ABSTRACT
Age-dependent changes in reactive oxygen species (ROS) levels are critical in leaf senescence. While H2O2-reducing enzymes such as catalases and cytosolic ASCORBATE PEROXIDASE1 (APX1) tightly control the oxidative load during senescence, their regulation and function are not specific to senescence. Previously, we identified the role of ASCORBATE PEROXIDASE6 (APX6) during seed maturation in Arabidopsis (Arabidopsis thaliana). Here, we show that APX6 is a bona fide senescence-associated gene. APX6 expression is specifically induced in aging leaves and in response to senescence-promoting stimuli such as abscisic acid (ABA), extended darkness, and osmotic stress. apx6 mutants showed early developmental senescence and increased sensitivity to dark stress. Reduced APX activity, increased H2O2 level, and altered redox state of the ascorbate pool in mature pre-senescing green leaves of the apx6 mutants correlated with the early onset of senescence. Using transient expression assays in Nicotiana benthamiana leaves, we unraveled the age-dependent post-transcriptional regulation of APX6. We then identified the coding sequence of APX6 as a potential target of miR398, which is a key regulator of copper redistribution. Furthermore, we showed that mutants of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7), the master regulator of copper homeostasis and miR398 expression, have a higher APX6 level compared with the wild type, which further increased under copper deficiency. Our study suggests that APX6 is a modulator of ROS/redox homeostasis and signaling in aging leaves that plays an important role in developmental- and stress-induced senescence programs.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Ascorbate Peroxidases/metabolism , DNA-Binding Proteins/metabolism , Signal Transduction , Transcription Factors/metabolism , Abscisic Acid/metabolism , Arabidopsis/genetics , Arabidopsis/physiology , Arabidopsis Proteins/genetics , Ascorbate Peroxidases/genetics , Copper/deficiency , DNA-Binding Proteins/genetics , Darkness , Homeostasis , Hydrogen Peroxide/metabolism , MicroRNAs/genetics , Oxidation-Reduction , Plant Growth Regulators/metabolism , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Leaves/physiology , Reactive Oxygen Species/metabolism , Time Factors , Nicotiana/enzymology , Nicotiana/genetics , Nicotiana/physiology , Transcription Factors/geneticsABSTRACT
Phenotypic plasticity is an adaptive feature of all organisms, which, in land plants, entails changes in orientation of growth (tropism), patterns of development, organ architecture, timing of developmental processes, and resource allocation. However, little is known about the molecular components that integrate exogenous environmental cues with internal hormonal signaling pathways. This addendum describes a role for calcium-regulated calmodulin-binding transcription 1 (CAMTA1) in auxin signaling and stress responses. We discuss possible mechanisms that may underlie this role of CAMTA1, and speculate on the more general roles of CAMTAs in auxin responses and phenotypic plasticity.
Subject(s)
Calmodulin-Binding Proteins/metabolism , Indoleacetic Acids/metabolism , Trans-Activators/metabolism , Arabidopsis/metabolism , Biological Transport , Environment , Homeostasis , Hypocotyl/growth & development , Hypocotyl/metabolism , Models, Biological , Multigene Family , Signal TransductionABSTRACT
The past two decades revealed a plethora of Ca2+-responsive proteins and downstream targets in plants, of which several are unique to plants. More recent high-throughput 'omics' approaches and bioinformatics are exposing Ca2+-responsive cis-elements and the corresponding Ca2+-responsive genes. Here, we review the current knowledge on Ca2+-signaling pathways that regulate gene expression in plants, and we link these to mechanisms by which plants respond to biotic and abiotic stresses.
