Diversity of Ralstonia pseudosolanacearum, the causal agent of bacterial wilt on Eucalyptus pellita in Indonesia




Abstract. Siregar BA, Giyanto, Hidayat SH, Siregar IZ, Tjahjono B. 2021. Diversity of Ralstonia pseudosolanacearum, the causal agent of bacterial wilt on Eucalyptus pellita in Indonesia. Biodiversitas 22: 2538-2545. The Ralstonia species complex was initially classified into five races and five biovars but the classification could not accommodate the isolates' phylogenetic history or geographic origins. A phylotype and sequevar system is based on the geographic distribution and characteristics of endoglucanase (egl) and hypersensitive response and pathogenicity (hrp) gene sequences. This study aims to describe pathogen diversity of the causal agent of bacterial wilt on Eucalyptus pellita F. Muell. Pathogens were isolated from wilting seedlings and trees in several Eucalyptus plantations. The phenotypic diversity analysis included biovar, exopolysaccharide quantification and virulence tests, while genotypic diversity included phylotypes and sequevar determination based on egl gene sequences. A total of 35 strains were isolated from the field and nurseries of Eucalyptus in various locations. All isolates were confirmed as Ralstonia species complex based on morphological characteristics and molecular studies using species-specific primers. These isolates were dominantly classified as biovar 3 and 4 and had a high variation on virulence and EPS production. Based on the egl sequence's alignment, 29 strains of Phylotype I are grouped into four sequevar references (sequevars 14, 17, 18, 30) and new a sequevar 58. This study shows that strains of R. pseudosolanacearum causing bacterial wilt on E. pellita in Indonesia have high phenotypic and genotypic diversities.


