A large number of Gram-negative, motile, mesophilic, violacein-producing bacteria were isolated from the soils and roots of Vaccinium macrocarpon Ait. and Kalmia angustifolia L. plants and from irrigation ponds associated with wild and cultivated cranberry bogs in Massachusetts, USA. Phylogenetic analyses of 16S rRNA gene sequences placed these isolates in a clade with Chromobacterium species, but the specialized environment from which they were isolated, their low genomic DNA relatedness with Chromobacterium violaceum ATCC 12472T and C. subtsugae PRAA4-1T, significant differences in fatty acid composition and colony morphology indicate that the cranberry and Kalmia isolates comprise a separate species of Chromobacterium , for which the name Chromobacterium vaccinii sp. nov. is proposed. Strain MWU205T ( = ATCC BAA-2314T = DSM 25150T) is proposed as the type strain for the novel species. Phenotypic analysis of 26 independent isolates of C. vaccinii sp. nov. indicates that, despite close geographical and biological proximity, there is considerable metabolic diversity among individuals within the population.
The study of chemical bioactivity in the rhizosphere has recently broadened to include microbial metabolites, and their roles in niche construction and competition via growth promotion, growth inhibition, and toxicity. Several prior studies have identified bacteria that produce volatile organic compounds (VOCs) with antifungal activities, indicating their potential use as biocontrol organisms to suppress phytopathogenic fungi and reduce agricultural losses. We sought to expand the roster of soil bacteria with known antifungal VOCs by testing bacterial isolates from wild and cultivated cranberry bog soils for VOCs that inhibit the growth of four common fungal and oomycete plant pathogens, and Trichoderma sp. Twenty one of the screened isolates inhibited the growth of at least one fungus by the production of VOCs, and isolates of Chromobacterium vaccinii had broad antifungal VOC activity, with growth inhibition over 90% for some fungi. Fungi exposed to C. vaccinii VOCs had extensive morphological abnormalities such as swollen hyphal cells, vacuolar depositions, and cell wall alterations. Quorum-insensitive cviR − mutants of C. vaccinii were significantly less fungistatic, indicating a role for quorum regulation in the production of antifungal VOCs. We collected and characterized VOCs from co-cultivation assays of Phoma sp. exposed to wild-type C. vaccinii MWU328, and its cviR − mutant using stir bar sorptive extraction and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (SBSE-GC×GC-TOFMS). We detected 53 VOCs that differ significantly in abundance between microbial cultures and media controls, including four candidate quorum-regulated fungistatic VOCs produced by C. vaccinii. Importantly, the metabolomes of the bacterial-fungal co-cultures were not the sum of the monoculture VOCs, an emergent property of their VOC-mediated interactions. These data suggest semiochemical feedback loops between microbes that have co-evolved for sensing and responding to exogenous VOCs.
Most plants synthesize antimicrobial compounds as part of normal plant development (i.e., phytoanticipins) or synthesize such compounds de novo when challenged by microorganisms (i.e., phytoalexins). The presumed role of these plant antibiotics is to protect the plant from disease. However, many phytopathogenic fungi have enzymes that can detoxify the phytoanticipins or phytoalexins produced by their host. This may be a means that these pathogens have evolved to circumvent resistance mechanisms based on the production of plant antibiotics. Many of the phytoanticipin- and phytoalexin-detoxifying enzymes produced by phytopathogenic fungi have biochemical and regulatory properties that would indicate the phytoanticipins and phytoalexins produced by their host are their normal substrates. In addition, their activity, enzymatic products, or transcripts can be detected in infected plant tissue suggesting that they are functioning in planta during pathogenesis. Specific mutations have been made by transformation-mediated gene-disruption procedures that eliminate the ability of Gaeumannomyces graminis var. avenae, Gloeocercospora sorghi, and Nectria haematococca to detoxify the phytoanticipins or phytoalexins produced by their hosts. The effect of these mutations on pathogenicity indicates a requirement for detoxifying enzymes in G. graminis var. avenae but not in G. sorghi or N. haematococca. Key words: disease resistance, pathogenicity mechanisms, isoflavonoids, saponins, cyanide.
The spreading of Rhizobium meliloti strains in various autoclaved soils was measured by using simple modifications of established techniques. Behavioral mutants were used to demonstrate that efficient spreading requires active motility and chemotaxis. The rate of spreading was affected by physical or chemical differences among soils and by changes in water content.
The precipitous drop in the cost of genomic sequencing and the concomitant availability of computational methods for comparing genome-level data has made the accurate taxonomic placement of bacteria affordable and relatively rapid. Inaccurate taxonomic placement of bacteria has serious implications in clinical, environmental, and regulatory microbiology, but it can also adversely affect interpretation of research results. The quorum biosensor strain CV026 was derived from an isolate of Chromobacterium that was labeled as C. violaceum ATCC 31532, and is catalogued by the ATCC under that species name. Nearly 200 papers have been published that use CV026 as an indicator for quorum sensing activity in many Gram negative bacteria, but the inability of C. violaceum strains to complement the quorum sensing mutation in CV026 has called the taxonomic placement of the parent strain into question. We used molecular phylogeny and a large number of metabolic and phenotypic characters to demonstrate that Chromobacterium strain ATCC 31532 is a member of species Chromobacterium subtsugae.
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