Post-flowering biotic and abiotic stresses impact nitrogen use efficiency and seed filling in Arabidopsis thaliana.

Nitrogen (N) is an essential nutrient that plants require for the synthesis of amino acids, proteins, and many other important metabolites. Plant metabolism and growth are consequently dependent on the amount of N that is assimilated and distributed from source leaves to developing sinks, such as fruits and seeds. The environmental stresses enhanced by climate change deeply influence seed yield and seed composition, and may disturb N use efficiency (NUE) in pants. We aimed to investigate plant responses to extreme climates with regard to NUE, N remobilization efficiency, and seed composition. By studying a collection of Arabidopsis genotypes showing a range of C:N ratios in seeds, we investigated the impact of different post-flowering growth conditions (control, heat, drought, low nitrate availability, induced senescence, and induced plant defense) on seed yield, N allocation in organs, NUE, and N remobilization efficiency. We analysed how post-flowering stresses could change seed filling and showed that post-flowering stresses change both the range of N and C concentrations and the C:N stoichiometry in seeds. Using a new trait, called delta seed composition, we measured the deviation in C:N stoichiometry of each genotype and revealed the genetic determinism of the C:N stoichiometry. Altogether, the results indicate that extreme climate impacts NUE dramatically in plants and generates different bottlenecks in N fluxes during seed filling.

Increased expression of a phloem membrane protein encoded by NHL26 alters phloem export and sugar partitioning in Arabidopsis.

The complex process of phloem sugar transport involves symplasmic and apoplasmic events. We characterized Arabidopsis thaliana lines ectopically expressing a phloem-specific gene encoding NDR1/HIN1-like26 (NHL26), a putative membrane protein. NHL26 overexpressor plants grew more slowly than wild-type plants, accumulated high levels of carbohydrates in mature leaves, and had a higher shoot biomass, contrasting with slower root growth and a lower seed yield. Similar effects were observed when NHL26 was overexpressed in companion cells, under the control of a companion cell-specific promoter. The soluble sugar content of the phloem sap and sink organs was lower than that in the wild type, providing evidence of a sugar export defect. This was confirmed in a phloem-export assay with the symplastic tracer carboxyfluorescein diacetate. Leaf sugar accumulation was accompanied by higher organic acid, amino acid, and protein contents, whereas analysis of the metabolite profile of phloem sap exudate revealed no change in amino acid or organic acid content, indicating a specific effect on sugar export. NHL26 was found to be located in the phloem plasmodesmata and the endoplasmic reticulum. These findings reveal that NHL26 accumulation affects either the permeability of plasmodesmata or sugar signaling in companion cells, with a specific effect on sugar export.

ACCELERATED CELL DEATH 6 Acts on Natural Leaf Senescence and Nitrogen Fluxes in Arabidopsis.

As the last step of leaf development, senescence is a molecular process involving cell death mechanism. Leaf senescence is trigged by both internal age-dependent factors and environmental stresses. It must be tightly regulated for the plant to adopt a proper response to environmental variation and to allow the plant to recycle nutrients stored in senescing organs. However, little is known about factors that regulate both nutrients fluxes and plant senescence. Taking advantage of variation for natural leaf senescence between Arabidopsis thaliana accessions, Col-0 and Ct-1, we did a fine mapping of a quantitative trait loci for leaf senescence and identified ACCELERATED CELL DEATH 6 (ACD6) as the causal gene. Using two near-isogeneic lines, differing solely around the ACD6 locus, we showed that ACD6 regulates rosette growth, leaf chlorophyll content, as well as leaf nitrogen and carbon percentages. To unravel the role of ACD6 in N remobilization, the two isogenic lines and acd6 mutant were grown and labeled with 15N at the vegetative stage in order to determine 15N partitioning between plant organs at harvest. Results showed that N remobilization efficiency was significantly lower in all the genotypes with lower ACD6 activity irrespective of plant growth and productivity. Measurement of N uptake at vegetative and reproductive stages revealed that ACD6 did not modify N uptake efficiency but enhanced nitrogen translocation from root to silique. In this study, we have evidenced a new role of ACD6 in regulating both sequential and monocarpic senescences and disrupting the balance between N remobilization and N uptake that is required for a good seed filling.