Abstract

Central metabolism is the engine of plant biomass, supplying fruit growth with building blocks, energy, and biochemical cofactors. Among metabolic cornerstones, nicotinamide adenine dinucleotide (NAD) is particularly pivotal for electron transfer through reduction–oxidation (redox) reactions, thus participating in a myriad of biochemical processes. Besides redox functions, NAD is now assumed to act as an integral regulator of signaling cascades involved in growth and environmental responses. However, the regulation of NAD metabolism and signaling during fruit development remains poorly studied and understood. Here, we benefit from RNAseq and proteomic data obtained from nine growth stages of tomato fruit (var. Moneymaker) to dissect mRNA and protein profiles that link to NAD metabolism, including de novo biosynthesis, recycling, utilization, and putative transport. As expected for a cofactor synthesis pathway, protein profiles failed to detect enzymes involved in NAD synthesis or utilization, except for nicotinic acid phosphoribosyltransferase (NaPT) and nicotinamidase (NIC), which suggested that most NAD metabolic enzymes were poorly represented quantitatively. Further investigations on transcript data unveiled differential expression patterns during fruit development. Interestingly, among specific NAD metabolism-related genes, early de novo biosynthetic genes were transcriptionally induced in very young fruits, in association with NAD kinase, while later stages of fruit growth rather showed an accumulation of transcripts involved in later stages of de novo synthesis and in NAD recycling, which agreed with augmented NAD(P) levels. In addition, a more global overview of 119 mRNA and 78 protein significant markers for NAD(P)-dependent enzymes revealed differential patterns during tomato growth that evidenced clear regulations of primary metabolism, notably with respect to mitochondrial functions. Overall, we propose that NAD metabolism and signaling are very dynamic in the developing tomato fruit and that its differential regulation is certainly critical to fuel central metabolism linking to growth mechanisms.

Highlights

  • Plant metabolism is maintained by universal metabolic cornerstones including pyridine nucleotides such as nicotinamide adenine dinucleotide (NAD) (Noctor et al, 2006; Gakière et al, 2018b)

  • As a first attempt to clarify the importance of pyridine nucleotides for fruit growth, we measured total cellular NAD+, NADP+, NADH, and NADPH pools from nine growth stages of tomato fruit

  • Global changes in the pools of these pyridine nucleotides were statistically significant (ANOVA followed by binary Student’s t tests) and showed higher levels of both NAD(H) and NADP(H) in the very young fruit, with the highest pools observed at 8 days postanthesis for growth stage 1 (GS1) and the lowest for mature green (GS6, 41 dpa) and for red ripe (GS9, 53 dpa) stages of tomato fruit, respectively

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Summary

Introduction

Plant metabolism is maintained by universal metabolic cornerstones including pyridine nucleotides such as nicotinamide adenine dinucleotide (NAD) (Noctor et al, 2006; Gakière et al, 2018b). NAD and its phosphorylated form NADP are ubiquitous electron carriers modulating energy homeostasis through the transport of electrons within reduction–oxidation (redox) processes (Geigenberger and Fernie, 2014; Gakière et al, 2018a). While NAD is mostly found as oxidized NAD+, NADP mostly acts as a reductant (NADPH) (Noctor et al, 2006; Gakière et al, 2018b). Phosphorylation of NAD(H) to NADP(H) is catalyzed via highly conserved NAD+ kinases (E.C. 2.7.1.23) and NADH kinases (E.C. 2.7.1.86) playing essential roles in metabolic and redox reactions including photosynthesis performance and reactive oxygen species (ROS) homeostasis, which are both crucial for plant growth and responses to stress (Turner et al, 2004; Waller et al, 2010; Li et al, 2014; Li et al, 2018a)

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