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 The GenBank accession numbers for the sequences determined in this work are AF207891–AF207897.

Poussier, Stéphane; Trigalet-Demery, Danielle; Vandewalle, Peggy; Goffinet, Bruno; Luisetti, Jacques; Trigalet, André

doi:10.1099/00221287-146-7-1679

Cited by 118 publications

(93 citation statements)

References 40 publications

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“…Thus, in contrast to biovar 2 strains, a higher genetic diversity can be supposed for biovar 1 isolates. This observation confirms the finding reported by Poussier et al (27) on May 10, 2018 by guest http://aem.asm.org/ the approach previously described by Poussier et al (27) for differentiating this species from R. solanacearum. Although the limited number of isolates belonging to biovars 1, 3, and 4 that were included in the present study does not allow us to fully evaluate the discriminatory power of the RsolfliC PCR-DGGE approach, it might be particularly useful to analyze mixed infections with strains of different biovars of R. solanacearum in the plant or in the soil on the condition that sufficiently high numbers of R. solanacearum cells are present.…”

Section: Separation Of Flic Fragments In Dggesupporting

confidence: 83%

Specific and Sensitive Detection of Ralstoniasolanacearum in Soil on the Basis of PCR Amplification of fliC Fragments

Schonfeld¹,

Heuer

Elsas³

et al. 2003

Appl Environ Microbiol

122

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Ralstonia solanacearum is the causative agent of bacterial wilt in many important crops. A specific and sensitive PCR detection method that uses primers targeting the gene coding for the flagella subunit, fliC, was established. Based on the first fliC gene sequence of R. solanacearum strain K60 available at GenBank, the RalfliC PCR primer system was designed; this system yielded a single 724-bp product with the DNAs of all of the R. solanacearum strains tested. However, R. pickettii and four environmental Ralstonia isolates also yielded amplicons. The RalfliC PCR products obtained with 12 strains (R. solanacearum, R. pickettii, and environmental isolates) were sequenced. By sequence alignment, RsolfliC primers specific for R. solanacearum were designed. With this primer system, a specific 400-bp PCR product was obtained from all 82 strains of R. solanacearum tested. Six strains of R. pickettii and several closely related environmental isolates yielded no PCR product; however, a product was obtained with one Pseudomonas syzygii strain. A GC-clamped 400-bp fliC product could be separated in denaturing gradient gels and allowed us to distinguish P. syzygii from R. solanacearum. The RsolfliC PCR system was applied to detect R. solanacearum in soil. PCR amplification, followed by Southern blot hybridization, allowed us to detect about one target DNA molecule per PCR, which is equivalent to 10 3 CFU g of bulk soil ؊1 . The system was applied to survey soils from different geographic origins for the presence of R. solanacearum.Ralstonia solanacearum is the causal agent of bacterial wilt in solanaceous crops but has also been recorded to infect a large range of more than 200 species representing over 50 families of plants (17). Traditionally, the pathogen has been classified in five biovars according to carbon source utilization (16, 18) and in six races based on host range (8,26). R. solanacearum is supposed to be a soil-borne bacterium originating from the tropics, subtropics, and warm temperate regions (15), but strains causing brown rot of potato in geographic regions with a temperate climate are possibly adapted to lower temperatures (20,24). In recent years, the increasing number of sites infested with potentially cold-adapted strains of R. solanacearum in several places in Europe dramatically enhanced the threat posed to European potato crops (20,24,43). Thus, reliable methods to detect the pathogen not only in tubers but also in soil or soil-related habitats are required. Several PCRbased methods for the detection of R. solanacearum have been described in the literature. These approaches are usually based on the amplification of ribosomal gene sequences (i.e., 16S or 16S-23S intergenic spacer region of the ribosomal DNA [rDNA]) (5,12,25,35,42,44). However, due to the high degree of conservation of the ribosomal genes within the genus Ralstonia, 16S rDNA sequence similarities between species can be as high as 98% (30,35,38). This can lead to positive signals with related species, such as R. picketti, and thus rRN...

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Section: Separation Of Flic Fragments In Dggesupporting

confidence: 83%

Specific and Sensitive Detection of Ralstoniasolanacearum in Soil on the Basis of PCR Amplification of fliC Fragments

Schonfeld¹,

Heuer

Elsas³

et al. 2003

Appl Environ Microbiol

122

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“…The investigation first focused on cold-tolerant phylotype IIB-1 strains, which represent a serious threat for potato production in Europe and temperate regions of the world. These strains were formerly known to present a clonal structure (20,23,27,38), and considerable diversity associated with the geographical distribution of strains was inferred in this study. Analysis showed two major clusters, but surprisingly, whereas the inference of phylogeny reflected a separation between sequevar 1 and 2 strains (phylotype II), the Bayesian population structure assessment method grouped strains from the Andean region together.…”

