DOI QR코드

DOI QR Code

RNAseq-based Transcriptome Analysis of Burkholderia glumae Quorum Sensing

  • Kim, Sunyoung (Department of Microbiology, Pusan National University) ;
  • Park, Jungwook (Department of Microbiology, Pusan National University) ;
  • Kim, Ji Hyeon (Department of Microbiology, Pusan National University) ;
  • Lee, Jongyun (Department of Microbiology, Pusan National University) ;
  • Bang, Bongjun (Department of Microbiology, Pusan National University) ;
  • Hwang, Ingyu (Department of Agricultural Biotechnology, Seoul National University) ;
  • Seo, Young-Su (Department of Microbiology, Pusan National University)
  • Received : 2013.04.29
  • Accepted : 2013.05.16
  • Published : 2013.09.01

Abstract

Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::${\Omega}$) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::${\Omega}$). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.

Keywords

References

  1. Cother, E. J., Noble, D. H., de Ven, R. J. van., Lanoiselet, V., Ash, G., Vuthy, N., Visarto, P. and Stodart, B. 2010. Bacterial pathogens of rice in the Kingdom of Cambodia and description of a new pathogen causing a serious sheath rot disease. Plant Pathol. 5:944-953.
  2. Chun, H., Choi, O., Goo, E., Kim, N., Kim, H., Kang, Y., Kim, J., Moon, J. S. and Hwang, I. 2009. The quorum sensing-dependent gene katG of Burkholderia glumae is important for protection from visible light. J. Bacteriol. 191:4152-4157. https://doi.org/10.1128/JB.00227-09
  3. Deng, Y., Wu, J., Tao, F. and Zhang, L. H. 2011. Listening to a new language: DSF-based quorum sensing in Gram-negative bacteria. Chem. Rev. 111:160-173. https://doi.org/10.1021/cr100354f
  4. Devescovi, G., Bigirimana, J., Degrassi, G., Cabrio, L., LiPuma, J. J., Kim, J., Hwang, I. and Venturi, V. 2007. Involvement of a quorum-sensing-regulated lipase secreted by a clinical isolate of Burkholderia glumae in severe disease symptoms in rice. Appl. Environ. Microbiol. 73:4950-4958. https://doi.org/10.1128/AEM.00105-07
  5. Fuqua, W. C., Winans, S. C. and Greenberg, E. P. 1994. Quorum sensing in bacteria: The LuxR-LuxI family of cell densityresponsive transcriptional regulators. J. Bacteriol. 176:269-275. https://doi.org/10.1128/jb.176.2.269-275.1994
  6. Goo, E., Majerczyk, C. D., An, J. H., Chandler, J. R., Seo, Y.-S., Ham, H., Lim, J. Y., Kim, H., Lee, B., Jang, M. S., Greenberg, E. P. and Hwang, I. 2012. Bacterial quorum sensing, cooperativity, and anticipation of stationary-phase stress. Proc. Natl. Acad. Sci. USA 109:19775-19780. https://doi.org/10.1073/pnas.1218092109
  7. Jimenez, P. N., Koch, G., Thompson, J. A., Xavier, K. B., Cool, R. H. and Quax, W. J. 2012. The multiple signaling systems regulating virulence in Pseudomonas aeruginosa. Microbiol. Mol. Biol. Rev. 76:46-65. https://doi.org/10.1128/MMBR.05007-11
  8. Kapatral, V., Campbell, J. W., Minnich, S. A., Thomson, N. R., Matsumura, P. and Pruss, B. M. 2004. Gene array analysis of Yersinia enterocolitica FlhD and FlhC: regulation of enzymes affecting synthesis and degradation of carbamoylphosphate. Microbiology 150:2289-2300. https://doi.org/10.1099/mic.0.26814-0
  9. Kim, J., Kim, J. G., Kang, Y., Jang, J. Y., Jog, G. J., Lim, J. Y., Kim, S., Suga, H., Nagamatsu, T. and Hwang, I. 2004. Quorum sensing and the LysR-type transcriptional activator ToxR regulate toxoflavin biosynthesis and transport in Burkholderia glumae. Mol. Microbiol. 54:921-934. https://doi.org/10.1111/j.1365-2958.2004.04338.x
