Bacterial Soft Rots

Pérombelon, M. C. M.; Salmond, George P. C.

doi:10.1016/b978-0-08-042510-8.50008-x

Cited by 41 publications

(31 citation statements)

References 88 publications

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“…The Erwinia genus includes a diverse group of pathogens that cause disease on a wide variety of plants (28). However, visual disease symptoms are not always sufficient to make an unequivocal identification of the pathogen involved.…”

Section: Fig 1 Its-pcr Amplification Patterns Of Species Belonging Tomentioning

confidence: 99%

“…carotovora, and subsp. wasabiae and E. chrysanthemi all cause similar diseases on potato (11,28). Thus, to ensure that a correct diagnosis is made and that steps are taken toward reducing disease spread, identification systems are required.…”

Section: Fig 1 Its-pcr Amplification Patterns Of Species Belonging Tomentioning

confidence: 99%

“…One group within the genus, the soft rot erwinias, causes soft rot diseases of many plant species worldwide (28). The most important of the soft rot erwinias commercially are E. chrysanthemi, E. carotovora subsp.…”

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confidence: 99%

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Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses

Toth¹,

Avrova²,

Hyman³

2001

Appl Environ Microbiol

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Current identification methods for the soft rot erwinias are both imprecise and time-consuming. We have used the 16S-23S rRNA intergenic transcribed spacer (ITS) to aid in their identification. Analysis by ITS-PCR and ITS-restriction fragment length polymorphism was found to be a simple, precise, and rapid method compared to current molecular and phenotypic techniques. The ITS was amplified from Erwinia and other genera using universal PCR primers. After PCR, the banding patterns generated allowed the soft rot erwinias to be differentiated from all other Erwinia and non-Erwinia species and placed into one of three groups (I to III). Group I comprised all Erwinia carotovora subsp. atroseptica and subsp. betavasculorum isolates. Group II comprised all E. carotovora subsp. carotovora, subsp. odorifera, and subsp. wasabiae and E. cacticida isolates, and group III comprised all E. chrysanthemi isolates. To increase the level of discrimination further, the ITS-PCR products were digested with one of two restriction enzymes. Digestion with CfoI identified E. carotovora subsp. atroseptica and subsp. betavasculorum (group I) and E. chrysanthemi (group III) isolates, while digestion with RsaI identified E. carotovora subsp. wasabiae, subsp. carotovora, and subsp. odorifera/carotovora and E. cacticida isolates (group II). In the latter case, it was necessary to distinguish E. carotovora subsp. odorifera and subsp. carotovora using the ␣-methyl glucoside test. Sixty suspected soft rot erwinia isolates from Australia were identified as E. carotovora subsp. atroseptica, E. chrysanthemi, E. carotovora subsp. carotovora, and non-soft rot species. Ten "atypical" E. carotovora subsp. atroseptica isolates were identified as E. carotovora subsp. atroseptica, subsp. carotovora, and subsp. betavasculorum and non-soft rot species, and two "atypical" E. carotovora subsp. carotovora isolates were identified as E. carotovora subsp. carotovora and subsp. atroseptica.

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Section: Fig 1 Its-pcr Amplification Patterns Of Species Belonging Tomentioning

confidence: 99%

Section: Fig 1 Its-pcr Amplification Patterns Of Species Belonging Tomentioning

confidence: 99%

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Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses

Toth¹,

Avrova²,

Hyman³

2001

Appl Environ Microbiol

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“…Consequently, it causes important losses within cool temperate regions, where potatoes have traditionally been grown. The optimal temperature for pathogenicity, estimated to be around 20°C (19,21), is a good compromise allowing both a fast multiplication (optimal at 24°C) and an efficient production of lytic enzymes, which is optimal at temperatures ranging between 12 and 24°C (26). As the thermoregulation of bacterial multiplication differs from that of exoenzyme production, we investigated the thermoregulation of QS that triggers exoenzyme synthesis.…”

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confidence: 99%

Thermoregulation of N -Acyl Homoserine Lactone-Based Quorum Sensing in the Soft Rot Bacterium Pectobacterium atrosepticum

