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Mitochondrial beta-cyanoalanine synthase is essential for root hair formation in Arabidopsis thaliana

ABSTRACT: Cyanide is stoichiometrically produced as a co-product of the ethylene biosynthesis pathway, and it is detoxified by the b-cyanoalanine synthase enzyme. The molecular and phenotypical analysis of T-DNA insertional mutants of the mitochondrial b-cyanoalanine synthase CYS-C1 suggests that discrete accumulation of cyanide is not toxic for the plant and does not alter mitochondrial respiration rates, but does act as a strong inhibitor of root hair development. The cys-c1 null allele is defective in root hair formation and accumulates cyanide in root tissues. The root hair defect is phenocopied in wild type plants by the exogenous addition of cyanide to the growth medium and is reversed by the addition of hydroxocobalamin. Hydroxocobalamin not only recovers the root phenotype of the mutant, but also the formation of ROS at the initial step of the root hair tip. Transcriptional profile analysis of the cys-c1 mutant reveals that cyanide accumulation acts as a repressor signal for several genes encoding enzymes involved in cell wall rebuilding and the formation of the root hair tip, as well as genes involved in ethylene signaling and metabolism. Our results demonstrate that mitochondrial b-cyanoalanine synthase activity is essential to maintain a low level of cyanide for proper root hair development.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE19242 | GEO | 2010/10/01

SECONDARY ACCESSION(S): PRJNA120775

REPOSITORIES: GEO

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Mitochondrial beta-cyanoalanine synthase is essential for root hair formation in Arabidopsis thaliana

Project description:Cyanide is stoichiometrically produced as a co-product of the ethylene biosynthesis pathway, and it is detoxified by the b-cyanoalanine synthase enzyme. The molecular and phenotypical analysis of T-DNA insertional mutants of the mitochondrial b-cyanoalanine synthase CYS-C1 suggests that discrete accumulation of cyanide is not toxic for the plant and does not alter mitochondrial respiration rates, but does act as a strong inhibitor of root hair development. The cys-c1 null allele is defective in root hair formation and accumulates cyanide in root tissues. The root hair defect is phenocopied in wild type plants by the exogenous addition of cyanide to the growth medium and is reversed by the addition of hydroxocobalamin. Hydroxocobalamin not only recovers the root phenotype of the mutant, but also the formation of ROS at the initial step of the root hair tip. Transcriptional profile analysis of the cys-c1 mutant reveals that cyanide accumulation acts as a repressor signal for several genes encoding enzymes involved in cell wall rebuilding and the formation of the root hair tip, as well as genes involved in ethylene signaling and metabolism. Our results demonstrate that mitochondrial b-cyanoalanine synthase activity is essential to maintain a low level of cyanide for proper root hair development. Using Affymetrix ATH1 GeneChips, we performed a comparative transcriptomic analysis of roots of the cys-c1 and wild type plants. Total RNA was extracted from roots of 14-days-old plants grown under identical conditions on MS medium (three biological replicates for each genotype), and these samples were used to prepare complementary RNA and and analyzed using the Affymetrix-Arabidopsis ATH1GeneChip array.

2010-10-01 | E-GEOD-19242 | biostudies-arrayexpress

New insights on the role of HCN in root hair elongation through single cell proteomics.

Project description:In order to identify the possible protein targets of cyanide in the regulation of root hair, we have carried out a single cell proteomic approach to characterize the protein atlas in wild type root hair cells compared to the cas-c1 mutant cells. Root hair specific cells were obtained from isolated protoplast from root tissues of Arabidopsis wt-pCOBL9:GFP and cas-c1-pCOBL9:GFP lines. To analyze the effect of the CAS-C1 mutation, we isolated 1 x 106 root hair cells by Fluorescence-activated cell sorting (FACS) from three independent replicate of wt-pCOBL9:GFP and cas-c1-pCOBL9:GFP roots. The extracted proteins from each sample were trypsin-digested and the digested peptides were analyzed by liquid chromatography-high resolution mass spectrometry (LC-MS/MS) for protein identification. In wild type samples, we have identified 3829 unique proteins at a false discovery rate below 1% (FDR<1%), that represent almost 10% of the total Arabidopsis proteome. In protein extract of the cas-c1-pCOBL9:GFP roots samples, we have identified 3972 proteins of which 3515 were commons in both cell types, 310 were only identified y wild type and 457 only in cas-c1 mutant (Fig. 5).

