Summary
Barley (Hordeum vulgare L.) has both NADH-specific and NAD(P)H-bispecific nitrate reductases. Genomic and cDNA clones of the NADH nitrate reductase have been sequenced. In this study, a genomic clone (pMJ4.1) of a second type of nitrate reductase was isolated from barley by homology to a partial-length NADH nitrate reductase cDNA and the sequence determined. The open reading frame encodes a polypeptide of 891 amino acids and its interrupted by two small introns. The deduced amino acid sequence has 70% identity to the barley NADH-specific nitrate reductase. The non-coding regions of the pMJ4.1 gene have low homology (ca. 40%) to the corresponding regions of the NADH nitrate reductase gene. Expression of the pMJ4.1 nitrate reductase gene is induced by nitrate in root tissues which corresponds to the induction of NAD(P)H nitrate reductase activity. The pMJ4.1 nitrate reductase gene is sufficiently different from all previously reported higher plant nitrate reductase genes to suggest that it encodes the barley NAD(P)H-bispecific nitrate reductase.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andrews M, de Faria SM, Mclnroy SG, Sprent JI (1990) Constitutive nitrate reductase in the Leguminosae. Phytochemistry 29:49–54
Cheng C-L, Dewdney J, Kleinhofs A, Goodman HM (1986) Cloning and nitrate induction of nitrate reductase mRNA. Proc Natl Acad Sci USA 83:6825–6828
Cheng C-L, Dewdeny J, Nam HG, den Boer BGW, Goodman HM (1988) A new locus (NIA1) in Arabidopsis thaliana encoding nitrate reductase. EMBO J; 7:3309–3314
Choi HK, Kleinhofs A, An G (1989) Nucleotide sequence of rice nitrate reductase genes. Plant Mol Biol 13:731–733
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Cove DJ (1979) Genetic studies of nitrate assimilation in Aspergillus nidulans. Biol Rev 54:291–327
Crawford NM, Campbell WH (1990) Fertile Fields. Plant Cell 2:829–835
Crawford NM, Smith M, Bellissimo D, Davis RW (1988) Sequence and nitrate regulation of the Arabidopsis thaliana mRNA encoding nitrate reductase, a metalloflavoprotein with three functional domains. Proc Natl Acad Sci USA 85:5006–5010
Curtis LT, Smarrelli J Jr (1986) Metabolite control of nitrate reductase activity in soybean seedlings. J Plant Physiol 126:135–143
Dailey FA, Kuo T, Warner RL (1982a) Pyridine nucleotide specificity of barley nitrate reductase. Plant Physiol 69:1196–1199
Dailey FA, Warner RL, Somers DA, Kleinhofs A (1982b) Characteristics of a nitrate reductase in a barley mutant deficient in NADH nitrate reductase. Plant Physiol 69:1200–1204
Daniel-Vedele F, Dorbe M-F, Caboche M, Rouzé P (1989) Cloning and analysis of the tomato nitrate reductase-encoding gene: protein domain structure and amino acid homologies in higher plants. Gene 85:371–380
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Filner F (1966) Regulation of nitrate reductase in cultured tobacco cells. Biochim Biophys Acta 118:299–310
Gerlach WL, Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res 7:1869–1885
Gowri G, Campbell WH (1989) cDNA clones for corn leaf NADH:nitrate reductase and chloroplast NAD(P)+:glyceral-dehyde-3-phosphate dehydrogenase. Plant Physiol 90:792–798
Guerrero MG, Vega JM, Losada M (1981) The assimilatory nitrate-reducing system and its regulation. Annu Rev Plant Physiol 32:169–204
Hanley BA, Schuler MA (1988) Plant intron sequences: evidence for distinct groups of introns. Nucleic Acids Res 16:7159–7177
Harker AR, Narayanan KR, Warner RL, Kleinhofs A (1986) NAD(P)H bispecific nitrate reductase in barley leaves: partial purification and characterization. Phytochemistry 25:1275–1279
Kinghorn JR, Campbell EI (1989) Amino acid sequence relationships between bacterial, fungal, and plant nitrate reductase and nitrite reductase proteins. In: Wray JL, Kinghorn JR (eds) Molecular and Genetic Aspects of Nitrate Assimilation. Oxford University Press, Oxford, pp 385–403
Kleinhofs A, Warner RL (1990) Advances in nitrate assimilation. In: Stumpf PK, Conn EE (eds) The Biochemistry of Plants. Miflin BJ, Lea PJ (eds) Intermediary Nitrogen Metabolism, vol 16. Academic Press, New York, pp 89–120
Kleinhofs A, Chao S, Sharp PJ (1988) Mapping of nitrate reductase genes in barley and wheat. In: Miller TE, Koebner RMD (eds) Proceedings Seventh International Wheat Genetics Symposium, vol 1. Institute of Plant Science Research, Cambridge, pp 541–546
Kleinhofs A, Warner RL, Lawrence JM, Melzer JM, Jeter JM, Kudrna DA (1989) Molecular genetics of nitrate reductase in barley. In: Wray JL, Kinghorn JR (eds) Molecular and Genetic Aspects of Nitrate Assimilation. Oxford University Press, Oxford, pp 197–211
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Martino SJ, Smarrelli J Jr (1989) Nitrate reductase synthesis in squash cotyledons. Plant Sci 61:61–67
Melzer JM, Kleinhofs A, Warner RL (1989) Nitrate reductase regulation: effects of nitrate and light on nitrate reductase mRNA accumulation. Mot Gen Genet 217:341–346
Prosser IM, Lazarus CM (1990) Nucleotide sequence of a spinach nitrate reductase cDNA. Plant Mot Biol 15:187–190
Rychlik W, Rhoades R (1989) A computer program for choosing optimal oligonucleotides for filter hybridization, sequencing and in vitro amplification of DNA. Nucleic Acids Res 17:8543–8551
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schnorr KM, Juricek J, Huang C, Culley D, Kleinhofs A (1991) Analysis of barley nitrate reductase cDNA and genomic clones. Mol Gen Genet, in press
Stewart GR, Orebamjo TO (1979) Some unusual characteristics of nitrate reduction in Erythrina senegalensis DC. New Phytol 83:311–319
Vaucheret H, Kronenberger J, Rouzé P, Caboche M (1989) Complete nucleotide sequence of the two homeologous tobacco nitrate reductase genes. Plant Mot Biol 12:597–600
Warner RL, Huffaker RC (1989) Nitrate transport is independent of NADH and NAD(P)H nitrate reductases in barley seedlings. Plant Physiol 91:947–953
Warner RL, Narayanan KR, Kleinhofs A (1987) Inheritance and expression of NAD(P)H nitrate reductase in barley. Theor Appl Genet 74:714–717
Author information
Authors and Affiliations
Additional information
Communicated by H. Böhme
Scientific Paper No. 9101-14. College of Agriculture and Home Economics Research Center, Washington State University, Research Project Nos. 0233 and 0745
Rights and permissions
About this article
Cite this article
Miyazaki, J., Juricek, M., Angelis, K. et al. Characterization and sequence of a novel nitrate reductase from barley. Molec. Gen. Genet. 228, 329–334 (1991). https://doi.org/10.1007/BF00260624
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00260624