Nitrogen Partitioning and Remobilization in Arabidopsis Under Sufficient and Depleted Conditions

  • Adel ZayedEmail author
  • Robert Crosby


Nitrogen is the essential nutrient most limiting to plant productivity. Two of the critical steps limiting the efficient use of nitrogen are the ability of plants to acquire it from applied fertilizers followed by the efficient remobilization of accumulated nitrogen from source to sink tissues. In order to characterize the contributions of these processes to nitrogen utilization by plants and how the expression of certain genes may influence these steps, a hydroponic-based nitrogen depletion method was developed to evaluate genetic efficacy of both phases of nitrogen metabolism, accumulation, and remobilization, in Arabidopsis thaliana. The method was tested under short and long days and it was found that short-day (10 h) conditions were effective in increasing nitrogen storage in source tissues more than long day, allowing differences in as low as 1 week of treatment initiation of nitrogen depletion to show exponential changes in seed yield. Under these conditions, plants were grown for seven weeks in sufficient N solution, where a subset of the population was harvested to determine basal N allocation. Half of the remaining plants remained in N sufficient conditions, while the rest were subjected to progressive N depletion conditions to determine the ability to remobilize nitrogen when supply is eliminated. At growth stage (GS) 9.7, effect of N treatments on total N, nitrate, seed yield, and biomass allocation was evaluated under N sufficient and N depletion conditions. Plants transformed with one of two genes [12S seed storage globulin precursor as antisense orientation (At1g07750) and a hypothetical protein as sense orientation (At3g49550)], that were previously identified among others to impact Arabidopsis tolerance to insufficient nitrogen levels, were characterized by the hydroponic method for their impact on nitrogen accumulation and remobilization in Arabidopsis. Phenotypic and metabolic analysis showed that the gene encoding the 12S seed storage globulin precursor has improved accumulation, assimilation, and remobilization, leading to increased seed yield under N depletion conditions. Conversely, the hypothetical protein has improved accumulation in vegetative tissues, but reduced assimilation and remobilization, leading to reduced seed yield. The results from this research help improve our understanding of how plants improve their efficiency to assimilate and translocate nitrogen when they face environmental stress factors that affect their ability to accumulate nitrogen from the surrounding environment.


Arabidopsis thaliana Hydroponics Nitrogen Remobilization Accumulation Assimilation Screen Assay High throughput 







12S seed storage globulin protein


Hypothetical protein


BASF, Bayer, Ciba-Geigy, Hoechst (BBCH) scale


Days after sowing


Days after transplanting


Reverse osmosis


  1. Boyes DC, Zayed AM, Ascenzi R, McCaskill AJ, Hoffman NE, Davis KR, Görlach J (2001) Growth stage-based phenotypic analysis of Arabidopsis: a model for high throughput functional genomics in plants. Plant Cell 13:1499–1510CrossRefPubMedPubMedCentralGoogle Scholar
  2. Cameron KC, Di HJ, Moir JL (2013) Nitrogen losses from the soil/plant system: a review. Ann Appl Biol 162(2013):145–173CrossRefGoogle Scholar
  3. Cooper AJ (1975) Crop production in recirculating nutrient solution. Sci Hortic 3:251–258CrossRefGoogle Scholar
  4. Guan P, Wang R, Nacry P, Breton G, Kay SA, Pruneda-Paz JL, Davani A, Crawford NM (2014) Nitrate foraging by Arabidopsis roots is mediated by the transcription factor TCP20 through the systemic signaling pathway. Proc Natl Acad Sci USA 111:15267–15272CrossRefPubMedGoogle Scholar
  5. Higuchi K, Iwase J, Tsukiori Y, Nakura D, Kobayashi N, Ohashi H, Saito A, Miwa E (2014) Early senescence of the oldest leaves of Fe-deficient barley plants may contribute to phytosiderophore release from the roots. Physiol Plant 151:313–322CrossRefPubMedGoogle Scholar
  6. 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):2369–2387CrossRefPubMedGoogle Scholar
  7. Hoagland DR (1950) The water-culture method for growing plants without soil. University of California, College of Agriculture, Agricultural Experiment Station, Berkeley, Calif, p 39:ill. (Circular; 347)Google Scholar
  8. Hou X, Tong H, Selby J, Dewitt J, Peng X, He ZH (2005) Involvement of a cell wall-associated kinase, WALK4, in Arabidopsis mineral responses. Plant Physiol 139:1704–1716CrossRefPubMedPubMedCentralGoogle Scholar
  9. Kant S, Bi YM, Rothstein SJ (2011) Understanding plant response to nitrogen limitation for the improvement of crop nitrogen use efficiency. J Exp Bot 62(4):1499–1509Google Scholar
  10. Lancashire PD, Bleiholder H, Boom TVD, Langelu¨ddeke P, Stauss R, Weber E, Witzenberger A (1991) A uniform decimal code for growth stages of crops and weeds. Ann Appl Biol 119:561–601CrossRefGoogle Scholar
  11. Lemaitre T, Gaufichon L, Boutet-Mercey S, Christ A, Masclaux-Daubresse C (2008) Enzymatic and metabolic diagnostic of nitrogen deficiency in Arabidopsis thaliana Wassileskija accession. Plant Cell Physiol 49(7):1056–1065CrossRefPubMedGoogle Scholar
  12. Masclaux-Daubresse C, Reisdorf-Cren M, Orsel M (2008). Leaf nitrogen remobilisation for plant development and grain filling. Plant Biol (Stuttg) 10 Suppl 1:23–36Google Scholar
  13. Masclaux-Daubresse C, Daniel-Vedele F, Dechorgnat J, Chardon F, Gaufichon L, Suzuki A (2010). Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. Ann Bot 105: 1141–1157Google Scholar
  14. Noodén LD, Penney JP (2001) Correlative controls of senescence and plant death in Arabidopsis thaliana (Brassicaceae). J Exp Bot 52:2151–2159CrossRefPubMedGoogle Scholar
  15. Norén H, Svensson P, Andersson B (2004) A convenient and versatile hydroponic cultivation system for Arabidopsis thaliana. Physiol Plant 121:343–348CrossRefGoogle Scholar
  16. Richard-Molard C, Krapp A, Brun F, Ney B, Daniel-Vedele F, Chaillou S (2008) Plant response to nitrate starvation is determined by N storage capacity matched by nitrate uptake capacity in two Arabidopsis genotypes. J Exp Bot 59(4):779–791CrossRefPubMedGoogle Scholar
  17. Sawyer JE (2015) Nitrogen use in Iowa corn production. Iowa State University Extension and Outreach, Ames, IowaGoogle Scholar
  18. Schippers JHM, Schmidt R, Wagstaff C, and Jing HC (2015). Living to die and dying to live: the survival strategy behind leaf senescence. Plant Physiol 169(2): 914–930Google Scholar
  19. Thomas H (2013) Senescence, ageing and death of the whole plant. New Phytol 197:696–711CrossRefPubMedGoogle Scholar
  20. Zayed A, Jones L, Tierney C, Miller M (2012) Hydroponics apparatus and methods of use. United States Patent Application # 20120277117. November 1, 2012Google Scholar
  21. Zhang X, Henriques R, Lin SS, Niu QW, Chua NH (2006) Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat Protoc 1(2):641–646Google Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.BiotechnologyMonsanto CompanySt. LouisUSA
  2. 2.108 T.W. Alexander Drive, Research Triangle ParkDurhamUSA

Personalised recommendations