Section: Discussionmentioning

confidence: 99%

“…Also, since the ancestral Ralstonia prototype is assumed to be a plant pathogen (15,19), study of the extent of the diversity of this genus, along with the evolutionary pathways involved, is of major importance for a broad understanding of pathogen evolution. We hypothesized that there was much more biodiversity to discover from genomic analysis of phylotype II strains of R. solanacearum, namely, cold-tolerant potato brown rot strains (IIB-1), previously recognized to be clonal after various neutral marker approaches (5,27,34), tropical Moko disease-causing strains (IIB-4), and emerging strains (IIB-4NPB). We thus focus on phylogenetically closely related phylotype II groups of strains with well-characterized and diverging ecological and phenotypical traits, in an attempt to reconstruct their epidemiological pathways along with the acquisition of their lifestyles.…”

mentioning

confidence: 99%

Phylogeny and Population Structure of Brown Rot- and Moko Disease-Causing Strains of Ralstonia solanacearum Phylotype II

Cellier

Remenant

Chiroleu

et al. 2012

Appl Environ Microbiol

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The ancient soilborne plant vascular pathogen Ralstonia solanacearum has evolved and adapted to cause severe damage in an unusually wide range of plants. In order to better describe and understand these adaptations, strains with very similar lifestyles and host specializations are grouped into ecotypes. We used comparative genomic hybridization (CGH) to investigate three particular ecotypes in the American phylotype II group: (i) brown rot strains from phylotypes IIB-1 and IIB-2, historically known as race 3 biovar 2 and clonal; (ii) new pathogenic variants from phylotype IIB-4NPB that lack pathogenicity for banana but can infect many other plant species; and (iii) Moko disease-causing strains from phylotypes IIB-3, IIB-4, and IIA-6, historically known as race 2, that cause wilt on banana, plantain, and Heliconia spp. We compared the genomes of 72 R. solanacearum strains, mainly from the three major ecotypes of phylotype II, using a newly developed pangenomic microarray to decipher their population structure and gain clues about the epidemiology of these ecotypes. Strain phylogeny and population structure were reconstructed. The results revealed a phylogeographic structure within brown rot strains, allowing us to distinguish European outbreak strains of Andean and African origins. The pangenomic CGH data also demonstrated that Moko ecotype IIB-4 is phylogenetically distinct from the emerging IIB-4NPB strains. These findings improved our understanding of the epidemiology of important ecotypes in phylotype II and will be useful for evolutionary analyses and the development of new DNA-based diagnostic tools.

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“…To improve our understanding of the genetic relationship among the R. solanacearum species complex, DNA-based analyses were used, such as restriction fragment length polymorphisms (RFLP), polymerase chain reaction (PCR) -RFLP, 16S rRNA gene sequence analysis, repetitive extragenic palindromic (REP)-PCR, and amplified fragment length polymorphism (AFLP) (Cook et al 1989;Li et al 1993;Frey et al 1996;Poussier et al 2000b;Horita and Tsuchiya 2001). As examination of the diversity of the strains increased, it became clear that the R. solanacearum strains had four major phylotype subdivisions .…”

Section: Introductionmentioning

confidence: 99%

The sequevar distribution of Ralstonia solanacearum in tobacco-growing zones of China is structured by elevation

Liu

et al. 2016

Eur J Plant Pathol

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Bacterial wilt, caused by Ralstonia solanacearum, is a devastating disease resulting in tremendous losses of economic crops such as plants in the Solanaceae. Recent studies showed that R. solanacearum is spreading from the lowlands to the highlands in China. We studied 97 Chinese R. solanacearum strains that were isolated from four tobacco-growing zones over a wide range of elevations using phylotype specific multiplex polymerase chain reaction (Pmx-PCR) and phylogenetic relationships (egl and mutS). The results showed that all isolates belonged to phylotype I, which were further clustered into eight egl-sequence type groups (egl-group, sequevar): sequevars 13, 14, 15, 17, 34, 44, 54, and 55. In addition, Sequevar 55, found from the highlands, was a new/unknown one. Southeast China (Z3) had the largest number of egl-groups, containing six sequevars. The basin of the Yangzi River (Z1) and southwestern China (Z2) contained five egl-groups. The basin of the Huai River (Z4), near the north of China, where slight bacterial wilt occurred recently, contained a single group, sequevar 15. The distribution of sequevars was associated with elevation. Sequevar 15 was over-represented in lowland elevations, while sequevar 54 and the new/unknown one were only found in areas of moderate to high elevations. This finding suggested that the phylotype I strains infecting tobacco were diverse in China and regional integrated control strategies should be considered.

show abstract

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 The GenBank accession numbers for the sequences determined in this work are AF207891–AF207897.

Cited by 118 publications

References 40 publications

Specific and Sensitive Detection of Ralstoniasolanacearum in Soil on the Basis of PCR Amplification of fliC Fragments

Specific and Sensitive Detection of Ralstoniasolanacearum in Soil on the Basis of PCR Amplification of fliC Fragments

Phylogeny and Population Structure of Brown Rot- and Moko Disease-Causing Strains of Ralstonia solanacearum Phylotype II

The sequevar distribution of Ralstonia solanacearum in tobacco-growing zones of China is structured by elevation

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