Latour

Diallo

Chevalier

et al. 2007

Appl Environ Microbiol

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The psychrotolerant bacterium Pectobacterium atrosepticum produces four N-acyl homoserine lactones under a wide range of temperatures. Their thermoregulation differs from that of the exoenzyme production, described as being under quorum-sensing control. A mechanism involved in this thermoregulation consists of controlling N-acyl homoserine lactones synthase production at a transcriptional level.Several gram-negative bacteria synthesize N-acyl homoserine lactone (HSL) signal molecules that serve for cell-to-cell communication called for the first time quorum sensing (QS) by Fuqua et al. (7). Such regulatory systems operate to allow bacteria to sense cell density and to synchronize the functions of the entire population (reviewed in references 15, 23, and 29). In the Pectobacterium atrosepticum species (formerly Erwinia carotovora subsp. atroseptica) (8), QS regulation is involved in pectinolytic, cellulase, and protease activities, tuber maceration, and harpin synthesis leading to a hypersensitive response in nonhost plant (18,25). In contrast, QS via HSL production does not regulate growth and mobility during initial infection events or during liquid culture in a synthetic medium (25).Pectobacterium atrosepticum is a psychrotrophic bacterium involved in the soft rot of Solanum tuberosum (17, 26). Consequently, it causes important losses within cool temperate regions, where potatoes have traditionally been grown. The optimal temperature for pathogenicity, estimated to be around 20°C (19, 21), is a good compromise allowing both a fast multiplication (optimal at 24°C) and an efficient production of lytic enzymes, which is optimal at temperatures ranging between 12 and 24°C (26). As the thermoregulation of bacterial multiplication differs from that of exoenzyme production, we investigated the thermoregulation of QS that triggers exoenzyme synthesis. Therefore, HSL production and accumulation were measured during the three growth phases of a typical P. atrosepticum strain (Table 1) cultured at six temperatures. The role of the expI gene encoding the HSL synthase was determined by cloning this gene in Escherichia coli and by engineering a mutated strain of P. atrosepticum. Finally, the transcription of expI was estimated by relative reverse transcription-PCR (RT-PCR) under the same range of temperatures.Effect of temperature on N-acyl HSL diversity and levels. Bacterial cultures were grown at 8,12,15,20,24, and 28°C in minimal medium with polygalacturonic acid (PGA) as the sole source of carbon. This vegetable compound induces synthesis of virulence factors involved in plant disease or resistance that have been demonstrated to be under QS control (25). Characterization of HSLs was performed by high-performance liquid chromatography coupled with mass spectrometry (16). In these conditions, P. atrosepticum 6276 produces four HSLs: mainly N-3-oxo-octanoyl-L-HSL (3-oxo-C8-HSL) and minor quantities of N-octanoyl-L-HSL (C8-HSL), N-3-oxo-hexanoyl-L-HSL (3-oxo-C6-HSL), and N-3-oxo-decanoyl-L-HSL (3-oxo-C10-HSL). The HSL level...

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“…Almost all genes known to influence virulence in E. carotovora have been identified by either direct gene cloning or transposon mutagenesis, and demonstration of their role in virulence has largely depended on simple plant assays, usually involving stem or tuber inoculation tests (Andersson et al, 1999 ;Hinton et al, 1989 ;Pe! rombelon & Salmond, 1995).…”

Section: Introductionmentioning

confidence: 99%

Sample sequencing of a selected region of the genome of Erwinia carotovora subsp. atroseptica reveals candidate phytopathogenicity genes and allows comparison with Escherichia coli The GenBank accession numbers for the 424 sequences determined in this work are BH614193 to BH614616.

et al. 2002

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Genome sequencing is making a profound impact on microbiology. Currently, however, only one plant pathogen genome sequence is publicly available and no genome-sequencing project has been initiated for any species of the genus Erwinia, which includes several important plant pathogens. This paper describes a targeted sample sequencing approach to study the genome of Erwinia carotovora subsp. atroseptica (Eca), a major soft-rot pathogen of potato. A large insert DNA (approx. 115 kb) library of Eca was constructed using a bacterial artificial chromosome (BAC) vector. Hybridization and endsequence data revealed two overlapping BAC clones that span an entire hrp gene cluster. Random subcloning and one-fold sequence coverage (S 200 kb) across these BACs identified 25 (89 %) of 28 hrp genes predicted from the orthologous hrp cluster of Erwinia amylovora. Regions flanking the hrp cluster contained orthologues of known or putative pathogenicity operons from other Erwinia species, including dspEF (E. amylovora), hecAB and pecSM (E. chrysanthemi), sequences similar to genes from the plant pathogen Xylella fastidiosa, including haemagglutinin-like genes, and sequences similar to genes involved in rhizobacterium-plant interactions. Approximately 10 % of the sequences showed strongest nucleotide similarities to genes in the closely related model bacterium and animal pathogen Escherichia coli. However, the positions of some of these genes were different in the two genomes. Approximately 30 % of sequences showed no significant similarity to any database entries. A physical map was made across the genomic region spanning the hrp cluster by hybridization to the BAC library and to digested BAC clones, and by PCR between sequence contigs. A multiple genome coverage BAC library and one-fold sample sequencing are an effective combination for extracting useful information from important regions of the Eca genome, providing a wealth of candidate novel pathogenicity genes for functional analyses. Other genomic regions could be similarly targeted.

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Bacterial Soft Rots

Cited by 41 publications

References 88 publications

Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses

Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses

Thermoregulation of N -Acyl Homoserine Lactone-Based Quorum Sensing in the Soft Rot Bacterium Pectobacterium atrosepticum

Sample sequencing of a selected region of the genome of Erwinia carotovora subsp. atroseptica reveals candidate phytopathogenicity genes and allows comparison with Escherichia coli The GenBank accession numbers for the 424 sequences determined in this work are BH614193 to BH614616.

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