Plant Physiology Preview
Published on September 29, 2006; 10.1104/pp.106.085209
Received June 16, 2006
Accepted September 19, 2006
Characterisation of a Two Component High Affinity Nitrate Uptake System in Arabidopsis; Physiology and Protein-Protein Interaction
Mathilde Orsel , Franck Chopin , Olivier Leleu , Susan J Smith , Anne Krapp , Françoise Daniel-Vedele , and Anthony J Miller *
Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK
Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK; Unité de la Nutrition Azotée des Plantes, Institut National de la Recherche Agronomique, Route de St. Cyr, F-78026 Versailles cedex, France
* Corresponding author; email: tony.miller{at}bbsrc.ac.uk.
The identification of a family of the NAR2-type genes in higher plants showed that there was a homologue in Arabidopsis, AtNAR2.1 (Tong et al., 2005, Plant J 41: 442-450). These genes encode part of a two component nitrate high affinity transport system (HATS). As the Arabidopsis NRT2 gene family of nitrate transporters has been characterized we tested the idea that AtNAR2.1 and AtNRT2.1 are partners in a two component HATS. Results using the yeast split ubiquitin system and Xenopus oocyte expression showed that the two proteins interacted to give a functional high affinity transport system. The growth and nitrogen physiology of two Arabidopsis gene knock out mutants, atnrt2.1-1 and atnar2.1-1 one for each partner protein were compared. Both types of plants had lost HATS activity at 0.2 mM nitrate, but the effect was more severe in atnar2.1-1 plants. The relationship between plant N status and nitrate transporter expression revealed a pattern that was characteristic of nitrogen deficiency that was again stronger in atnar2.1-1. Plants resulting from a cross between both mutants (atnrt2.1-1 x atnar2.1-1) showed a phenotype like that of the atnar2.1-1 mutant when grown in 0.5 mM nitrate. Lateral root assays also revealed growth differences between the two mutants confirming that atnar2.1-1 had a stronger phenotype. To show that the impaired HATS did not result from the decreased expression of AtNRT2.1 we tested if constitutive root expression of a tobacco gene, NpNRT2.1 that has previously been shown to complement the atnrt2.1-1, can restore HATS to the atnar2.1-1 mutant. These plants did not recover wild type nitrate HATS. Taken together these results all show that AtNAR2.1 is essential for HATS of nitrate in Arabidopsis.
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