紫花苜蓿对铜的吸收、积累和耐受机制研究

采用原位微区X射线荧光分析法(In-situ micro-X-ray fluorescence spectrometry, μ-XRF)和分步提取技术, 研究了苜蓿幼苗中Cu原位微区分布特征及其根茎叶中Cu的不同结合形态.结果表明, 苜蓿根部富集Cu浓度高达12.06 mg/g, 是茎的8倍, 叶的4.9倍.微区分布结果表明, 过量Cu暴露下的苜蓿根部是富集Cu的主要部位, 且在根茎交界处存在Cu的阻隔屏障, 以减轻过量Cu对地上部分的毒害作用.过量Cu(大于50 μmol/L时)还抑制了苜蓿对Zn和Ca的向上吸收, 增强了苜蓿对Fe的吸收, 但对K和Mn的吸收影响不明显.亚细胞分步提取结果表明, 根细胞中主要通过形成难溶的残渣态(41%)和细胞壁螯合态(20%)实现对过量Cu的固定;而在茎中则以疏水蛋白质结合态、细胞壁结合态、残渣态、水溶态等4种形式存在, 以进一步减少过量Cu向叶中输送;在叶中, 进入叶细胞的过量Cu主要以液泡区隔和难溶残渣态形式存在, 以实现对Cu的耐受和解毒.

Study on Copper Absorption,Accumulation and Tolerance Mechanism of Alfalfa

To reveal the mechanism of Cu enrichment of alfalfa, in-situ micro-X-ray fluorescence spectrometry (μ-XRF) and fractional extraction were used to explore in-situ Cu distribution information in alfalfa seedlings and different combination forms of Cu in organs.The results showed that alfalfa roots were enriched with Cu up to 12.06 mg/g, which was 8 times of stem and 4.9 times of leaves.The in-situ μ-XRF result showed that the root of alfalfa was the main site of enrichment of Cu, and there was a barrier of Cu at the rhizome junction to alleviate the toxic effect of excess Cu on the shoots.Excess Cu (more than 50 μmol/L) also inhibited the uptake of Zn and Ca in alfalfa, enhanced the uptake of Fe in alfalfa, but had no obvious effect on the uptake of K and Mn.Plant fractional extraction showed that the Cu in root cells was mainly fixed in the form of insoluble residues (41%) and cell wall chelate (20%), while in the stem, Cu existed as four forms including hydrophobic protein binding, cell wall binding state, residual state and water-soluble state, which further reduce the excess transport of Cu to the leaves.In the leaves, excess Cu in the leaf cells was mainly in the vacuole and insoluble residue to achieve tolerance and detoxification to Cu.