宁夏枸杞幼苗叶片中抗氧化酶及游离氨基酸对镉胁迫的响应
Response of Antioxidative Enzymes Activities and Amino Acids Concentrations in Leaf Tissues of Lycium barbarum L. Seedlings under Cadmium Stress
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摘要: 为探讨镉（cadmium，Cd）胁迫对宁夏枸杞幼苗的叶片细胞、抗氧化酶活性、茎叶游离氨基酸及其关键基因表达模式的影响。以宁杞1号的组培苗为试验材料，采用水培处理方式，分别设置0 μmol·L-1和100 μmol·L-1的CdCl2胁迫，根据石蜡切片观察第72小时、第144小时叶片细胞的变化，测定过氧化酶的活性，通过高效液相色谱技术测定第72小时枸杞地上部组织中游离氨基酸含量，利用转录组测序及分析挖掘与氨基酸转运代谢相关的功能基因，结合实时荧光定量PCR技术进行验证，以期为游离氨基酸对植物适应外源Cd胁迫中的功能提供理论依据。结果表明，宁夏枸杞的地上组织在Cd胁迫下，叶片细胞、抗氧化酶活性以及氮代谢均发生显著变化，并且Cd胁迫还促使茎叶累积了一些与抗逆性有关的氨基酸，如脯氨酸、丙氨酸和瓜氨酸等，同时诱导了一些氨基酸转运及吸收相关基因的表达。Abstract: This study aimed to investigate the effects of cadmium stress on leaf cytoarchitecture, antioxidant enzyme activities, free amino acids, and key gene expression patterns of Lycium barbarum L. seedlings. The tissue culture plantlet of Lycium barbarum L. was used as the research material and was treated with 0 μmol·L-1 and 100 μmol·L-1 CdCl2 respectively in Hoagland solutions. The changes of leaf cytoarchitecture on the 72 h and 144 h were observed by paraffin section method, the antioxidative enzymes activities were determined, the contents of free amino acids in the tissue of Lycium barbarum L. on the 72 h were determined by high-performance liquid chromatography (HPLC) and the functional genes related to amino acid transport and metabolism were verified by real-time fluorescent quantitative PCR technology. The results showed that leaf cytoarchitecture, antioxidant enzyme activities, and nitrogen metabolism of Lycium barbarum L. were changed significantly under cadmium stress. We found that Cd stress also promoted the accumulation of some amino acids related to stress resistance, such as proline, alanine, citrulline. At the same time, Cd induced the expression of some genes related to amino acid transport and absorption, which provided a theoretical basis to adapt to exogenous Cd stress.
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