▪ Abstract Quorum sensing is the regulation of gene expression in response to fluctuations in cell-population density. Quorum sensing bacteria produce and release chemical signal molecules called autoinducers that increase in concentration as a function of cell density. The detection of a minimal threshold stimulatory concentration of an autoinducer leads to an alteration in gene expression. Gram-positive and Gram-negative bacteria use quorum sensing communication circuits to regulate a diverse array of physiological activities. These processes include symbiosis, virulence, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. In general, Gram-negative bacteria use acylated homoserine lactones as autoinducers, and Gram-positive bacteria use processed oligo-peptides to communicate. Recent advances in the field indicate that cell-cell communication via autoinducers occurs both within and between bacterial species. Furthermore, there is mounting data suggesting that ba...

Quorum Sensing in Bacteria

It has long been appreciated that certain groups of bacteria exhibit cooperative behavioral patterns. For example, feeding and sporulation of both myxobacteria and actinomycetes seem optimized for large populations of cells behaving almost as a single multicellular organism. The swarming motility of microorganisms such as Vibrio parahaemolyticus and Proteus mirabilis provides another excellent example of multicellular behavior among bacteria (2). Intercellular communication likewise has been appreciated for several years in Vibrio fischeri, Myxococcus xanthus, Bacillus subtilis, Streptomyces spp., the eukaryotic slime mold Dictyostelium discoideum, and other species (44). Here we first review how the marine luminescent bacterium V. fischeri uses the LuxR and LuxI proteins for intercellular communication and then describe a newly discovered family of LuxR and LuxI homologs in diverse bacterial species.

Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators.

Many plant bacteriologists, if not all, feel that their particular microbe should appear in any list of the most important bacterial plant pathogens. However, to our knowledge, no such list exists. The aim of this review was to survey all bacterial pathologists with an association with the journal Molecular Plant Pathology and ask them to nominate the bacterial pathogens they would place in a 'Top 10' based on scientific/economic importance. The survey generated 458 votes from the international community, and allowed the construction of a Top 10 bacterial plant pathogen list. The list includes, in rank order: (1) Pseudomonas syringae pathovars; (2) Ralstonia solanacearum; (3) Agrobacterium tumefaciens; (4) Xanthomonas oryzae pv. oryzae; (5) Xanthomonas campestris pathovars; (6) Xanthomonas axonopodis pathovars; (7) Erwinia amylovora; (8) Xylella fastidiosa; (9) Dickeya (dadantii and solani); (10) Pectobacterium carotovorum (and Pectobacterium atrosepticum). Bacteria garnering honourable mentions for just missing out on the Top 10 include Clavibacter michiganensis (michiganensis and sepedonicus), Pseudomonas savastanoi and Candidatus Liberibacter asiaticus. This review article presents a short section on each bacterium in the Top 10 list and its importance, with the intention of initiating discussion and debate amongst the plant bacteriology community, as well as laying down a benchmark. It will be interesting to see, in future years, how perceptions change and which bacterial pathogens enter and leave the Top 10.

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Top 10 plant pathogenic bacteria in molecular plant pathology

Proteins encoded by plasmid DNA are specifically labeled in uv-irradiated cells of Escherichia coli carrying recA and uvrA mutations because extensive degradation of the chromosome DNA occurs concurrently with amplification of plasmid DNA.

Simple method for identification of plasmid-coded proteins. [Escherichia coli, uv radiation]

Soft rot Enterobacteriaceae (SRE) are bacterial plant pathogens that cause blackleg, wilt and soft rot diseases on a broad range of important crop and ornamental plants worldwide. These organisms (spanning the genera Erwinia, Pectobacterium, Dickeya, and Pantoea) cause significant economic and yield losses in the field, and in storage. They are transmissible through surface water, by trade and other movement of plant material and soil, and in some cases are subject to international legislative and quarantine restrictions. Effective detection and diagnosis in support of food security legislation and epidemiology is dependent on the ability to classify pathogenic isolates precisely. Diagnostics and classification are made more difficult by the influence of horizontal gene transfer on phenotype, and historically complex and sometimes inaccurate nomenclatural and taxonomic assignments that persist in strain collections and online sequence databases. Here, we briefly discuss the relationship between taxonomy, genotype and phenotype in the SRE, and their implications for diagnostic testing and legislation. We present novel whole-genome classifications of the SRE, illustrating inconsistencies between the established taxonomies and evidence from completely sequenced isolates. We conclude with a perspective on the future impact of widespread whole-genome sequencing and classification methods on detection and identification of bacterial plant pathogens in support of legislative and policy efforts in food security.

