, Volume 231, Issue 5, pp 1145–1157 | Cite as

Conditional modulation of NAD levels and metabolite profiles in Nicotiana sylvestris by mitochondrial electron transport and carbon/nitrogen supply

  • Jutta Hager
  • Till K. Pellny
  • Caroline Mauve
  • Caroline Lelarge-Trouverie
  • Rosine De Paepe
  • Christine H. Foyer
  • Graham Noctor
Original Article


Environmental controls on leaf NAD status remain poorly understood. Here, we analyzed the effects of two key environmental variables, CO2 and nitrogen, on leaf metabolite profiles, NAD status and the abundance of key transcripts involved in de novo NAD synthesis in wild-type (WT) Nicotiana sylvestris and the CMSII mutant that lacks respiratory complex I. High CO2 and increased N supply both significantly enhanced NAD+ and NADH pools in WT leaves. In nitrogen-sufficient conditions, CMSII leaves were enriched in NAD+ and NADH compared to the WT, but the differences in NADH were smaller at high CO2 than in air because high CO2 increased WT NADH/NAD+. The CMSII-linked increases in NAD+ and NADH status were abolished by growth with limited nitrogen, which also depleted the nicotine and nicotinic acid pools in the CMSII leaves. Few statistically significant genotype and N-dependent differences were detected in NAD synthesis transcripts, with effects only on aspartate oxidase and NAD synthetase mRNAs. Non-targeted metabolite profiling as well as quantitative amine analysis showed that NAD+ and NADH contents correlated tightly with leaf amino acid contents across all samples. The results reveal considerable genotype- and condition-dependent plasticity in leaf NAD+ and NADH contents that is not linked to modified expression of NAD synthesis genes at the transcript level and show that NAD+ and NADH contents are tightly integrated with nitrogen metabolism. A regulatory two-way feedback circuit between nitrogen and NAD in the regulation of N assimilation is proposed that potentially links the nutritional status to NAD-dependent signaling pathways.


Nucleotides NAD synthesis Photorespiration Redox GC–MS 



Analysis of variance


Aspartate oxidase


Alternative oxidase


Cytoplasmic male sterile


Cytochrome oxidase


Glyceraldehyde-3-phosphate dehydrogenase


Gas chromatography–time of flight-mass spectrometry


High nitrogen


High-performance liquid chromatography


Low nitrogen


Nicotinic acid adenine dinucleotide


Nicotinamide adenine dinucleotide


NAD synthetase


Nicotinic acid mononucleotide


Nicotinic acid mononucleotide adenyl transferase


Nicotinic acid phosphoribosyl transferase


Nitrate reductase


5′-Phosphoribosyl 1-pyrophosphate


Quinolinate phosphoribosyl transferase


Quinolinate synthase


Retention index


Ribulose 1,5-bisphosphate


Wild type



This work was partly funded by the French Agence Nationale de la Recherche-Genoplante initiative, project no. GNP0508G and the UK Biotechnology and Biological Sciences Research Council grant BB/C51508X/1. Rothamsted Research receives grant-aided support from the BBSRC (UK).

Supplementary material

425_2010_1117_MOESM1_ESM.ppt (110 kb)
Supplementary material 1 (PPT 110 kb)
425_2010_1117_MOESM2_ESM.doc (38 kb)
Supplementary material 2 (DOC 38 kb)
425_2010_1117_MOESM3_ESM.xls (26 kb)
Supplementary material 3 (XLS 26 kb)
425_2010_1117_MOESM4_ESM.xls (26 kb)
Supplementary material 4 (XLS 26 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Jutta Hager
    • 1
  • Till K. Pellny
    • 3
  • Caroline Mauve
    • 2
  • Caroline Lelarge-Trouverie
    • 2
  • Rosine De Paepe
    • 1
  • Christine H. Foyer
    • 4
  • Graham Noctor
    • 1
  1. 1.Institut de Biotechnologie des PlantesUniversité de Paris sud XI, UMR CNRS 8618Orsay CedexFrance
  2. 2.Plate-forme Métabolisme-Métabolome, IFR87, Institut de Biotechnologie des PlantesUniversité de Paris sud XI, UMR CNRS 8618Orsay CedexFrance
  3. 3.Department of Plant SciencesRothamsted ResearchHertsUK
  4. 4.Center for Plant Sciences, Faculty of Biological SciencesUniversity of LeedsLeedsUK

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