Skip to main content

Advertisement

Log in

Genetic diversity of Ralstonia solanacearum infecting solanaceous vegetables from India reveals the existence of unknown or newer sequevars of Phylotype I strains

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Bacterial wilt is one of the important constraints in the cultivation of solanaceous vegetables in India. The disease is caused by Ralstonia solanacearum, a soil bacterium. We have collected 232 isolates of R. solanacearum infecting solanaceous vegetables (eggplant, tomato and chilli) and other crops from different parts of India. Pathogenicity of the isolates was tested on eggplant, tomato and chilli and the pathogen was confirmed by PCR. Multiplex PCR and biochemical tests indicated that all the isolates were phylotype I and biovar 3. Ninety-five representative isolates selected based on geographical region, host range and pathogenicity were subjected to further phylogenetic and diversity analysis. Sequence analysis of egl, pga and hrpB genes of 95 isolates and genetic diversity of 50 representative isolates was reported and discussed. Indian isolates within the Phylotype I did not group based on the host or geographical location, except clustering of isolates from the Andaman Islands. Indian isolates clustered into two sub groups based on egl and pga trees indicating the presence of two major population groups. Sub group 1 is the dominant group in the data set and consists of unknown/newer sequevars, and sub group 2 consist of mainly the isolates which are designated with sequevar numbers based on egl sequences. In the hrpB based tree, the sub group 2 is the dominant group in the data set and it is the same for the sub group 1 of the egl tree. Indian phylotpe I R. solanacearum strains are phenotypically diverse including the previously described sequevars 14, 17, 44, 47 and 48. Our studies indicated the existence of R. solanacearum isolates with unknown/newer sequevars; the diversity existing among the phylotype I isolates might be due to a continuous evolutionary process. To our knowledge this is the first detailed report on the diversity of phylotype I R. solanacearum strains infecting solanaceous vegetables and the existence of unknown/newer sequevars in India.

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

Access this article

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

Similar content being viewed by others

References

  • Bertolla, F., Frostegård, A., Brito, B., Nesme, X., & Simonet, P. (1999). During infection of its hosts, the plant pathogen Ralstonia solanacearum naturally develops a state of competence and exchanges genetic material. Molecular Plant-Microbe Interactions, 12, 467–472.

    Article  CAS  Google Scholar 

  • Cook, D., & Sequeira, L. (1994). Strain differentiation of Pseudomonas solanacearum by molecular genetic methods. In A. C. Hayward & G. L. Hartman (Eds.), Bacterial Wilt: The disease and its causative agent, Pseudomonas solanacearum (pp. 77–93). Wallingford: CAB International.

    Google Scholar 

  • Cook, D., Barlow, E., & Sequeira, L. (1989). Genetic diversity of Pseudomonas solanacearum: detection of restriction fragment length polymorphisms with DNA probes that specify virulence and the hypersensitive response. Molecular Plant-Microbe Interactions, 2, 113–121.

    Article  Google Scholar 

  • Fegan, M., & Prior, P. (2005). How complex is the Ralstonia solanacearum species complex? In C. Allen, P. Prior, & A. C. Hayward (Eds.), Bacterial wilt disease and the Ralstonia solanacearum species complex (pp. 449–461). St. Paul: American Phytopathological Society.

    Google Scholar 

  • Gillings, M., Fahy, P., & Davies, C. (1993). Restriction analysis of an amplified polygalacturonase gene fragment differentiates strains of the phytopathogenic bacterium Pseudomonas solanacearum. Letters in Applied Microbiology, 17, 44–48.

    Article  PubMed  CAS  Google Scholar 

  • Grover, A., Azmi, W., Gadewar, A. V., Pattanayak, D., Naik, P. S., Shekhawat, G. S., & Chakrabarti, S. K. (2006). Genotypic diversity in a localized population of Ralstonia solanacearum as revealed by random amplified polymorphic DNA markers. Journal of Applied Microbiology, 101, 798–806.

    Article  PubMed  CAS  Google Scholar 

  • Guidot, A., Prior, P., Schoenfeld, J., Carrere, S., Genin, S., & Boucher, C. (2007). Genomic structure and phylogeny of the plant pathogen Ralstonia solanacearum inferred from gene distribution analysis. Journal of Bacteriology, 189, 377–387.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Hayward, A. C. (1991). Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum. Annual Review of Phytopathology, 29, 65–87.

