and that the high Cd tolerance of CjNRAMP1 is due to its strong Fe uptake activity, even in the presence of high Cd concentrations in the rhizosphere.
大量的铁和镉在金属积累豆科植物豇豆的叶子中积累。从豇豆中克隆出一种属于金属转运体 NRAMP 家族的成员。该克隆物的氨基酸序列被命名为 CjNRAMP1,与参与铁和镉转运的拟南芥 AtNRAMP1 的序列相似。组织特异性分析表明,CjNRAMP1 mRNA 主要在豇豆植株的叶片中表达,在茎和根中也有表达。用绿色荧光蛋白融合 CjNRAMP1 表明其定位于植物细胞的质膜上。利用金属转运系统受损的酵母菌株进行的互补实验表明,CjNRAMP1 在细胞内向内运输铁和镉。过表达 CjNRAMP1 的转基因拟南芥植物对 Cd 具有高耐受性,转基因植物中 Cd 从根部向叶片的转运明显大于野生型植物。CjNRAMP1 的过表达导致地上部和根中 Fe 的积累增加,表明 CjNRAMP1 将 Fe 和 Cd 识别为底物,并且 CjNRAMP1 的高 Cd 耐受性归因于其对 Fe 的强烈摄取活性,即使在根际存在高浓度的 Cd 时也是如此。