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Plant Cell. Aug 1999; 11(8): 1381–1392.
PMCID: PMC144300

Cloning and functional characterization of an Arabidopsis nitrate transporter gene that encodes a constitutive component of low-affinity uptake.

Abstract

The Arabidopsis CHL1 (AtNRT1) gene encodes an inducible component of low-affinity nitrate uptake, which necessitates a "two-component" model to account for the constitutive low-affinity uptake observed in physiological studies. Here, we report the cloning and characterization of a CHL1 homolog, AtNRT1:2 (originally named NTL1), with data to indicate that this gene encodes a constitutive component of low-affinity nitrate uptake. Transgenic plants expressing antisense AtNRT1:2 exhibited reduced nitrate-induced membrane depolarization and nitrate uptake activities in assays with 10 mM nitrate. Furthermore, transgenic plants expressing antisense AtNRT1:2 in the chl1-5 background exhibited an enhanced resistance to chlorate (7 mM as opposed to 2 mM for the chl1-5 mutant). Kinetic analysis of AtNRT1:2-injected Xenopus oocytes yielded a K(m) for nitrate of approximately 5.9 mM. In contrast to CHL1, AtNRT1:2 was constitutively expressed before and after nitrate exposure (it was repressed transiently only when the level of CHL1 mRNA started to increase significantly), and its mRNA was found primarily in root hairs and the epidermis in both young (root tips) and mature regions of roots. We conclude that low-affinity systems of nitrate uptake, like high-affinity systems, are composed of inducible and constitutive components and that with their distinct functions, they are part of an elaborate nitrate uptake network in Arabidopsis.

