Phenotypic and genetic properties of Ralstonia solanacearum strains, the causal agent of bacterial wilt of potato isolated from Kurdistan province*

Document Type : Research Article

Authors

Abstract

Phenotypic properties and genetic diversity of 24 strains of Ralstonia solanacearum causing bacterial wilt of potato plants isolated from various locations in Kurdistan province was investigated. According to physiological and biochemical properties, 19 and 5 of isolates were identified as R. solanacearum biovar 2 and 3, respectively. Identification of isolates was further confirmed by PCR using 759/760 primers specific to R. Solanasearum. Based on multiplex PCR, all strains identified as biovar 2 were belonging to phylotype II. In multiplex PCR, strains identified as biovar 3 produced R. solanacearum species-specific fragment but failed to produce expected phylotype-specific amplicon. Genetic diversity of selected strains was investigated by rep-PCR fingerprinting using REP, ERIC and BOX primers. UPGMA analysis of combined data obtained from rep-PCR fingerprint pattern using Dice,s coefficient revealed the strains could be separated into 5 groups at a similarity level of approximately 43%. There was no direct relationship between geographic locations of strains and grouping. Obtained results demonstrated the existence of a genetic diversity among R. solanacearum strains causing bacterial wilt disease of potato in Kurdistan province.

Keywords


Azadvar M., Rahimian H. 2006. Studies on bacteriological characteristics of casual agent of bacterial wilt of potato in Southern Kerman. Pajouhesh and Sazandegi, 19:94-102.
Bagheri A. and S.M. Taghavi. 2001. Characteristics of strains of the causal agent of bacterial wilt of potato and tomato in Fars province and reaction of some potato and tomato cultivars to the pathogen. Iranian Journal of Plant Pathology 36:69–72.
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 Interaction, 2:113-121.
Cook D., Sequeira L. 1994. Strain differentiation of Pseudomonas solanacearum by molecular genetic methods. In: Hayward AC, Hartman GL, eds, Bacterial wilt, the  disease and its causative agent, Pseudomonas solanacearum. Wallingford, UK: CAB International, 77-93.
Danesh D., Bahar M. 1984. Occurrence of bacterial wilt of potato in Iran. In: Proceedings of the ninth triennial conference of the European association for potato research, Interlaken, Switzerland, 407-408.
Denny T.P. 2006. Plant pathogenic Ralstonia species. In: Gnanamanickam S.S., ed. Plant associated bacteria. Dordrecht, The Netherlands: Springer Publishing, 573-644.
Dice L.R. 1945. Measurement of the amount of ecologic association between species. Ecology, 26:297–302.
Elphinstone J.G. 2005. The current bacterial wilt situation: a global overview. Pages 9-28 in: Bacterial Wilt: The Disease and the Ralstonia solanacearum Species Complex. C. Allen, Prior P., and A. C. Hayward, eds. American Phytopathological Society, St. Paul, MN.
Fegan M., Prior P. 2005. How complex is the Ralstonia solanacearum species complex? Pages 449-461 in: Bacterial wilt disease and the Ralstonia solanacearum species complex. C. Allen, P. Prior, and A. C. Hayward, eds. American Phytopathological Society, St. Paul, MN.
Fegan M., Taghavi M., Sly L., Hayward A.C. 1998. Phylogeny, diversity and molecular diagnostic of Ralstonia solanacearum. Pages 19-33 in: Bacterial wilt disease: Molecular and Ecological aspects. P. Prior, C. Allen, and J. Elphinstone, eds. INRA Edition, Paris.
Gillings M., Fahy P. 1993. Genetic diversity of Pseudomonas solanacearum biovar 2 and N2 assessed using restriction endonuclease analysis of total genomic DNA. Plant Pathology 42:744-753.
Gillings M., Fahy P., Davies C. 1993. Restriction analysis of an amplified polygalactoronase gene fragment differentiates strains of the phytopathogenic bacterium Pseudomonas solanacearum. Letters in  Applied Microbiology 17:44-48.
Hayward A.C. 1964. Characterization of Pseudomonas solanacearum. Journal Applied Bacteriology 27:265-277.
Hayward A.C. 1991. Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum. Annual Review Phytopathology 29:65-87.
Hayward A.C. 1994. The hosts of Pseudomonas solanacearum. In Bacterial wilt: The disease and its causative agent, Pseudomonas solanacearum, eds. A.C. Hayward and G.L. Hartman, 9-23. Wallingford: CAB International.
Hayward A.C. 1994. Systematic and phylogeny of Pseudomonas solanacearum and related bacteria. In: Hayward AC, Hartman GL, eds. Bacterial wilt: the disease and its causative agent, Pseudomonas solanacearum. Wallingford, UK: CAB International, 123-135.
