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. 2022 Sep 11;10(9):1818.
doi: 10.3390/microorganisms10091818.

Emergence and Transfer of Plasmid-Harbored rmtB in a Clinical Multidrug-Resistant Pseudomonas aeruginosa Strain

Jiacong Gao  1 , Xiaoya Wei  1 , Liwen Yin  1 , Yongxin Jin  1 , Fang Bai  1 , Zhihui Cheng  1 , Weihui Wu  1
Affiliations

Emergence and Transfer of Plasmid-Harbored rmtB in a Clinical Multidrug-Resistant Pseudomonas aeruginosa Strain

Jiacong Gao et al. Microorganisms. .

Abstract

Multidrug-resistant (MDR) Pseudomonas aeruginosa poses a great challenge to clinical treatment. In this study, we characterized a ST768 MDR P. aeruginosa strain, Pa150, that was isolated from a diabetic foot patient. The minimum inhibitory concentration (MIC) assay showed that Pa150 was resistant to almost all kinds of antibiotics, especially aminoglycosides. Whole genome sequencing revealed multiple antibiotic resistant genes on the chromosome and a 437-Kb plasmid (named pTJPa150) that harbors conjugation-related genes. A conjugation assay verified its self-transmissibility. On the pTJPa150 plasmid, we identified a 16S rRNA methylase gene, rmtB, that is flanked by mobile genetic elements (MGEs). The transfer of the pTJPa150 plasmid or the cloning of the rmtB gene into the reference strain, PAO1, significantly increased the bacterial resistance to aminoglycoside antibiotics. To the best of our knowledge, this is the first report of an rmtB-carrying conjugative plasmid isolated from P. aeruginosa, revealing a novel possible transmission mechanism of the rmtB gene.

Keywords: Pseudomonas aeruginosa; conjugative plasmid; mobile genetic element; multidrug-resistance; rmtB.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Graphical circular schematic map of the clinical isolate Pa150 from circle 1 (outmost) to circle 7 (innermost). Circles 1 and 2, coding sequences (CDS), forward and reverse frames, respectively; circle 3, tRNAs (orange) and rRNAs (purple); circle 4, CRISPR-related genes (blue); circle 5, GC content; circle 6, GC-Skew; circle 7, coverage.
Figure 2
Figure 2
Schematic map of the pTJPa150 plasmid. The innermost circle is the scale. The GC content is illustrated in the second circle. The third circle shows GC skew (+: green, −: purple). The outermost circle indicates the CDs, in which antibiotic resistance genes are highlighted in red and mobile genetic elements in blue.
Figure 3
Figure 3
Phylogenetic tree and genetic information of rmtB. (A) Phylogenetic tree of RmtB and RmtB-like proteins. Neighbor-joining tree based on the amino acid sequences of RmtB and RmtB-like proteins (obtained from the NCBI databases) generated using MEGA7; the bootstrap was 1000. The bacteria species are indicated by the colors as follows: P. aeruginosa, green; K. pneumonia, orange; E. coli, blue; others, black. (B) Comparison of the genetic environment of rmtB with those of closely related sequences. The mobile genetic elements are shown in blue.
Figure 4
Figure 4
Prevalence and distribution of rmtB-positive isolates. (A) Species classification of rmtB positive isolates. (B) Source statistics of rmtB-positive isolates (C) Geographical distribution of rmtB-positive isolates in China. The color depth correlates with the quantity. (D) Distribution of rmtB-positive isolates worldwide. (E) Isolation of rmtB-positive isolates from 2003 to 2021.
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