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Nitrate is a major nitrogen source for land plants and also acts as a signaling molecule that induces changes in growth and gene expression. To identify the cis‐acting DNA element involved in nitrate‐responsive gene expression, we analyzed the promoter of the Arabidopsis gene encoding nitrite reductase (NIR1). A region from positions −188 to −1, relative to the translation start site, was found to contain at least one cis‐element necessary for the nitrate‐dependent activation of the promoter, in which the activity of nitrate transporter NRT2.1 and/or NRT2.2 plays a critical role. To define this nitrate‐responsive cis‐element (NRE), we compared the sequences of several nitrite reductase gene promoters from various higher plants and identified a conserved sequence motif as the putative NRE. A synthetic promoter in which the four copies of a 43‐bp sequence containing the motif were fused to the 35S minimal promoter was found to direct nitrate‐responsive transcription. Furthermore, mutations within this conserved motif in the native NIR1 promoter markedly reduced the nitrate‐responsive activity of the promoter, indicating that the 43‐bp sequence is an NRE that is both necessary and sufficient for nitrate‐responsive transcription. We also show that both the native NIR1 promoter and the synthetic promoter display a similar level of sensitivity to nitrate, but respond differentially to exogenously supplied glutamine, indicating independent modulation of NIR1 expression by NRE‐mediated nitrate induction and feedback repression mediated by other cis‐element(s). These findings thus define the presence of multiple cis‐elements involved in the nitrogen response in Arabidopsis.
The Plant Journal – Wiley
Published: Jul 1, 2010
Keywords: ; ; ; ; ;
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