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Detailed in the Operation Manual here. Currently viewing:. Technical support. While the production of other deleterious volatiles is obvious from the plant growth inhibition observed upon exposure to the HCN-negative S. Moreover, the symptoms observed upon contact with the volatiles of cyanogenic bacteria chlorosis, reduced growth, or even death were mimicked when plants were challenged with pure HCN.
Similar observations were made in the field, where external application of HCN proved much more plant growth inhibitory than cyanogenesis by inoculated rhizosphere pseudomonad populations In addition to the putative production of growth promoting volatiles by the bacterial strains, which could counteract the negative effect of HCN, the different timing dynamics of application is likely to account in great part for the differences observed in plant responses. Quorum sensing has been shown to regulate many phenotypes associated with virulence reviewed in references 2 and 8.
We were therefore interested in assessing whether volatile-mediated phytotoxicity would also be quorum sensing dependent. If the main compound responsible for volatile-mediated phytotoxicity were indeed HCN, we would expect the quorum-sensing mutants to lose their volatile-mediated virulence to A. This is precisely what we observed with P. In contrast to P. We show that in PUPa3, disruption of both quorum-sensing systems was also necessary to abolish cyanogenesis and to reduce phytotoxicity.
Although the mechanisms underlying virulence have not yet been assessed, it is tempting to speculate that reduced virulence might be due, at least in part, to the loss of cyanogenesis in the PUPa3 double mutant. This contrasts with observations made in PAO1, where single mutants showed reduced cyanogenesis relative to the wild type.
We provide here quantitative data showing that inactivation of the AHL synthase in C. Surprisingly, the quorum-sensing mutant CV promoted plant growth, suggesting the production of plant-growth-promoting volatiles, whose beneficial effects on plants were masked by HCN in the wild type. We are currently investigating the chemical nature of these volatiles. Finally, we discovered that P.
Two quorum-sensing systems have been identified in P. Both systems are needed for proteolytic activity and rhizosphere competence 32 , Cyanogenesis in this organism has been reported to be independent of quorum sensing 55 , a finding that was supported by the results of the quorum-quenching experiment performed on a closely related strain in the present study. Besides the inhibitory effect of HCN when applied exogenously in high amounts to plants, recent studies suggest roles for HCN in planta , far beyond its being solely a by-product of ethylene biosynthesis 29 : in addition to its function in plant defense against herbivory through the wound-induced hydrolysis of cyanogenic glucosides , HCN has been shown to break seed dormancy in various species and to be involved in induced resistance against viruses 12 , 28 , It has further been shown that the gene expression changes occurring upon treatment with either KCN or with the ROS-generating methyl viologen were very similar 6.
We therefore hypothesized that the phytotoxicity of bacterial volatiles might be linked to oxidative stress. The results we obtained upon overexpression of the alternative oxidase AOX1 and concomitant supplementation of ascorbate as a ROS-trapping compound confirmed this hypothesis, and indicated that oxidative stress is a key process in the volatile-mediated negative impact of cyanogenic bacteria on plants.
Interestingly, in a study investigating the impact of truffle volatiles on plant growth, deleterious fungal volatiles other than HCN were shown to induce oxidative stress in Arabidopsis This suggests that oxidative stress might be a general response of plants to detection of microbial volatiles 25 , In summary, we present evidence that HCN, when produced in high amounts by bacteria, can kill plants. We suggest that this accounts to a great extent for the deleterious impact of bacterial volatiles observed in previous studies using cocultivation of Arabidopsis and bacterial strains.
Moreover, we report for the first time that cyanogenesis is not restricted to members of the Pseudomonas , Chromobacterium , and Rhizobium genera but that Serratia species, e. We further demonstrate that the volatile-mediated phytotoxicity of C. The environmental strain PUPa3 showed less phytotoxicity and lower cyanogenesis than the clinical isolate PAO1, but in both strains, cyanogenesis was dependent on functional quorum-sensing systems.
In contrast, quorum-sensing was not required for HCN production in P. Our data also provide initial insights into the mechanism of action of these bacterial volatiles leading to growth reduction or plant death: supplying ascorbate to alternative oxidase overexpressing lines of Arabidopsis increased their tolerance to cyanogenic pseudomonads drastically, while silenced lines showed higher susceptibility.
This indicates that volatile-mediated phytotoxicity involves oxidative stress and that the perception of deleterious bacterial volatiles causes, like many other biotic and abiotic stresses, an oxidative burst in the plant. Future studies are required for a more detailed understanding of the metabolic changes occurring in plants upon exposure to toxic microbial volatiles.
We thank Thomas Boller for constructive discussions, Thomas Kost for his help in setting up the cyanide measurement methodology, and Kirsty Agnoli for English corrections. National Center for Biotechnology Information , U. Journal List Appl Environ Microbiol v. Appl Environ Microbiol. Published online Nov Author information Article notes Copyright and License information Disclaimer.
Phone: 41 44 82 Fax: 41 44 82 E-mail: hc. Received Aug 19; Accepted Nov This article has been cited by other articles in PMC. Associated Data Supplementary Materials [Supplemental material].
Abstract The volatile-mediated impact of bacteria on plant growth is well documented, and contrasting effects have been reported ranging from 6-fold plant promotion to plant killing. TABLE 1. Strains used in this study. Species Strain name Strain no. Open in a separate window. TABLE 2. Arabidopsis thaliana cell lines used in this study. Background ID no. Chemicals, culture media, and growth conditions. Plant-bacterial dual growth experiments.
Chemical exposure of plants to HCN. Collection and measurement of HCN. Statistical analyses. An HCN-negative mutant is attenuated in A. The plant growth inhibitory effect of bacterial volatiles can be mimicked by supplying HCN. Role of quorum-sensing in HCN-mediated plant killing. Bacterial volatiles induce oxidative stress in A.
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