Cubas, L. A. | bioRxiv Science

bioRxiv2025

Sub-optimal temperature leads to tighter coupling between photosynthetic electron transport and CO2 assimilation under fluctuating light in maize.

The C4 carbon concentrating pathway promotes high CO2 assimilation rates. To keep C4 photosynthesis energetically efficient, electron transport reactions and downstream biochemistry need to be carefully balanced. Here we use a combination of non-invasive measurements and metabolic profiling to study the efficiency of C4 photosynthesis in maize under two conditions that can lead to decoupling between electron transport and carbon assimilation: fluctuating light and suboptimal temperature. Measurements were performed for three fluctuating light regimes and three temperatures, providing the most detailed study to date of the interaction between fluctuating light and suboptimal temperature on the photosynthetic performance of maize, an important global crop. At room temperature, CO2 assimilation rates were decoupled from photosynthetic electron transport under fluctuating light regimes, in contrast to tight coordination observed under constant light. This decoupling was underpinned by metabolic flexibility and buffering by large pools of C4 transfer metabolites. Surprisingly, at sub-optimal temperatures, CO2 assimilation rates became more tightly coupled to photosynthetic electron transport rates under fluctuating light regimes. This appeared to be caused by strong feedback downregulation of electron transport and a stronger degree of light-saturation of CO2 assimilation at low temperature. Low temperature impacted carbon assimilation rates more strongly than metabolite pools or intercellular metabolite distribution, which could reflect negative effects on diffusional metabolite transfer through plasmodesmata. Altogether, these results show that maize is able to maintain energetic efficiency by buffering light transitions under room temperature, as well as avoid oxidative damage by strongly downregulating electron transfer under short-term exposure to low temperature. One-sentence summaryAnalysis of maize CO2 assimilation under fluctuating light shows significant decoupling from photosynthetic electron transport at room temperature, supported by metabolic flexibility and buffering by large pools of C4 transfer metabolites, but tight coordination is restored under suboptimal temperature due to enhanced feedback regulation of electron transport and a stronger degree of light saturation of CO2 assimilation.

plant biology↗

bioRxiv2025

Faster relaxation of nonphotochemical quenching (NPQ) in C4 than in C3 species

Acceleration of photoprotective non-photochemical quenching (NPQ) responses to changes in light intensity has been suggested as a strategy to enhance crop yield. Despite many key crops utilising C4 photosynthesis, our current understanding of NPQ overwhelmingly comes from C3 species. Using a series of experiments on three phylogenetically controlled C3 and C4 comparisons, we show that NPQ relaxation is faster in C4 species. Temporal analysis of NPQ relaxation in leaves infiltrated with inhibitors to block proton motive force formation or xanthophyll de-epoxidation showed that the faster relaxation observed in C4 species is driven by a greater contribution of energy-dependent quenching (qE) to overall NPQ. We show that the C4-associated enhancement of qE is linked to altered regulation of lumen pH in C4 species, reflecting increases in cyclic electron flow and membrane proton conductivity to meet the increased ATP demands of the C4 pathway. Indeed, in two of the three tested C4 species, NPQ relaxation became significantly slower and statistically indistinguishable from paired C3 species when ATP and NADPH consumption was suppressed by performing measurements in CO2-free air. Altogether, our results suggest that NPQ responses in C4 species may already be optimised to maintain high photosynthetic efficiency in the fluctuating light conditions typically found within C4 canopies. Given the intrinsically faster NPQ in C4 photosynthesis, further acceleration of NPQ may have limited scope to enhance crop photosynthetic efficiency. Significance StatementAcceleration of non-photochemical quenching has been proposed as a means to enhance crop photosynthetic efficiency in C3 species but whether this strategy has potential in C4 species, which include several major crops, remains unclear. We use three phylogenetically paired C3 and C4 species to show that NPQ relaxation is significantly faster in species with the C4 pathway, possibly aiding the maintenance of photosynthetic efficiency in fluctuating light environments. As a result, accelerating the rate of NPQ relaxation in C4 crops may have a more limited scope to enhance photosynthesis.

plant biology↗