Cell type-specific auxin responses in the Arabidopsis thaliana root
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ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE35580 | GEO | 2013/08/06
SECONDARY ACCESSION(S): PRJNA152235
REPOSITORIES: GEO
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Project description:We performed an analysis of transcriptomic responses to auxin within four distinct tissues of the Arabidopsis thaliana root. This high-resolution dataset shows how different cell types are predisposed to react to auxin with discrete transcriptional responses. The sensitivity provided by the analysis lies in the ability to detect cell-type specific responses diluted in organ-level analyses. This dataset provides a novel resource to examine how auxin, a widespread signal in plant development, influences differentiation and patterning in the plant through tissue-specific transcriptional regulation. To analyze the effect of auxin in separate spatial domains of the root, early transcriptional changes in response to auxin treatment were assayed by means of fluorescence activated cell sorting (FACS) and microarray analysis in four tissues of the Arabidopsis root (wild type Col-0). The samples covered inner and outer as well as proximal and distal cell populations; including the stele (reporter line pWOL::GFP), xylem-pole (xp) pericycle (enhancer trap line E3754), epidermis/lateral root cap (reporter line pWER::GFP) and columella (enhancer trap line PET111). One-week-old seedlings of the individual lines were treated with auxin (two hours, 5µM indole-3-acetic acid [IAA]) or mock treated, after which roots were harvested and cells were dissociated by cell wall digestion (1 hour; including 5uM IAA) . GFP-positive cells were sorted and used for microarray transcriptome analysis (as in Bargmann and Birnbaum, Plant Phys. 2010). For comparison, transcriptional responses to auxin were also assayed in intact (undigested) roots.
2013-08-06 | E-GEOD-35580 | biostudies-arrayexpress
Project description:The plant signaling molecule auxin triggers both fast and slow cellular responses across the plant lineage, including both land plants and algae. We discovered an ultra-fast proteome-wide phosphorylation response to auxin across 5 land plant and algal species, converging on a core group of shared target proteins. We find conserved rapid physiological responses to auxin in the same species and identified a RAF-like protein kinase as a central mediator of auxin-triggered phosphorylation across species.
2024-01-26 | PXD038363 | Pride
Project description:Proper functioning of the nuclear auxin pathway is essential for regulating plant growth and development by maintaining auxin homeostasis. To understand better physiological mechanisms involved in auxin signaling pathways we investigated the localization and effect of accumulation of auxin coreceptor IAA17/AXR3 in root. We demonstrate that the accumulation of stable nuclear AXR3-1 protein interferes with auxin homeostasis, causing auxin insensitivity and increased rapid root cell elongation followed by detained growth. This growth pattern is associated with changes in phytohormone gene expression. Data from transcriptomic screen combined with reporter lines and mutant studies declare essential role of auxin homeostasis in maintaining optimal root growth rate and development. We proposed a model in which rapid cell elongation is caused by combination of AXR3-1-dependent auxin insensitivity associated with unblocked gibberellin effect on root. This study demonstrate that plants coordinate gibberellin homeostasis by the auxin signaling pathway, contributing to avoid excessive root elongation.
2023-04-01 | GSE227820 | GEO
Project description:Understanding how developmental and environmental signals are integrated to produce specific responses is one of the main challenges of modern biology. Hormones and, most importantly, interactions between different hormones serve as crucial regulators of plant growth and development, playing central roles in the coordination of internal developmental processes with the environment. Herein, a combination of physiological, genetic, cellular, and whole-genome expression profiling approaches has been employed to investigate the mechanisms of interaction between two key plant hormones, ethylene and auxin. Quantification of the morphological effects of ethylene and auxin in a variety of mutant backgrounds indicates that auxin biosynthesis, transport, signaling and response are required for the ethylene-induced growth inhibition in roots but not in hypocotyls. Analysis of the activation of early auxin and ethylene responses at cellular level, as well as of global changes in gene expression in the wild type versus auxin and ethylene mutants, suggests a simple mechanistic model for the interaction between these two hormones in roots. This model not only implies existence of several levels of interaction but also provides a likely explanation for the strong ethylene response defects observed in auxin mutants. Experiment Overall Design: The effect of ethylene on gene expression in Col-0 (wild-type) and aux1-7 (an auxin resistant mutant) seedlings' roots was investigated. Hydrocarbon-free air was used as control for the ethylene treatment. Two biological replicates were examined. Experiment Overall Design: Similarly, the effect of auxin (IAA) on gene expression in Col (wild-type) and ein2-5 (an ethylene insensitive mutant) seedlings' roots was studied. Two biological replicates were employed.
