The importance of genome sequence quality to microbial comparative genomics

The quality of microbial genome sequences has been a concern ever since the emergence of genome sequencing. The quality of the genome assemblies is dependent on the sequencing technology used and the aims for which the sequence was generated. Novel sequencing and bioinformatics technologies are not intrinsically better than the older technologies, although they are generally more efficient. In this correspondence, the importance for comparative genomics of additional manual assembly efforts over autoassembly and careful annotation is emphasized

Erwinia species identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry

Rapid and reliable identification of plant pathogenic bacteria is critical for effective implementation of phytosanitary measures. The genus Erwinia includes a number of economically important plant pathogens such as fire blight agent Erwinia amylovora or Asian pear pathogen Erwinia pyrifoliae, together with closely related plant epiphytes of unknown pathogenicity or even with a potential use for biological control like Erwinia tasmaniensis or Erwinia billingiae, respectively. Current laboratory methods to achieve satisfactory identification and discrimination between species within the Erwinia genus are based on the isolation on semi-selective media, serology, specific PCR and gene locus sequencing: these approaches are complicated and time-consuming, often requiring a priori assumptions over the identity of the isolates. Here we present a streamlined approach based on whole-cell Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS) based on the AXIMA mass spectrometer of Shimadzu-Biotech Corp that demonstrates the potential of this technology for quick species identification in plant diagnostics within the genus Erwinia

Transcriptional profile of Salmonella enterica subsp. enterica serovar Weltevreden during alfalfa sprout colonization

Sprouted seeds represent a great risk for infection by human enteric pathogens because of favourable growth conditions for pathogens during their germination. The aim of this study was to identify mechanisms of interactions of Salmonella enterica subsp. enterica Weltevreden with alfalfa sprouts. RNA-seq analysis of S. Weltevreden grown with sprouts in comparison with M9-glucose medium showed that among a total of 4158 annotated coding sequences, 177 genes (4.3%) and 345 genes (8.3%) were transcribed at higher levels with sprouts and in minimal medium respectively. Genes that were higher transcribed with sprouts are coding for proteins involved in mechanisms known to be important for attachment, motility and biofilm formation. Besides gene expression required for phenotypic adaption, genes involved in sulphate acquisition were higher transcribed, suggesting that the surface on alfalfa sprouts may be poor in sulphate. Genes encoding structural and effector proteins of Salmonella pathogenicity island 2, involved in survival within macrophages during infection of animal tissue, were higher transcribed with sprouts possibly as a response to environmental conditions. This study provides insight on additional mechanisms that may be important for pathogen interactions with sprouts

The histidine-reversible antibiotic herbicolin O produced by Pantoea vagans C9-1 is pantocin A

Pantoea vagans C9-1 is one of the most effective and reliable biocontrol agents against fire blight, and has been commercialized as Blight Ban C9-1. Production of multiple antibiotics contributes to its antagonism of Erwinia amylovora. Here we describe the genetics, chemical isolation and structure of herbicolin O, the histidine-reversible antibiotic produced by P. vagans C9-1. Mutational analyses indicated that biosynthesis of herbicolin O depends on paaAB and a sequence encoding the peptide precursor of pantocin A. The paaABC gene cluster encoding biosynthesis and autoresistance was located within a 28-kb chromosomal genomic island of the complete genome sequence of P. vagans C9-1. The cluster was cloned and expressed in E. coli and purified antibiotic was isolated using improved methods for small peptides. The 1H NMR spectra of the C9-1 antibiotic closely resembled those of pantocin A produced by P. agglomerans Eh318. Detailed analysis of the proton spin systems showed that the chemical shift values and coupling constants of the protons in C9-1 herbicolin O correspond exactly to those of pantocin A. Based on these genetic and chemical analyses, herbicolin O and pantocin A are confirmed to be the same antibiotic

Host–pathogen interactions between Xanthomonas fragariae and its host Fragaria × ananassa investigated with a dual RNA-Seq analysis

Strawberry is economically important and widely grown, but susceptible to a large variety of phytopathogenic organisms. Among them, Xanthomonas fragariae is a quarantine bacterial pathogen threatening strawberry productions by causing angular leaf spots. Using whole transcriptome sequencing, the gene expression of both plant and bacteria in planta was analyzed at two time points, 12 and 29 days post inoculation, in order to compare the pathogen and host response between the stages of early visible and of well-developed symptoms. Among 28,588 known genes in strawberry and 4046 known genes in X. fragariae expressed at both time points, a total of 361 plant and 144 bacterial genes were significantly differentially expressed, respectively. The identified higher expressed genes in the plants were pathogen-associated molecular pattern receptors and pathogenesis-related thaumatin encoding genes, whereas the more expressed early genes were related to chloroplast metabolism as well as photosynthesis related coding genes. Most X. fragariae genes involved in host interaction, recognition, and pathogenesis were lower expressed at late-phase infection. This study gives a first insight into the interaction of X. fragariae with its host. The strawberry plant changed gene expression in order to consistently adapt its metabolism with the progression of infection

