基于16S rRNA测序技术分析桶装水中野生铜绿假单胞菌的系统发育

Phylogenetic analysis of wild Pseudomonas aeruginosa in bottled water based on the 16S rRNA sequencing technology

目的以铜绿假单胞菌16S rRNA为研究对象,利用16S rRNA测序分析技术构建系统发育树,研究湖南省内28株野生水源性铜绿假单胞菌的系统发育。方法收集28株野生阳性铜绿假单胞菌,利用27F/1492R通用引物进行16S rRNA PCR扩增,将胶回收产物进行测序,测序结果利用MEGA 11.0构建系统发育树,并进行遗传距离计算。结果28株铜绿假单胞菌可分为3支,一株来源于永州的菌株单独为一支,3株分别来源于常德、永州、郴州的菌株聚为一支,其余24株聚为一支;28株铜绿假单胞菌平均遗传距离为0.0064。结论28株野生铜绿假单胞菌之间具有高度同源性,通过实验进一步完善了水源性铜绿假单胞菌溯源数据库,为桶装水铜绿假单胞菌污染源监测提供了理论依据。

Objective To study the phylogeny of 28 wild water source Pseudomonas aeruginosa strains in Hunan Province,a phylogenetic tree was constructed using 16S rRNA sequencing analysis technology,with Pseudomonas aeruginosa 16S rRNA as the research object.Methods Twenty-eight wild-type Pseudomonas aeruginosa strains were collected and amplified using 27F/1492R universal primers for 16S rRNA.The PCR products obtained from the gels were sequenced.The phylogenetic tree was constructed using MEGA 11.0 and the genetic distance was calculated.Results The 28 strains of Pseudomonas aeruginosa could be divided into three branches,one strain from Yongzhou was a single strain,three strains from Changde,Yongzhou and Chenzhou were combined into one strain,and the other 24 strains were combined into one strain.The average genetic distance of 28 Pseudomonas aeruginosa strains was 0.0064.Conclusion There is a high degree of homology between 28 wild Pseudomonas aeruginosa strains.The traceability database of waterborne Pseudomonas aeruginosa is further improved by the experiments and the theoretical basis for monitoring the source of Pseudomonas aeruginosa contamination in bottled water has been provided.