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Sci Rep
2015 Jan 22;5:7962. doi: 10.1038/srep07962.
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AtNPF5.5, a nitrate transporter affecting nitrogen accumulation in Arabidopsis embryo.
Léran S
,
Garg B
,
Boursiac Y
,
Corratgé-Faillie C
,
Brachet C
,
Tillard P
,
Gojon A
,
Lacombe B
.
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Dipeptide (Leu-Leu) and nitrate transport activities of 26 Arabidopsis NPF (NRT1/PTR Family) proteins were screened in Saccharomyces cerevisiae and Xenopus laevis oocytes, respectively. Dipeptide transport activity has been confirmed for 2 already known dipeptide transporters (AtNPF8.1 and AtNPF8.3) but none of the other tested NPFs displays dipeptide transport. The nitrate transport screen resulted in the identification of two new nitrate transporters, AtNPF5.5 and AtNPF5.10. The localization of the mRNA coding for NPF5.5 demonstrates that it is the first NPF transporter reported to be expressed in Arabidopsis embryo. Two independent homozygous npf5.5 KO lines display reduced total nitrogen content in the embryo as compared to WT plants, demonstrating an effect of NPF5.5 function on the embryo nitrogen content. Finally, NPF5.5 gene produces two different transcripts (AtNPF5.5a and AtNPF5.5b) encoding proteins with different N-terminal ends. Both proteins are able to transport nitrate in xenopus oocytes.
Figure 1. Screen for dipeptide transport complementation by NPFs in S. cerevisiae.Control (C, non-transformed) and NPF-transformed ΔPTR2 yeast mutants were grown for 20 hours on SC minimal media (complemented with uracil for control yeast) containing His, Leu and Met (black bars), SC minimal media containing Met (red bars) and SC minimal media containing His, Met and Leu-Leu (green bars). (inset) Growth kinetics in the 3 different media for yeasts expressing NPF8.1. Results from one experiment representative of 2–3 independent repeats.
Figure 2. Screen for nitrate transport activity of NPFs in xenopus oocytes.Control (non-injected) and NPF-injected oocytes were bathed in 30â mM K15NO3 for 2â hours.15N accumulation is expressed as % of accumulation in NPF6.3/NRT1.1-expressing oocytes. Values are mean +/â SEM from 3 experiments. *** indicate significant difference with control oocytes at p < 0.001 (t-test).
Figure 3. Characterization of npf5.5 KO lines.(a) Schematic representation of NPF5.5 insertion lines. Blue arrows represent the sens and antisens oligonucleotides used for Q-PCR analysis. (b) Q-RT-PCR analysis were performed on mRNA isolated from embryo at the bent cotyledon stage. Values are mean +/â SEM from 3 experiments. *** indicate significant difference with control oocytes at p < 0.001 (t-test). (c) Nitrogen content in embryos of two NPF5.5 KO lines. The total nitrogen content was determined in embryos of two NPF5.5 KO lines (npf5.5-1 and npf5.5-2). Values are expressed in percent of N content decrease compared to total N content in WT embryos. The average was made on 5 batches of 75 embryos, n = 5.
Figure 4. Alternative NPF5.5 transcripts.(a) Schematic representation of NPF5.5 5â² genomic and RNA sequences. Blue arrows represent the sens oligonucleotides used for identification of NPF5.5a and b. (b) Amino-acid alignment of the N-terminal region of the 3 different NPF5.5 proteins. (c) Influx in control (non-injected) oocytes (white bar), in NPF6.3-expresssing oocytes (black bar) and in the 3 forms NPF5.5-expressing oocytes (grey bars) after 2â hours of incubation in 30â mM15NO3 solution. P: predicted sequence, a and b: 2 NPF5.5 forms. Influx in control oocytes (white bar), in NPF6.3-expresssing oocytes (black bar) and in NPF5.5-expressing oocytes (grey bars) after 2â hours of incubation in 30â mM15NO3 solution. *** and ** indicate significant difference with control oocytes at p < 0.001 and p < 0.005 respectively (t-test).
Almagro,
Characterization of the Arabidopsis nitrate transporter NRT1.6 reveals a role of nitrate in early embryo development.
2008, Pubmed,
Xenbase
Almagro,
Characterization of the Arabidopsis nitrate transporter NRT1.6 reveals a role of nitrate in early embryo development.
2008,
Pubmed
,
Xenbase
Barbier-Brygoo,
Anion channels/transporters in plants: from molecular bases to regulatory networks.
2011,
Pubmed
Bouguyon,
Nitrate sensing and signaling in plants.