Comparative genomic analysis of the biotechnological potential of the novel species Pseudomonas wadenswilerensis CCOS 864T and Pseudomonas reidholzensis CCOS 865T

In recent years, the use of whole-cell biocatalysts and biocatalytic enzymes in biotechnological applications originating from the genus Pseudomonas has greatly increased. In 2014, two new species within the Pseudomonas putida group were isolated from Swiss forest soil. In this study, the high quality draft genome sequences of Pseudomonas wadenswilerensis CCOS 864T and Pseudomonas reidholzensis CCOS 865T were used in a comparative genomics approach to identify genomic features that either di ered between these two new species or to selected members of the P. putida group. The genomes of P. wadenswilerensis CCOS 864T and P. reidholzensis CCOS 865T were found to share genomic features for the degradation of aromatic compounds or the synthesis of secondary metabolites. In particular, genes encoding for biocatalytic relevant enzymes belonging to the class of oxidoreductases, proteases and isomerases were found, that could yield potential applications in biotechnology. Ecologically relevant features revealed that both species are probably playing an important role in the degradation of soil organic material, the accumulation of phosphate and biocontrol against plant pathogens

Draft genome sequence of the commercial biocontrol strain Pantoea agglomerans P10c

We report here the draft genome sequence of the biocontrol strain Pantoea agglomerans P10c, composed of a draft chromosome and two plasmids: the 559-kb large Pantoea plasmid 1 (pPag3) and a 182-kb plasmid (pPag1). A genomic island containing pantocin A biosynthesis genes was identified

Phylogenomic, pan-genomic, pathogenomic and evolutionary genomic insights into the agronomically relevant enterobacteria Pantoea ananatis and Pantoea stewartii

Pantoea ananatis is ubiquitously found in the environment and causes disease on a wide range of plant hosts. By contrast, its sister species, Pantoea stewartii subsp. stewartii is the host-specific causative agent of the devastating maize disease Stewart's wilt. This pathogen has a restricted lifecycle, overwintering in an insect vector before being introduced into susceptible maize cultivars, causing disease and returning to overwinter in its vector. The other subspecies of P. stewartii subsp. indologenes, has been isolated from different plant hosts and is predicted to proliferate in different environmental niches. Here we have, by the use of comparative genomics and a comprehensive suite of bioinformatic tools, analyzed the genomes of ten P. stewartii and nineteen P. ananatis strains. Our phylogenomic analyses have revealed that there are two distinct clades within P. ananatis while far less phylogenetic diversity was observed among the P. stewartii subspecies. Pan-genome analyses revealed a large core genome comprising of 3,571 protein coding sequences is shared among the twenty-nine compared strains. Furthermore, we showed that an extensive accessory genome made up largely by a mobilome of plasmids, integrated prophages, integrative and conjugative elements and insertion elements has resulted in extensive diversification of P. stewartii and P. ananatis. While these organisms share many pathogenicity determinants, our comparative genomic analyses show that they differ in terms of the secretion systems they encode. The genomic differences identified in this study have allowed us to postulate on the divergent evolutionary histories of the analyzed P. ananatis and P. stewartii strains and on the molecular basis underlying their ecological success and host range

Draft genome sequences for the onion center roth pathogen Pantoea ananatis PA4 and maize brown stalk rot pathogen P. ananatis BD442

Pantoea ananatis is an emerging phytopathogen that infects a broad spectrum of plant hosts. Here, we present the genomes of two South African isolates, P. ananatis PA4, which causes center rot of onion, and BD442, isolated from brown stalk rot of maize

Volcanic impacts on peatland microbial communities: A tephropalaeoecological hypothesis-test

Volcanic eruptions affect peatlands around the world, depositing volcanic ash (tephra) and a variety of chemicals including compounds of sulphur. These volcanic impacts may be important for many reasons, in particular sulphur deposition has been shown to suppress peatland methane flux, potentially reinforcing climatic cooling. Experiments have shown that sulphur deposition also forces changes in testate amoeba communities, potentially relating to the reduced methane flux. Large volcanic eruptions in regions with extensive peatlands are relatively rare so it is difficult to assess the extent to which volcanic eruptions affect peatland microbial communities; palaeoecological analyses across tephra layers provide a means to resolve this uncertainty. In this study, testate amoebae were analysed across multiple monoliths from a peatland in southern Alaska containing two tephras, probably representing the 1883 eruption of Augustine Volcano and a 20th Century eruption of Redoubt Volcano. Results showed relatively distinct and often statistically significant changes in testate amoeba community coincident with tephra layers which largely matched the response found in experimental studies of sulphur deposition. The results suggest volcanic impacts on peatland microbial communities which might relate to changes in methane flux