ABSTRACT
Production of reactive oxygen species (ROS) by NADPH oxidases (NOXs) impacts many processes in animals and plants, and many plant receptor pathways involve rapid, NOX-dependent increases of ROS. Yet, their general reactivity has made it challenging to pinpoint the precise role and immediate molecular action of ROS. A well-understood ROS action in plants is to provide the co-substrate for lignin peroxidases in the cell wall. Lignin can be deposited with exquisite spatial control, but the underlying mechanisms have remained elusive. Here, we establish a kinase signaling relay that exerts direct, spatial control over ROS production and lignification within the cell wall. We show that polar localization of a single kinase component is crucial for pathway function. Our data indicate that an intersection of more broadly localized components allows for micrometer-scale precision of lignification and that this system is triggered through initiation of ROS production as a critical peroxidase co-substrate.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Lignin/metabolism , Protein Kinases/metabolism , Reactive Oxygen Species/metabolism , Gene Expression Regulation, Plant , NADPH Oxidases/metabolism , Peroxidases/metabolism , Plant Roots/metabolismABSTRACT
Plants use leucine-rich repeat receptor kinases (LRR-RKs) to sense sequence diverse peptide hormones at the cell surface. A 3.0-Å crystal structure of the LRR-RK GSO1/SGN3 regulating Casparian strip formation in the endodermis reveals a large spiral-shaped ectodomain. The domain provides a binding platform for 21 amino acid CIF peptide ligands, which are tyrosine sulfated by the tyrosylprotein sulfotransferase TPST/SGN2. GSO1/SGN3 harbors a binding pocket for sulfotyrosine and makes extended backbone interactions with CIF2. Quantitative biochemical comparisons reveal that GSO1/SGN3-CIF2 represents one of the strongest receptor-ligand pairs known in plants. Multiple missense mutations are required to block CIF2 binding in vitro and GSO1/SGN3 function in vivo. Using structure-guided sequence analysis we uncover previously uncharacterized CIF peptides conserved among higher plants. Quantitative binding assays with known and novel CIFs suggest that the homologous LRR-RKs GSO1/SGN3 and GSO2 have evolved unique peptide binding properties to control different developmental processes. A quantitative biochemical interaction screen, a CIF peptide antagonist and genetic analyses together implicate SERK proteins as essential coreceptor kinases required for GSO1/SGN3 and GSO2 receptor activation. Our work provides a mechanistic framework for the recognition of sequence-divergent peptide hormones in plants.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Peptides/metabolism , Protein Kinases/metabolism , Amino Acid Sequence , Arabidopsis/chemistry , Arabidopsis/enzymology , Arabidopsis/genetics , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/genetics , Kinetics , Ligands , Peptides/chemistry , Plant Growth Regulators/chemistry , Plant Growth Regulators/metabolism , Protein Binding , Protein Kinases/chemistry , Protein Kinases/geneticsABSTRACT
The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) enzyme catalyzes the major rate-limiting step of the mevalonic acid (MVA) pathway from which sterols and other isoprenoids are synthesized. In contrast with our extensive knowledge of the regulation of HMGR in yeast and animals, little is known about this process in plants. To identify regulatory components of the MVA pathway in plants, we performed a genetic screen for second-site suppressor mutations of the Arabidopsis thaliana highly drought-sensitive drought hypersensitive2 (dry2) mutant that shows decreased squalene epoxidase activity. We show that mutations in SUPPRESSOR OF DRY2 DEFECTS1 (SUD1) gene recover most developmental defects in dry2 through changes in HMGR activity. SUD1 encodes a putative E3 ubiquitin ligase that shows sequence and structural similarity to yeast Degradation of α factor (Doα10) and human TEB4, components of the endoplasmic reticulum-associated degradation C (ERAD-C) pathway. While in yeast and animals, the alternative ERAD-L/ERAD-M pathway regulates HMGR activity by controlling protein stability, SUD1 regulates HMGR activity without apparent changes in protein content. These results highlight similarities, as well as important mechanistic differences, among the components involved in HMGR regulation in plants, yeast, and animals.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Hydroxymethylglutaryl CoA Reductases/metabolism , Ubiquitin-Protein Ligases/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Humans , Membrane Proteins/genetics , Mevalonic Acid/metabolism , Mutation , Phenotype , Plant Roots/genetics , Plant Roots/growth & development , Plant Roots/metabolism , Plant Shoots/genetics , Plant Shoots/metabolism , Plants, Genetically Modified , Saccharomyces cerevisiae Proteins/genetics , Sequence Homology, Amino Acid , Sterols/metabolism , Ubiquitin-Protein Ligases/geneticsABSTRACT
