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目的:分析CYP81A6基因在苯达松及甲磺隆处理下的诱导表达模式,解释该基因与两种除草剂代谢相关的可能原因。创新点:从两种除草剂降解途径中产生的小分子物质的结构相似性出发,通过基因诱导表达的特点分析,解释CYP81A6和两种除草剂降解相关的原因。方法:通过实时定量聚合酶链反应(PCR)来分析基因表达的特点;利用CYP81A6启动子与GUS报告基因构建的载体来分析组织特异性表达;通过亚细胞定位来确定CYP81A6发挥功能的场所。结论:CYP81A6基因受苯达松及甲磺隆诱导,在不同的时间点开始上调,说明了甲磺隆的降解中间产物可以诱导这个基因的表达;CYP81A6是组成型表达,在根、茎、叶中均有表达;亚细胞定位结果证明CYP81A6是一个内质网上的蛋白。
OBJECTIVE: To analyze the induced expression pattern of CYP81A6 in the presence of bentasone and metsulfuron-methyl, and to explain the possible causes of its association with the metabolism of two herbicides. Innovative point: Starting from the structural similarity of small molecules produced by the two herbicide-degrading pathways, we explain why CYP81A6 is associated with the degradation of two herbicides by characterizing gene-induced expression. Methods: The characteristics of gene expression were analyzed by real-time quantitative polymerase chain reaction (PCR). The tissue-specific expression was analyzed by using the vector constructed with the promoter of CYP81A6 and the GUS reporter gene. The location of CYP81A6 function was determined by subcellular localization. CONCLUSION: The CYP81A6 gene was induced by bendazon and metsulfuron and began to be up-regulated at different time points, indicating that the degradation intermediate of mesotrione could induce the expression of this gene. CYP81A6 was constitutively expressed in roots, stems and leaves The results of subcellular localization proved that CYP81A6 is an endoplasmic reticulum protein.