Skip to main content

Advertisement

SpringerLink
Log in

Properties of nitrate reductase from seedling leaves of selected barley genotypes

  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

Crude extracts from leaves of 6-day barley seedlings of parental genotypes (cv. Aramir and primitive line R567) and selected doubled haploid (DH) lines were not found to have significant differences in the NADH:NR activity, while considerable differences between these genotypes were shown by the NAD(P)H:NR activity. The cv. Aramir and DH lines did not differ by nitrate accumulation in the leaves. However, the primitive line R567, as compared to the remaining genotypes, was characterized by an appreciably lower ability to accumulate nitrates.

In partially purified leaf extracts, significant differences in total NADH:NR activity and in distal activity dependent on methyl viologen (MV:NR) were found between the parental genotypes and selected DH lines. The studied genotypes differed also in dehydrogenase NR activity, i.e. cytochrome c reductase activity in crude extracts.

In the studied genotypes, the NADH:NR activity in partially purified leaf extracts did not substantially differ by Km values for nitrates. Calculated Vmax values for NADH:NR in these genotypes were similar to total NR activity in partially purified extracts.

Significant differences between the parental genotypes and selected DH lines were found in the thermal NADH:NR stability in crude and partially purified leaf extracts.

From the performed studies it follows that different NR stability was one of the reasons of revealed differences in total activity and in partial NR activities in the leaf extracts between the studied genotypes of spring barley. Besides, it is suggested that varied NR gene expression in the leaves of these barley genotypes could also influence NR activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DH:

doubled haploid

NR:

nitrate reductase

References

  • Aryan A.P., Batt R.G., Wallace W. 1983. Reversible inactivation of nitrate reductase by NADH and the occurrence of partially inactive enzyme in the wheat leaf. Plant Physiol. 71: 582–587.

    PubMed  CAS  Google Scholar 

  • Barbacki S., Caliński T., Surma M., Kurhańska G., Adamski T., Kaczmarek Z., Dobek A., Karczewska A., Jeżowski S. 1978. Transgression in barley (Hordeum sativum Jess.) 7a. Transgressions of F6 and F7 hybrids from the cross Burea x Brown. Gen. Pol. 19(4): 403–421.

    Google Scholar 

  • Barber M.J., Notton B.A. 1990. Spinach nitrate reductase: effect of ionic strength and pH on the full and partial enzyme activities. Plant Physiol. 93: 537–540.

    PubMed  CAS  Google Scholar 

  • Buczek J., Marciniak J. 1990. Nitrate reductase and nitrite reductase a key enzymes of nitrate assimilation in higher plants. Wiad. Bot. 34(1): 19–32.

    Google Scholar 

  • Caboche M. and Rouze P. 1990. Nitrate reductase: a target for molecular and cellular studies in higher plants. Trends in Gen. 6(6): 187–192.

    Article  CAS  Google Scholar 

  • Campbell W.H. 1989. Structure and regulation of nitrate reductase in higher plants. In Wray J. and Kinghorn J.R. eds., Molecular and genetic aspects of nitrate assimilation. Oxford Science, Oxford, UK, pp 125–154.

    Google Scholar 

  • Crawford N.M. 1995. Nitrate: nutrient and signal of plant growth. The Plant Cell. 7: 859–868.

    Article  PubMed  CAS  Google Scholar 

  • Dailey F.A., Kuo T., Warner R.L. 1982. Pyridine nucleotide specifity of barley nitrate reductase. Plant Physiol. 69: 1196–1199.

    PubMed  CAS  Google Scholar 

  • Hamano T, Oji Y., Mitsuhashi Y., Matsuki Y., Okamoto S. 1984. Action of thiol proteinase on nitrate reductase in leaves of Hordeum distichum L. Plant Cell Physiol. 25(8): 1469–1475.

    CAS  Google Scholar 

  • Hoff T., Truong H.N. and Caboche M. 1994. The use of mutants and transgenic plants to study nitrate assimilation. Plant, Cell Environ. 17: 489–506.

    Article  CAS  Google Scholar 

  • Jones P.W. and Mhuimhneachain M.N. 1985. The activity and stability of wheat nitrate reductase in vitro. Phytochem. 24: 385–392.

    Article  CAS  Google Scholar 

  • Kaiser W., Brendle-Behnisch E. 1991. Rapid modulation of spinach leaf nitrate reductase activity by photosynthesis. I Modulation in vivo by CO2 avialibility. Plant Physiol. 96: 363–367.

    PubMed  CAS  Google Scholar 

  • Kaiser W.M. and Huber S.C. 1994. Posttranslational regulation of nitrate reductase in higher plants. Plant Physiol. 106: 817–821.

    PubMed  CAS  Google Scholar 

  • Kaiser W.M. and Spill D. 1991. Rapid modulation of spinach leaf nitrate reductase by photosynthesis. II. in vitro modulation by ATP and AMP. Plant Physiol. 96: 368–375.

    PubMed  CAS  Google Scholar 

  • Kasha K.J., Kao K.N. 1970. High frequency haploid production in barley (Hordeum vulgare L.) Nature 225: 874–875.

