Abstract
Ammonium transport is a key process in nitrogen metabolism. In the green alga Chlamydomonas, we have characterized molecularly the largest family of ammonium transporters (AMT1) so far described consisting of eight members. CrAmt1 genes have an interesting transcript structure with some very small exons. Differential expression patterns were found for each CrAmt1 gene in response to the nitrogen source by using Real Time PCR. These expression patterns were similar under high and low CO2 atmosphere. CrAmt1;1 expression was characterized in detail. It was repressed in both ammonium and nitrate medium, and strongly expressed in nitrogen-free media. Treatment with a Glutamine synthetase inhibitor released partially repression in ammonium and nitrate suggesting that ammonium and its derivatives participate in the observed repressing effects. By studying CrAmt1;1 expression in mutants deficient at different steps of the nitrate assimilation pathway, it has been shown that nitrate has a double negative effect on this gene expression; one related to its reduction to ammonium, and a second one by itself. This second effect of nitrate was dependent on the functionality of the regulatory gene Nit2, specific for nitrate assimilation. Thus, NIT2 would have a dual role on gene expression: the well-known positive one on nitrate assimilation and a novel negative one on Amt1;1 regulation.
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References
W.D. Benton R.W. Davis (1977) ArticleTitleScreening lambda gt recombinant clones by hybridization to single plaques in situ Science 196 180–182
A.J. Bloom S.S. Sukrapanna R.L. Warner (1992) ArticleTitleRoot respiration associated with ammonium and nitrate absorption and assimilation by barley Plant Physiol. 99 1294–1301
R. Breathnach P. Chambon (1981) ArticleTitleOrganization and expression of eukaryotic split genes coding for proteins Annu. Rev. Biochem. 50 349–383
D.T. Britto A.D.M. Glass H.J. Kronzucker M.Y. Siddiqi (2001a) ArticleTitleCytosolic concentrations and transmembrane fluxes of NH+4/NH3. An evaluation of recent proposals Plant Physiol. 125 523–526
D.T. Britto M.Y. Siddiqi A.D.M. Glass H.J. Kronzucker (2001b) ArticleTitleFutile transmembrane NH+4 cycling: a cellular hypothesis to explain ammonium toxicity in plants Proc. Natl. Acad. Sci. USA 98 4255–4258
T.E. Byrne M.R. Wells C.H. Johnson (1992) ArticleTitleCircadian rhythms of chemotaxis to ammonium and of methylammonium uptake in Chlamydomonas Plant Physiol. 93 879–886
F. Córdoba J. Cárdenas E. Fernández (1987) ArticleTitleCooperative regulation by ammonium and ammonium derivatives of nitrite uptake in Chlamydomonas reinhardtii Biochim. Biophys. Acta. 902 287–292
Crawford, N.M. and Forde, B.G. 2002. Molecular and developmental biology of inorganic nitrogen nutrition. In: C.R. Somerville and E.M. Meyerowitz (Eds.), The Arabidopsis Book Rockville, American Society of Plant Biologists, MD, doi/10.1199/tab.0011, http://www.aspb.org/publications/arabidopsis/
E. Fernández J. Cárdenas (1989) Genetic and regulatory aspects of nitrate assimilation in algae J.L. Wray J.R. Kinghorn (Eds) Molecular and Genetic Aspects of Nitrate Assimilation Oxford University Press Oxford 101–124
E. Fernández R.F. Matagne (1984) ArticleTitleGenetic analysis of nitrate reductase-deficient mutants in Chlamydomonas reinhardii Curr. Genet. 8 635–640
E. Fernández A. Galván A. Quesada (1998) Nitrogen assimilations and its regulation J-D. Rochaix M. Goldschmidt-Clermont S. Merchant (Eds) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas Kluwer Acadamic Publishers Dordrecht 637–659
B.G. Forde D.T. Clarkson (1999) ArticleTitleNitrate and ammonium nutrition of plants: Physiological and molecular perspectives Adv. Bot. Res. 30 1–90
A.R. Franco J. Cárdenas E. Fernández (1987) ArticleTitleA mutant of Chlamydomonas reinhardtii altered in the transport of ammonium and methylammonium Mol. Gen. Genet. 206 414–418
A.R. Franco J. Cárdenas E. Fernández (1988) ArticleTitleTwo different carriers transport both ammonium and methylammonium in Chlamydomonas reinhardtii J. Biol. Chem. 263 14039–14043