2008-06-15 | E-GEOD-7432 | biostudies-arrayexpress
Project description:Analysis of brassinosteroid (BR) and auxin effects on gene expression in Arabidopsis roots. Our genomic results indicate that BR and auxin induce largely opposite gene expression responses in primary roots.
2015-04-30 | GSE52966 | GEO
Project description:Analysis of brassinosteroid (BR) and auxin effects on gene expression in Arabidopsis roots. Our genomic results indicate that BR and auxin induce largely opposite gene expression responses in primary roots. RNA-Seq for 7-day-old Arabidopsis Col-0, dwf4, bri1-116, and bri1-116;bzr1-1D roots grown on regular medium and treated with brassinolide, auxin or mock solution for 4 hr.
2015-04-30 | E-GEOD-52966 | biostudies-arrayexpress
Project description:Auxin plays a pivotal role in virtually every aspect of plant morphogenesis. It simultaneously orchestrates a wide variety of processes, including cell wall biogenesis, transition through the cell cycle, and metabolism of a wide range of chemical substances, among others. The coordination principles for such complex regulation in a cell at a system level are poorly understood. Here, we apply a whole-genome approach to estimate the coherence of the auxin-regulated transcriptional response in gene groups, united by a certain biological process, molecular function or cell component, based on the fold change of the expression level. The corresponding analysis of RNA-Seq data for Arabidopsis roots treated with auxin for 6 h revealed that the intervals for the fold change of gene expression levels are quite narrow for many functional gene groups. The fold-specific manner of auxin response in most of these groups was confirmed using independent microarray data. To predict the mechanisms underlying such a coordinated expression, we performed motif enrichment analysis, which revealed TBX1-3, SBX, REG and TCP/site 2 as the candidates associated with fold-specific responses to auxin.
2017-06-01 | GSE97258 | GEO
Project description:TRH1, also classified as KUP4, belongs in the family of plant HAK/KUP/KT (High-Affinity K+/ K+ UPtake/ K+ Transporter) transporters, homologous to the bacterial KUP and fungal HAK transporters, which most likely as K+/H+ exchangers function as electrochemical potential-driven transporters. Arabidopsis contains 13 genes, classified into five major clades (Clades I-V). Members of Clade II, including TRH1, are associated with developmental processes. Targeted expression experiments demonstrated that K+ transporter TRH1 is a coupler of two auxin-mediated developmental processes that determine Root System Architecture, root hair morphogenesis and root gravitropism. Targeting TRH1 expression in central cylinder cells that coincide with the acropetal route of auxin transport, rescued trh1 root agravitropism, whereas positional TRH1 expression in peripheral cell layers that define the basipetal route of auxin transport, restored trh1 root hair defects. To dissect the two auxin-mediated developmental responses, a transcriptome analysis was performed identifying the differentially expressed genes involved in either root bending or root hair development.
