, Volume 199, Issue 1–2, pp 155–166

Genetic diversity for nitrogen use efficiency in spinach (Spinacia oleracea L.) cultivars using the Ingestad model on hydroponics

  • Rafael Chan-Navarrete
  • Asako Kawai
  • Oene Dolstra
  • Edith T. Lammerts van Bueren
  • C. Gerard van der Linden


Spinach is a leafy vegetable that requires a high N fertilization to have a satisfactory yield and quality, in part because it has poor nitrogen use efficiency (NUE). Therefore, there is a need to breed for cultivars with an excellent NUE. To this end the genetic diversity for NUE-related traits was studied in a diverse set of commercial cultivars. This set was evaluated in a hydroponic system using the Ingestad model; the system was set at a relative growth rate of 0.14 and 0.18 g g−1 day−1 (low and high N, respectively). Experiments were performed at low and high plant density. Traits monitored for single plants included fresh and dry weight, leaf area, specific leaf area, dry weight ratio between root and shoot, and chlorophyll content. The high density experiment showed more genotypic variation for the observed traits than the low density one. Biomass production was considerably lower at low than at high N. Path analysis revealed that leaf area had the highest direct effect on NUE, while specific leaf area was an important trait determining variation in NUE at low N. Slow and fast growing genotypes were shown to use different strategies to utilize N, and these strategies are expressed differently at high and low N availability. This indicates that improving spinach for NUE is feasible using the analysed genotypes as source material, and different strategies can be targeted for adaptation of spinach cultivars to low N conditions.


Spinach Hydroponics Ingestad model NUE Nitrogen use efficiency 

Supplementary material

10681_2014_1186_MOESM1_ESM.docx (61 kb)
Supplementary material 1 (DOC 280 kb)


