Skip to main content
SpringerLink
Log in

Identification of virulence factors and type III effectors of phylotype I, Indian Ralstonia solanacearum strains Rs-09-161 and Rs-10-244

  • Research Article
  • Published:
Journal of Genetics Aims and scope Submit manuscript

Abstract

Ralstonia solanacearum is a well-known phytopathogen causing bacterial wilt in a large number of agriculturally important crops. The pathogenicity of R. solanacearum is expressed due to the presence of various virulence factors and effector proteins. In this study, various virulence factors and type III effector proteins of R. solanacearum that are present in the strains Rs-09-161 and Rs-10-244 were identified through bioinformatics approach and compared with other reference strains. R. solanacearum strains, Rs-09-161 and Rs-10-244 belong to the phylotype I, biovar3, and are the only sequenced strains from India infecting solanaceous vegetables. Similarity matrix obtained by comparing the sequences of virulence genes of Rs-09-161 and Rs-10-244 with other reference strains indicated that Rs-09-161 and Rs-10-244 share more than 99% similarity between them and are closely related to GMI1000. The virulence factors in R. solanacearum appear to be highly conserved in the R. solanacearum species complex. Rs-09-161 has 72 type III effectors whereas Rs-10-244 has 77. Comparison of the complete genes of type III effectors of Rs-09-161, Rs-10-244 and GMI1000 revealed the presence of 60 common effectors within them. Further, Rs-09-161 has two unique effectors and Rs-10-244 has four unique effectors. Phylogenetic trees of RipA, RipG, RipH and RipS effector sequences resulted in the grouping of the isolates based on their phylotypes. Group 1 consists of strains that belong to phylotype I including Rs-09-161 and Rs-10-244. Phylotype III strain CMR15 forms a group closely associated with phylotype I. The strains belonging to phylotypes II and IV have separated to form two different groups.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Alfano J. R. and Collmer A. 2004 Type III secretion system effector proteins: double agents in bacterial disease and plant defense. Annu. Rev. Phytopathol. 42, 385–414.

    Article  CAS  PubMed  Google Scholar 

  • Alvarez B., Biosca E. and Lopez M. 2010 On the life of Ralstonia solanacearum, a destructive bacterial plant pathogen. In Current research, technology and education topics in applied microbiology and microbial biotechnology. (ed. A. Mendez Vilas), vol. 1, pp. 267–279. Formatex, Badajoz.

  • Boucher C. A., Van Gijsegem F., Barberies P. A., Arlat M. and Zischek C. 1987 Pseudomanas solanacearum genes controlling both pathogenicity on tomato and hypersensitivity on tobacco are clustered. J. Bacteriol. 169, 5626–5633.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chandrashekara K. N., Kumar M. P. and Saroja S. 2012 Aggressiveness of Ralstonia solanacearum isolates on tomato. J. Exp. Sci. 3, 05-09.

    CAS  Google Scholar 

  • Chen L., Shirota M., Zhang Y., Kiba A., Hikichi Y. and Ohnishi, K. 2014 Involvement of HLK effectors in Ralstonia solanacearum disease development in tomato. J. Gen. Plant Pathol. 80, 79–84.

    Article  CAS  Google Scholar 

  • Cunnac S., Occhialini A., Barberis P., Boucher C. and Genin S. 2004 Inventory and functional analysis of the large Hrp regulon in Ralstonia solanacearum: identification of novel effector proteins translocated to plant host cells through the type III secretion system. Mol. Microbiol. 53, 115–128.

    Article  CAS  PubMed  Google Scholar 

  • Fegan M. and Prior P. 2005. How complex is the Ralstonia solanacearum species complex? In bacterial wilt disease and the Ralstonia solanacearum species complex (ed. C. Allen, P. Prior and A. C. Hayward), pp. 449–461. APS Press, St Paul.

    Google Scholar 

  • Gabriel D. W., Allen C., Schell M., Denny T. P., Greenberg J. T., Duan Y. P. et al. 2006 Identification of open reading frames unique to a select agent: Ralstonia solanacearum race 3 biovar 2. Mol. Plant Microbe Interact. 19, 69–79.

    Article  CAS  PubMed  Google Scholar 

  • Genin S., Gough C. L., Zischek C. and Boucher C. A. 1992 Evidence that the hrpB gene encodes a positive regulator of pathogenicity genes from Pseudomonas solanacearum. Mol. Microbiol. 6, 3065–3076.

    Article  CAS  PubMed  Google Scholar 

  • Genin S. and Denny T. P. 2012 Pathogenomics of the Ralstonia solanacearum species complex. Annu. Rev. Phytopathol. 50, 67–89.

    Article  CAS  PubMed  Google Scholar 

  • Hajri A., Pothier J. F., Fischer-Le Saux M., Bonneau S., Poussier S. et al. 2011 Type three effector genes distribution and sequence analysis provides new insights into pathogenicity of plant pathogenic Xanthomonas arboricola. Appl. Environ. Microbiol. 78, 371–384.

