Illuminated leaves assimilate CO 2 via gross photosynthesis and liberate CO 2 via photorespiration and ‘day respiration’, often denoted as R d. Day respiration is a minor CO 2-exchange component of net photosynthesis but is important for carbon use efficiency, computations of internal conductance, or the interpretation of net photosynthetic 12C/ 13C fractionation. Unfortunately, there is no simple method to measure R d and tracing the origin of C-atoms found in day-respired CO 2 is difficult. As a result, a common misconception is that day respiration is simply a catabolic, CO 2-producing flux through ordinary catabolism (glycolysis and Krebs ‘cycle’). In the past few years, considerable progress has been made in our understanding of day respiration. It appears that R d is a net flux resulting from several CO 2-generating and CO 2-fixing reactions, not only related to catabolism but also to anabolism (biosyntheses). In addition, there is now direct evidence that decarboxylating reactions are partly fed by carbon sources disconnected from current photosynthesis and this effect has consequences for isotopic mass-balance. Therefore, leaf day ‘respiration’ is much more than just CO 2 production by respiratory catabolism. Rather, it reflects whole metabolic orchestration of leaves from N fixation to secondary metabolism and it perhaps deserves another name, such as “day decarboxylations”.

Day respiration ( R d) is the metabolic, non-photorespiratory process by which illuminated leaves liberate CO 2 during photosynthesis. R d is used routinely in photosynthetic models and is thus critical for calculations. However, metabolic details associated with R d are poorly known, and this can be problematic to predict how R d changes with environmental conditions and relates to night respiration. It is often assumed that day respiratory CO 2 release just reflects ‘ordinary’ catabolism (glycolysis and Krebs ‘cycle’). Here, we carried out a pulse-chase experiment, whereby a 13CO 2 pulse in the light was followed by a chase period in darkness and then in the light. We took advantage of non-targeted, isotope-assisted metabolomics to determine non-‘ordinary’ metabolism, detect carbon remobilisation, and compare light and dark 13C utilisation. We found that several concurrent metabolic pathways (‘ordinary’ catabolism, oxidative pentose phosphates pathway, amino acid production, nucleotide biosynthesis, and secondary metabolism) took place in the light and participate in net CO 2 efflux associated with day respiration. Flux reconstruction from metabolomics leads to an underestimation of R d, further suggesting the contribution of a variety of CO 2-evolving processes. Also, the cornerstone of the Krebs ‘cycle’, citrate, is synthetised de novo from photosynthates mostly in darkness, and remobilised or synthesised from stored material in the light. Collectively, our data provides direct evidence that leaf day respiration ( i) involves several CO 2-producing reactions and ( ii) is fed by different carbon sources, including stored carbon disconnected from current photosynthates.

D-amino acids are the D stereoisomers of common L-amino acids found in proteins. In the past two decades, the occurrence of D-amino acids in plants has been reported and circumstantial evidence for a role in several processes has been provided, including the interaction with soil microorganisms or an interference with cellular signalling. However, examples are relatively scarce and D-amino acids can also be detrimental, some of them inhibiting growth and development. Thus, the persistence of a D-amino acid metabolism in plants is rather surprising and evolutive origins of D-amino acid metabolism is presently unclear. Systemic analysis of sequences associated with enzymes of D-amino acid metabolism shows that they are not simply inherited from cyanobacterial metabolism. In effect, the history of enzymes of plant D-amino acid metabolism likely involves several steps, cellular compartments, gene transfers and losses. Regardless of evolutive steps, enzymes of D-amino acid metabolism like D-amino acid transferases or racemases have been kept by higher plants and not simply eliminated, hence it is likely that they fulfil important metabolic roles, which can be illustrated with serine, tryptophan, and folate metabolism. We suggest that D-amino acid metabolism was perhaps crucial to support metabolic functions required during land plants evolution.

Phloem sap transport, velocity and allocation have been proposed to play a role in physiological limitations of crop yield, along with photosynthetic activity or water use efficiency. Although there is clear evidence that carbon allocation to grains effectively drives yield in cereals like wheat (as reflected by the harvest index), the influence of phloem transport rate and velocity is less clear. Here, we took advantage of previously published data on yield, respiration, carbon isotope composition, nitrogen content and water consumption in winter wheat cultivars grown across several sites with or without irrigation, to express grain production in terms of phloem sucrose transport and compare with xylem water transport. Our results suggest that phloem sucrose transport rate follows the same relationship with phloem N transport regardless of irrigation conditions and cultivars, and seems to depend mostly on grain weight (i.e. mg per grain). When compared to xylem sap water movement, phloem sap velocity (in m s -1) was 5.8 to 7.7 times lower. Depending on the assumption made for phloem sap sucrose concentration, either phloem sap velocity or its proportionality coefficient to xylem velocity change little with environmental conditions. Taken as a whole, phloem transport from leaves to grains seems to be homeostatic within a narrow range of values and following relationships with other plant physiological parameters across cultivars and conditions. This suggests that phloem transport per se is not a limitation for yield in wheat but rather, is controlled to sustain grain filling.