  1. Bais HP, Weir TL, Perry LG, Gilroy S, Vivanco JM (2006) The role of root exudates in rhizosphere interactions with plants and other organisms. Ann Revi Plant Biol 57:233–266CrossRefGoogle Scholar
  2. Baresel JP, Zimmermann G, Reents HJ (2008) Effects of genotype and environment on N uptake and N partition in organically grown winter wheat (Triticum aestivum L.) in Germany. Euphytica 163:347–354CrossRefGoogle Scholar
  3. Barker AV, Peck NH, MacDonald GE (1971) Nitrate accumulation in vegetables. 1. Spinach grown in upland soils. Agron J 63:126–129CrossRefGoogle Scholar
  4. Benincasa P, Guiducci M, Tei F (2011) The nitrogen use efficiency: meaning and source of variation—case studies on three vegetable crops in central Italy. HortTechnology 21(3):266–273Google Scholar
  5. Biemond H (1995) Effects of nitrogen on development and growth of the leaves of vegetables. 3. Appearance and expansion growth of leaves of spinach. Neth J Agric Sci 43:247–260Google Scholar
  6. Biemond H, Vos J, Struik PC (1996) Effects of nitrogen on accumulation and partitioning of dry matter and nitrogen of vegetables. 3. Spinach. Neth J Agric Sci 44:227–239Google Scholar
  7. Boese SR, Huner NPA (1990) Effect of growth temperature and temperature shifts on spinach leaf morphology and photosynthesis. Plant Physiol 94:1830–1836PubMedCentralPubMedCrossRefGoogle Scholar
  8. Breimer T (1982) Environmental factors and cultural measures affecting the nitrate content in spinach. Fertil Res 3:191–292CrossRefGoogle Scholar
  9. Cantliffe DJ (1973) Nitrate accumulation in table beets and spinach as affected by nitrogen, phosporus, and potassium nutrition and light intensity. Agron J 65:563–565CrossRefGoogle Scholar
  10. Dewey DR, Lu KH (1959) A correlation and path coefficient analysis of components of crested wheatgrass seed production. Agron J 51:515–518CrossRefGoogle Scholar
  11. Dolstra O, Denneboom C, de Vos ALF, van Loo EN (2007) Marker-assisted selection. In: Guimaraes EP, Ruane J, Scherf BD, Sonino A, Dargie JD (eds) Marker-assisted selection for improving quantitative traits of forage crops., Current status and future perspectives in crops, livestock, forestry and fishFAO, Rome, pp 59–65Google Scholar
  12. European Commission (2010) European Commission: report from the commission to the Council and the European Parliament on implementation of the Council Directive 91/676/EEC concerning the protection of water against pollution caused by s from agricultural sources for the period 2004-2007 SEC(2010)118, COM(2007)47 final/2, Brussels, 2011Google Scholar
  13. Evans JR (1989) Photosynthesis and nitrogen relationships in leaves of C3 plants. Oecologia 78:9–19CrossRefGoogle Scholar
  14. Evans JR, Poorter H (2001) Photosynthetic acclimation of plants to growth irradiance: the relative importance of specific leaf area and nitrogen partitioning in maximizing carbon gain. Plant Cell Environ 24:755–767CrossRefGoogle Scholar
  15. Fageria NK, Baligar VC (2005) Enhancing nitrogen use efficiency in crop plants. Adv Agron 88:97–185CrossRefGoogle Scholar
  16. FAOSTAT (2013) Food and Agriculture Organization of the United Nations. Accessed 12 May 2013
  17. Glimskär A, Ericsson T (1999) Relative nitrogen limitation at steady-state nutrition as a determinant of plasticity in five grassland plant species. Ann Bot 84:413–420CrossRefGoogle Scholar
  18. Gourley CJP, Allan DL, Russelle MP (1994) Plant nutrient efficiency: a comparison of definitions and suggested improvement. Plant Soil 158:29–37CrossRefGoogle Scholar
  19. Grindlay DJC (1997) Towards an explanation of crop nitrogen demand based on the optimization of leaf nitrogen per unit leaf area. Journal of Agric Sci 128:196–377CrossRefGoogle Scholar
  20. Hermans C, Hammond JP, White PJ, Verbruggen N (2006) How do plants respond to nutrient shortage by biomass allocation? Trends Plant Sci 11:610–617PubMedCrossRefGoogle Scholar
  21. Hirel B, Le Gouis J, Ney B, Gallais A (2007) The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches. J Exp Bot 58(9):2369–2387PubMedCrossRefGoogle Scholar
  22. Hirose T (1984) Nitrogen use efficiency in growth of Polygonum cuspidatum Sieb. et Zucc. Ann Bot 54:695–704Google Scholar
