Erwinia amylovora Novel Plasmid pEI70: Complete Sequence, Biogeography, and Role in Aggressiveness in the Fire Blight Phytopathogen

Llop, Pablo; Cabrefiga, Jordi; Smits, Theo H. M.; Dreo, T.; Barbé, Silvia; Puławska, Joanna; Bultreys, Alain; Blom, Jochen; Duffy, Brion; Seguí, Emilio Montesinos; López, Marı́a M.

doi:10.1371/journal.pone.0028651

Cited by 41 publications

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References 47 publications

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“…We sequenced six plasmids comprising 4.7% of the pan-genome of E. amylovora but found only five of the fourteen currently known plasmids [45] within our 12 genomes. The nearly ubiquitous and diagnostic plasmid pEA29 ( Figure 1 – PL29) which encodes genes for thiamine biosynthesis [46] was present in all strains except for UPN527 ( Table 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…The genome sequence of strain ACW 56400 from Switzerland contained the recently described plasmid pEI70, which contains an ICE as a major feature and has thus far only been reported in European E. amylovora populations [45]. The precise function of pEI70, which has high sequence similarity to pEB102 from the epiphyte E. billingiae Eb661, is to a large extent unknown and it is thought that the ICE is unable to integrate into the chromosome of E. amylovora [45].…”

Section: Resultsmentioning

confidence: 99%

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Comparative Genomics of 12 Strains of Erwinia amylovora Identifies a Pan-Genome with a Large Conserved Core

et al. 2013

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The plant pathogen Erwinia amylovora can be divided into two host-specific groupings; strains infecting a broad range of hosts within the Rosaceae subfamily Spiraeoideae (e.g., Malus, Pyrus, Crataegus, Sorbus) and strains infecting Rubus (raspberries and blackberries). Comparative genomic analysis of 12 strains representing distinct populations (e.g., geographic, temporal, host origin) of E. amylovora was used to describe the pan-genome of this major pathogen. The pan-genome contains 5751 coding sequences and is highly conserved relative to other phytopathogenic bacteria comprising on average 89% conserved, core genes. The chromosomes of Spiraeoideae-infecting strains were highly homogeneous, while greater genetic diversity was observed between Spiraeoideae- and Rubus-infecting strains (and among individual Rubus-infecting strains), the majority of which was attributed to variable genomic islands. Based on genomic distance scores and phylogenetic analysis, the Rubus-infecting strain ATCC BAA-2158 was genetically more closely related to the Spiraeoideae-infecting strains of E. amylovora than it was to the other Rubus-infecting strains. Analysis of the accessory genomes of Spiraeoideae- and Rubus-infecting strains has identified putative host-specific determinants including variation in the effector protein HopX1Ea and a putative secondary metabolite pathway only present in Rubus-infecting strains.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Comparative Genomics of 12 Strains of Erwinia amylovora Identifies a Pan-Genome with a Large Conserved Core

et al. 2013

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“…Besides the production of the siderophore desferrioxamine for the acquisition of iron molecules from the host tissue (Dellagi et al, 1998;Expert, 1999; and the presence of other virulence factors such as metalloproteases (Zhang et al, 1999), the presence of plasmids (Llop et al, 2011(Llop et al, , 2012McGhee & Jones, 2000;Mohammadi, 2010), two-component signal transduction systems Zhao et al, 2009) and histone-like proteins (Hildebrand et al, 2006) are also important factors in pathogenesis. However, probably the most essential reasons for differences in virulence between different strains of E. amylovora are due to a variation in synthesis of exopolysaccharides and the mechanism of the type III secretion system (T3SS) and associated proteins.…”

Section: Pathogenicity and Virulence Of E Amylovoramentioning

confidence: 99%

Pathogenicity and infection strategies of the fire blight pathogen Erwinia amylovora in Rosaceae: State of the art

Vrancken

Holtappels

Schoofs³

et al. 2013

Microbiology

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Plants are host to a large amount of pathogenic bacteria. Fire blight, caused by the bacterium Erwinia amylovora, is an important disease in Rosaceae. Pathogenicity of E. amylovora is greatly influenced by the production of exopolysaccharides, such as amylovoran, and the use of the type III secretion system, which enables bacteria to penetrate host tissue and cause disease. When infection takes place, plants have to rely on the ability of each cell to recognize the pathogen and the signals emanating from the infection site in order to generate several defence mechanisms. These mechanisms consist of physical barriers and the production of antimicrobial components, both in a preformed and an inducible manner. Inducible defence responses are activated upon the recognition of elicitor molecules by plant cell receptors, either derived from invading microorganisms or from pathogen-induced degradation of plant tissue. This recognition event triggers a signal transduction cascade, leading to a range of defence responses [reactive oxygen species (ROS), plant hormones, secondary metabolites, ...] and redeployment of cellular energy in a fast, efficient and multiresponsive manner, which prevents further pathogen ingress. This review highlights the research that has been performed during recent years regarding this specific plantpathogen interaction between Erwinia amylovora and Rosaceae, with a special emphasis on the pathogenicity and the infection strategy of E. amylovora and the possible defence mechanisms of the plant against this disease.

