Project description:Ascorbate is a major plant metabolite that plays crucial roles in various processes, from reactive oxygen scavenging to epigenetic regulation. However, to what extent and how ascorbate modulates metabolism is largely unknown. To address this, we investigated the consequences of chloroplastic and total cellular ascorbate-deficiencies by studying chloroplastic ascorbate transporter pht4;4 mutant lines, and the ascorbate-deficient vtc2-4 mutant of Arabidopsis thaliana. Under regular growth conditions, both ascorbate-deficiencies caused minor alterations in photosynthesis, with no apparent signs of oxidative damage. In contrast, metabolomics analysis revealed a global and largely overlapping metabolome rewiring in both ascorbate deficiencies, suggesting that chloroplastic ascorbate modulates plant metabolism. We observed significant alterations in amino acid metabolism, particularly in arginine metabolism, activation of nucleotide salvage pathways, and changes in secondary metabolism. In addition, proteome wide analysis of thermostability revealed that ascorbate may interact with enzymes involved in arginine metabolism, the Calvin-Benson cycle, and several photosynthetic electron transport components. Overall, our results suggest that, independently of oxidative stress, chloroplastic ascorbate interconnects and coordinates diverse metabolic pathways in vascular plants and thus acts as a regulatory hub.
Project description:VITAMIN C DEFECTIVE 3 (VTC3) is a distinctive protein with both kinase and phosphatase domains and is essential for ascorbate accumulation in Arabidopsis. In the vtc3 mutant, ascorbate levels are approximately 50% of those in the wild type, and light-induced increases in the ascorbate pool size are markedly suppressed. The regulation of the ascorbate pool size by light involves GDP-L-galactose phosphorylase (VTC2), a key biosynthetic enzyme. To elucidate the physiological function of VTC3, we compared the transcriptomes of vtc3, wild-type, and vtc2 plants. Contrary to expectations, the loss of VTC3 had minimal impact on the transcriptome. Among the few genes upregulated in the vtc3 mutant, several were related to pathogen responses.
Project description:In this study we explain the physiological, biochemical and gene expression mechanisms adopted by ammonium nitrate-fed Arabidopsis thaliana plants growing under elevated [CO2], highlighting the importance of root-to-shoot interactions in these responses A transcriptomic analysis enabled the identification of photoassimilate allocation and remobilization as fundamental process used by the plants to maintain the outstanding photosynthetic performance. Moreover, based on the relationship between plant carbon status and hormone functioning, the transcriptomic analyses provided an explanation of why phenology accelerates in elevated [CO2] conditions.
Project description:In this study we explain the physiological, biochemical and gene expression mechanisms adopted by ammonium nitrate-fed Arabidopsis thaliana plants growing under elevated [CO2], highlighting the importance of root-to-shoot interactions in these responses A transcriptomic analysis enabled the identification of photoassimilate allocation and remobilization as fundamental process used by the plants to maintain the outstanding photosynthetic performance. Moreover, based on the relationship between plant carbon status and hormone functioning, the transcriptomic analyses provided an explanation of why phenology accelerates in elevated [CO2] conditions.
Project description:Phosphate is an essential macronutrient required for plant growth and development. However, it is present at suboptimal levels in many terrestrial ecosystems. To ameliorate this limitation, plants have evolved developmental and physiological mechanisms known as phosphate starvation responses (PSR). One of the main PSR in Arabidopsis thaliana is a deep restructuration of the root system architecture, which includes a reduction in primary root growth resulting in a shallower root system better adapted to explore the nutrient-rich topsoil. Intense research over the last years has shown that this developmental change is dependent on the accumulation and redistribution of iron (Fe) at the root tip, which in turn, participates in Fenton reactions and generates reactive oxygen species that affect meristem function and cell elongation. We have recently identified and characterized a cytochrome-containing protein in A. thaliana, named CRR, which is involved in the primary root growth response to phosphate starvation. Our results showed that CRR is an ascorbate-dependent ferric-reductase whose expression levels modulates iron distribution pattern in the root, affecting meristem function and cell elongation. Moreover, this activity also has shown to be critical for iron toxicity tolerance since CRR determines the transport rate of iron from root to shoot.
Project description:Phosphate is an essential macronutrient required for plant growth and development. However, it is present at suboptimal levels in many terrestrial ecosystems. To ameliorate this limitation, plants have evolved developmental and physiological mechanisms known as phosphate starvation responses (PSR). One of the main PSR in Arabidopsis thaliana is a deep restructuration of the root system architecture, which includes a reduction in primary root growth resulting in a shallower root system better adapted to explore the nutrient-rich topsoil. Intense research over the last years has shown that this developmental change is dependent on the accumulation and redistribution of iron (Fe) at the root tip, which in turn, participates in Fenton reactions and generates reactive oxygen species that affect meristem function and cell elongation. We have recently identified and characterized a cytochrome-containing protein in A. thaliana, named CRR, which is involved in the primary root growth response to phosphate starvation. Our results showed that CRR is an ascorbate-dependent ferric-reductase whose expression levels modulates iron distribution pattern in the root, affecting meristem function and cell elongation. Moreover, this activity also has shown to be critical for iron toxicity tolerance since CRR determines the transport rate of iron from root to shoot.
Project description:We explore where transcriptional regulation of ascorbate concentration lies in plants. Is it in biosynthesis, recycling, regulation or consumption? Arabidopsis thaliana plants were grown under controlled environment at four photon flux density levels (PFD). Rosettes from plants were harvested at the four PFD levels and over a diurnal cycle and after a step change in PFD and analysed for ascorbate concentration and transcript levels measured by RNAseq. Ascorbate concentrations and expression of genes in the L-galactose ascorbate biosynthesis, recycling, consumption pathways and regulation are presented to provide a full analysis of the control of ascorbate by environmentally modulated gene expression. Ascorbate concentration responded to PFD levels but not to time of day and showed only a small response to change of PFD after 2 days. Of the L-galactose pathway genes, only GDP galactose phosphorylase (GGP) showed a significant response in to different PFDs, time of day and to change in PFD. Other genes also showed limited responses. This study compares gene expression of a range of ascorbate related genes to changes in environment in a unified way and supports the concept that GGP is the key regulatory gene in ascorbate biosynthesis and that post transcriptional regulation is also important.