Plant metabolomics has been used widely in plant physiology, in particular to analyse metabolic responses to environmental parameters. Derivatization (via trimethylsilylation-methoximation) followed by GC-MS metabolic profiling is a major technique to quantify low molecular weight, common metabolites of primary carbon, sulphur and nitrogen metabolism. There are now excellent opportunities for new generation analyses, using high resolution, exact mass GC-MS spectrometers that are progressively becoming relatively cheap. However, exact mass GC-MS analyses for routine metabolic profiling are not common, there is no dedicated available database. Also, exact mass GC-MS is usually dedicated to structural resolution of targeted secondary metabolites. Here, we present a curated database for exact mass metabolic profiling (made of 336 analytes, 1,064 characteristic exact mass fragments) focused on molecules of primary metabolism. We show advantages of exact mass analyses, in particular to resolve isotopic patterns, localise S-containing metabolites, and avoid identification errors when analytes have common nominal mass peaks in their spectrum. We provide a practical example using leaves of different Arabidopsis ecotypes and show how exact mass GC-MS analysis can be applied to plant samples and identify metabolic profiles.

The natural 13C abundance (δ13C) in plant leaves has been used for decades with great success in agronomy to monitor water use efficiency and select modern cultivars adapted to dry conditions. However, in wheat, breeding also implies looking for genotypes with high carbon allocation to spikes and grains, and thus with a high harvest index and/or low carbon losses via respiration. Finding isotope-based markers of optimal carbon partitioning to grains would be extremely useful since isotope analyses are inexpensive and can be performed routinely at high throughput. Here, we took advantage of a set of field trials made of more than 600 plots with several wheat cultivars and measured agronomic parameters as well as δ13C values in leaves and grains. We find a linear relationship between the apparent isotope discrimination between leaves and grain (denoted as Δδcorr), and the respiration use efficiency-to-harvest index ratio. It means that overall, efficient carbon allocation to grains is associated with a small isotopic difference between leaves and grains. Our results show that 13C natural abundance in grains has some potential to help finding genotypes with better carbon allocation properties and assisting current wheat breeding technologies

Since the first description of phloem sap composition nearly 60 years ago, it is generally assumed that phloem sap does not contain nitrate and that there is little or no backflow of nitrate from shoots to roots. While it is true that nitrate can occasionally be absent from phloem sap, there is now substantial evidence that phloem can carry nitrate and furthermore, transporters involved in nitrate redistribution to shoot sink organs and roots have been found. This raises the question of why nitrate may or may not be present in phloem sap, why its concentration is generally kept low, and whether plant shoot-root nutrient cycling also involves nitrate. We propose here that phloem sap nitrate is not only an essential component of plant nutritional signaling but also contributes to physical properties of phloem sap and as such, its concentration is controlled to ensure proper coordination of plant development and nutrient transport.

Since the first description of phloem sap composition nearly 60 years ago, it is generally assumed that phloem sap does not contain nitrate and that there is little or no backflow of nitrate from shoots to roots. While it is true that nitrate can occasionally be absent from phloem sap, there is now substantial evidence that phloem can carry nitrate and furthermore, transporters involved in nitrate redistribution to shoot sink organs and roots have been found. This raises the question of why nitrate may or may not be present in phloem sap, why its concentration is generally kept low, and whether plant shoot-root nutrient cycling also involves nitrate. We propose here that phloem sap nitrate is not only an essential component of plant nutritional signaling but also contributes to physical properties of phloem sap and as such, its concentration is controlled to ensure proper coordination of plant development and nutrient transport.

Effects of K deficiency have been investigated for several decades and recently, progress has been made in identifying metabolomics signatures thereby offering potential to monitor the K status of crops in the field. However, effects of low K conditions could also be due to the antagonism with other nutrients like calcium (Ca) and the well-known biomarker of K deficiency, putrescine, could be a response to Ca/K imbalance rather than K deficiency. We carried out experiments in sunflower grown at either low or high K, at high or low Ca, with or without putrescine added to the nutrient solution. Using metabolomics and proteomics analysis, we show that a significant part of the low-K response such as lower photosynthesis and N assimilation, is due to calcium and can be suppressed by low Ca conditions. Putrescine addition tends to restore photosynthesis and N assimilation but but aggravates the impact of low-K conditions on catabolism. We conclude that (i) effects of K deficiency can be partly alleviated by the use of low Ca and not only by K fertilization, and (ii) in addition to its role as a metabolite, putrescine participates in the regulation of the content in enzymes involved in carbon primary metabolism.