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. 2022 Nov 26;10(12):2342.
doi: 10.3390/microorganisms10122342.

PafS Containing GGDEF-Domain Regulates Life Activities of Pseudomonas glycinae MS82

Xianyi Chen  1 , Shaoxuan Qu  1 , Xin Luo  1 , Shi-En Lu  2 , Youzhou Liu  3 , Huiping Li  1 , Lijuan Hou  1 , Jinsheng Lin  1 , Ning Jiang  1 , Lin Ma  1
Affiliations

PafS Containing GGDEF-Domain Regulates Life Activities of Pseudomonas glycinae MS82

Xianyi Chen et al. Microorganisms. .

Abstract

Cyclic dimeric guanosine monophosphate (c-di-GMP) is synthesized by diguanylate cyclase (DGC) with the GGDEF domain. As a ubiquitous bacterial second messenger, it regulates diverse life-activity phenotypes in some bacteria. Although 38 genes encoding GGDEF-domain-containing proteins have been identified in the genome of the Pseudomonas glycinae strain MS82, whether c-di-GMP functions as a facilitator or repressor of life-activity phenotypes is poorly understood. In this study, one of the 38 genes containing a GGDEF domain in MS82, PafS was investigated to explore its regulatory function in bacterial life activities. The PafS-deletion mutant ΔPafS and reversion mutant PafS-comp were constructed by the method of biparental conjugation and homologous recombination. The life activities of the mutants, such as antifungal activity, biofilm formation ability, polysaccharide content, and motor behavior, were explored. The results showed that all life-activity phenotypes were significantly reduced after knocking out PafS, whereas all were significantly restored to a similar level to that of MS82 after the complementation of PafS. These results suggested that PafS plays an important role in the regulation of a range of cellular activities by c-di-GMP in P. glycinae MS82.

Keywords: GGDEF domain; PafS; Pseudomonas glycinae MS82; life activities.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The PCR confirmation of PafS-deletion and reversion mutants of P. glycinae strain MS82. M, marker 5000; 1, wild-type MS82 strain with the primer pair pafS-compF/R; 2, deletion mutant ΔPafS with the primer pair ΔPafS-F1/R2; 3, reversion mutant PafS-comp with the primer pair pafS-compF/R; 4, the upstream fragment of PafS with the primer pair ΔPafS-F1/R1; 5, the downstream fragment of PafS with the primer pair ΔPafS-F2/R2.
Figure 2
Figure 2
Inhibition of T. virens by PafS-deletion and reversion mutants of P. glycinae strain MS82. Different lowercase letters above bars indicate a significant difference compared with the control (p < 0.05). The error bars indicate the SD and the statistical test used Student’s t-test.
Figure 3
Figure 3
Biofilm formation by PafS-deletion and reversion mutants of P. glycinae strain MS82. ‘*’ under bars indicate a significant difference compared with the control (p < 0.05). The error bars indicate the SD value.
Figure 4
Figure 4
Extracellular polysaccharides (EPS) content of PafS-deletion and reversion mutants of P. glycinae strain MS82. Different lowercase letters above bars for a specific timepoint indicate a significant difference compared with the control (p < 0.05). The error bars indicate the SD and the statistical test used Student’s t-test.
Figure 5
Figure 5
Motility assay of PafS-deletion and reversion mutants of P. glycinae strain MS82. Different lowercase letters above bars for a motility type indicate a significant difference compared with the control (p < 0.05). The error bars indicate the SD and the statistical test used Student’s t-test.
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