  10. Kim, J., Kang, Y., Choi, O., Jeong, Y., Jeong, J. E., Lim, J. Y., Kim, M., Moon, J. S., Suga, H. and Hwang, I. 2007. Regulation of polar flagellum genes is mediated by quorum sensing and FlhDC in Burkholderia glumae. Mol. Microbiol. 64:165-179. https://doi.org/10.1111/j.1365-2958.2007.05646.x
  11. Kim, J., Oh, J., Choi, O., Kang, Y., Kim, H., Goo, E., Ma, J., Nagamatsu, T., Moon, J. S. and Hwang, I. 2009. Biochemical evidence for ToxR and ToxJ binding to the tox operons of Burkholderia glumae and mutational analysis of ToxR. J. Bacteriol. 191:4870-4878. https://doi.org/10.1128/JB.01561-08
  12. Kim, J., Kang, Y., Kim, J.-G., Choi, O. and Hwang, I. 2010. Occurrence of Burkholderia glumae on rice and field crops in Korea. Plant Pathol. J. 26:271-272. https://doi.org/10.5423/PPJ.2010.26.3.271
  13. Labbate, M., Queck, S. Y., Koh, K. S., Rice, S. A., Givskov, M. and Kjelleberg, S. 2004. Quorum sensing-controlled biofilm development in Serratia liquefaciens MG1. J. Bacteriol. 186:692-698. https://doi.org/10.1128/JB.186.3.692-698.2004
  14. Milton, D. L. 2006. Quorum sensing in vibrios: complexity for diversification. Int. J. Med. Microbiol. 296:61-71. https://doi.org/10.1016/j.ijmm.2006.01.044
  15. Moreira, C. G., Weinshenker, D. and Sperandio, V. 2010. QseC mediates Salmonella enterica serovar typhimurium virulence in vitro and in vivo. Infect. Immun. 78:914-926. https://doi.org/10.1128/IAI.01038-09
  16. Mortazavi, A., Williams, B. A., McCue, K., Schaeffer, L. and Wold, B. 2008. Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat. Methods 5:621-628. https://doi.org/10.1038/nmeth.1226
  17. Nandakumar, R., Shahjahan, A. K. M., Yuan, X. L., Dickstein, E. R., Groth, D. E., Clark, C. A., Cartwright, R. D. and Rush, M. C. 2009. Burkholderia glumae and B. gladioli cause bacterial panicle blight in rice in the southern United States. Plant Dis. 93:896-905. https://doi.org/10.1094/PDIS-93-9-0896
  18. Pai, A., Tanouchi, Y. and You, L. 2012. Optimality and robustness in quorum sensing (QS)-mediated regulation of a costly public good enzyme. Proc. Natl. Acad. Sci. USA 109:19810-19815. https://doi.org/10.1073/pnas.1211072109
  19. Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor laboratory Press, Cold Srping Harbor, NY.
  20. Sato, Z., Koiso, Y., Iwasaki, S. and Matsuda, I. 1989. Toxins produced by Pseudomonas glumae. Ann. Phytopathol. Soc. Jpn. 55:356-359.
  21. Storey, J. D. and Tibshirani, R. 2003. Statistical significance for genomewide studies. Proc. Natl. Acad. Sci. USA. 100:9440-9445. https://doi.org/10.1073/pnas.1530509100

Cited by

  1. N -acylhomoserine lactone-regulation of genes mediating motility and pathogenicity in Pseudomonas syringae pathovar tabaci 11528 vol.6, pp.3, 2017, https://doi.org/10.1002/mbo3.440
  2. Understanding pathogenic Burkholderia glumae metabolic and signaling pathways within rice tissues through in vivo transcriptome analyses vol.547, pp.1, 2014, https://doi.org/10.1016/j.gene.2014.06.029
  3. Susceptibility of Opportunistic Burkholderia glumae to Copper Surfaces Following Wet or Dry Surface Contact vol.19, pp.7, 2014, https://doi.org/10.3390/molecules19079975
  4. Functional and genomic insights into the pathogenesis of B urkholderia species to rice vol.18, pp.3, 2016, https://doi.org/10.1111/1462-2920.13189
  5. (Meta)genomic insights into the pathogenome of Cellulosimicrobium cellulans vol.6, pp.1, 2016, https://doi.org/10.1038/srep25527
  6. Genome-Wide RNA Sequencing Analysis of Quorum Sensing-Controlled Regulons in the Plant-Associated Burkholderia glumae PG1 Strain vol.81, pp.23, 2015, https://doi.org/10.1128/AEM.01043-15