TETRATRICOPEPTIDE THIOREDOXIN-LIKE (TTL) proteins are characterized by the presence of six tetratricopeptide repeats in conserved positions and a carboxyl-terminal region known as the thioredoxin-like domain with homology to thioredoxins. In Arabidopsis (Arabidopsis thaliana), the TTL gene family is composed by four members, and the founder member, TTL1, is required for osmotic stress tolerance. Analysis of sequenced genomes indicates that TTL genes are specific to land plants. In this study, we report the expression profiles of Arabidopsis TTL genes using data mining and promoter-reporter ß-glucuronidase fusions. Our results show that TTL1, TTL3, and TTL4 display ubiquitous expression in normal growing conditions but differential expression patterns in response to osmotic and NaCl stresses. TTL2 shows a very different expression pattern, being specific to pollen grains. Consistent with the expression data, ttl1, ttl3, and ttl4 mutants show reduced root growth under osmotic stress, and the analysis of double and triple mutants indicates that TTL1, TTL3, and TTL4 have partially overlapping yet specific functions in abiotic stress tolerance while TTL2 is involved in male gametophytic transmission.
Subject(s)
Adaptation, Physiological , Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Stress, Physiological , Arabidopsis/drug effects , Arabidopsis/metabolism , Arabidopsis/physiology , Arabidopsis Proteins/classification , Arabidopsis Proteins/genetics , Computational Biology , Data Mining , Gene Expression Profiling , Gene Expression Regulation, Plant , Genes, Plant , Genes, Reporter , Glucuronidase/genetics , Glucuronidase/metabolism , Multigene Family , Mutation , Phylogeny , Plant Roots/genetics , Plant Roots/metabolism , Plant Roots/physiology , Plants, Genetically Modified/drug effects , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/physiology , Pollen/genetics , Pollen/metabolism , Pollen/physiology , Promoter Regions, Genetic , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Sodium Chloride/pharmacologyABSTRACT
The growth of plants, like that of other walled organisms, depends on the ability of the cell wall to yield without losing its integrity. In this context, plant cells can sense the perturbation of their walls and trigger adaptive modifications in cell wall polymer interactions. Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) THESEUS1 (THE1) was previously shown in Arabidopsis to trigger growth inhibition and defense responses upon perturbation of the cell wall, but so far, neither the ligand nor the role of the receptor in normal development was known. Here, we report that THE1 is a receptor for the peptide rapid alkalinization factor (RALF) 34 and that this signaling module has a role in the fine-tuning of lateral root initiation. We also show that RALF34-THE1 signaling depends, at least for some responses, on FERONIA (FER), another RALF receptor involved in a variety of processes, including immune signaling, mechanosensing, and reproduction [1]. Together, the results show that RALF34 and THE1 are part of a signaling network that integrates information on the integrity of the cell wall with the coordination of normal morphogenesis.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Peptide Hormones/genetics , Plant Roots/growth & development , Protein Kinases/genetics , Receptors, Cell Surface/genetics , Signal Transduction , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Cell Wall/metabolism , Peptide Hormones/metabolism , Plant Roots/genetics , Protein Kinases/metabolism , Receptors, Cell Surface/metabolismABSTRACT
Plant roots acquire nutrients from the soil and transport them upwards to the aerial parts. To reach the central vasculature of the root, water and nutrients radially cross all external cell layers. The endodermis surrounds the vascular tissues and forms diffusion barriers. It thereby compartmentalizes the root and allows control of nutrient transport from the soil to the vasculature, as well as preventing backflow of nutrients from the stele. To achieve this role, endodermal cells undergo two specialized differentiations states consisting of deposition of two impermeable polymers in the cell wall: lignin, forming the Casparian strips, and suberin lamellae. Recent publications showed that endodermal barrier formation is not a hard-wired, irreversible process. Synthesis and degradation of suberin lamellae is highly regulated by plant hormones in response to nutrient stresses. Moreover, Casparian strip continuity seems to be constantly checked by two small peptides produced in the vasculature that diffuse into the apoplastic space in order to test endodermal barrier integrity. This review discusses the recent understanding of endodermal barrier surveillance and plasticity and its role in plant nutrition.
