BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon
ABSTRACT An efficient nitrate uptake system contributes to the improvement of crop nitrogen use efficiency under low nitrogen availability. The High Affinity nitrate Transport System (HATS) in plants is active in low range of external nitrate and is mediated by a two‐component system (high affinity...
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Wiley
2025-06-01
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| Series: | Plant Direct |
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| Online Access: | https://doi.org/10.1002/pld3.70075 |
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| author | Laure C. David Mathilde Grégoire Patrick Berquin Anne Marmagne Marion Dalmais Abdelhafid Bendahmane Tony J. Miller Anne Krapp Françoise Daniel‐Vedele Thomas Girin Sylvie Ferrario‐Méry |
| author_facet | Laure C. David Mathilde Grégoire Patrick Berquin Anne Marmagne Marion Dalmais Abdelhafid Bendahmane Tony J. Miller Anne Krapp Françoise Daniel‐Vedele Thomas Girin Sylvie Ferrario‐Méry |
| author_sort | Laure C. David |
| collection | DOAJ |
| description | ABSTRACT An efficient nitrate uptake system contributes to the improvement of crop nitrogen use efficiency under low nitrogen availability. The High Affinity nitrate Transport System (HATS) in plants is active in low range of external nitrate and is mediated by a two‐component system (high affinity transporters NRT2 associated to a partner protein NRT3 (NAR2)). In Brachypodium, the model plant for C3 cereals, we investigated the role of BdNRT2A and BdNRT3.2 through various experimental approaches. Expression profile of BdNRT2.A and BdNRT3.2 genes in response to nitrate availability fits perfectly with the characteristics of the HATS components. 15Nitrate influx measurements decreased in bdnrt2a mutants (one NaN3 induced mutant with a truncated NRT2A protein and two amiRNA mutants). In addition, the N limited phenotype of the mutant with a truncated NRT2A protein confirmed that BdNRT2A is a major contributor of the HATS in Brachypodium. An effective nitrate transport in the heterologous expression system Xenopus oocytes required the coexpression of BdNRT2A and BdNRT3.2 that characterizes two‐component system of the HATS. Functional interaction between BdNRT2A‐GFP and BdNRT3.2‐RFP fusion proteins was observed at the plasma membrane in Arabidopsis protoplasts in transient expression experiments with BdNRT3.2 being necessary for the plasma membrane localization of BdNRT2A. The role of a conserved Ser residue in BdNRT2A (S461) specific to monocotyledons was evaluated in the BdNRT2A and BdNRT3.2 interaction leading to plasma membrane targeting. Assuming that S461 could be regulated by phosphorylation, a directed mutagenesis was performed to mimic a nonphosphorylated (S461A) or a constitutively phosphorylated (S461D), However, the mimicking the phosphorylation status of S461 by mutagenesis did not modify the BdNRT2A and BdNRT3.2 interaction, suggesting a more complex regulating mechanism. In conclusion, our data show that BdNRT2A and BdNRT3.2 are the main components of the nitrate HATS activity in Brachypodium (Bd21‐3) and allow an optimal growth in low N conditions. |
| format | Article |
| id | doaj-art-4af7682dbc8046d6a1532a40b303eb81 |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-06-01 |
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| series | Plant Direct |
| spelling | doaj-art-4af7682dbc8046d6a1532a40b303eb812025-08-20T03:56:41ZengWileyPlant Direct2475-44552025-06-0196n/an/a10.1002/pld3.70075BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyonLaure C. David0Mathilde Grégoire1Patrick Berquin2Anne Marmagne3Marion Dalmais4Abdelhafid Bendahmane5Tony J. Miller6Anne Krapp7Françoise Daniel‐Vedele8Thomas Girin9Sylvie Ferrario‐Méry10Université Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, CNRS, INRAE, Université Evry, Institute of Plant Sciences Paris‐Saclay (IPS2) Gif sur Yvette FranceUniversité Paris‐Saclay, CNRS, INRAE, Université Evry, Institute of Plant Sciences Paris‐Saclay (IPS2) Gif sur Yvette FranceBiochemistry and Metabolism Department, John Innes Center Norwich UKUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceUniversité Paris‐Saclay, INRAE, AgroParisTech, Institut Jean‐Pierre Bourgin‐Sciences du Végétal (IJPB), Versailles, France Versailles FranceABSTRACT An efficient nitrate uptake system contributes to the improvement of crop nitrogen use efficiency under low nitrogen availability. The High Affinity nitrate Transport System (HATS) in plants is active in low range of external nitrate and is mediated by a two‐component system (high affinity transporters NRT2 associated to a partner protein NRT3 (NAR2)). In Brachypodium, the model plant for C3 cereals, we investigated the role of BdNRT2A and BdNRT3.2 through various experimental approaches. Expression profile of BdNRT2.A and BdNRT3.2 genes in response to nitrate availability fits perfectly with the characteristics of the HATS components. 15Nitrate influx measurements decreased in bdnrt2a mutants (one NaN3 induced mutant with a truncated NRT2A protein and two amiRNA mutants). In addition, the N limited phenotype of the mutant with a truncated NRT2A protein confirmed that BdNRT2A is a major contributor of the HATS in Brachypodium. An effective nitrate transport in the heterologous expression system Xenopus oocytes required the coexpression of BdNRT2A and BdNRT3.2 that characterizes two‐component system of the HATS. Functional interaction between BdNRT2A‐GFP and BdNRT3.2‐RFP fusion proteins was observed at the plasma membrane in Arabidopsis protoplasts in transient expression experiments with BdNRT3.2 being necessary for the plasma membrane localization of BdNRT2A. The role of a conserved Ser residue in BdNRT2A (S461) specific to monocotyledons was evaluated in the BdNRT2A and BdNRT3.2 interaction leading to plasma membrane targeting. Assuming that S461 could be regulated by phosphorylation, a directed mutagenesis was performed to mimic a nonphosphorylated (S461A) or a constitutively phosphorylated (S461D), However, the mimicking the phosphorylation status of S461 by mutagenesis did not modify the BdNRT2A and BdNRT3.2 interaction, suggesting a more complex regulating mechanism. In conclusion, our data show that BdNRT2A and BdNRT3.2 are the main components of the nitrate HATS activity in Brachypodium (Bd21‐3) and allow an optimal growth in low N conditions.https://doi.org/10.1002/pld3.70075Brachypodium distachyon (Bd21‐3)high affinity nitrate transport system (HATS)low nitrogen supplynitrate influxNRT2/NRT3 |
| spellingShingle | Laure C. David Mathilde Grégoire Patrick Berquin Anne Marmagne Marion Dalmais Abdelhafid Bendahmane Tony J. Miller Anne Krapp Françoise Daniel‐Vedele Thomas Girin Sylvie Ferrario‐Méry BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon Plant Direct Brachypodium distachyon (Bd21‐3) high affinity nitrate transport system (HATS) low nitrogen supply nitrate influx NRT2/NRT3 |
| title | BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon |
| title_full | BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon |
| title_fullStr | BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon |
| title_full_unstemmed | BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon |
| title_short | BdNRT2A and BdNRT3.2 Are the Major Components of the High‐Affinity Nitrate Transport System in Brachypodium distachyon |
| title_sort | bdnrt2a and bdnrt3 2 are the major components of the high affinity nitrate transport system in brachypodium distachyon |
| topic | Brachypodium distachyon (Bd21‐3) high affinity nitrate transport system (HATS) low nitrogen supply nitrate influx NRT2/NRT3 |
| url | https://doi.org/10.1002/pld3.70075 |
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