恶臭假单胞菌的正丁醇耐受性驯化及比较基因组学分析

Adapting n-Butanol Tolerance of Pseudomonas putida and Comparative Genomics Analysis

恶臭假单胞菌(Pseudomonas putida)是一类非常重要的工业微生物,广泛用于环境修复、生物活性物质合成等方面。作者采用常压室温等离子体诱变技术(atmospheric and room temperature plasma,ARTP)对实验室保藏的恶臭假单胞菌901进行随机诱变,以正丁醇作为筛选压力进行驯化培养,最终筛选得到一株能耐受12 g/L正丁醇的突变株。通过Illumina HiSeq平台进行全基因组重测序和生物信息学分析。结果显示共获得大约1 900万对reads,平均读长为150 nt,与参考基因组的片段配对率为92.03%,覆盖率为90.34%。共检测到40 704个单核苷酸位点变异(singlenucleotide variation,SNV),其中纯合的为39 319个,杂合的为1 385个;存在2 153个基因小片段插入/缺失序列(insert&deletions,InDels),其中纯合的突变位点数量为1 871个,杂合的突变位点数量为282个。最后,将所有突变进行GO/KEGG数据库功能注释,发现突变数目最多的是代谢相关基因,其中以氨基酸代谢和糖代谢路径相关基因为主。

Pseudomonas putida is one of the most important industrial microorganisms,widely applied in environmental remediation,production of bioactive natural products and secondary metabolites,etc.To enhance the n-butanol tolerance of Pseudomonas putida 901,atmospheric and room temperature plasma(ARTP)was performed to generate mutant library.After gradient adaptation in n-butanol,a mutant capable of tolerating 12 g/L n-butanol was obtained.The whole genome was re-sequenced and analyzed through Illumina HiSeq platform.About 19 million reads was obtained with an average read of 150 nt.Compared with the reference genome,the fragment pairing rate was 92.03%,while the coverage rate was 90.34%.A total of 40704 single nucleotide variations was detected,with isozygoty of 39319 and heterozygote of 40704.There were 2153 inserts and deletions,with homozygous mutation sites of 1871 and heterozygous mutation sites of 282. All the mutations were functionally annotated according to GO/KEGG database.