UC Berkeley

Although apparent light inhibition of leaf day respiration is a widespread reported phenomenon, the mechanisms involved, including utilization of alternate respiratory pathways and substrates and light inhibition of TCA cycle enzymes are under active investigation. Recently, acetate fermentation was highlighted as a key drought survival strategy mediated through protein acetylation and jasmonate signaling. Here, we evaluate the light-dependence of acetate transport and assimilation in Populus trichocarpa trees using the dynamic xylem solution injection (DXSI) method developed here for continuous studies of C1 and C2 organic acid transport and light-dependent metabolism. Over 7 days, 1.0 L of [¹³C]formate and [¹³C₂]acetate solutions were delivered to the stem base of 2-year old potted poplar trees, while continuous diurnal observations were made in the canopy of CO₂, H₂O, and isoprene gas exchange together with δ¹³CO₂. Stem base injection of 10 mM [¹³C₂]acetate induced an overall pattern of canopy branch headspace ¹³CO₂ enrichment (δ¹³CO₂ +27‱) with a diurnal structure in δ¹³CO₂ reaching a mid-day minimum followed by a maximum shortly after darkening where δ¹³CO₂ values rapidly increased up to +12‱. In contrast, 50 mM injections of [¹³C]formate were required to reach similar δ¹³CO₂ enrichment levels in the canopy with δ¹³CO₂ following diurnal patterns of transpiration. Illuminated leaves of detached poplar branches pretreated with 10 mM [¹³C₂]acetate showed lower δ¹³CO₂ (+20‱) compared to leaves treated with 10 mM [¹³C]formate (+320‱), the opposite pattern observed at the whole plant scale. Following dark/light cycles at the leaf-scale, rapid, strong, and reversible enhancements in headspace δ¹³CO₂ by up to +60‱ were observed in [¹³C₂]acetate-treated leaves which showed enhanced dihydrojasmonic acid and TCA cycle intermediate concentrations. The results are consistent with acetate in the transpiration stream as an effective activator of the jasmonate signaling pathway and respiratory substrate. The shorter lifetime of formate relative to acetate in the transpiration stream suggests rapid formate oxidation to CO₂ during transport to the canopy. In contrast, acetate is efficiently transported to the canopy where an increased allocation towards mitochondrial dark respiration occurs at night. The results highlight the potential for an effective integration of acetate into glyoxylate and TCA cycles and the light-inhibition of citrate synthase as a potential regulatory mechanism controlling the diurnal allocation of acetate between anabolic and catabolic processes.