ZHANG, Youjun; KRAHNERT, Ina; BOLZE, Antje; GIBON, Yves; FERNIE, Alisdair

doi:10.1002/cppb.20115

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ZHANG, Youjun
Max-Planck-Institut für Molekulare Pflanzenphysiologie [MPI-MP]
Center of Plant Systems Biology and Biotechnology [Plovdiv] [CPSBB]

KRAHNERT, Ina
Max-Planck-Institut für Molekulare Pflanzenphysiologie [MPI-MP]

BOLZE, Antje
Max-Planck-Institut für Molekulare Pflanzenphysiologie [MPI-MP]

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Article de revue

Ce document a été publié dans

Current Protocols in Plant Biology. 2020, vol. 5, n° 3, p. e20115

Résumé en anglais

As the principal co-factors of many metabolic pathways, the measurement of both adenine nucleotides and nicotinamide adenine dinucleotide provides important information about cellular energy metabolism. However, given their rapid and reversible conversion as well as their relatively low concentration ranges, it is difficult to measure these compounds. Here, we describe a highly sensitive and selective ion-pairing HPLC method with fluorescence detection to quantify adenine nucleotides in plants. In addition, nicotinamide adenine dinucleotide is a crucially important redox-active substrate for multiple catabolic and anabolic reactions with the ratios of NAD + /NADH and NADP + /NADPH being suggested as indicators of the general intracellular re-dox potential and hence metabolic state. Here, we describe highly sensitive enzyme cycling−based colorimetric assays (with a detection limit in the pmol range) performed subsequent to a simple extraction procedure involving acid or base extraction to allow the measurement of the cellular levels of these metabo-lites. © 2020 The Authors. Basic Protocol 1: Preparation of plant material for the measurement Basic Protocol 2: Measurement of ATP, ADP, and AMP via HPLC Basic Protocol 3: NAD + /NADP + measurements Basic Protocol 4: NADH/NADPH measurements Basic Protocol 5: Data analysis and quality control approaches Keywords: ADP r ATP r NAD + /NADP + r NADH/NADPH< Réduire

Mots clés

Métabolisme

Plante

Physiologie végétale

Mots clés en anglais

ADP

ATP

NAD+/NADP+

NADH/NADPH

DOI

http://dx.doi.org/10.1002/cppb.20115

Project ANR

Centre français de phénomique végétale - ANR-11-INBS-0012

Origine

Importé de hal

Unités de recherche

Biologie du Fruit & Pathologie (BFP) - UMR 1332