Addy HS, Askora A, Kawasaki T, Fujie M, Yamada T. 2012. Loss of virulence of the phytopathogen Ralstonia solanacearum through infection by ?RSM filamentous phages. Phytopathol. 102: 469-477
Albuquerque GMR, Santos LA, Felix KCS, Rollemberg CL, Silva AMF, Souza EB, Cellier G, Prior P, Mariano RL. 2014. Moko disease-causing strains of Ralstonia solanacearum from Brazil extend known diversity in paraphyletic phylotype II. Phytopathol. 104: 1175-1182.
Álvarez B, Biosca EG, López MM. 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. Méndez-Vilas, (Badajoz: Formatex), 267–279.
Araud-Razou I, Vasse J, Montrozier H, Etchebar C, Trigalet A. 1998. Detection and visualization of the major acidic exopolysaccharide of Ralstonia solanacearum and its role in tomato root infection and vascular colonization. Eur J Plant Pathol 104(8): 795-809.
Cao JD. 1982. Investigation of bacterial wilt in E. saligna and E. grandis introduced from Brazil. Guangxi For. Sci. Technol. 4: 30–31.
Carstensen G, Venter S, Chen S, Wingfield M, Coutinho T. 2017. Two Ralstonia species associated with bacterial wilt of Eucalyptus. Plant Pathol. 66: 393-403
Coutinho T, Jolanda R, Karl-Heinz R, Terblanche J. Wingfield M. 2008. First report of bacterial wilt caused by Ralstonia solanacearum on eucalypts in South Africa. Forest Pathol. 30: 205-210
Coutinho TA, Roux J, Riedel K, Terblanche J, Wingfield MJ. 2000. First report of bacterial wilt caused by Ralstonia solanacearum on eucalypts in South Africa. Forest Pathol. 30: 205-210
Denny TP, Hayward AC. 2001. Gram-negative bacteria Ralstonia. In: Schaad NW, Jones JB. Chun W (eds) Laboratory Guide for Identifcation of Plant Pathogenic Bacteria, 3rd Edition. APS Press, St. Paul
Denny TP, Makini FW, Brumbley SM. 1988. Characterization of Pseudomonas solanacearum Tn5 mutants deficient in extracellular polysaccharide. Mol. Plant-Microbe Interact. 1: 215-223.
Fegan M, Prior P. 2005. How complex is the Ralstonia solanacearum species complex? In: Allen C, Prior P, Hayward AC (eds) Bacterial Wilt Disease and The Ralstonia solanacearum Species Complex. Amer Phytopathological Society, St. Paul
Fonseca NR, Guimarães LMS, Hermenegildo PS, Teixeira RU, Lopes CA, Alfenas AC, 2014. Molecular characterization of Ralstonia solanacearum infecting Eucalyptus spp. in Brazil. Forest Pathol 44: 107–16
Fonseca NR, Oliveira LSS, Guimarães LMS, Teixeira RU, Lopes CA, Alfenas AC. 2015. An efficient inoculation method of Ralstonia solanacearum to test wilt resistance in Eucalyptus spp. Trop Plant Pathol. 41: 42–47
Hikichi, Y. 2016. Interactions between plant pathogenic bacteria and host plants during the establishment of susceptibility. J. Gen. Plant Pathol. 82: 326–331.
Ji P, Allen C, Sanchez-Perez A, Yao J, Elphinstone JG, Jones JB, Momol MT. 2007. New Diversity of Ralstonia solanacearum Strains Associated with Vegetable and Ornamental Crops in Florida. Plant Disease 91: 195-203.
Machmud M. 1985. Bacterial wilt in Indonesia. In: Craswell ET, Pushperajah E (eds) Bacterial Wilt Disease in Asia and The South Pacific. ACIAR, Canberra
Marques E, Uesugi CH, Ferreira MASV, Rezende D. 2012. Characterization of isolates of Ralstonia solanacearum biovar 2, pathogenic to Eucalyptus "urograndis" hybrids. Tropical Plant Pathology. 37: 399-408
McGarvey JA, Bell CJ, Denny TP, Schell MA. 1998. Analysis of extracellular polysaccharide I in culture and in planta using immunological methods: New insights and implications. In: Prior P, Allen C, Elphinstone J (eds) Bacterial Wilt Disease: Molecular and Ecological Aspects. Springer, New York
Negishi H, Yamada T, Shiraishi T, Oku H, Tanaka H. 1993. Pseudomonas solanacearum plasmid pJTPS1 mediates a shift from the pathogenic to nonpathogenic phenotype. Mol. Plant-Microbe Interact. 6: 203-209.
Peyraud R, Denny TP, Genin S. 2017. Exopolysaccharide quantification for the plant pathogen Ralstonia solanacearum. Bio-protocol 7: 1-8
Safni I, Cleenwerck I, De Vos P, Fegan M, Sly L, Kappler U. 2014. Polyphasic taxonomic revision of the Ralstonia solanacearum species complex: proposal to emend the descriptions of R. solanacearum and R. 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. celebesensis subsp. nov., and R. solanacearum phylotypes I and III strains as Ralstonia pseudosolanacearum sp. nov. Int J Syst Evol Microbiol. 64: 3087–3103
Saile E, McGarvey JA, Schell MA, Denny TP. 1997. Role of extracellular polysaccharide and endoglucanase in root invasion and colonization of tomato plants by Ralstonia solanacearum. Phytopathology 87: 1264-1271.
Santiago TR, Grabowski C, Mizubuti ESG, 2014. First report of bacterial wilt caused by Ralstonia solanacearum on Eucalyptus sp. in Paraguay. New Disease Reports 29: 2.
Schell MA. 2000. Control of virulence and pathogenicity genes of Ralstonia solanacearum by an elaborate sensory network. Annu Rev Phytopathol 38: 263-292.
Seal SE, Jackson LA, Young JPW, Daniels MJ. 1993. Detection of Pseudomonas solanacearum, Pseudomonas syzygii, Pseudomonas pickettii and Blood Disease Bacterium by partial 16S rRNA sequencing: construction of oligonucleotide primers for sensitive detection by polymerase chain reaction. J. Gen. Microbiol. 139: 1587-1594.
She X, Yu L, Lan G, Tang Y and He Z. 2017. Identification and genetic characterization of Ralstonia solanacearum species complex isolates from Cucurbita maxima in China. Front. Plant Sci. 8: 1794
She XM, He ZF, Luo FF. 2013. Pathogen identification of Ageratum conyzoides bacterial wilt disease and its biological characteristics. Acta Phytophyl. Sin. 40: 533–539
Siregar BA, Giyanto, Hidayat SH, Siregar IZ, Tjahjono B. 2020. Epidemiology of bacterial wilt disease on Eucalyptus pellita F. Muell. in Indonesia. IOP Conf. Ser.: Earth Environ. Sci. 468 012033
Siregar BA, Indrayadi H, Mardai. 2012. Ancaman penyakit layu bakteri terhadap produktivitas Eucalyptus pellita di dataran rendah tropis. In: Plant Protection (eds) Laporan Tahunan R&D PT. Arara Abadi. PT. Arara Abadi, Perawang
Sudo S, Oliveira GHN, Perreira AC. 1983. Eucalipto (Eucalyptus sp) e Bracatinga (Mimosa escabrela), novos hospedeiros de Pseudomonas solanacearum EF Smith. Fitopathol. Bras. 8: 631.
Tjahjono B. 2015. Isolasi dan Identifikasi Penyakit Layu Bakteri pada Eukaliptus. In: Sinarmas Forestry (eds). Instruksi Kerja R&D PT. Arara Abadi. PT. Arara Abadi, Perawang

Most read articles by the same author(s)