黑藻(Hydrilla verticillat)在铅、锌胁迫下的代谢组学研究
Metabolomics Study of Hydrilla verticillata under Heavy Metal Stress of Lead and Zinc
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摘要: 黑藻（Hydrilla verticillat）作为我国淡水水域常见的沉水植物，对水环境重金属污染修复有着重要作用。通过超高效液相色谱串联质谱（UPLC-MS/MS）技术检测了黑藻中342个初级代谢产物，并采用单变量、主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS-DA）多元统计分析方法分析代谢物，以探究在不同浓度铅、锌胁迫下黑藻初级代谢产物的变化特性，弄清铅、锌对生物体代谢活动的影响。结果表明，铅单一胁迫主要促进黑藻机体酚酸类和糖类物质的生成，抑制脂类的生成，N-苯乙酰基-L-谷氨酰胺和异水杨酸-O-葡萄糖显著上调，相比单一铅胁迫，铅锌复合胁迫主要促进黑藻氨基酸、糖类、有机酸和脂质等代谢物的合成，抑制酚酸类、维生素和脂质等代谢物的合成；锌单一胁迫主要促进氨基酸、核苷酸、糖类和有机酸等代谢物的生成，抑制部分酚酸类和脂质代谢物的生成，相比单一锌胁迫，高浓度铅锌复合胁迫（Pb0.10+Zn2.00）可主要促进酚酸类、核苷酸、有机酸和脂质等代谢物的合成，抑制以谷胱甘肽为代表的氨基酸等代谢物的合成；超高浓度铅锌复合胁迫（Pb0.20+Zn4.00）可主要促进对苯二甲酸（C8H6O4）和去鼠李糖异洋丁香酚苷B （C23H26O11）2种酚酸类代谢物的合成，抑制氨基酸、核苷酸、维生素和有机酸等代谢物的合成。另外，N-苯乙酰基-L-谷氨酰胺（C13H16N2O4）、酪氨（C8H11NO）、对香豆酰咖啡酰酒石酸（C22H18O11）、没食子鞣质（C13H16O10）和异水杨酸-O-葡萄糖（C13H16O8）可作为铅胁迫的代谢标记物，5-氨基戊酸（C5H11NO2）、L-（+）-精氨酸（C6H14N4O2）和L-焦谷氨酸（C5H7NO3）等可作为锌胁迫的代谢标记物。Abstract: Hydrilla verticillata, a common submerged plant in freshwater of China, has a certain adsorption effect on heavy metals, which made it play an important role in water environment protection. Excessive lead accumulation could affect the growth and physiological metabolism in plants, meanwhile, even cause plant death. As the associated metal of lead, zinc is one of the necessary elements of organisms, but when its content exceeds the micro-demand required, it will cause a certain degree of damage to the body. Three hundred and forty-two primary metabolites in H. verticillata were detected by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) detection technology. We explored the changes of the first metabolites and the metabolism activities in H. verticillata under 28-day stress of different concentrations of lead-zinc by univariate, principal part-related analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The results showed that single lead stress mainly increased the production of phenolic acids and saccharides in H. verticillata and inhibited the production of lipids, which involved physiological processes such as phenylalanine metabolism, carbohydrate metabolism and biosynthesis of secondary metabolites. It indicated phenolic acids and carbohydrates played a major role in resistance under lead stress. While lead-zinc combined stress promoted the yield of amino acids, sugars, organic acids and lipids, and depressed the production of phenolic acids, vitamins and lipids. Single stress of zinc mainly increased the production of amino acids, nucleotides, saccharides and organic acids, and inhibited the content of phenolic acids and lipid metabolites. The physiological processes relates to citrate cycle (TCA cycle), arginine and proline metabolism, carbohydrate metabolism, biosynthesis of secondary metabolites, carbon metabolism and ABC transporter metabolism. Amino acids, nucleotides, carbohydrates and other substances take protective action on the body after subjected to external environmental stress, indicating that the body has carried out metabolic regulation to combat stress. Compared with single zinc stress, high-concentration lead-zinc combined stress (Pb0.10+Zn2.00) mainly promoted to increase the content of phenolic acids, nucleotides, organic acids and lipids, and depressed the production of amino acids represented by oxidized glutathione. In addition, the ultra-high concentration of lead and zinc combined stress (Pb0.20+Zn4.00) mainly increased the production of phenolic acid (terephthalic acid and calceorioside B), and inhibited the content of amino acids, nucleotides, vitamins and organic acids. Consequently, phenylacetyl-L-glutamine, L-tyramine, p-coumaroylcaffeoyl tartaric acid, glucogallin and isosalicylic acid O-glycoside could be used as metabolism markers in H. verticillata under lead stress, while 5-aminovaleric acid, L-(+)-arginine and L-pyroglutamic acid could be used as metabolism markers in H. verticillata under zinc stress.
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Key words:
- lead /
- zinc /
- Hydrilla verticillata /
- hytotoxicity /
- metabolomics
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