Skip to main content

Advertisement

SpringerLink
Log in

A physiological and molecular study of the effects of nickel deficiency and phenylphosphorodiamidate (PPD) application on urea metabolism in oilseed rape (Brassica napus L.)

  • Regular Article
  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Background and aims

Urea is the major nitrogen (N) form supplied as fertilizer in agriculture. However, urease, a nickel-dependent enzyme, allows plants to use external or internally generated urea as a nitrogen source. Since a urease inhibitor is frequently applied in conjunction with urea fertilizer, the N-metabolism of plants may be affected. The aim of this study was to determine physiological and molecular effects of nickel deficiency and a urease inhibitor on urea uptake and assimilation in oilseed rape.

Methods

Plants were grown on hydroponic solution with urea as the sole N source under three treatments: plants treated with nickel (+Ni) as a control, without nickel (−Ni) and with nickel and phenylphosphorodiamidate (+Ni+PPD). Urea transport and assimilation were investigated.

Results

The results show that Ni-deficiency or PPD supply led to reduced growth and reduced 15N-uptake from urea. This effect was more pronounced in PPD-treated plants, which accumulated high amounts of urea and ammonium. Thus, Ni-deficiency or addition of PPD, limit the availability of N and decreased shoot and root amino acid content. The up-regulation of BnDUR3 in roots indicated that this gene is a component of the stress response to nitrogen-deficiency. A general decline of glutamine synthetase (GS) activity and activation of glutamate dehydrogenase (GDH) and increases in its expression level were observed in control plants. At the same time, in (−N) or (+Ni+PPD) treated plants, no increases in GS or GDH activities and expression level were found.

Conclusions

Overall results showed that plants require Ni as a nutrient (while most widely used nutrient solutions are devoid of Ni), whether they are grown with or without a urea supply, and that urease inhibitors may have deleterious effects at least in hydroponic grown oilseed rape.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

Ni:

Nickel

PPD:

Phenylphosphorodiamidate

NBPT:

N-(n-butyl) thiophosphoric triamide

GS:

Glutamine synthetase

GDH:

Glutamate dehydrogenase

References

  • Artola E, Cruchaga S, Ariz I, Moran JF, Garnica M, Houdusse F, Garcia Mina JM, Irigoyen I, Lasa B, Aparicio-Tejo PM (2011) Effect of N-(n-butyl) thiophosphoric triamide on urea metabolism and the assimilation of ammonium by Triticum aestivum L. Plant Growth Regul 63:73–79

    Article  CAS  Google Scholar 

  • Bremner JM (1995) Recent research on problems in the use of urea as a nitrogen fertilizer. Fertilizer Res 42:321–329

    Article  CAS  Google Scholar 

  • Brown PH, Welch RM, Madison JT (1990) Effect of nickel deficiency on soluble anion, amino acid, and nitrogen levels in barley. Plant Soil 125:19–27

    Article  CAS  Google Scholar 

  • Chen YG, Ching TM (1988) Induction of barley leaf urease. Plant Physiol 86:941–945

    Article  PubMed  CAS  Google Scholar 

  • Cruchaga S, Artola E, Lasa B, Ariz I, Irigoyen I, Moran JF, Aparicio-Tejo PM (2011) Short term physiological implications of NBPT application on the N metabolism of Pisum sativum and Spinacea oleracea. J Plant Physiol 168:329–336

    Article  PubMed  CAS  Google Scholar 

  • Eskew DL, Welch RM, Norvall WA (1984) Nickel in higher plants. Further evidence for an essential role. Plant Physiol 76:691–693

    Article  PubMed  CAS  Google Scholar 

  • Galili G, Amir R, Galili G (2008) Genetic engineering of amino acid metabolism in plants. Adv Plant Biochem Mol Biol 1:48–80

    Google Scholar 

  • Gerendás J, Sattelmacher B (1997) Significance of Ni supply for growth, urease activity and the contents of urea, amino acids and mineral nutrients of urea-grown plants. Plant Soil 190:153–162

    Article  Google Scholar 

  • Gerendás J, Sattelmacher B (1999) Influence of Ni supply on growth and nitrogen metabolism of Brassica napus L. grown with NH4NO3 or urea as N source. Ann Bot London 83:65–71

    Article  Google Scholar 

  • Gerendás J, Zhu Z, Sattelmacher B (1998) Influence of N and Ni supply on nitrogen metabolism and urease activity in rice (Oryza sativa L.). J Exp Bot 49:1545–1554

    Google Scholar 

  • Hogan ME, Swift IE, Done J (1983) Urease assay and ammonia release from leaf tissues. Phytochemistry 22:663–667

    Article  CAS  Google Scholar 

  • Kojima S, Bohner A, Gassert B, Yuan L, Von Wirén N (2007) AtDUR3 represents the major transporter for high-affinity urea transport across the plasma membrane of nitrogen-deficient Arabidopsis roots. Plant J 52:30–40

    Article  PubMed  CAS  Google Scholar 

  • Krogmeier MJ, McCarty GW, Bremner JM (1989) Potential phytotoxicity associated with the use of soil urease inhibitors. Proc Natl Acad Sci U S A 86:1110–1112

