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水稻条斑病菌(Xanthomonas oryzae pv.oryzicola,Xoc)为稻黄单胞菌种下的致病变种,引起细菌性条斑病(bacterial leaf streak,BLS),对水稻安全生产构成严重威胁。为准确在水稻条斑病菌中进行致病相关基因的转录表达和调控分析,本研究构建了包含终止子、gus A报道基因和多克隆位点等启动子探针元件的载体p UTG01和p UTG14。选取Xoc的hrp F启动子,构建在p UTG01载体上,将其导入hrp X突变体RΔhrp X中,GUS活性测定结果显示,hrp F基因的表达显著减低,验证了hrp F受Hrp X正调控,证实该载体可有效进行基因的转录表达分析;通过双质粒兼容共存策略,在hrp X突变体中同时实现了hrp X基因的功能互补和通过GUS活性定量测定hrp F基因的转录表达分析。该载体系统的建立,为后续分析稻黄单胞菌致病相关基因的表达调控提供了有效的工作系统。
Xanthomonas oryzae pv.oryzicola (Xoc), a disease-causing strain under the species of yellow rice, causes bacterial leaf streak (BLS), which poses a serious threat to the safe production of rice. In order to accurately analyze the transcriptional expression and regulation of the pathogenicity-related genes in leaf spot pathogen, the vectors p UTG01 and p UTG14 containing the promoter elements of the promoter, gus A reporter gene and multiple cloning sites were constructed . The hrp F promoter of Xoc was constructed and transformed into hrp X mutant RΔhrp X. The result of GUS activity assay showed that the expression of hrp F gene was significantly reduced, which confirmed that hrp F was positively regulated by Hrp X, It was confirmed that this vector can effectively analyze the transcriptional expression of hrp gene. By the strategy of compatible double plasmids, the hrp X gene complements the hrp X mutant functionally and the transcriptional expression analysis of hrp F gene is quantitatively determined by GUS activity. The establishment of the vector system provides an effective working system for the subsequent analysis of the expression regulation of pathogenicity-related genes of the bacteria.