    Article  PubMed  Google Scholar 

  • Heesemann J., Algermissen B. and Laufs R. 1984 Genetically manipulated virulence of Yersinia enterocolitica. Infect. Immun. 46, 105–110.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hua Z. and Vierstra R. D. 2011 The cullin-RING ubiquitin-protein ligases. Annu. Rev. Plant Biol. 62, 299–334.

    Article  CAS  PubMed  Google Scholar 

  • Hueck C. J. 1998 Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiol. Mol. Biol. Rev. 62, 379–433.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Jukes T. H. and Cantor C. R. 1969 Evolution of protein molecules. InMammalian protein metabolism (ed. H. N. Munro), pp. 21–132. Academic Press, New York.

    Chapter  Google Scholar 

  • Kang Y., Liu H., Genin S., Schell M. A. and Denny T. P. 2002 Ralstonia solanacearum requires type 4 pili to adhere to multiple surfaces and for natural transformation and virulence. Mol. Microbiol. 46, 427–437.

    Article  CAS  PubMed  Google Scholar 

  • Kumar A., Sarma Y. R. and Anandaraj M. 2004 Evaluation of genetic diversity of Ralstonia solanacearum causing bacterial wilt of ginger using REP-PCR and PCR-RFLP. Curr. Sci. 87, 1555–1561.

    Google Scholar 

  • Mansfield J., Genin S., Magori S., Citovsky V., Sriariyanum M., Ronald P. et al. 2012 Top 10 plant pathogenic bacteria in molecular plant pathology. Mol. Plant Pathol. 13, 614–629.

    Article  PubMed  Google Scholar 

  • Meng F., Yao J. and Allen C. 2011 A MotN mutant of Ralstonia solanacearum is hypermotile and has reduced virulence. J. Bacteriol. 193, 2477–2486.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mukaihara T., Tamura N., Murata Y. and Iwabuchi M. 2004 Genetic screening of Hrp type III-related pathogenicity genes controlled by the HrpB transcriptional activator in Ralstonia solanacearum. Mol. Microbiol. 54, 863–875.

    Article  CAS  PubMed  Google Scholar 

  • Mukaihara T. and Tamura N. 2009 Identification of novel Ralstonia solanacearum type III effector proteins through translocation analysis of hrpB-regulated gene products. Microbiology 155, 2235–2244.

    Article  CAS  PubMed  Google Scholar 

  • Mukaihara T., Tamura N. and Iwabuchi M. 2010 Genome-Wide identification of a large repertoire of Ralstonia solanacearum type III effector proteins by a new functional screen Mol. Plant Microbe Interact. 23, 251–262.

    Article  CAS  Google Scholar 

  • Peeters N., Carrere S., Anisimova M., Plener L., Cazale A. C. and Genin S. 2013 Repertoire, unified nomenclature and evolution of the type III effector gene set in the Ralstonia solanacearum species complex. BMC Genomics 14, 859–877.

    Article  PubMed  PubMed Central  Google Scholar 

  • Poueymiro M. and Genin S. 2009 Secreted proteins from Ralstonia solanacearum: a hundred tricks to kill a plant. Curr. Opin. Microbiol. 12, 44–52.

    Article  CAS  PubMed  Google Scholar 

  • Ramesh R. and Phadke G. S. 2012 Rhizosphere and endophytic bacteria for the suppression of eggplant wilt caused by Ralstonia solanacearum. Crop Prot. 37, 35–41.

    Article  Google Scholar 

  • Ramesh R., Gaitonde S., Achari G., Asolkar T., Singh N. P., Carrere S. et al. 2014a Genome sequencing of Ralstonia solanacearum Biovar 3, phylotype I, strains Rs-09-161 and Rs-10-244, ssolated from eggplant and chili in India. Genome Announc. 2, 1–2.

    Article  Google Scholar 

  • Ramesh R., Achari G. A. and Gaitonde S. 2014b Genetic diversity of Ralstonia solanacearum infecting solanaceous vegetables from India reveals the existence of unknown or newer sequevars of Phylotype I strains. Eur. J. Plant Pathol. 140, 543–562.

    Article  CAS  Google Scholar 

  • Remenant B., Coupat-Goutaland B., Guidot A., Cellier G., Wicker E., Allen, C. et al. 2010 Genomes of three tomato pathogens within the Ralstonia solanacearum species complex reveal significant evolutionary divergence. BMC Genomics. 11, 379–395.

