High nitrogen insensitive 9 (HNI9)-mediated systemic repression of root NO3- uptake is associated with changes in histone methylation.

In plants, root nitrate uptake systems are under systemic feedback repression by the N satiety of the whole organism, thus adjusting the N acquisition capacity to the N demand for growth; however, the underlying molecular mechanisms are largely unknown. We previously isolated the Arabidopsis high nitrogen-insensitive 9-1 (hni9-1) mutant, impaired in the systemic feedback repression of the root nitrate transporter NRT2.1 by high N supply. Here, we show that HNI9 encodes Arabidopsis INTERACT WITH SPT6 (AtIWS1), an evolutionary conserved component of the RNA polymerase II complex. HNI9/AtIWS1 acts in roots to repress NRT2.1 transcription in response to high N supply. At a genomic level, HNI9/AtIWS1 is shown to play a broader role in N signaling by regulating several hundred N-responsive genes in roots. Repression of NRT2.1 transcription by high N supply is associated with an HNI9/AtIWS1-dependent increase in histone H3 lysine 27 trimethylation at the NRT2.1 locus. Our findings highlight the hypothesis that posttranslational chromatin modifications control nutrient acquisition in plants.

Multiple FtsZ2 isoforms involved in chloroplast division and biogenesis are developmentally associated with thylakoid membranes in Arabidopsis.

Seed plants and algae have two distinct FtsZ protein families, FtsZ1 and FtsZ2, involved in plastid division. Distinctively, seed plants and mosses contain two FtsZ2 family members (FtsZ2-1 and FtsZ2-2) thus raising the question of the role of these FtsZ2 paralogs in plants. We show that both FtsZ2 paralogs, in addition to being present in the stroma, are associated with the thylakoid membranes and that association is developmentally regulated. We also show that several FtsZ2-1 isoforms are present with distinct intra-plastidial localization. Mutant analyses show that FtsZ2-1 is essential for chloroplast division and that FtsZ2-2 plays a specific role in chloroplast morphology and internal organisation in addition to participating in chloroplast partition.

Identification of Arabidopsis mutants impaired in the systemic regulation of root nitrate uptake by the nitrogen status of the plant.

Nitrate uptake by the roots is under systemic feedback repression by high nitrogen (N) status of the whole plant. The NRT2.1 gene, which encodes a NO(3)(-) transporter involved in high-affinity root uptake, is a major target of this N signaling mechanism. Using transgenic Arabidopsis (Arabidopsis thaliana) plants expressing the pNRT2.1::LUC reporter gene (NL line), we performed a genetic screen to isolate mutants altered in the NRT2.1 response to high N provision. Three hni (for high nitrogen insensitive) mutants belonging to three genetic loci and related to single and recessive mutations were selected. Compared to NL plants, these mutants display reduced down-regulation of both NRT2.1 expression and high-affinity NO(3)(-) influx under repressive conditions. Split-root experiments demonstrated that this is associated with an almost complete suppression of systemic repression of pNRT2.1 activity by high N status of the whole plant. Other mechanisms related to N and carbon nutrition regulating NRT2.1 or involved in the control of root SO(4)(-) uptake by the plant sulfur status are not or are slightly affected. The hni mutations did not lead to significant changes in total N and NO(3)(-) contents of the tissues, indicating that hni mutants are more likely regulatory mutants rather than assimilatory mutants. Nevertheless, hni mutations induce changes in amino acid, organic acid, and sugars pools, suggesting a possible role of these metabolites in the control of NO(3)(-) uptake by the plant N status. Altogether, our data indicate that the three hni mutants define a new class of N signaling mutants specifically impaired in the systemic feedback repression of root NO(3)(-) uptake.

Developmentally regulated association of plastid division protein FtsZ1 with thylakoid membranes in Arabidopsis thaliana.

FtsZ is a key protein involved in bacterial and organellar division. Bacteria have only one ftsZ gene, while chlorophytes (higher plants and green alga) have two distinct FtsZ gene families, named FtsZ1 and FtsZ2. This raises the question of why chloroplasts in these organisms need distinct FtsZ proteins to divide. In order to unravel new functions associated with FtsZ proteins, we have identified and characterized an Arabidopsis thaliana FtsZ1 loss-of-function mutant. ftsZ1-knockout mutants are impeded in chloroplast division, and division is restored when FtsZ1 is expressed at a low level. FtsZ1-overexpressing plants show a drastic inhibition of chloroplast division. Chloroplast morphology is altered in ftsZ1, with chloroplasts having abnormalities in the thylakoid membrane network. Overexpression of FtsZ1 also induced defects in thylakoid organization with an increased network of twisting thylakoids and larger grana. We show that FtsZ1, in addition to being present in the stroma, is tightly associated with the thylakoid fraction. This association is developmentally regulated since FtsZ1 is found in the thylakoid fraction of young developing plant leaves but not in mature and old plant leaves. Our results suggest that plastid division protein FtsZ1 may have a function during leaf development in thylakoid organization, thus highlighting new functions for green plastid FtsZ.

The plastid division proteins, FtsZ1 and FtsZ2, differ in their biochemical properties and sub-plastidial localization.

Plastid division in higher plants is morphologically similar to bacterial cell division, with a process termed binary fission involving constriction of the envelope membranes. FtsZ proteins involved in bacterial division are also present in higher plants, in which the ftsZ genes belong to two distinct families: ftsZ1 and ftsZ2. However, the roles of the corresponding proteins FtsZ1 and FtsZ2 in plastid division have not been determined. Here we show that the expression of plant FtsZ1 and FtsZ2 in bacteria has different effects on cell division, and that distinct protein domains are involved in the process. We have studied the assembly of purified FtsZ1 and FtsZ2 using a chemical cross-linking approach followed by PAGE and electron microscopy analyses of the resulting polymers. This has revealed that FtsZ1 is capable of forming long rod-shaped polymers and rings similar to the bacterial FtsZ structures, whereas FtsZ2 does not form any organized polymer. Moreover, using purified sub-plastidial fractions, we show that both proteins are present in the stroma, and that a subset of FtsZ2 is tightly bound to the purified envelope membranes. These results indicate that FtsZ2 has a localization pattern distinct from that of FtsZ1, which can be related to distinct properties of the proteins. From the results presented here, we propose a model for the sequential topological localization and functions of green plant FtsZ1 and FtsZ2 in chloroplast division.