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Biological functions of nirS in Pseudomonas aeruginosa ATCC 9027 under aerobic conditions

  • Gang Zhou
  • Hong Peng
  • Ying-si Wang
  • Cai-ling Li
  • Peng-fei Shen
  • Xiao-mo Huang
  • Xiao-bao XieEmail author
  • Qing-shan ShiEmail author
Genetics and Molecular Biology of Industrial Organisms - Original Paper

Abstract

Through our previous study, we found an up-regulation in the expression of nitrite reductase (nirS) in the isothiazolone-resistant strain of Pseudomonas aeruginosa. However, the definitive molecular role of nirS in ascribing the resistance remained elusive. In the present study, the nirS gene was deleted from the chromosome of P. aeruginosa ATCC 9027 and the resulting phenotypic changes of ΔnirS were studied alongside the wild-type (WT) strain under aerobic conditions. The results demonstrated a decline in the formations of biofilms but not planktonic growth by ΔnirS as compared to WT, especially in the presence of benzisothiazolinone (BIT). Meanwhile, the deletion of nirS impaired swimming motility of P. aeruginosa under the stress of BIT. To assess the influence of nirS on the transcriptome of P. aeruginosa, RNA-seq experiments comparing the ΔnirS with WT were also performed. A total of 694 genes were found to be differentially expressed in ΔnirS, of which 192 were up-regulated, while 502 were down-regulated. In addition, these differently expressed genes were noted to significantly enrich the carbon metabolism along with glyoxylate and dicarboxylate metabolisms. Meanwhile, results from RT-PCR suggested the contribution of mexEF-oprN to the development of BIT resistance by ΔnirS. Further, c-di-GMP was less in ΔnirS than in WT, as revealed by HPLC. Taken together, our results confirm that nirS of P. aeruginosa ATCC 9027 plays a role in BIT resistance along with biofilm formation and further affects several metabolic patterns under aerobic conditions.

Keywords

Pseudomonas aeruginosa Nitrite reductase Isothiazolones Biofilm formation and dispersal RNA-seq technology 

Notes

Acknowledgements

This work was funded by the GDAS’ Project of Science and Technology Development (No. 2019GDASYL-0104006), the National Natural Science Foundation of China (Nos. 31770091 and 31500036), and Natural Science Foundation of Guangdong Province (No. 2015A030313713). We are grateful to Prof. Hai-hong Wang of South China Agricultural University for generously providing us the plasmids of pK18-GM and pSRK-GM.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

10295_2019_2232_MOESM1_ESM.xls (768 kb)
Additional file 1. Detection of differently expressed genes between WT and ΔnirS transcriptomes using RNA-seq technique. Differences with FDR ≤ 0.05 and log2FC absolute value ≥ 1 were set as the threshold for significant differences in gene expression. (XLS 767 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2019

Authors and Affiliations

  • Gang Zhou
    • 1
  • Hong Peng
    • 1
  • Ying-si Wang
    • 1
  • Cai-ling Li
    • 1
  • Peng-fei Shen
    • 1
  • Xiao-mo Huang
    • 1
  • Xiao-bao Xie
    • 1
    Email author
  • Qing-shan Shi
    • 1
    Email author
  1. 1.Guangdong Open Laboratory of Applied Microbiology, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of MicrobiologyGuangdong Academy of SciencesGuangzhouPeople’s Republic of China

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