    Article  PubMed  PubMed Central  Google Scholar 

  • Remenant B., de Cambiaire J-C., Cellier G., Jacobs J. M., Mangenot S., Barbe V. et al. 2011 Ralstonia syzygii, the blood disease bacterium and some Asian R. Solanacearum strains form a single genomic species despite divergent lifestyles. PLoS One 6, e24356.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Remigi P., Anisimova M., Guidot A., Genin S. and Peeter N. 2011 Functional diversification of the GALA type III effector family contributes to Ralstonia solanacearum adaptation on different plant hosts. New Phytol. 192, 976–987.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saile E., McGarvey J., Schell M. and Denny T. 1997 Role of extracellular polysaccharide and endoglucanase in root invasion and colonization of tomato plants by Ralstonia solanacearum. Phytopathology 87, 1264–1271.

    Article  CAS  PubMed  Google Scholar 

  • Salanoubat M., Genin S., Artiguenave F., Gouzy J., Mangenot S., Arlat M. et al. 2002 Genome sequence of the plant pathogen Ralstonia solanacearum. Nature 415, 497–502.

    Article  CAS  PubMed  Google Scholar 

  • Schell M. A. 2000 Control of virulence and pathogenicity genes of Ralstonia solanacearum by an elaborate sensory network. Annu. Rev. Phytopathol. 38, 263–292.

    Article  CAS  PubMed  Google Scholar 

  • Sole M., Popa C., Mith O., Sohn K. H., Jones J. D., Deslandes L. et al. 2012 The awr gene family encodes a novel class of Ralstonia solanacearum type III effectors displaying virulence and avirulence activities. Mol. Plant Microbe Interact. 25, 941–953.

    Article  CAS  PubMed  Google Scholar 

  • Tamura K., Stecher G., Peterson D., Filipski A. and Kumar S. 2013 MEGA6: molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30, 2725–2729.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tans-Kersten J., Huang H. and Allen C. 2001 Ralstonia solanacearum needs motility for invasive virulence on tomato. J. Bacteriol. 183, 3597–3605.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thompson J. D., Higgins D. G. and Gibson T. J. 1994 CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673–4680.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Van Gijsegem F., Vasse J., Camus J.-C., Marenda M. and Boucher C. 2000 Ralstonia solanacearum produces Hrp-dependent pili that are required for PopA secretion but not for attachment of bacteria to plant cells. Mol. Microbiol. 36, 249–260.

    Article  PubMed  Google Scholar 

  • Van Gijsegem F., Vasse J., Rycke R. D., Castello P. and Boucher C. 2002 Genetic dissection of the Ralstonia solanacearum hrp gene cluster reveals that the HrpV and HrpX proteins are required for Hrp pilus assembly. Mol. Microbiol. 44, 935–946.

    Article  PubMed  Google Scholar 

  • Vos M. and Velicer, G. J. 2006 Genetic population structure of the soil bacterium Myxococcus xanthus at the centimeter scale. Appl. Environ. Microbiol. 72, 3615–3625.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu J., Zheng H. J., Liu L., Pan Z. C., Prior P., Tang B. et al. 2011 Complete genome sequence of the plant pathogen Ralstonia solanacearum strain Po82. J. Bacteriol. 193, 4261–4262.

  • Yao J. and Allen C. 2006 Chemotaxis is required for virulence and competitive fitness of the bacterial wilt pathogen Ralstonia solanacearum. J. Bacteriol. 188, 3697–3708.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zumaquero A., Macho A. P., Rufian J. S. and Beuzon C. R. 2010 Analysis of the role of the type III effector inventory of Pseudomonas syringae pv. phaseolicola 1448a in interaction with the plant. J. Bacteriol. 192, 4474–4488.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge the financial support by Indian Council of Agricultural Research, New Delhi, India through ‘Outreach project on Phytophthora, Fusarium and Ralstonia diseases of horticultural and field crops (PhytoFuRa)’ and Director of ICAR-CCARI for providing necessary support. We acknowledge Nemo Peeters, Sebastien Carrere and Stephane Genin-INRA, Laboratoire des Interactions Plantes-Microorganismes (LIPM), Castanet-Tolosan, France for annotation of the sequences of R. solanacearum strains Rs-09-161 and Rs-10-244.

Author information

Authors and Affiliations

  1. Department of Microbiology, Goa University, Taleigao Plateau, 403 206, India

    Trupti Asolkar

  2. ICAR-Central Coastal Agricultural Research Institute, Old Goa, 403 402, India

    Trupti Asolkar & Raman Ramesh

Authors
  1. Trupti Asolkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raman Ramesh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Raman Ramesh.

Additional information

Corresponding editor: Umesh Varshney

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (docx 26 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Asolkar, T., Ramesh, R. Identification of virulence factors and type III effectors of phylotype I, Indian Ralstonia solanacearum strains Rs-09-161 and Rs-10-244. J Genet 97, 55–66 (2018). https://doi.org/10.1007/s12041-018-0894-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12041-018-0894-z

Keywords

Search

Navigation