Genomics and taxonomy in diagnostics for food security: soft-rotting enterobacterial plant pathogens

Three soft rot erwinias, Erwinia carotovora ssp. carotovora, E. carotovora ssp. atroseptica and E. chrysanthemi are associated with potatoes causing tuber soft rot and blackleg (stem rot). Latent infection of tubers and stems is widespread. As opportunistic pathogens, the bacteria tend to cause disease when potato resistance is impaired. Pathogenesis or disease development in potato tubers and stems is discussed in terms of the interaction between pathogen, host and environment, microbial competition and recent findings on the molecular basis of pathogenicity. Emphasis is placed on the role of free water and anaerobiosis in weakening tuber resistance and in providing nutrient for erwinias to multiply. Blackleg symptoms are expressed when erwinias predominate in rotting mother tubers, invade the stems and multiply in xylem vessels under favourable weather conditions. Soft rot erwinias tend to out-compete other bacteria in tuber rots because of their ability to produce larger quantities of a wider range of cell wall-degrading enzymes. However, despite extensive studies on their induction, regulation and secretion, little is known about the precise role of the different enzymes in pathogenesis. The putative role of quorum-sensing regulation of these enzymes in disease development is evaluated. The role certain pathogenicity-related characters, including motility, adhesion, siderophores, detoxifying systems and the hrp gene complex, common to most bacteria including symbionts and saprophytes, could play in latent and active infections is also discussed.

https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.0032-0862.2001.Shorttitle.doc.x

Potato diseases caused by soft rot erwinias : an overview of pathogenesis

Two modifications of the selective-diagnostic crystal violet pectate (CVP) medium for soft rot erwinias were developed using a new formulation of sodium polypectate; these were a single layer medium and a double layer medium consisting of an agar base with a pectate overlayer. Both media contained 0.1% tryptone to promote growth and cavity formation by erwinias and 40 μg ml−1 novobiocin instead of sodium lauryl sulphate to increase recovery rate and selectivity. Colonies of the three erwinias commonly associated with potatoes formed characteristic deep cavities in the single layer modified CVP. Cavities in the double layer CVP although deep were smaller but remained distinct longer. Recovery rate and selectivity with both media were better than with the original CVP.

Two modified crystal violet pectate (CVP) media for the detection, isolation and enumeration of soft rot erwinias

A PCR-based kit, ProbeliaTM, for the detection ofErwinia carotovora subsp.atroseptica (Eca) on potatoes was evaluated at five laboratories in four countries. The kit is based on DNA-specific PCR amplification followed by detection of amplicons by hybridization to a peroxidase-labelled DNA probe in a microplate. Specificity of the PCR primers for Eca, regardless of serogroups, was confirmed by testing against 246 bacterial, fungal and plant species. Detection limits of the assay varied little between six Eca strains in pure cultures (1.3×102 to 1.5×103 cells ml−1). When Eca-free tuber peel extract from four cultivars was inoculated with known numbers of 15 Eca strains, detection limits were more variable (1.0×101 to 6.2×103 cells ml−1 peel extract), attributed probably to inconsistency in the recovery of DNA during extraction. When the PCR assay was compared with three current commercial Eca detection methods, using naturally contaminated tubers, results matched most closely those from viable counts on a selective medium, the most sensitive method (88%), followed by enrichment ELISA (72%) and last ELISA (30%), the least sensitive method.

Evaluation of a PCR kit for the detection of Erwinia carotovora subsp. atroseptica on potato tubers

The selective-diagnostic crystal violet pectate (CVP) medium for the detection and isolation of soft rot erwinias was modified and improved to allow the use of a new source of sodium polypectate (Slendid type 440), as the previous polypectate (Bulmer) is no longer available. Two formulations were developed: CVP-S1 medium was less transparent but otherwise similar to the Bulmer polypectate-based CVP medium (CVP-B) except that NaOH was added and CaCl2 concentration reduced. CVP-S2 medium was prepared by mixing equal volumes of two double strength preparations containing 1) polypectate and NaOH, and 2) all other ingredients, both sterilised separately. Although erwinia cavity formation was slower, it showed a number of advantages over CVP-B and CVP-S1 media, thereby facilitating colony/cavity detection and enumeration. These included the absence of precipitate, greater firmness and a reduced risk of liquefaction in the presence of large erwinia numbers, and a reduction in the number of saprophytic bacteria.

Modified crystal violet pectate medium (CVP) based on a new polypectate source (Slendid) for the detection and isolation of soft rot erwinias.

The blackleg pathogen Erwinia carotovora subsp. atroseptica (Eca) causes an economically important disease of potatoes. We selected a genetically amenable Eca strain for the genetic analysis of virulence. Tn5 mutagenesis was used to generate nine mutants which exhibited reduced virulence (Rvi-) of strain SCRI1043. Following physiological characterisation, mutants were divided into three classes: (1) auxotrophs; (2) extracellular enzyme mutants; and (3) a growth rate mutant. The isolation of these Rvi- mutants has allowed us to consider some factors that affect Eca virulence.

M. C. M. Pérombelon

Papers

Potato diseases caused by soft rot erwinias : an overview of pathogenesis

Two modified crystal violet pectate (CVP) media for the detection, isolation and enumeration of soft rot erwinias

Evaluation of a PCR kit for the detection of Erwinia carotovora subsp. atroseptica on potato tubers

Modified crystal violet pectate medium (CVP) based on a new polypectate source (Slendid) for the detection and isolation of soft rot erwinias.

Isolation and characterisation of transposon-induced mutants of Erwinia carotovora subsp. atroseptica exhibiting reduced virulence.