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Int. J. Mol. Sci. 2018, 19(4), 1049; https://doi.org/10.3390/ijms19041049

Metabolomics and Transcriptomics Identify Multiple Downstream Targets of Paraburkholderia phymatum σ54 During Symbiosis with Phaseolus vulgaris

1
Department of Plant and Microbial Biology, University of Zurich, CH-8057 Zurich, Switzerland
2
Agroscope, Research Group Molecular Diagnostics, Genomics and Bioinformatics & Swiss Institute of Bioinformatics (SIB), CH-8820 Wädenswil, Switzerland
3
Institute of Molecular Systems Biology, ETH Zurich, CH-8093 Zurich, Switzerland
*
Author to whom correspondence should be addressed.
Received: 28 February 2018 / Revised: 28 March 2018 / Accepted: 28 March 2018 / Published: 1 April 2018
(This article belongs to the Special Issue Metabolomics in the Plant Sciences 2017)
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Abstract

RpoN (or σ54) is the key sigma factor for the regulation of transcription of nitrogen fixation genes in diazotrophic bacteria, which include α- and β-rhizobia. Our previous studies showed that an rpoN mutant of the β-rhizobial strain Paraburkholderia phymatum STM815T formed root nodules on Phaseolus vulgaris cv. Negro jamapa, which were unable to reduce atmospheric nitrogen into ammonia. In an effort to further characterize the RpoN regulon of P. phymatum, transcriptomics was combined with a powerful metabolomics approach. The metabolome of P. vulgaris root nodules infected by a P. phymatum rpoN Fix mutant revealed statistically significant metabolic changes compared to wild-type Fix+ nodules, including reduced amounts of chorismate and elevated levels of flavonoids. A transcriptome analysis on Fix and Fix+ nodules—combined with a search for RpoN binding sequences in promoter regions of regulated genes—confirmed the expected control of σ54 on nitrogen fixation genes in nodules. The transcriptomic data also allowed us to identify additional target genes, whose differential expression was able to explain the observed metabolite changes in numerous cases. Moreover, the genes encoding the two-component regulatory system NtrBC were downregulated in root nodules induced by the rpoN mutant, and contained a putative RpoN binding motif in their promoter region, suggesting direct regulation. The construction and characterization of an ntrB mutant strain revealed impaired nitrogen assimilation in free-living conditions, as well as a noticeable symbiotic phenotype, as fewer but heavier nodules were formed on P. vulgaris roots. View Full-Text
Keywords: rhizobia; papilionoid; legumes; rpoN; sigma factor; metabolome; RNA-sequencing; nitrogen fixation rhizobia; papilionoid; legumes; rpoN; sigma factor; metabolome; RNA-sequencing; nitrogen fixation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Lardi, M.; Liu, Y.; Giudice, G.; Ahrens, C.H.; Zamboni, N.; Pessi, G. Metabolomics and Transcriptomics Identify Multiple Downstream Targets of Paraburkholderia phymatum σ54 During Symbiosis with Phaseolus vulgaris. Int. J. Mol. Sci. 2018, 19, 1049.

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