    Article  PubMed  CAS  Google Scholar 

  • Krogmeier MJ, McCarty GW, Shogren DR, Bremner JM (1991) Effect of nickel deficiency in soybeans on the phytotoxicity of foliar applied urea. Plant Soil 135:283–286

    Article  CAS  Google Scholar 

  • Kyllingsbæk A (1975) Extraction and colorimetric determination of urea in plants. Acta Agricult Scand B Soil Plant Sci 25:109–112

    Google Scholar 

  • Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Liu LH, Ludewig U, Frommer WB, von Wirén N (2003) AtDUR3 encodes a new type of high-affinity urea/H+ symporter in Arabidopsis. Plant Cell 15:790–800

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Loulakakis KA, Roubelakis-Angelakis KA (1991) Plant NAD(H)-glutamate dehydrogenase consists of two subunit polypeptides and their participation in the seven isoenzymes occurs in an ordered ratio. Plant Physiol 97:104–111

    Article  PubMed  CAS  Google Scholar 

  • Matsumoto H, Yasuda T, Koayashi M, Takahashi E (1966) The inducible formation of urease in rice plants. Soil Sci Plant Nutr 12:239–244

    Article  CAS  Google Scholar 

  • Matsumoto H, Hasegawa Y, Kobayashi M, Takahashi E (1968) Inducible formation of urease in Canavalia ensiformis. Physiol Plant 21:872–881

    Article  CAS  Google Scholar 

  • Mérigout P, Lelandais M, Bitton F, Renou JP, Briand X, Meyer C, Daniel-Vedele F (2008) Physiological and transcriptomic aspects of urea uptake and assimilation in Arabidopsis plants. Plant Physiol 147:1225–1238

    Article  PubMed  Google Scholar 

  • Mobley HL, Hausinger RP (1989) Microbial ureases: significance, regulation and molecular characterization. Microbiol Rev 53:85–108

    PubMed  CAS  Google Scholar 

  • Mobley HL, Island MD, Hausinger RP (1995) Molecular biology of microbial ureases. Microbiol Rev 59:451–480

    PubMed  CAS  Google Scholar 

  • O’Neal D, Joy KW (1973) Glutamine synthetase of pea leaves. I. Purification, stabilisation and pH optima. Arch Biochem Biophys 159:113–122

    Article  PubMed  Google Scholar 

  • Purnell MP, Botella JR (2007) Tobacco isoenzyme 1 of NAD(H)-dependent glutamate dehydrogenase catabolizes glutamate in vivo. Plant Physiol 143:530–539

    Article  PubMed  CAS  Google Scholar 

  • Purnell MP, Skopelitis DS, Roubelakis-Angelakis KA, Botella JR (2005) Modulation of higher-plant NAD(H)-dependent glutamate dehydrogenase activity in transgenic tobacco via alteration of β subunit levels. Planta 222:167–180

    Article  PubMed  CAS  Google Scholar 

  • Rhodes D, Brunk OG, Magalhaes JR (1989) Assimilation of ammonium by glutamate dehydrogenase? In: Poulten TE, Romeo JT, Conn EE (eds) Recent advances in phytochemistry. Plenum, New York, pp 191–226

    Google Scholar 

  • Skopelitis DS, Paranychiankis NV, Paschalidis KA, Plianokis ED, Delis ID, Yakoumakis DI et al (2006) Abiotic stress generates ROS that signal expression of anionic glutamate dehydrogenase to form glutamate for proline synthesis in tobacco and grapevine. Plant Cell 18:2767–2781

    Article  PubMed  CAS  Google Scholar 

  • Skopelitis DS, Paranychiankis NV, Kouvarakis A, Spyros A, Stephanou EG, Roubelakis-Angelakis KA (2007) The isoenzyme 7 of tobacco NAD(H)-dependent glutamate dehydrogenase exhibits high deaminating and low aminating activity. Plant Physiol 145:1726–1734

    Article  PubMed  CAS  Google Scholar 

  • Tercé-Laforgue T, Dubois F, Ferrario-Méry S, Pou de Crecenzo MA, Sangwan R, Hirel B (2004) Glutamate dehydrogenase of tobacco (Nicotiana tabacum L.) is mainly induced in the cytosol of phloem companion cells when ammonia is provided either externally or released during photorespiration. Plant Physiol 136:4308–4317

    Article  PubMed  Google Scholar 

  • Terman GL (1979) Volatilization losses of nitrogen as ammonia from surface-applied fertilizers, organic amendments, and crop residues. Adv Agron 31:189–223

    Article  CAS  Google Scholar 

  • Turano FJ, Dashner R, Upadhayaya A, Caldwell CR (1996) Purification of mitochondrial glutamate dehydrogenase from dark-grown soybean seedlings. Plant Physiol 112:1357–1364

    PubMed  CAS  Google Scholar 

  • Walker CD, Graham RD, Madison JT, Cary EE, Welch RM (1985) Effects of nickel deficiency on some nitrogen metabolites in cowpeas (Vigna unguiculata L. Walp). Plant Physiol 79:474–479

