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目的通过检测绿脓假单胞菌生物被膜形成过程中藻酸盐合成相关基因的表达,研究藻酸盐在绿脓假单胞菌生物被膜形成过程中的作用。方法改良平板培养法建立黏液型绿脓假单胞菌PA17和非黏液型绿脓假单胞菌PAO1生物被膜模型,半定量RT-PCR测定生物被膜形成不同时间点algD、algU的表达,并进行统计学分析。结果PAO1和PA17分别于第3天和第6天形成成熟生物被膜。algD和algU均在微菌落分化为成熟生物被膜的阶段表达增高。单因素方差分析示同一菌株的相同基因在生物被膜形成过程的不同时间点表达的差异有统计学意义;两独立样本秩和检验示同一基因在不同菌株生物被膜形成过程中表达的差异无统计学意义。结论藻酸盐在绿脓假单胞菌生物被膜形成过程中微菌落分化为成熟生物被膜的阶段发挥作用,不受绿脓假单胞菌表型影响。
Objective To investigate the role of alginate in the biofilm formation of Pseudomonas aeruginosa by detecting the expression of alginate-related genes in the biofilm formation of Pseudomonas aeruginosa. Methods The mucosal Pseudomonas aeruginosa PA17 and non-mucoid Pseudomonas aeruginosa PAO1 biofilms were established by modified plate culture method. The expression of algD and algU at different time points was measured by semi-quantitative RT-PCR. Statistical analysis. Results PAO1 and PA17 formed mature biofilms on day 3 and day 6, respectively. Both algD and algU were up-regulated during microcolony differentiation into mature biofilms. One-way analysis of variance showed that there was significant difference in expression of the same genes of the same strain at different time points in biofilm formation. Two independent samples rank sum test showed that there was no statistical difference in the expression of the same gene in biofilm formation of different strains significance. Conclusion Alginate plays a role in the differentiation of microcolon into mature biofilm during the formation of Pseudomonas aeruginosa biofilm, and is not affected by Pseudomonas aeruginosa phenotype.