Genome-wide association study unveils ascorbate regulation by PAS/LOV PROTEIN during high light acclimation

Abstract

Varying light conditions elicit metabolic responses as part of acclimation with changes in ascorbate levels being an important component. Here, we adopted a genome-wide association-based approach to characterize the response in ascorbate levels on high light acclimation in a panel of 315 Arabidopsis (Arabidopsis thaliana) accessions. These studies revealed statistically significant SNPs for total and reduced ascorbate under high light conditions at a locus in chromosome 2. Ascorbate levels under high light and the region upstream and within PAS/LOV PROTEIN (PLP) were strongly associated. Intriguingly, subcellular localization analyses revealed that the PLPA and PLPB splice variants co-localized with VITAMIN C DEFECTIVE2 (VTC2) and VTC5 in both the cytosol and nucleus. Yeast 2-hybrid and Bimolecular fluorescence complementation analyses revealed that PLPA and PLPB interact with VTC2 and that blue light diminishes this interaction. Furthermore, PLPB knockout mutants were characterized by 1.5- to 1.7- fold elevations in their ascorbate levels, whereas knockout mutants of the cry2 cryptochromes displayed 1.2- to 1.3- fold elevations compared to WT. Our results collectively indicate that PLP plays a critical role in the elevation of ascorbate levels, which is a signature response of high light acclimation. The results strongly suggest that this is achieved via release of the inhibitory effect of PLP on VTC2 upon blue light illumination, as the VTC2-PLPB interaction is stronger under darkness. The conditional importance of the cryptochrome receptors under different environmental conditions suggests a complex hierarchy underpinning the environmental control of ascorbate levels. However, the data we present here clearly demonstrate that PLP dominates during high light acclimation.

The D‐mannose/L‐galactose pathway plays a predominant role in ascorbate biosynthesis in the liverwort Marchantia polymorpha but is not regulated by light and oxidative stress

“…In Arabidopsis plants, the CONSTITUTIVE PHOTOMORPHOGENIC9-Signalosome subunit 5B (CSN5B) has been reported to negatively regulate ascorbate biosynthesis by being involved in the degradation of GMP proteins during the dark period (Wang et al, 2013). Recently, Aarabi et al (2023) reported the PAS/LOV protein (PLP) as a novel ascorbate regulator, which suppresses ascorbate biosynthesis by interacting with GGP in the dark, and light irradiation resolves this association and allows GGP to function, thereby increasing ascorbate levels. In M. polymorpha, both CSN5B and PLP orthologs are present in the genome (Mapoly0006s0112.1 and Mapoly0022s0166.1, respectively), and their involvement in ascorbate biosynthesis will need to be investigated in the future.…”