    Article  PubMed  CAS  Google Scholar 

  • Wang WH, Kohler B, Cao FQ, Liu LH (2008) Molecular and physiological aspects of urea transport in higher plants. Plant Sci 175:467–477

    Article  CAS  Google Scholar 

  • Watson CJ, Miller H (1996) Short-term effects of urea amended with the urease inhibitor N-(n-butyl) thiophosphoric triamide on perennial grass. Plant Soil 184:33–45

    Article  CAS  Google Scholar 

  • Watson CJ, Miller H, Poland P, Kilpatrick DJ, Allen MDB, Garret MK, Christianson CB (1994) Soil properties and the ability of the urease inhibitor N-(N-butyl)thiophosphoric triamide (NBTPT) to reduce ammonia volatilization from surface-applied urea. Soil Biol Biochem 26:1165–1171

    Article  CAS  Google Scholar 

  • Witte CP, Tiller SA, Taylor MA, Davies HV (2002) Leaf urea metabolism in potato. Urease activity profile and patterns of recovery and distribution of 15N after foliar urea application in wild-type and urease-antisense transgenics. Plant Physiol 128:1129–1136

    Article  PubMed  CAS  Google Scholar 

  • Witte CP, Rosso MG, Romeis T (2005) Identification of three urease accessory proteins that are required for urease activation in Arabidopsis. Plant Physiol 139:1155–1162

    Article  PubMed  CAS  Google Scholar 

  • Yamaya T, Oaks A, Rhodes D, Matsumoto H (1986) Synthesis of [15N]glutamate from [2–15N]glutamate and [15N]glycine by mitochondria isolated from pea and corn shoots. Plant Physiol 81:754–757

    Article  PubMed  CAS  Google Scholar 

  • Yuan L, Loqué D, Ye F, Frommer WB, Von Wirèn N (2007) Nitrogen-dependent posttranscriptional regulation of the ammonium transporter AtAMT1;1. Plant Physiol 143:732–744

    Article  PubMed  CAS  Google Scholar 

  • Zonia LE, Stebbins NE, Polacco JC (1995) Essential role of urease in germination of nitrogen limited Arabidopsis thaliana seeds. Plant Physiol 107:1097–1103

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Marie-Paule Bataillé and Raphaël Ségura for IRMS analyses. We acknowledge Xavier Sarda for helping with plant culture and harvest and Laurence Cantrill for kindly improving the English of the manuscript. The authors thank the «Pôle de compétitivité Mer-Bretagne» and the «Fond Unique Inter-ministériel» which supported this work conducted through the AZOSTIMER project.

Author information

Authors and Affiliations

  1. UMR 950 Ecophysiologie Végétale, Agronomie et nutritions N, C, S, Université de Caen Basse-Normandie, Esplanade de la Paix, 14032, Caen, Cedex, France

    Mustapha Arkoun, Laëtitia Jannin, Philippe Laîné, Philippe Etienne & Alain Ourry

  2. INRA, UMR 950 Ecophysiologie Végétale, Agronomie et nutritions N, C, S, Esplanade de la Paix, 14032, Caen, Cedex, France

    Mustapha Arkoun, Laëtitia Jannin, Philippe Laîné, Philippe Etienne & Alain Ourry

  3. Institut Jean-Pierre Bourgin, IJPB, UMR1318 INRA-AgroParisTech Bâtiment 2 INRA Centre de Versailles-Grignon Route de St-Cyr (RD10), 78026, Versailles, Cedex, France

    Céline Masclaux-Daubresse

  4. Plateau Technique de Chimie du Végétal, Institut Jean-Pierre Bourgin, UMR1318 INRA-AgroParisTech Bâtiment 1, INRA Centre de Versailles-Grignon, Route de St-Cyr (RD10), 78026, Versailles, Cedex, France

    Sylvie Citerne

  5. Timac Agro Spain, Poligono de Arazuri-Orcoyen Calle C n°32, 31160, Orcoyen, Spain

    Maria Garnica & José-Maria Garcia-Mina

  6. Timac Agro International, 27 avenue Franklin Roosevelt, 35408, Saint-Malo, France

    Jean-Claude Yvin

Authors
  1. Mustapha Arkoun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laëtitia Jannin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Philippe Laîné
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Philippe Etienne
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Céline Masclaux-Daubresse
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sylvie Citerne
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Maria Garnica
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. José-Maria Garcia-Mina
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jean-Claude Yvin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Alain Ourry
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alain Ourry.

Additional information

Responsible Editor: A.C. Borstlap.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOC 44 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arkoun, M., Jannin, L., Laîné, P. et al. A physiological and molecular study of the effects of nickel deficiency and phenylphosphorodiamidate (PPD) application on urea metabolism in oilseed rape (Brassica napus L.). Plant Soil 362, 79–92 (2013). https://doi.org/10.1007/s11104-012-1227-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11104-012-1227-2

Keywords

Search

Navigation