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采用溶液培养的方法,以Al敏感型黑大豆(SB)为材料,研究外源添加水杨酸(SA)对Al胁迫下SB根生理生化指标和Al胁迫相关基因表达的影响,探讨外源SA缓解黑大豆Al毒害的分子机理。结果表明:低浓度(10,20!mol·L-1)SA缓解了Al毒引起的根伸长抑制,其中20!mol·L-1SA的缓解效果更明显;而高浓度SA和SA合成抑制剂多效唑(PAC)处理加重了Al毒引起的根伸长抑制;外源添加20!mol·L-1SA使Al胁迫下SB根尖MDA和H2O2含量下降,根系分泌物中柠檬酸的含量约是单独Al处理的2倍;基因表达分析表明外源添加20!mol·L-1SA促进Al胁迫下SB根中12个14-3-3亚型、质膜H+-ATP酶和柠檬酸通道蛋白(MATE)基因的表达;免疫共沉淀的结果表明外源添加20!mol·L-1SA提高了Al胁迫下质膜H+-ATP酶蛋白磷酸化水平及其与14-3-3蛋白结合能力;外源添加PAC的效果与外源添加SA的相反。外源SA可能通过诱导Al胁迫下MATE表达,提高14-3-3蛋白和质膜H+-ATP酶基因蛋白水平和互作,增加柠檬酸的分泌,增强黑大豆对Al毒害的耐受性。
The effects of exogenous salicylic acid (SA) on the physiological and biochemical indexes of SB root and the expression of Al stress-related genes under Al stress were studied by solution culture using Al-sensitive black soybean (SB) as material. The effects of exogenous SA Alleviating the molecular mechanism of Al toxicity in black soybean. The results showed that low concentration (10,20 mol·L-1) SA alleviated Al toxicity-induced root elongation inhibition, of which 20 mol·L-1 SA was more effective; The treatment with paclobutrazol (PAC) aggravated the inhibition of root elongation induced by Al toxicity. The content of MDA and H2O2 in root tip of SB decreased with the exogenous addition of 20! Mol·L-1 SA, the content of citric acid in root exudates was about The results of gene expression analysis showed that exogenous addition of 20! Mol·L-1 SA promoted 12 subunits 14-3-3, plasma membrane H + -ATPase and citrate channel protein MATE). The results of co-immunoprecipitation showed that the exogenous addition of 20! Mol·L-1 SA increased the phosphorylation of plasma membrane H + -ATPase and the binding ability to 14-3-3 protein under Al stress. The effect of source addition of PAC is opposite to that of exogenous addition of SA. Exogenous SA may increase the protein level and interaction of 14-3-3 protein and plasma membrane H + -ATPase gene, increase the secretion of citric acid, and enhance the tolerance of black soybean to Al toxicity by inducing MATE expression under Al stress.