2022-10-11 | GSE159476 | GEO
Project description:Protein phosphorylation is instrumental to early signaling events. Studying system-wide phosphorylation in relation to processes under investigation requires a quantitative proteomics approach. In Arabidopsis, auxin application can induce pericycle cell divisions and lateral root formation. Initiation of lateral root formation requires transcriptional reprogramming following auxin-mediated degradation of transcriptional repressors. The immediate early signaling events prior this derepression are virtually uncharacterized. To identify the signal molecules responding to auxin application we used the Lateral Root Inducible System which was previously developed to trigger synchronous division of pericycle cells. To identify and quantify the early signaling events following this induction we combined 15N based metabolic labeling and phospho-enrichment and applied a mass spectrometry based approach. In total, 3068 phosphopeptides were identified from auxin treated root tissue. This experiment represents one of the largest quantitative phosphoprotein dataset on Arabidopsis to date. Key proteins responding to auxin treatment included the MDR and PIN2 auxin carriers, auxin response factor2, suppressor of auxin resistance3 and sorting nexin1 (SNX1). Mutational analysis of Serine16 of SNX1 showed that overexpression of the mutated forms of SNX1led to retarded growth and reduction of lateral root formation due to the reduced outgrowth of the primodium, showing proof of principle for our approach. Data Processing: All MS/MS spectra were centroided and merged to a single peak list file using MaxQuant (version 1.0.13.13), which was searched using the Mascot search engine (version 2.2.0, Matrix Science) containing a total 67344 entries (which includes contaminants and an equivalent number of decoy sequences) generated from the publicly available Arabidopsis database (The Arabidopsis Information Resource (TAIR9); June 2009; file name, TAIR9_pep_20090619) with carbamidomethylcysteine as a fixed modification. Oxidized methionine and phosphorylation (serine, threonine, and tyrosine) were searched as variable modifications. Searches were done with tryptic specificity allowing two miscleavages and an initial tolerance on mass measurement of 50 ppm in MS mode and 0.6 Da for MS/MS ions and score cutoff of 20. The resulting .dat files were exported and filtered for a <1% false discovery rate (FDR) at the peptide level using in-house developed software Rockerbox (Version1.1.0) (35). Relative quantification ratios of the identified phosphopeptides and protein were derived by MSQuant (MSQ2.0b4, 2010-02-25) (36). Proteins were quantified with at least two unique non phosphopeptides or single peptide with a score no less than 60. The phospho peptide ratio was normalized by dividing the ratio of its protein of origin. To determine whether phosphorylation ratios for normalized peptides were significantly different a p-value was generated using significance B which is part of the MaxQuant package. The changes were considered significant at a p-value of 0.05 or less. For comparison at the phosphopeptide level the most recent the full dataset with experimental sites was downloaded from PhosPhAt 4.0. Phosphopeptides with ambiguous residues denoted as (s), (t), etc., were converted to S and T, respectively. In addition we used Supplementary Table S1 from Lan et al., (2012) for phosphopeptide comparison. Modifications other than nonambiguous phosphorylation were ignored resulting in a reduction from the original 879 phosphopeptides to 556 peptides. We used pep2pro 'TAIR10 wos' dataset for comparison at the protein level.
2014-02-25 | PXD000177 | Pride
Project description:Wild tobacco flowers wave rhythmically to facilitate specific pollinator interactions. This movement behavior is controlled by a regulatory network that involves the circadian clock- and auxin-signaling pathways. The plant hormone auxin, similarly to its function in tropic movements, acts as growth regulator in the circadian regulation of floral movement. Dorsoventral asymmetry in auxin levels and auxin transcriptional responses mediate the growth responses in the floral peduncle that make flowers move. Multiple components of the auxin-signaling pathway and auxin responses are under the control of circadian clock. However, it is unclear where these two pathways intersect and how collectively contribute to regulate specific rhythmic outputs. Here we found that the blue light photoreceptor and circadian clock component ZEITLUPE (ZTL) controls auxin responses through the regulation of the auxin-signaling pathway in a time-of-day and blue light specific manner. Abrogation of ZTL expression abolishes flower movement and the temporal gating of auxin-induced growth responses in the floral peduncle. ZTL regulates transcription and directly interacts with indole-3-acetic acid inducible 19 (IAA19), a circadian controlled gene that regulates development of curvature in moving organs. Indicating that ZTL modulates auxin sensitivity in part through the regulation of AUX/IAA transcriptional repressors. At night, growth responses in the peduncle to the synthetic auxin 2,4-D revealed that ZTL additionally controls auxin responses regulating auxin homeostasis. These results indicate that ZTL conveys temporal and environmental information, at multiple levels, into the auxin signaling-pathway and in this way sculpts the temporal gating of auxin responses that allow flowers to move. To gain further insight into the molecular basis of temporal regulation of the movement of flowers we used a whole genome microarray as a discovery platform to identify genes differentially expressed in a RNAi knockdown line silenced in the expression of the circadian clock component ZEITLUPE (irZTL-314).
2019-06-30 | GSE116648 | GEO