PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family
Purchase on Springer.com
$39.95 / €34.95 / £29.95*
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.
A family of high-affinity nitrate transporters has been identified in Aspergillus nidulans and Chlamydomonas reinhardtii, and recently homologues of this family have been cloned from a higher plant (barley). Based on six of the peptide sequences most strongly conserved between the barley and C. reinhardtii polypeptides, a set of degenerate primers was designed to permit amplification of the corresponding genes from other plant species. The utility of these primers was demonstrated by RT-PCR with cDNA made from poly(A)+ RNA from barley, C. reinhardtii and Nicotiana plumbaginifolia. A PCR fragment amplified from N. plumbaginifolia was used as probe to isolate a full-length cDNA clone which encodes a protein, NRT2;1Np, that is closely related to the previously isolated crnA homologue from barley. Genomic Southern blots indicated that there are only 1 or 2 members of the Nrt2 gene family in N. plumbaginifolia. Northern blotting showed that the Nrt2 transcripts are most strongly expressed in roots. The effects of external treatments with different N sources showed that the regulation of the Nrt2 gene(s) is very similar to that reported for nitrate reductase and nitrite reductase genes: their expression was strongly induced by nitrate but was repressed when reduced forms of N were supplied to the roots.
- Brownlee AG, Arst HN: Nitrate uptake in Aspergillus nidulans and involvement of the third gene of the nitrate assimilation cluster. J Bact 155: 1138–1146 (1983).
- Caboche M, Rouz é P: Nitrate reductase: a target for molecular and cellular studies in higher plants. Trends Genet Sci 6: 187–192 (1990).
- Caboche M, Campbell W, Crawford NM, Fernández E, Kleinhofs A, Ida S, Mendel R, Omata T, Rothstein S, Wray J: Genes involved in nitrate assimilation. Plant Mol Biol Rep 12: 45–49 (1994).
- Crawford NM: Nitrate: nutrient and signal for plant growth. Plant Cell 7: 859–868 (1995).
- Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).
- Devereux J: The GCG sequence analysis software package, version 7.0. Genetic Computer Group, University Research Park, Madison, WI (1991).
- Dorbe MF, Caboche M, Daniele-Vedele F: The tomato nia gene complements a Nicotiana plumbaginifolia nitrate reductasedeficient mutant and is properly regulated. Plant Mol Biol 18: 363–375 (1992).
- Fauré J-D, Vincentz M, Kronenberger J, Caboche M: Coregulated expression of nitrate and nitrite reductases. Plant J 1: 107–113 (1991).
- Frommer WB, Hummel S, Riesmeier JW: Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporter. FEBS Lett 347: 185–189 (1994).
- Galván A, Quesada A, Fernández E: Nitrate and nitrite are transported by different specific transport systems and by a bispecific transporter in Chlamydomonas reinhardtii. J Biol Chem 271: 2088–2092 (1996).
- Harris EH: The Chlamydomonas sourcebook. Academic Press, New York (1989).
- Hoff T, Truong HN, Caboche M: The use of mutants and transgenic plants to study nitrate assimilation. Plant Cell Environ 17: 489–506 (1994).
- Johnstone JL, McCabe PC, Greaves P, Gurr SJ, Cole GE, Brow MAD, Unkles SE, Clutterbuck AJ, Kinghorn JR, Innis MA: Isolation and characterization of the crnA-niiA-niaD gene cluster for nitrate assimilation in Aspergillus nidulans. Gene 90: 181–192 (1990).
- Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucl Acids Res 15: 9627–9640 (1987).
- Lauter FR, Ninnemann O, Bucher M, Riesmeier J, Frommer W: Preferential expression of an ammonium transporter and two putative nitrate transporters in root hairs of tomato. Proc Natl Acad Sci USA 93: 8139–8144 (1996).
- Larsson CM, Ingemarsson B: Molecular aspects of nitrate uptake in higher plants. In: Wray J, Kinghorn JR (eds) Molecular and Genetic Aspects of Nitrate Assimilation, pp. 3–14. Oxford Science Publications, Oxford (1989).
