Planta

, 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

Abstract

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.

Keywords

Nucleotides NAD synthesis Photorespiration Redox GC–MS 

Abbreviations

ANOVA

Analysis of variance

AO

Aspartate oxidase

AOX

Alternative oxidase

CMS

Cytoplasmic male sterile

COX

Cytochrome oxidase

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

GC–TOF-MS

Gas chromatography–time of flight-mass spectrometry

HN

High nitrogen

HPLC

High-performance liquid chromatography

LN

Low nitrogen

NaAD

Nicotinic acid adenine dinucleotide

NAD

Nicotinamide adenine dinucleotide

NADS

NAD synthetase

NaMN

Nicotinic acid mononucleotide

NaMNAT

Nicotinic acid mononucleotide adenyl transferase

NaPRT

Nicotinic acid phosphoribosyl transferase

NR

Nitrate reductase

PRPP

5′-Phosphoribosyl 1-pyrophosphate

QPRT

Quinolinate phosphoribosyl transferase

QS

Quinolinate synthase

RI

Retention index

RuBP

Ribulose 1,5-bisphosphate

WT

Wild type

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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