Horita M., Tsuchiya K. 2001. Genetic Diversity of Japanese strains of Ralstonia solanacearum. Phytopathology 91:399-407.
Horita M., Tsuchiya K., Ooshiro A. 2005. Characteristics of Ralstonia Solanacearum biovar N2 strains in Asia. Journal of Phytopathology 153:209-213.
Horita M., Tsuchiya K., Suga Y., Yano K., Waki T., Kurose D., Furuya N. 2014. Current classification of Ralstonia solanacearum and genetic diversity of the strains in Japan. Journal of General Plant Pathology 80:455-465.
Ivey M.L., Gardener B.B., Opina N., Miller S.A. 2007. Diversity of Ralstonia solanacearum Infecting Eggplant in the Philippines. Phytopathology 97:1467-75.
Izadiyan M., Taghavi S.M. 2011. Diversity of Iranian isolates of Ralstonia solanacearum. Phytopathologia mediterranea 50:236-244.
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.
Kelman A. 1954. The relationship of pathogenicity in Pseudomonas solanacearum to colony appearance on a tetrazolium chloride medium. Phytopathology 44:693-695.
Louws F.J., Fulbright D.W., Stephens C.T., de Bruijn F.J. 1994. Specific genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR. Applied and Environmental Microbiology 60:2286-2295.
Maghooli M., Hayati J., Taghavi M., Mostafazadeh G.R. 2004. Characteristics of Ralstonia solanacearum in Khuzestan. In: Proceedings of the Sixteenth plant protection congress, Tabriz, Iran,218.
Melanie L., Lewis I., Brian B., Gardner M., Opina N., Miller S.A. 2007. Diversity of Ralstonia solanacearum infecting eggplant in the Philippines. Phytopathology 97:1467-1475.
Montanelli C.A., Chiari A., Chiari T., Stefanini F., Nascari G. 1995. Evaluation of resistance to Pseudomonas solanacearum in potato under controlled conditions. Euphytica 81:35-43.
Norman D.J., Zapata M., Gabriel D.W., Duan Y.P., Yuen J.M.F., Mangravita-Novo A., Donahoo R.S. 2009. Genetic diversity and host range variation of Ralstonia solanacearum strains entering North America. Phytopathology 99:1070-1077.
Nouri S., Bahar M., Fegan M. 2009. Diversity of Ralstonia solanacearum causing potato bacterial wilt in Iran and the first record of phylotype II/biovar 2T strains outside South America. Plant pathology 58:243-249.
Opina N., Tavner F., Hollway G., Wang J.F., Li T.H., Maghirang R., Fegan M., Hayward A.C., Krishnapillai V., Hong W.F., Holloway B.W., Timmis J. 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 Molecular Biology and Biotechnology 5:19-30.
Prior P., Fegan M. 2005. Recent developments in the phylogeny and classification of Ralstonia solanacearum. Acta Horticaltura 695:127-136.
Prior P., Ailloud F., Dalsing B.L., Remenant B., Sanchez B., Allen C. 2016. Genomic and proteomic evidence support the division of the plant pathogen Ralstonia solanacearum into three species. BioMed Central Genomics [http://bmcgenomics.biomedcentral.com] 10.1186/s12864-016-2413-z.
Poussier S., Prior P., Luisetti J., Hayward C., Fegan M. 2000. Partial sequencing of the hrpB and endoglucanase genes confirmed and expands the known diversity within the Ralstonia solanacearum species complex. Systematic and Applied Microbiology 23:479-486.
Poussier S., Trigalet-Demery D., Wandewalle P., Goffinet B., Luisetti J., Trigalet A. 2000. Genetic diversity of Ralstonia solanacearum as assessed by PCR-RFLP of the hrp gene region, AFLP and 16S rRNA sequence analysis, and identification of an African subdivision. Micrbiology 146:1679-1692.
Robertson A.E., Fortnum B.A., Wood T.C., Kluepfel D.A. 2001. Diversity of Ralstonia solanacearum in the Southeastern United States. Contributions to Tobacco Research 19:323-331.
Safni I., Cleenwerck I., De Vos P., Fegan M., Sly L., Kappler U. Polyphasic taxonomic revision of the Ralstonia solanacearum species complex: proposal to emend the descriptions of Ralstonia solanacearum and Ralstonia syzygii and reclassify current R. syzygii strains as Ralstonia syzygii subsp. syzygii subsp. nov., R. solanacearum phylotype IV strains as Ralstonia syzygii subsp. indonesiensis subsp. nov., banana blood disease bacterium strains as Ralstonia syzygii subsp. celebensis subsp. nov., and R. solanacearum phylotype I and III strains as Ralstinia pseudosolanacearum sp. nov. International Journal of Systematic and Evolutionary Microbiology 64:3087-3103.
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 IVof Ralstonia solanacearum. European Journal of Plant Pathology, 138:51–65.
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.
Thwaites R., Mansfield J., Eden-Green S., Seal S. 1999. RAPD and rep PCR-based fingerprinting of vascular bacterial pathogens of Musa spp. Plant Pathology 48:121-128. 
Winstead N.N., Kelman A. 1952. Inoculation techniques for evaluating resistance to Pseudomonas solanacearum. Phytopathology 42:628-634.