  23. Ingestad T (1982) Relative addition rate and external concentration; driving variables used in plant nutrition research. Plant, Cell Environ 5:443–453CrossRefGoogle Scholar
  24. Ingestad T, Ågren GI (1995) Plant nutrition and growth: basic principles. Plant Soil 168–169(15):20Google Scholar
  25. Koh E, Charoenprasert S, Mitchell AE (2012) Effect of organic and conventional cropping systems on ascorbic acid, vitamin C, flavonoids, nitrate and oxalate in 27 varieties of spinach (Spinacia oleracea L.). J Agric Food Chem 60(12):3144PubMedCrossRefGoogle Scholar
  26. Lambers H (1987) Does variation in photosynthetic rate explain variation in growth rate and yield ? Neth J Agric Sci 35:505–519Google Scholar
  27. Lawlor DW, Kontturi M, Kendal AC (1989) Photosynthesis by flag leaves of wheat in relation to protein, ribulose bisphosphate carboxylase activity and nitrogen supply. J Exp Bot 40:43–52CrossRefGoogle Scholar
  28. Liu Y, Tong Y, Zhu H, Ding E, Smith A (2006) Leaf chlorophyll readings as an indicator for spinach yield and nutritional quality with different nitrogen fertilizer applications. J Plant Nutr 29:1207–1217CrossRefGoogle Scholar
  29. Noguchi K, Terashima I (2006) Response to spinach leaf mitochondria to low N availability. Plant Cell Environ 29:710–719PubMedCrossRefGoogle Scholar
  30. Nunes-Nesi A, Fernie AR, Stitt M (2010) Metabolic and signalling aspects underpinning the regulation of plant carbon nitrogen interactions. Mol Plant 3:973–996PubMedCrossRefGoogle Scholar
  31. Remans T, Nacry P, Pervent M, Filleur S, Diatoff E, Mounier E, Tillard P, Forde BG, Gojon A (2006) The Arabidopsis NRT1.1 transporter participates in the signalling pathway triggering root colonization of nitrate rich patches. Proc Natl Acad Sci USA 103:19206–19211PubMedCentralPubMedCrossRefGoogle Scholar
  32. Santamaria P (2006) Nitrate in vegetables: toxicity, content, intake and EC regulation. J Sci Food Agric 86:10–17CrossRefGoogle Scholar
  33. Scheible WR, Gonzalez-Fontes A, Lauerer M, Muller-Rober B, Caboche M, Stitt M (1997a) Nitrate acts as a signal to induce organic acid metabolism and repress starch metabolism in tobacco. Plant Cell 9:783–798PubMedCentralPubMedCrossRefGoogle Scholar
  34. Scheible WR, Lauerer M, Schulze ED, Caboche M, Stitt M (1997b) Accumulation of nitrate in the shoot act as a signal to regulate shoot-root allocation in tobacco. Plant J 11(4):671–691CrossRefGoogle Scholar
  35. Smolders E, Merckx R (1992) Growth and shoot:root partitioning of spinach plants as affected by nitrogen supply. Plant Cell Environ 15:795–807CrossRefGoogle Scholar
  36. Smolders E, Buysse J, Merckx R (1993) Growth analysis of soil-grown spinach plants at different N-regimes. Plant Soil 154:73–80CrossRefGoogle Scholar
  37. Spiertz JHJ (2010) Nitrogen, sustainable agriculture and food security:a review. Agron Sustain Dev 30:43–55CrossRefGoogle Scholar
  38. Stagnari F, Di Bitetto V, Pisante M (2007) Effects of N fertilizers and rates on yield, safety and nutrients in processing spinach genotypes. Sci Hortic 114:225–233CrossRefGoogle Scholar
  39. Steingrover EG (1986) Nitrate accumulation in spinach. PhD thesis, RU GroningenGoogle Scholar
  40. Stitt M, Krapp A (1999) The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background. Plant Cell Environ 22(6):583–621CrossRefGoogle Scholar
  41. Van Loo EN, Schapendonk AHCM, de Vos ALF (1992) Effects of nitrogen supply on tillering dynamics and regrowth of perennial ryegrass populations. Neth J Agric Sci 40:381–400Google Scholar
  42. Wang JF, Zhou Y, Dong CX, Shen QR, Putheti R (2009) Effects of NH4 +-N/NO−3 N ratios on growth, nitrate uptake and organic acid levels of spinach (Spinacia oleracea L.). Afr J Biotechnol 8:3597–3602Google Scholar
  43. Wolfe A, Patz JA (2002) Reactive nitrogen and human health: acute and long-term implications. AMBIO 31(2):120–125PubMedGoogle Scholar
  44. Xu G, Fan X, Miller AJ (2012) Plant nitrogen assimilation and use efficiency. Annu Rev Plant Biol 63:153–182PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Rafael Chan-Navarrete
    • 1
  • Asako Kawai
    • 1
  • Oene Dolstra
    • 1
  • Edith T. Lammerts van Bueren
    • 1
  • C. Gerard van der Linden
    • 1
  1. 1.Wageningen UR Plant BreedingWageningen University and Research CentreWageningenThe Netherlands

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