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Int. J. Mol. Sci. 2014, 15(4), 6328-6342; doi:10.3390/ijms15046328
Article

Isolation of the Autoinducer-Quenching Strain that Inhibits LasR in Pseudomonas aeruginosa

1,* , 2
, 3
 and 2,*
Received: 4 July 2013; in revised form: 21 March 2014 / Accepted: 28 March 2014 / Published: 14 April 2014
(This article belongs to the Special Issue Quorum Sensing Research in Microbial Systems)
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Abstract: Quorum sensing (QS) has been recognized as a general phenomenon in microorganisms and plays an important role in many pathogenic bacteria. In this report, we used the Agrobacterium tumefaciens biosensor strain NT1 to rapidly screen for autoinducer-quenching inhibitors from bacteria. After initial screening 5389 isolates obtained from land and beach soil, 53 putative positive strains were identified. A confirmatory bioassay was carried out after concentrating the putative positive culture supernatant, and 22 strains were confirmed to have anti-LasR activity. Finally, we determined the strain JM2, which could completely inhibit biofilm formation of Pseudomonas aeruginosa PAO1, belonged to the genus Pseudomonas by analysis of 16S rDNA. Partially purified inhibitor factor(s) F5 derived from culture supernatants specifically inhibited LasR-controlled elastase and protease in wild type P. aeruginosa PAO1 by 68% and 73%, respectively, without significantly affecting growth; the rhl-controlled pyocyanin and rhamnolipids were inhibited by 54% and 52% in the presence of 100 µg/mL of F5. The swarming motility and biofilm of PAO1 were also inhibited by F5. Real time RT-PCR on samples from 100 µg/mL F5-treated P. aeruginosa showed downregulation of autoinducer synthase (LasRI and rhlI) and cognate receptor (lasR and rhlR) genes by 50%, 28%, 48%, and 29%, respectively. These results provide compelling evidence that the F5 inhibitor(s) interferes with the las system and significantly inhibits biofilm formation.
Keywords: quorum sensing; Pseudomonas aeruginosa; Agrobacterium tumefaciens NT1; biofilm; virulence factor quorum sensing; Pseudomonas aeruginosa; Agrobacterium tumefaciens NT1; biofilm; virulence factor
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Weng, L.; Zhang, Y.; Yang, Y.; Wang, L. Isolation of the Autoinducer-Quenching Strain that Inhibits LasR in Pseudomonas aeruginosa. Int. J. Mol. Sci. 2014, 15, 6328-6342.

AMA Style

Weng L, Zhang Y, Yang Y, Wang L. Isolation of the Autoinducer-Quenching Strain that Inhibits LasR in Pseudomonas aeruginosa. International Journal of Molecular Sciences. 2014; 15(4):6328-6342.

Chicago/Turabian Style

Weng, Lixing; Zhang, Yuqian; Yang, Yuxiang; Wang, Lianhui. 2014. "Isolation of the Autoinducer-Quenching Strain that Inhibits LasR in Pseudomonas aeruginosa." Int. J. Mol. Sci. 15, no. 4: 6328-6342.


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