    Article  PubMed  CAS  Google Scholar 

  • Hayward, A. C. (2000). Ralstonia solanacearum. In J. Lederberg (Ed.), Encyclopedia of microbiology (pp. 32–42). San Diego: Academic.

    Google Scholar 

  • Horita, M., & Tsuchiya, K. (2000). Comparative analysis of Japanese and foreign strains of Ralstonia solanacearum based on 16S ribosomal RNA gene sequences. Journal of General Plant Pathology, 66, 132–137.

    Article  CAS  Google Scholar 

  • Ivey, M. L. L., Gardener, B. B. M., Opina, N., & Miller, S. A. (2007). Diversity of Ralstonia solanacearum infecting eggplant in the Philippines. Phytopathology, 97, 1467–1475.

    Article  PubMed  CAS  Google Scholar 

  • Jeong, Y., Kim, J., Kang, Y., Lee, S., & Hwang, I. (2007). Genetic diversity and distribution of Korean isolates of Ralstonia solanacearum. Plant Disease, 91, 1277–1287.

    Article  CAS  Google Scholar 

  • Jukes, T. H., & Cantor, C. R. (1969). Evolution of protein molecules. In H. N. Munro (Ed.), Mammalian protein metabolism (pp. 21–132). New York: Academic.

    Chapter  Google Scholar 

  • Kelman, A. (1954). The relationship of pathogenicity in Pseudomonas solanacearum to colony appearance on a tetrazolium medium. Phytopathology, 44, 693–695.

    Google Scholar 

  • Kelman, A., & Person, L. H. (1961). Strains of Pseudomonas solanacearum differing in pathogenicity to tobacco and peanut. Phytopathology, 51, 158–161.

    Google Scholar 

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

    Google Scholar 

  • Kumar, A., Prameela, T. P., Suseela Bhai, R., Siljo, A., Biju, C. N., Anandaraj, M., & Vinatzer, B. A. (2012). Small cardamom (Elettaria cardamomum Maton.) and ginger (Zingiber officinale Roxb) bacterial wilt is caused by same strain of Ralstonia solanacearum: a result revealed by multilocus sequence typing (MLST). European Journal of Plant Pathology, 132, 477–482.

    Article  CAS  Google Scholar 

  • Kumar, A., Prameela, T. P., & Suseelabhai, R. (2013). A unique DNA repair and recombination gene (recN) sequence for identification and intraspecific molecular typing of bacterial wilt pathogen Ralstonia solanacearum and its comparative analysis with ribosomal DNA sequences. Journal of Biosciences, 38, 1–12.

    Article  Google Scholar 

  • Liu, Y., Kanda, Y., Yano, K., Kiba, A., Hikichi, Y., Aino, M., Kawaguchi, A., Mizoguchi, S., Nakaho, K., Shiomi, H., Takikawa, Y., & Ohnishi, K. (2009). Molecular typing of Japanese strains of Ralstonia solanacearum in relation to the ability to induce a hypersensitive reaction in tobacco. Journal of General Plant Pathology, 75, 369–380.

    Article  CAS  Google Scholar 

  • Mondal, B., Bhattacharya, I., & Khatua, D. C. (2011). Crop and weed hosts of Ralstonia solanacearum in West Bengal. Journal of Crop and Weed, 7, 195–199.

    Google Scholar 

  • Opina, N., Tavner, F., Holloway, G., Wang, J. F., Li, T. H., Maghirang, R., Fegan, M., Hayward, A. C., Krishnapillai, V., Hong, W. F., Holloway, B. W., & Timmis, J. N. (1997). A novel method for development of species and strain-specific DNA probes and PCR primers for identifying Burkholderia solanacearum (formerly Pseudomonas solanacearum). Asia Pacific Journal of Molecular Biology and Biotechnology, 5, 19–33.

    Google Scholar 

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

    Article  Google Scholar 

  • Remenant, B., Coupat-Goutaland, B., Guidot, A., Cellier, G., Wicker, E., Allen, C., Fegan, M., Pruvost, O., Elbaz, M., Calteau, A., Salvignol, G., Mornico, D., Mangenot, S., Barbe, V., Médigue, C., & Prior, P. (2010). Genomes of three tomato pathogens within the Ralstonia solanacearum species complex reveal significant evolutionary divergence. BMC Genomics, 11, 379. doi:10.1186/1471-2164-11-379.