M.A. Frohman (1995) Rapid amplification of cDNA ends PCR primer C.W. Dieffenbach G.S. Dveksler (Eds) A laboratory manual Cold Spring Harbor Laboratory Press NY 381–409
X. Gansel S. Munos P. Tillard A. Gojon (2001) ArticleTitleDifferential regulation of the NO−3 and NH+4 transporter genes AtNrt2.1 and AtAmt1.1 in Arabidopsis: relation with long-distance and local controls by N status of the plant Plant J. 26 143–155
S. Gazzarrini L. Lejay A. Gojon O. Ninnemann W.B. Frommer N. Wirén Particlevon (1999) ArticleTitleThree functional transporters for constitutive, diurnally regulated and starvation-induced uptake of ammonium into Arabidopsis roots Plant Cell 11 937–948
A.D.M. Glass (2003) ArticleTitleNitrogen use efficiency of crop plants: physiological constraints upon nitrogen absorption Crit. Rev. Plant. Sci. 22 453–470
A.D.M. Glass M.Y. Siddiqi (1995) Nitrogen absorption in higher plants H.S. Srivastava R.P. Singh (Eds) Nitrogen nutrition in higher plants Association Publishing New Delhi 21–55
E. Harris (1989) The Chlamydomonas sourcebook Academic Press New York
S.M. Howitt M.K. Udvardi (2000) ArticleTitleStructure, function and regulation of ammonium transporters in plants Biochim. Biophys. Acta. 1465 152–170
H.J. Kronzucker M.Y. Siddiqi A.D.M. Glass G.J.D. Kirk (1999) ArticleTitleInhibition of nitrate uptake by ammonium in barley: Analysis of component fluxes Plant Physiol. 120 283–292
A. Llamas M.I. Igeño A. Galván E. Fernández (2002) ArticleTitleNitrate signalling on the nitrate reductase gene promoter depends directly on the activity of the nitrate transport systems in Chlamydomonas Plant J. 30 261–271
M.C. Lorenz J. Heitman (1998) ArticleTitleRegulators of pseudohyphal differentiation in Saccharomyces cerevisiae identified through multicopy suppressor analysis in ammonium permease mutant strains Genetics 150 1443–1457
A.M. Marini S. Soussi-Boudekou S. Vissers B. André (1997) ArticleTitleA family of ammonium transporters in Saccharomyces cerevisiae Mol. Cell. Biol. 17 4282–4293
A.M. Marini B. André (2000) ArticleTitleIn vivo N-glycosylation of the Mep2 high-affinity ammonium transporter of Saccharomyces cerevisiae reveals an extracytosolic N-terminus Mol. Microbiol. 38 552–564
H.L. Marschner (1995) Mineral Nutrition in Higher Plants Academic Press London
F. Merchán H. Ende ParticleVan der E. Fernández C.F. Beck (2001) ArticleTitleLow-expression genes induced by nitrogen starvation and subsequent sexual differentiation in Chlamydomonas reinhardtii, isolated by the differential display technique Planta 213 309–317 Occurrence Handle10.1007/s004250100567 Occurrence Handle11469597
O. Ninnemann J.C. Jauniaux W.B. Frommer (1994) ArticleTitleIdentification of a high affinity ammonium transporter from plants EMBO J. 13 3464–3471
J.N. Pearson J. Finnemann J.K. Schjoerring (2002) ArticleTitleRegulation of the high-affinity ammonium transporter (BnAMT1;2) in the leaves of Brassica napus by nitrogen status Plant Mol. Biol. 49 483–490
A. Quesada I. Gomez E. Fernandez (1998) ArticleTitleClustering of the nitrite reductase gene and a light-regulated gene with nitrate assimilation loci in Chlamydomonas reinhardtii Planta 206 259–265
A. Quesada A. Galván R.A. Schnell P.A. Lefebvre E. Fernández (1993) ArticleTitleFive nitrate assimilation related loci are clustered in Chlamydomonas reinhardtii Mol. Gen. Genet. 240 387–394
R. Rasmussen (2001) Quantification on the LightCycler S. Meuer C. Wittwer K. Nakagawara (Eds) Rapid Cycle Real-time PCR, Methods and Applications Springer Press Heidelberg 21–34
S.R. Rawat S.N. Silim H.J. Kronzucker M.Y. Siddiqi A.D.M. Glass (1999) ArticleTitleAtAMT1 gene expression and NH+4 uptake in roots of Arabidopsis thaliana. Evidence for regulation by root glutamine levels Plant J. 19 143–152
J. Rexach B. Montero E. Fernández A. Galván (1999) ArticleTitleDifferential regulation of the high affinity nitrite transport systems III and IV in Chlamydomonas reinhardtii J. Biol. Chem. 274 27801–27806
