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Multiple mechanisms of nitrate sensing by Arabidopsis nitrate transceptor NRT1.1

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Abstract

In Arabidopsis the plasma membrane nitrate transceptor (transporter/receptor) NRT1.1 governs many physiological and developmental responses to nitrate. Alongside facilitating nitrate uptake, NRT1.1 regulates the expression levels of many nitrate assimilation pathway genes, modulates root system architecture, relieves seed dormancy and protects plants from ammonium toxicity. Here, we assess the functional and phenotypic consequences of point mutations in two key residues of NRT1.1 (P492 and T101). We show that the point mutations differentially affect several of the NRT1.1-dependent responses to nitrate, namely the repression of lateral root development at low nitrate concentrations, and the short-term upregulation of the nitrate-uptake gene NRT2.1, and its longer-term downregulation, at high nitrate concentrations. We also show that these mutations have differential effects on genome-wide gene expression. Our findings indicate that NRT1.1 activates four separate signalling mechanisms, which have independent structural bases in the protein. In particular, we present evidence to suggest that the phosphorylated and non-phosphorylated forms of NRT1.1 at T101 have distinct signalling functions, and that the nitrate-dependent regulation of root development depends on the phosphorylated form. Our findings add to the evidence that NRT1.1 is able to trigger independent signalling pathways in Arabidopsis in response to different environmental conditions.

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Figure 1: Point mutations in the NRT1.1 nitrate transceptor differentially alter the nitrate regulation of lateral root growth.
Figure 2: P492L and T101A mutations alter the auxin transport activity of NRT1.1 and auxin gradients in lateral root primordia.
Figure 3: The NRT1.1 point mutant proteins are expressed in lateral root primordia and properly addressed at the plasma membrane except for NRT1.1P492L.
Figure 4: Point mutations in the NRT1.1 nitrate transceptor differentially alter the regulation of NRT2.1 expression in roots in response to nitrogen treatments.
Figure 5: Point mutations in the NRT1.1 nitrate transceptor affect different genome-wide responses to high nitrogen provision.
Figure 6: Schematic representation of the four signalling ‘modes’ of NRT1.1.

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Acknowledgements

This work was supported by the Institut National de la Recherche Agronomique (CJS PhD Fellowship to S.L. & Projet Département BAP, BAP2013-33-NITSE to B.L.), Agence Nationale de la Recherche (ANR-11-JSV6-002-01-NUTSE to B.L.), The Agropolis Foundation (RHIZOPOLIS project to A.G., E.G. and P.N.), the Région Languedoc-Roussillon (Chercheur d'Avenir to B.L.), the Grant Agency of the Czech Republic (GAP305/11/0797 to E.Z.) and the Knowledge Biobase Economy European project (KBBE-005-002 Root enhancement for crop improvement to M.P. and P.N.). The authors thank Jan Petrášek and Alexandre Martiniere-Delaunay for the help with analysis of confocal images, Véronique Santoni for sharing data on NRT1.1 phosphorylation, Gloria M. Coruzzi for providing access to transcriptome facilities and hosting E.B., Amy Marshall Colon for the help with transcriptome analyses and Nigel M. Crawford for critical reading and editing of the manuscript.

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E.B., F.B., M.K., D.M., M.P., S.L., K.H. and P.N. performed experimental work; E.B., M.K., B.L., G.K., K.H., P.N. and A.G. performed data analysis; E.B., E.G., E.Z., P.N. and A.G. oversaw project planning; E.B., G.K., K.H., P.N. and A.G. wrote the paper.

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Correspondence to A. Gojon.

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Bouguyon, E., Brun, F., Meynard, D. et al. Multiple mechanisms of nitrate sensing by Arabidopsis nitrate transceptor NRT1.1. Nature Plants 1, 15015 (2015). https://doi.org/10.1038/nplants.2015.15

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