Subject(s)
Plant Roots/metabolism , Plant Vascular Bundle/metabolismABSTRACT
The root endodermis forms its extracellular diffusion barrier by developing ringlike impregnations called Casparian strips. A factor responsible for their establishment is the SCHENGEN3/GASSHO1 (SGN3/GSO1) receptor-like kinase. Its loss of function causes discontinuous Casparian strips. SGN3 also mediates endodermal overlignification of other Casparian strip mutants. Yet, without ligand, SGN3 function remained elusive. Here we report that schengen2 (sgn2) is defective in an enzyme sulfating peptide ligands. On the basis of this observation, we identified two stele-expressed peptides (CASPARIAN STRIP INTEGRITY FACTORS, CIF1/2) that complement sgn2 at nanomolar concentrations and induce Casparian strip mislocalization as well as overlignification-all of which depend on SGN3. Direct peptide binding to recombinant SGN3 identifies these peptides as SGN3 ligands. We speculate that CIF1/2-SGN3 is part of a barrier surveillance system, evolved to guarantee effective sealing of the supracellular Casparian strip network.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Plant Roots/metabolism , Protein Kinases/metabolism , Sulfotransferases/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Diffusion , Ligands , Peptides/metabolism , Plant Roots/genetics , Protein Binding , Protein Kinases/genetics , Sulfotransferases/geneticsABSTRACT
Casparian strips are precisely localized and aligned ring-like cell wall modifications in the root of all higher plants. They set up an extracellular diffusion barrier analogous to animal tight junctions, and are crucial for maintaining the homeostatic capacity of plant roots. Casparian strips become localized because of the formation of a highly stable plasma membrane domain, consisting of a family of small transmembrane proteins called Casparian strip membrane domain proteins (CASPs). Here we report a large-scale forward genetic screen directly visualizing endodermal barrier function, which allowed us to identify factors required for the formation and integrity of Casparian strips. We present the identification and characterization of one of the mutants, schengen1 (sgn1), a receptor-like cytoplasmic kinase that we show localizes in a strictly polar fashion to the outer plasma membrane of endodermal cells and is required for the positioning and correct formation of the centrally located CASP domain.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Cell Membrane/metabolism , Cell Wall/metabolism , Membrane Proteins/genetics , Arabidopsis Proteins/metabolism , Diffusion , Membrane Proteins/metabolismABSTRACT
Increasing L-ascorbic acid (AsA, vitamin C) content in fruits is a common goal in current breeding programs due to its beneficial effect on human health. Attempts to increase AsA content by genetic engineering have resulted in variable success likely due to AsA's complex regulation. Here, we report the effect of ectopically expressing in tomato the D-galacturonate reductase (FaGalUR) gene from strawberry, involved in AsA biosynthesis, either under the control of the constitutive 35S or the tomato fruit-specific polygalucturonase (PG) promoters. Although transgenic lines showed a moderate increase on AsA content, complex changes in metabolites were found in transgenic fruits. Metabolomic analyses of ripe fruits identified a decrease in citrate, glutamate, asparagine, glucose, and fructose, accompanied by an increase of sucrose, galactinol, and chlorogenic acid. Significant metabolic changes also occurred in leaves of 35S-FaGalUR lines, which showed higher non-photochemical fluorescence quenching (NPQ), indicative of a higher constitutive photo-protective capacity. Overall, overexpression of FaGalUR increased total antioxidant capacity in fruits and the results suggest a tight control of AsA content, probably linked to a complex regulation of cellular redox state and metabolic adjustment.