Subject(s)
Calcium/metabolism , Plants/genetics , Plants/metabolism , Transcription, Genetic/genetics , Models, Biological , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
Auxin is a key plant hormone that regulates various aspects of plant development. However, the mechanisms integrating auxin growth effects with stress responses are not fully understood. In this study, we investigated the possible role of calmodulin-binding transcription activator 1 (CAMTA1), an Arabidopsis thaliana calcium/calmodulin-binding transcription activator, in auxin signaling and its responses to different stresses. Plants harboring the AtCAMTA1 promoter fused to the GUS reporter gene revealed cell-specific expression patterns reminiscent of auxin responses. The responsiveness of CAMTA1 to auxin was further assessed by chemical disturbances in polar auxin transport, and by RT-PCR analysis of gene expression of dissected leaf sections from plants exposed to the auxin transport inhibitor NPA. Furthermore, the intensity and cell-specific expression patterns of CAMTA1 changed significantly and differentially on exposure to increasing salt concentrations and heat. Transcriptome analysis of a camta1 T-DNA insertion mutant revealed 63 up-regulated genes, of which 17 are associated with auxin signaling. Finally, analysis of hypocotyl elongation in the presence and absence of auxin revealed that camta1 T-DNA insertion mutants and CAMTA1-repressor lines are hyper-responsive to auxin compared to wild-type seedlings. Thus, CAMTA1 participates in auxin signaling and responds to abiotic stresses.
Subject(s)
Arabidopsis/physiology , Calcium-Binding Proteins/physiology , Indoleacetic Acids/metabolism , Signal Transduction/physiology , Stress, Physiological , Arabidopsis/metabolism , Base Sequence , Calcium-Binding Proteins/genetics , DNA Primers , Gene Expression Regulation, Plant , Promoter Regions, Genetic , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Calmodulin-binding transcription activator (CAMTA) 3 (also called SR1) is a calmodulin-binding transcription factor in Arabidopsis. Two homozygous T-DNA insertion mutants (camta3-1, camta3-2) showed enhanced spontaneous lesions. Transcriptome analysis of both mutants revealed 6 genes with attenuated expression and 99 genes with elevated expression. Of the latter, 32 genes are related to defense against pathogens (e.g. WRKY33, PR1 and chitinase). Propagation of a virulent strain of the bacterial pathogen Pseudomonas syringae and the fungal pathogen Botrytis cinerea were attenuated in both mutants. Moreover, both mutants accumulated high levels of H2O2. We suggest that CAMTA3 regulates the expression of a set of genes involved in biotic defense responses.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Calmodulin-Binding Proteins/metabolism , Gene Expression Regulation, Plant , Plant Diseases/genetics , Transcription Factors/metabolism , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Botrytis/growth & development , Calmodulin-Binding Proteins/genetics , Gene Expression Profiling , Mutagenesis, Insertional , Plant Diseases/microbiology , Pseudomonas syringae/growth & development , Reactive Oxygen Species/metabolism , Transcription Factors/geneticsABSTRACT
The regulation of gene expression by cellular calcium is crucial for plant defense against biotic and abiotic stresses. However, the number of genes known to respond to specific transient calcium signals is limited, and as yet there is no definition of a calcium-responsive cis element in plants. Here, we generated specific cytosolic calcium transients in intact Arabidopsis thaliana seedlings and linked them to early transcriptome changes, followed by bioinformatic analysis of the responsive genes. A cytosolic calcium transient induced by calmodulin antagonists and blocked by lanthanides was characterized using aequorin-based luminometry and photon imaging. Analysis of transcriptome changes revealed 230 calcium-responsive genes, of which 162 were upregulated and 68 were downregulated. These include known early stress-responsive genes as well as genes of unknown function. Analysis of their upstream regions revealed, exclusively in the upregulated genes, a highly significant occurrence of a consensus sequence (P < 10(-13)) comprising two abscisic acid-specific cis elements: the abscisic acid-responsive element (ABRE; CACGTG[T/C/G]) and its coupling element ([C/A]ACGCG[T/C/G]) [corrected] Finally, we show that a tetramer of the ABRE cis element is sufficient to confer transcriptional activation in response to cytosolic Ca(2+) transients. Thus, at least for some specific Ca(2+) transients and motif combinations, ABREs function as Ca(2+)-responsive cis elements.