    Article  PubMed  CAS  Google Scholar 

  • Kleinhofs A., Kuo T. 1980. Characterization of nitrate reductase deficient barley mutants. Molec. Gen. Genet. 177: 421–425.

    Article  CAS  Google Scholar 

  • Kleinhofs A., Warner R.L., Lawrence J.M., Melzer J.M., Jeter J.M., Kudrna D.A. 1989. Molecular genetics of nitrate reductase in barley. In Wray J. and Kinghorn J. R. eds., Molecular and genetic aspects of nitrate assimilation. Oxford Science, Oxford, UK, pp 197–211.

    Google Scholar 

  • Kuo T.M., Kleinhofs A., Somers D.A., Warner R.L. 1984. Nitrate reductase- deficient mutants in barley: enzyme stability and peptide mapping. Biochem. 23: 229–232.

    CAS  Google Scholar 

  • Kuo T.M., Warner R.L., Kleinhofs A. 1982. in vitro stability of nitrate reductase from barley leaves. Biochem. 21: 531–533.

    CAS  Google Scholar 

  • Lillo C. and Ruoff P. 1992. Hysteretic behavior of nitrate reductase. Evidence of an allosteric binding site for reduced pyridine nucleotides. J. Biol. Chem. 267: 13456–13459.

    PubMed  CAS  Google Scholar 

  • Li X.Z., Oaks A. 1994. Induction and turnover of nitrate reductase in Zea mays. Influence of light. Plant. Physiol. 106(3): 1145–1149.

    PubMed  CAS  Google Scholar 

  • Mackintosh C., Douglas P., Lillo C. 1995. Identification of a protein that inhibits the phosphorylated form of nitrate reductase from spinach (Spinacia oleracea) leaves. Plant Physiol., 107: 451–457.

    PubMed  CAS  Google Scholar 

  • Salomonson L.P. and Barber M.J. 1990. Assimilatory nitrate reductase: functional properties and regulation. Ann. Rev. Plant Physiol. Plant Mol. Biol., 41: 225–253.

    Article  Google Scholar 

  • Sanderson G.W., Cocking E.C. 1964. Enzymatic assimilation of nitrate in tomato plants. I. Reduction of nitrate to nitrite. Plant Physiol. 53: 825–828.

    Google Scholar 

  • Sherrard J.H. and Dalling M.J. 1979. in vitro stability of nitrate reductase from wheat leaves. I. Stability of highly purified enzyme and its component activities. Plant Physiol. 63: 346–353.

    Article  PubMed  CAS  Google Scholar 

  • Skoczek H., Surma M., Adamski T. 1990. Inheritance of nitrate reductase activity in seedling leaves and roots of doubled haploid barley lines (Hordeum vulgare L.). I. Characteristics of doubled haploid lines with regard to nitrate reductase activity and grain protein yield. Gen. Pol. 31(1): 29–36. II. Estimation of gene effects. Gen. Pol 31(2): 83–88.

    Google Scholar 

  • Skoczek H. 1992. Nitrate reductase activity and protein content in leaves and roots of two doubled haploid barley lines depending on seedling age and nitrogen source. Acta Physiol. Plant. 14(2): 77–84.

    CAS  Google Scholar 

  • Spill D., Kaiser M. 1994. Partial purification of two proteins (100 kDa and 67 kDa) cooperating in the ATP-dependent inactivation of spinach leaf nitrate reductase. Planta 192: 183–188.

    Article  CAS  Google Scholar 

  • Srivastava H.S. 1992. Multiple functions and forms of higher plant nitrate reductase. Phytochem. 31: 2941–2947.

    Article  CAS  Google Scholar 

  • Sueyoshi K., Kleinhofs A., Warner R.L. 1995. Expression of NADH-specific and NAD(P)H-bispecific nitrate reductase genes in response to nitrate in barley. Plant Physiol. 107: 1303–1311.

    PubMed  CAS  Google Scholar 

  • Wray J.L. and Filner P. 1970. Structural and functional relationships of enzyme activities induced by nitrate in barley. Biochem.J. 119: 715–725.

    PubMed  CAS  Google Scholar 

  • Warner R.L. and Kleinhofs A. 1992. Genetics and molecular biology of nitrate metabolism in higher plants. Physiol. Plant. 85: 245–252.

    Article  CAS  Google Scholar 

  • Warner R.L., Kleinhofs A., Muehlbauer F.J. 1982. Characterization of nitrate reductase- deficient mutants in pea. Crop Science 22: 389–393.

    Article  Google Scholar 

  • Warner R.L., Warayanan K.R., Kleinhofs A. 1987. Inheritance and expression of NAD(P)H nitrate reductase in barley. Theor. Appl. Genet. 74: 714–717.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Physiology, Agricultural University of Poznań, Wołyńska 35, 60-637, Poznań, Poland

    Halina Gniazdowska-Skoczek

Authors
  1. Halina Gniazdowska-Skoczek
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gniazdowska-Skoczek, H. Properties of nitrate reductase from seedling leaves of selected barley genotypes. Acta Physiol Plant 19, 137–145 (1997). https://doi.org/10.1007/s11738-997-0030-9

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-997-0030-9

Key words

Search

Navigation