Abstract
Potato bacterial wilt is caused by the devastating bacterial pathogen Ralstonia solanacearum. Quantitative resistance to this disease has been and is currently introgressed from a number of wild relatives into cultivated varieties through laborious breeding programs. Here, we present two methods that we have developed to facilitate the screening for resistance to bacterial wilt in potato. The first one uses R. solanacearum reporter strains constitutively expressing the luxCDABE operon or the green fluorescent protein (gfp) to follow pathogen colonization in potato germplasm. Luminescent strains are used for nondestructive live imaging, while fluorescent ones enable precise pathogen visualization inside the plant tissues through confocal microscopy. The second method is a BIO-multiplex-PCR assay that is useful for sensitive and specific detection of viable R. solanacearum (IIB-1) cells in latently infected potato plants. This BIO-multiplex-PCR assay can specifically detect IIB-1 sequevar strains as well as strains belonging to all four R. solanacearum phylotypes and is sensitive enough to detect without DNA extraction ten bacterial cells per mL in complex samples.
The described methods allow the detection of latent infections in roots and stems of asymptomatic plants and were shown to be efficient tools to assist potato breeding programs.
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Mansfield J, Genin S, Magori S et al (2012) Top 10 plant pathogenic bacteria in molecular plant pathology. Mol Plant Pathol 13:614–629
Safni I, Cleenwerck I, De Vos P et al (2014) 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. celebesensis subsp. nov. and R. solanacearum phylotype I and III strains as Ralstonia pseudosolanacearum sp. nov. Int J Syst Evol Microbiol 64:3087–3103
Siri MI, Sanabria A, Pianzzola MJ (2011) Genetic diversity and aggressiveness of Ralstonia solanacearum strains causing bacterial wilt of potato in Uruguay. Plant Dis 95:1292–1301
Huet G (2014) Breeding for resistances to Ralstonia solanacearum. Front Plant Sci 715:1–5
Fock I, Collonnier C, Purwito A et al (2000) Resistance to bacterial wilt in somatic hybrids between Solanum tuberosum and Solanum phureja. Plant Sci 160:165–176
Carputo D, Aversano R, Barone A et al (2009) Resistance to Ralstonia solanacearum of sexual hybrids between Solanum commersonii and S. tuberosum. Am J Potato Res 86:196–202
Siri MI, Galván G, Quirici L et al (2009) Molecular marker diversity and bacterial wilt resistance in wild Solanum commersonii accessions from Uruguay. Euphytica 165:371–382
Gonzalez M, Galvan G, Siri MI et al (2013) Resistencia a la marchitez bacteriana de la papa en Solanum commersonii Dun. Agrociencia 17:45–54
Gutarra L, Kreuze J, Lindqvist-Kreuze H, De Mendiburu F (2014) Variation of resistance to different strains of Ralstonia solanacearum in highland tropics adapted potato genotypes. Am J Potato Res 92:258–265
Cruz APZ, Ferreira V, Pianzzola MJ et al (2014) A novel, sensitive method to evaluate potato germplasm for bacterial wilt resistance using a luminescent Ralstonia solanacearum reporter strain. Mol Plant-Microbe Interact 27:277–285
Ferreira V, Pianzzola MJ, Vilaró F et al (2017) Interspecific potato breeding lines display differential colonisation patterns and induced defense responses after Ralstonia solanacearum infection. Front Plant Sci 8:1424–1437
Guarischi-Sousa R, Puigvert M, Coll NS et al (2016) Complete genome sequence of the potato pathogen Ralstonia solanacearum UY031. Stand Genomic Sci 11:7–14
Elphinstone JG, Hennessy J, Wilson JK, Stead DE (1996) Sensitivity of different methods for the detection of Ralstonia solanacearum in potato tuber extracts. EPPO Bull 26:663–678
Opina N, Tavner F, Hollway G et al (1997) A novel method for development of species and strain-specific DNA probes and PCR primers for identifying Burkholderia solanacearum (formerly Pseudomonas solanacearum). Asia Pac J Mol Biol Biotechnol 5:19–30
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Ferreira, V., González, M., Pianzzola, M.J., Coll, N.S., Siri, M.I., Valls, M. (2021). Molecular Detection of Ralstonia solanacearum to Facilitate Breeding for Resistance to Bacterial Wilt in Potato. In: Dobnik, D., Gruden, K., Ramšak, Ž., Coll, A. (eds) Solanum tuberosum. Methods in Molecular Biology, vol 2354. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1609-3_18
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DOI: https://doi.org/10.1007/978-1-0716-1609-3_18
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