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“…The mixture was subjected to an initial denaturation step of 3 min at 94°C, followed by 40 cycles of 30 s at 94°C, 30 s at 56°C, 45 s at 72°C, and a final extension step of 10 min at 72°C. The amplification products obtained by conventional PCR were visualized by 2% (wt/vol) agarose gel electrophoresis in 0.5ϫ TAE buffer (40 Real-time PCR assays were performed in a LightCycler 480 thermocycler (Roche, USA) and in a SmartCycler system (Cepheid, USA). With culture suspensions and DNA extracted from plant samples, we used a final volume of 12 l (9 l of the mixture and 3 l of the sample).…”

Section: Methodsmentioning

confidence: 99%

Conventional and Real-Time PCRs for Detection of Erwinia piriflorinigrans Allow Its Distinction from the Fire Blight Pathogen, Erwinia amylovora

Barbé

Bertolini

Roselló

et al. 2014

Appl Environ Microbiol

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bErwinia piriflorinigrans is a new pathogenic species of the bacterial genus Erwinia that has been described recently in Spain. Accurate detection and identification of E. piriflorinigrans are challenging because its symptoms on pear blossoms are similar to those caused by Erwinia amylovora, the causal agent of fire blight. Moreover, these two species share phenotypic and molecular characteristics. Two specific and sensitive conventional and real-time PCR protocols were developed to identify and detect E. piriflorinigrans and to differentiate it from E. amylovora and other species of this genus. These protocols were based on sequences from plasmid pEPIR37, which is present in all strains of E. piriflorinigrans analyzed. After the stability of the plasmid was demonstrated, the specificities of the protocols were confirmed by the amplification of all E. piriflorinigrans strains tested, whereas 304 closely related pathogenic and nonpathogenic Erwinia strains and microbiota from pear trees were not amplified. In sensitivity assays, 10 3 cells/ml extract were detected in spiked plant material by conventional or real-time PCR, and 10 2 cells/ml were detected in DNA extracted from spiked plant material by real-time PCR. The protocols developed here succeeded in detecting E. piriflorinigrans in 102 out of 564 symptomatic and asymptomatic naturally infected pear samples (flowers, cortex stem tissue, leaves, shoots, and fruitlets), in necrotic Pyracantha sp. blossoms, and in necrotic pear and apple tissues infected with both E. amylovora and E. piriflorinigrans. Therefore, these new tools can be used in epidemiological studies that will enhance our understanding of the life cycle of E. piriflorinigrans in different hosts and plant tissues and its interaction with E. amylovora. Erwinia piriflorinigrans is a Gram-negative plant-pathogenic bacterium causing pear blossom necrosis (1). The disease affects pear production and can reduce fruit yield. It was first observed in Valencia, Spain, in 1999 (2, 3). After the first identification, the disease was observed at the same place in later years (1). Due to its recent discovery, the biology, ecology, and life cycle of this new species are still poorly understood.Unlike Erwinia amylovora, the causal agent of fire blight, E. piriflorinigrans affects blossoms and not other parts of plants (3). Nevertheless, E. piriflorinigrans colonies can be easily mistaken for E. amylovora in culture media, e.g., CCT medium (4), King's medium B (5), and sucrose nutrient agar (SNA) or Levan medium, which are usually utilized and recommended for the isolation of E. amylovora (6, 7). These two pathogens show similar metabolic profiles by use of the commercial API 20E, API 20NE, API ZYM, and API 50CH strips (bioMérieux, France), as well as close fatty acid profiles. E. piriflorinigrans can react with several antisera, and even with one of the monoclonal antibodies, used to detect E. amylovora (3). Moreover, E. piriflorinigrans reacts positively in PCR using the 23S rRNA gene sequence-based primers de...

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Erwinia amylovora Novel Plasmid pEI70: Complete Sequence, Biogeography, and Role in Aggressiveness in the Fire Blight Phytopathogen

Cited by 41 publications

References 47 publications

Comparative Genomics of 12 Strains of Erwinia amylovora Identifies a Pan-Genome with a Large Conserved Core

Comparative Genomics of 12 Strains of Erwinia amylovora Identifies a Pan-Genome with a Large Conserved Core

Pathogenicity and infection strategies of the fire blight pathogen Erwinia amylovora in Rosaceae: State of the art

Conventional and Real-Time PCRs for Detection of Erwinia piriflorinigrans Allow Its Distinction from the Fire Blight Pathogen, Erwinia amylovora

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