2012,
Pubmed
Boursiac,
ABA transport and transporters.
2013,
Pubmed
Chiang,
Mechanisms and functional properties of two peptide transporters, AtPTR2 and fPTR2.
2004,
Pubmed
,
Xenbase
Chiu,
Mutation of a nitrate transporter, AtNRT1:4, results in a reduced petiole nitrate content and altered leaf development.
2004,
Pubmed
,
Xenbase
Chopin,
The Arabidopsis ATNRT2.7 nitrate transporter controls nitrate content in seeds.
2007,
Pubmed
,
Xenbase
Dohmen,
An efficient transformation procedure enabling long-term storage of competent cells of various yeast genera.
1991,
Pubmed
Fan,
The Arabidopsis nitrate transporter NRT1.7, expressed in phloem, is responsible for source-to-sink remobilization of nitrate.
2009,
Pubmed
Frech,
Isolation of ion channel genes by expression cloning in Xenopus oocytes.
1992,
Pubmed
,
Xenbase
Frommer,
Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters.
1994,
Pubmed
Grillet,
Ascorbate efflux as a new strategy for iron reduction and transport in plants.
2014,
Pubmed
Hedrich,
Ion channels in plants.
2012,
Pubmed
Ho,
CHL1 functions as a nitrate sensor in plants.
2009,
Pubmed
Hsu,
Two phloem nitrate transporters, NRT1.11 and NRT1.12, are important for redistributing xylem-borne nitrate to enhance plant growth.
2013,
Pubmed
,
Xenbase
Huang,
Cloning and functional characterization of an Arabidopsis nitrate transporter gene that encodes a constitutive component of low-affinity uptake.
1999,
Pubmed
,
Xenbase
Kanno,
Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor.
2012,
Pubmed
Karim,
AtPTR3, a wound-induced peptide transporter needed for defence against virulent bacterial pathogens in Arabidopsis.
2007,
Pubmed
Krapp,
Nitrate transport and signalling in Arabidopsis.
2014,
Pubmed
Krouk,
Nitrate-regulated auxin transport by NRT1.1 defines a mechanism for nutrient sensing in plants.
2010,
Pubmed
,
Xenbase
Lacombe,
Evidence for a multi-ion pore behavior in the plant potassium channel KAT1.
1998,
Pubmed
,
Xenbase
Léran,
A unified nomenclature of NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER family members in plants.
2014,
Pubmed
Léran,
Arabidopsis NRT1.1 is a bidirectional transporter involved in root-to-shoot nitrate translocation.
2013,
Pubmed
Liman,
Subunit stoichiometry of a mammalian K+ channel determined by construction of multimeric cDNAs.
1992,
Pubmed
,
Xenbase
Mounier,
Auxin-mediated nitrate signalling by NRT1.1 participates in the adaptive response of Arabidopsis root architecture to the spatial heterogeneity of nitrate availability.
2014,
Pubmed
Nour-Eldin,
NRT/PTR transporters are essential for translocation of glucosinolate defence compounds to seeds.
2012,
Pubmed
,
Xenbase
Parker,
Molecular basis of nitrate uptake by the plant nitrate transporter NRT1.1.
2014,
Pubmed
Rentsch,
Structure and function of plasma membrane amino acid, oligopeptide and sucrose transporters from higher plants.
1998,
Pubmed
,
Xenbase
Rentsch,
NTR1 encodes a high affinity oligopeptide transporter in Arabidopsis.
1995,
Pubmed
Sasaki,
Closing plant stomata requires a homolog of an aluminum-activated malate transporter.
2010,
Pubmed
,
Xenbase
Sun,
Crystal structure of the plant dual-affinity nitrate transporter NRT1.1.
2014,
Pubmed
Tegeder,
Uptake and partitioning of amino acids and peptides.
2010,
Pubmed
Tsay,
Nitrate transporters and peptide transporters.
2007,
Pubmed
Tsay,
The herbicide sensitivity gene CHL1 of Arabidopsis encodes a nitrate-inducible nitrate transporter.
1993,
Pubmed
,
Xenbase
Wang,
Arabidopsis nitrate transporter NRT1.9 is important in phloem nitrate transport.
2011,
Pubmed
,
Xenbase
Winter,
An "Electronic Fluorescent Pictograph" browser for exploring and analyzing large-scale biological data sets.
2007,
Pubmed
Xiang,
Genome-wide analysis reveals gene expression and metabolic network dynamics during embryo development in Arabidopsis.
2011,
Pubmed
Yamada,
Empirical analysis of transcriptional activity in the Arabidopsis genome.
2003,
Pubmed