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Selected References

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  • Aslam M, Travis RL, Huffaker RC. Comparative kinetics and reciprocal inhibition of nitrate and nitrite uptake in roots of uninduced and induced barley (Hordeum vulgare L.) seedlings. Plant Physiol. 1992;99:1124–1133. [PMC free article] [PubMed]
  • Boorer KJ, Loo DD, Wright EM. Steady-state and presteady-state kinetics of the H+/hexose cotransporter (STP1) from Arabidopsis thaliana expressed in Xenopus oocytes. J Biol Chem. 1994 Aug 12;269(32):20417–20424. [PubMed]
  • Boorer KJ, Frommer WB, Bush DR, Kreman M, Loo DD, Wright EM. Kinetics and specificity of a H+/amino acid transporter from Arabidopsis thaliana. J Biol Chem. 1996 Jan 26;271(4):2213–2220. [PubMed]
  • Daniel-Vedele F, Filleur S, Caboche M. Nitrate transport: a key step in nitrate assimilation. Curr Opin Plant Biol. 1998 Jun;1(3):235–239. [PubMed]
  • Elledge SJ, Mulligan JT, Ramer SW, Spottswood M, Davis RW. Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1731–1735. [PMC free article] [PubMed]
  • Frommer WB, Hummel S, Rentsch D. Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters. FEBS Lett. 1994 Jun 27;347(2-3):185–189. [PubMed]
  • Hole DJ, Emran AM, Fares Y, Drew MC. Induction of nitrate transport in maize roots, and kinetics of influx, measured with nitrogen-13. Plant Physiol. 1990 Jun;93(2):642–647. [PMC free article] [PubMed]
  • Hopp TP, Woods KR. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. [PMC free article] [PubMed]
  • Huang NC, Chiang CS, Crawford NM, Tsay YF. CHL1 encodes a component of the low-affinity nitrate uptake system in Arabidopsis and shows cell type-specific expression in roots. Plant Cell. 1996 Dec;8(12):2183–2191. [PMC free article] [PubMed]
  • Lauter FR, Ninnemann O, Bucher M, Riesmeier JW, Frommer WB. Preferential expression of an ammonium transporter and of two putative nitrate transporters in root hairs of tomato. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):8139–8144. [PMC free article] [PubMed]
  • Liman ER, Tytgat J, Hess P. Subunit stoichiometry of a mammalian K+ channel determined by construction of multimeric cDNAs. Neuron. 1992 Nov;9(5):861–871. [PubMed]
  • Liu KH, Huang CY, Tsay YF. CHL1 is a dual-affinity nitrate transporter of Arabidopsis involved in multiple phases of nitrate uptake. Plant Cell. 1999 May;11(5):865–874. [PMC free article] [PubMed]
  • Mackenzie B, Loo DD, Fei Y, Liu WJ, Ganapathy V, Leibach FH, Wright EM. Mechanisms of the human intestinal H+-coupled oligopeptide transporter hPEPT1. J Biol Chem. 1996 Mar 8;271(10):5430–5437. [PubMed]
  • Meharg AA, Blatt MR. NO3- transport across the plasma membrane of Arabidopsis thaliana root hairs: kinetic control by pH and membrane voltage. J Membr Biol. 1995 May;145(1):49–66. [PubMed]
  • Paulsen IT, Skurray RA. The POT family of transport proteins. Trends Biochem Sci. 1994 Oct;19(10):404–404. [PubMed]
  • Quesada A, Krapp A, Trueman LJ, Daniel-Vedele F, Fernández E, Forde BG, Caboche M. PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family. Plant Mol Biol. 1997 May;34(2):265–274. [PubMed]
  • Rentsch D, Laloi M, Rouhara I, Schmelzer E, Delrot S, Frommer WB. NTR1 encodes a high affinity oligopeptide transporter in Arabidopsis. FEBS Lett. 1995 Aug 21;370(3):264–268. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Siddiqi MY, Glass AD, Ruth TJ, Rufty TW. Studies of the Uptake of Nitrate in Barley: I. Kinetics of NO(3) Influx. Plant Physiol. 1990 Aug;93(4):1426–1432. [PMC free article] [PubMed]
  • Song W, Steiner HY, Zhang L, Naider F, Stacey G, Becker JM. Cloning of a second Arabidopsis peptide transport gene. Plant Physiol. 1996 Jan;110(1):171–178. [PMC free article] [PubMed]
  • Steiner HY, Naider F, Becker JM. The PTR family: a new group of peptide transporters. Mol Microbiol. 1995 Jun;16(5):825–834. [PubMed]
  • Thayer JR, Huffaker RC. Determination of nitrate and nitrite by high-pressure liquid chromatography: comparison with other methods for nitrate determination. Anal Biochem. 1980 Feb;102(1):110–119. [PubMed]
  • Touraine B, Glass AD. NO3- and ClO3- fluxes in the chl1-5 mutant of Arabidopsis thaliana. Does the CHL1-5 gene encode a low-affinity NO3- transporter? Plant Physiol. 1997 May;114(1):137–144. [PMC free article] [PubMed]
  • Trueman LJ, Richardson A, Forde BG. Molecular cloning of higher plant homologues of the high-affinity nitrate transporters of Chlamydomonas reinhardtii and Aspergillus nidulans. Gene. 1996 Oct 10;175(1-2):223–231. [PubMed]
  • Tsay YF, Schroeder JI, Feldmann KA, Crawford NM. The herbicide sensitivity gene CHL1 of Arabidopsis encodes a nitrate-inducible nitrate transporter. Cell. 1993 Mar 12;72(5):705–713. [PubMed]
  • Wang R, Crawford NM. Genetic identification of a gene involved in constitutive, high-affinity nitrate transport in higher plants. Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9297–9301. [PMC free article] [PubMed]
  • Wang R, Liu D, Crawford NM. The Arabidopsis CHL1 protein plays a major role in high-affinity nitrate uptake. Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):15134–15139. [PMC free article] [PubMed]
  • West CE, Waterworth WM, Stephens SM, Smith CP, Bray CM. Cloning and functional characterisation of a peptide transporter expressed in the scutellum of barley grain during the early stages of germination. Plant J. 1998 Jul;15(2):221–229. [PubMed]
  • Wilkinson JQ, Crawford NM. Identification of the Arabidopsis CHL3 gene as the nitrate reductase structural gene NIA2. Plant Cell. 1991 May;3(5):461–471. [PMC free article] [PubMed]
  • Yamashita T, Shimada S, Guo W, Sato K, Kohmura E, Hayakawa T, Takagi T, Tohyama M. Cloning and functional expression of a brain peptide/histidine transporter. J Biol Chem. 1997 Apr 11;272(15):10205–10211. [PubMed]
  • Zhou JJ, Theodoulou FL, Muldin I, Ingemarsson B, Miller AJ. Cloning and functional characterization of a Brassica napus transporter that is able to transport nitrate and histidine. J Biol Chem. 1998 May 15;273(20):12017–12023. [PubMed]

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