- Lee RB, Rudge KA: Effects of nitrogen deficiency on the absortion of nitrate and ammonium by barley plants. Ann Bot 57: 471–486 (1986).
- Lutcke HA, Chow KC, Michel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG codons differs in plants and animals. EMBO J 6: 43–48 (1987).
- McClure BA, Gray JE, Anderson MA, Clarke AE: Selfincompatibility in Nicotiana alata involves degradation of pollen rRNA. Nature 347: 757–760 (1990).
- Malik NSA: Senescence in oat leaves: changes in translatable mRNAs. Physiol Plant 70: 438–446 (1987).
- Marin E, Nussaume L, Quesada A, Gonneau M, Sotta B, Hugueney P, Frey A, Marion-Poll A: Molecular identification of zeaxanthin epoxidase of Nicotiana plumbaginifolia, a gene involved in abcisic acid biosynthesis and corresponding to ABA locus of Arabidopsis thaliana. EMBO J 15: 2331–2342 (1996).
- Marger MD, Saier MH: A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends Biochem Sci 18: 13–20 (1993).
- Muller JF, Missonier C, Caboche M: Low density growth of cells derived from Nicotiana and Petunia protoplasts. Influence of the source of protoplasts and comparison of the growthpromoting activity of various auxins. Physiol Plant 57: 35–40 (1983).
- Noji S, Nohno T, Saito T, Taniguchi S: The narK gene product participates in nitrate transport induced in Escherichia coli nitrate-respiring cells. FEBS Lett 252: 139–143 (1989).
- Quesada A, Galván A, Schnell RA, Lefebvre PA, Fernández E: Five nitrate assimilation genes are clustered in Chlamydomonas reinhardtii. Mol Gen Genet 240: 387–394 (1993).
- Quesada A, Fernández E: Expression of nitrate assimilation genes in Chlamydomonas reinhardtii. Plant Mol Biol 24: 185–194 (1994).
- Quesada A, Galván A, Fernández E: Identification of nitrate transporter genes in Chlamydomonas reinhardtii. Plant J 5: 407–419 (1994).
- Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor, Laboratory, Cold Spring Harbor, NY (1989).
- Schloss JA, Silflow CD, Rosenbaum JL: mRNA abundance changes during flagellar regeneration in Chlamydomonas reinhardtii. Mol Cell Biol 4: 424–434 (1984).
- Steiner HY, Song W, Zhang L, Naider F, Becker JM, Stacey G: An Arabidopsis peptide transporter is a member of a new class of membrane transport proteins. Plant Cell 6: 1289–1299 (1995).
- 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 175: 223–231 (1996).
- Trueman LJ, Onyeocha I, Forde BG: Recent advances in the molecular biology of a family of eukaryotic high affinity nitrate transporters. Plant Physiol Biochem 34: 621–627 (1996).
- Tsay YF, Schroeder J, Feldman KA, Crawford NM: The herbicide sensitivity gene CHL1 of Arabidopsis encodes a nitrateinducible nitrate transporter. Cell 72: 705–713 (1993).
- Unkles SE, Hawker KL, Grieve C, Campbell EI, Van Montague P, Kinghorn JR: crnA encodes a nitrate transporter in Aspergillus nidulans. Proc Natl Acad Sci USA 88: 204–208 (1991).
- Wang R, Crawford NM: Genetic identification of a gene involved in constitutive, high-affinity nitrate transport in higher plants. Proc Natl Acad Sci USA 93: 9297–9301 (1996).
- PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family
Plant Molecular Biology
Volume 34, Issue 2 , pp 265-274
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- Chlamydomonas reinhardtii
- Nicotiana plumbaginifolia
- nitrate transport
- Nrt2 expression
- Industry Sectors
- Author Affiliations
- 1. Laboratoire de Biologie Cellulaire, I.N.R.A., 78026, Versailles Cedex, France
- 2. Departamento de Bioquímica y Biología Molecular. Facultad de Ciencias, Universidad de Córdoba, 14071, Córdoba, Spain
- 3. Biochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Herts, AL5 2JQ, UK