Arabidopsis lines with modified ascorbate concentrations reveal a link between ascorbate and auxin biosynthesis

Ascorbate is the most abundant water-soluble antioxidant in plants and an essential molecule for normal plant development. Although present in all green plants, ascorbate concentrations vary among plant species and tissues. While ascorbate accumulation is a trait of nutritional, and therefore, agronomical interest, the impact of different concentrations on cellular homeostasis remains elusive. To shed light on this question, we compared Arabidopsis (Arabidopsis thaliana) lines with very low (vtc2 mutant, 20% of wild-type (WT) levels), low (vtc4 mutant, 65% of WT levels), and high (vtc2/OE-VTC2, 165% of WT levels) ascorbate concentration in four-week-old rosette leaves. An 80% reduction of ascorbate increased the expression of genes implicated in defense against pathogens but repressed genes associated with abiotic stress responses. Unexpectedly, lines with increased (165% of WT) and decreased (65% of WT) ascorbate levels shared 85% of induced transcription factors and the gene ontology terms associated with their transcriptional programs. We identified TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA1), a gene encoding the enzyme that catalyzes the first step of auxin biosynthesis, among the group of genes whose expression was positively correlated with ascorbate content. Using a combination of genetic and pharmacological approaches in fluorescent and histochemical reporter lines for auxin biosynthesis and signaling activity, we revealed that TAA1- and TAA1 RELATED 2 (TAR2)-mediated auxin biosynthesis is necessary for plants to cope with increased ascorbate concentration in a light-dependent manner, revealing a layer of complexity in the regulatory landscape of redox homeostasis. Overall design: RNA-seq profilling of 4-week-old wild type (WT), vtc2-4 (vtc2), vtc4-4 (vtc4), and vtc2/OE-VTC2 L15 (OE) rosettes of Arabidopsis. Four-weeks old Arabidopsis rosettes grown under short day photoperiod (8-h light/16-h darkness cycle, 22±1?C, 300±70 µmol photons m-2 s-1) were collected 30 minutes after the lights turned on and flash-frozen in liquid nitrogen. RNA from three independent replicates per genotype (WT, vtc2, vtc4 and OE) was used in this experiment.