F. Salvemini A.-M. Marini. A. Riccio E.J. Patriarca M. Chiurazzi (2001) ArticleTitleFunctional characterization of an ammonium transporter gene from Lotus japonicus Gene 270 237–243
J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular cloning; A laboratory manual Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY
J.A. Schloss C.D. Silflow J.L. Rosenbaum (1984) ArticleTitlemRNA abundance changes during flagellar regeneration in Chlamydomonas reinhardtii Mol. Cell. Biol. 4 424–434
R.A. Schnell P.A. Lefebvre (1993) ArticleTitleIsolation of the Chlamydomonas regulatory gene NIT2 by transposon tagging Genetics 134 737–747 Occurrence Handle1:CAS:528:DyaK3sXkvVyitro%3D Occurrence Handle8394263
C.D. Silflow (1998) Organization of the nuclear genome J-D. Rochaix M. Goldschmidt-Clermont S. Merchant (Eds) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas Kluwer Academic Publishers Dordretch 25–40
U. Simon-Rosin C. Wood M.K. Udvardi (2003) ArticleTitleMolecular and cellular characterisation of LjAMT2;1, an ammonium transporter from the model legume Lotus japonicus Plant. Mol. Biol. 51 99–108
C. Sohlenkamp M. Shelden S. Howitt M. Udvardi (2000) ArticleTitleCharacterization of Arabidopsis AtAMT2, a novel ammonium transporter in plants FEBS Lett. 467 271–278
C. Sohlenkamp C.C. Wood G.W. Roeb M.K. Udvardi (2002) ArticleTitleCharacterization of Arabidopsis AtAMT2, a high-affinity ammonium transporter of the plasma membrane Plant Physiol. 130 1–9
E. Soupene W. Inwood S. Kustu (2004) ArticleTitleLack of the Rhesus protein Rh1 impairs growth of the green alga Chlamydomonas reinhardtii at high CO2 Proc. Natl. Acad. Sci. USA 101 7787–7792
E. Soupene N. King E. Field P. Liu K.K. Niyogi C.-H. Huang S. Kustu (2002) ArticleTitleRhesus expression in a green alga is regulated by CO2 Proc. Natl. Acad. Sci USA 99 7769–7773
J.M. Stark S.C. Hart (1977) ArticleTitleHigh rates of nitrification and nitrate turnover in undisturbed coniferous forests Nature 385 61–64
W.R. Ullrich M. Larsson C.-M. Larsson S. Lesch A. Novacky (1984) ArticleTitleAmmonium uptake in Lemna gibba G1, related membrane potential change and inhibition of anion uptake Physiol. Plant. 61 369–376
N. Wirén Particlevon A. Bergfeld O. Ninnemann W.B. Frommer (1997) ArticleTitleOsAMT1-1: a high-affinity ammonium transporter from rice (Oryza sativa cv. Nipponbare) Plant Mol. Biol. 35 681
N. Wirén Particlevon S. Gazzarrini A. Gojon W.B. Frommer (2000a) ArticleTitleThe molecular physiology of ammonium uptake and retrieval Curr. Opin. Plant Biol. 3 254–261
N. Wirén Particlevon F-R. Lauter O. Ninnemann B. Gillissen P. Walch-Liu C. Engels W. Jost W.B. Frommer (2000b) ArticleTitleDifferential regulation of three functional ammonium transporters genes by nitrogen in root hairs and by light in leaves of tomato Plant J. 21 167–175
R.M. Wallsgrove J.C. Turner N.P. Hall A.C. Kendall S.W.J. Bright (1987) ArticleTitleBarley mutants lacking chloroplast glutamine synthetase. Biochemical and genetic analysis Plant Physiol. 83 155–158
M.Y. Wang M.Y. Siddiqi T.J. Ruth A.D.M. Glass (1993a) ArticleTitleAmmonium uptake by rice roots (I. Fluxes and subcellular distribution of 13NH+4]) Plant Physiol. 103 1249–1258
M. Wang M.Y. Siddiqi T.J. Ruth A.D.M. Glass (1993b) ArticleTitleAmmonium uptake by rice roots. II. Kinetics of 13NH+4 influx across the plasmalemma Plant Physiol. 103 1259–1267
R. Wang K. Guegler S.T. LaBrie N.M. Crawford (2000) ArticleTitleGenomic analysis of a nutrient response in Arabidopsis reveals diverse expression patterns and novel metabolic and potential regulatory genes induced by nitrate Plant Cell 12 1491–1509
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González-Ballester, D., Camargo, A. & Fernández, E. Ammonium transporter genes in Chlamydomonas: the nitrate-specific regulatory gene Nit2 is involved in Amt1;1 expression. Plant Mol Biol 56, 863–878 (2004). https://doi.org/10.1007/s11103-004-5292-7
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DOI: https://doi.org/10.1007/s11103-004-5292-7