    Article  PubMed  PubMed Central  Google Scholar 

  • Sagar, V., Jeevalatha, A., Mian, S., Chakrabarti, S. K., Gurjar, M. S., Arora, R. K., Sharma, S., Bakade, R. R., & Singh, B. P. (2014). Potato bacterial wilt in India caused by strains of phylotype I, II and IV of Ralstonia solanacearum. European Journal of Plant Pathology, 138, 51–65.

    Article  Google Scholar 

  • Smith, J. J., Offord, L. C., Holderness, L. C., Holderness, M., & Saddler, G. S. (1995). Genetic diversity of Burkholderia solanacearum race 3 in Kenya. Applied and Environmental Microbiology, 61, 4263–4268.

    PubMed  CAS  PubMed Central  Google Scholar 

  • Stevens, P., & van Elsas, J. D. (2010). Genetic and phenotypic diversity of Ralstonia solanacearum biovar 2 strains obtained from Dutch waterways. Antonie van Leeuwenhoek, 97, 171–188.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Tamura, K., Dudley, J., Nei, M., & Kumar, S. (2007). MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24, 1596–1599.

    Article  PubMed  CAS  Google Scholar 

  • Thompson, J. D., Higgins, D. G., & 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 Research, 22, 4673–4680.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Villa, J. E., Tsuchiya, K., Horita, M., Natural, M., Opina, N., & Hyakumachi, M. (2005). Phylogenetic relationships of Ralstonia solanacearum species complex strains from Asia and other continents based on 16S rDNA, endoglucanase, and hrpB gene sequences. Journal of General Plant Pathology, 71, 39–46.

    Article  CAS  Google Scholar 

  • Wicker, E., Grassart, L., Coranson-Beaudu, R., Mian, D., Guilbaud, C., Fegan, M., & Prior, P. (2007). Ralstonia solanacearum strains from Martinique (French West Indies) exhibiting a new pathogenic potential. Applied and Environmental Microbiology, 73, 6790–6801.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Wicker, E., Lefeuvre, P., de Cambiaire, J. C., Lemaire, C., Poussier, S., & Prior, P. (2012). Contrasting recombination patterns and demographic histories of the plant pathogen Ralstonia solanacearum inferred from MLSA. International Society for Microbial Ecology Journal, 6, 961–974.

    CAS  Google Scholar 

  • Xu, J., Pan, Z. C., Prior, P., Xu, J. S., Zhang, Z., Zhang, H., Zhang, L. Q., He, L. Y., & Feng, J. (2009). Genetic diversity of Ralstonia solanacearum strains from China. European Journal of Plant Pathology, 125, 641–653.

    Article  CAS  Google Scholar 

  • Xue, Q. Y., Yin, Y. N., Yang, W., Heuer, H., Prior, P., Guo, J. H., & Smalla, K. (2011). Genetic diversity of Ralstonia solanacearum strains from China assessed by PCR-based fingerprints to unravel host plant- and site-dependent distribution patterns. FEMS Microbiology Ecology, 75, 507–519.

    Article  PubMed  CAS  Google Scholar 

  • Yabuuchi, E., Kosako, Y., Yano, I., Hotta, H., & Nishiuchi, Y. (1995). Transfer of two Burkholderia and an Alcaligenes species to Ralstonia gen. nov.: proposal of Ralstonia pickettii (Ralston, Palleroni and Doudoroff, 1973) comb. nov., Ralstonia solanacearum (Smith 1896) comb. nov. and Ralstonia eutropha (Davis 1969) comb. nov. Microbiology and Immunology, 39, 897–904.

    Article  PubMed  CAS  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). We are grateful to Director, CPRI, Shimla for providing sequencing facilities and to Director, ICAR Research Complex for Goa, Old Goa for providing other necessary facilities. We are thankful to Dr. Emmanuel Wicker, CIRAD, La ReUnion, France for analyzing egl sequence data to assign sequevar numbers.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Raman Ramesh.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ramesh, R., Achari, G.A. & Gaitonde, S. 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 (2014). https://doi.org/10.1007/s10658-014-0487-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-014-0487-5

Keywords

Navigation