2024-01-26 | PXD028991 | Pride

Proteomic analysis in Arabidopsis tissues reveals hydrogen cyanide regulation of intracellular processes through posttranslational modification of proteins by S-cyanylation

Project description:Hydrogen cyanide (HCN) is co-produced with ethylene in plant cell and enzymatically detoxified mainly by the mitochondrial ß-cyanoalanine synthase (CAS-C1). Permanent or transient depletion of CAS-C1 activity in Arabidopsis results in physiological alteration in the plant that point to the function of HCN as a gasotransmitter molecule. Label-free quantitative proteomic analysis of enriched mitochondrial samples isolated from wild type and cas-c1 mutant reveled significant changes in protein content, identifying 451 proteins that are absent or less abundant in cas-c1 and 353 proteins only present or more abundant in the mutant background. Gene onthology classification of these proteins highlights proteomic changes that explains the root hairless phenotype and the altered inmune response observed in cas-c1 mutant. The mechanism of action of cyanide as signaling molecule has been addressed by two proteomic approaches focused on identifying S-cyanylation of cysteine as a posttranslational modification of proteins. Both the 2-imino-thiazolidine chemical method and direct untargeted analysis of proteins by LC-MS/MS identified a set of 163 proteins susceptibles to be S-cyanylated that include sedoheptulose 1, 7 biphosphatase (SBPase), the peptidyl-prolyl cis-trans isomerase (CYP20-3) and enolase 2 (ENO2). In vitro analysis of these proteins identified this modification in SBPase Cys74, CYP20-3 Cys259 and ENO2 Cys346 residues that affected the enzymatic activity of the enzymes. GO classification and protein-protein interaction cluster analysis revealed the function of S-cyanylation in the regulation of primary metabolic pathways as glycolysis, and the Calvin and S-adenosylmethionine cicles.

2019-01-09 | PXD009812 | Pride

HCN regulates cellular processes through posttranslational modification of proteins by S-cyanylation

Project description:Hydrogen cyanide (HCN) is coproduced with ethylene in plant cells and primarily enzymatically detoxified by the mitochondrial ß-cyanoalanine synthase (CAS-C1). Permanent or transient depletion of CAS-C1 activity in Arabidopsis results in physiological alterations in the plant that suggest that the function of HCN is a gasotransmitter molecule. Label-free quantitative proteomic analysis of enriched mitochondrial samples isolated from the wild type and cas-c1 mutant revealed significant changes in protein content, identifying 451 proteins that are absent or less abundant in cas-c1 and 353 proteins that are only present or more abundant in the mutant background. Gene ontology classification of these proteins highlights proteomic changes that explains the root hairless phenotype and the altered immune response observed in the cas-c1 mutant. The mechanism of action of cyanide as a signaling molecule has been addressed using two proteomic approaches focused on identifying the S-cyanylation of cysteine as a posttranslational modification of proteins. Both the 2-imino-thiazolidine chemical method and the direct untargeted analysis of proteins using LC-MS/MS identified a set of 163 proteins susceptible to S-cyanylation that included sedoheptulose 1, 7-bisphosphatase (SBPase), the peptidyl-prolyl cis-trans isomerase (CYP20-3) and enolase 2 (ENO2). In vitro analysis of these proteins identified that this modification in the SBPase Cys74, CYP20-3 Cys259 and ENO2 Cys346 residues affected the enzymatic activity of the enzymes. GO classification and protein-protein interaction cluster analysis revealed the function of S-cyanylation in the regulation of primary metabolic pathways, such as glycolysis, and the Calvin and S-adenosylmethionine cycles.

2019-01-09 | PXD009929 | Pride

Arabidopsis thaliana

Project description:Mitochondrial beta-cyanoalanine synthase is essential for root hair formation in Arabidopsis thaliana

| PRJNA120775 | ENA

Ethylene promotes root hair growth through coordinated EIN3/EIL1 and RHD6/RSL1 activity in Arabidopsis

Project description:Root hairs are an extensive structure of root epidermal cells and are critical for nutrient acquisition, soil anchorage, and environ- mental interactions in sessile plants. The phytohormone ethylene (ET) promotes root hair growth and also mediates the effects of different signals that stimulate hair cell development. However, the molecular basis of ET-induced root hair growth remains poorly understood. Here, we show that ET-activated transcription factor ETHYLENE-INSENSITIVE 3 (EIN3) physically interacts with ROOT HAIR DEFECTIVE 6 (RHD6), a well-documented positive regulator of hair cells, and that the two factors directly coactivate the hair length-determining gene RHD6-LIKE 4 (RSL4) to promote root hair elongation. Transcriptome analysis further revealed the parallel roles of the regulator pairs EIN3/EIL1 (EIN3-LIKE 1) and RHD6/ RSL1 (RHD6-LIKE 1). EIN3/EIL1 and RHD6/RSL1 coordinately en- hance root hair initiation by selectively regulating a subset of core root hair (H) genes. Thus, our work reveals a key transcriptional complex consisting of EIN3/EIL1 and RHD6/RSL1 in the control of root hair initiation and elongation, and provides a molecular framework for the integration of environmental signals and in- trinsic regulators in modulating plant organ development.

2017-12-12 | GSE107699 | GEO

Timeseries microarray of Arabidopsis thaliana root tips treated with(out) ethylene followed by hypoxia and recovery

Project description:Preceding hypoxia by an ethylene treatment improves root tip survival. To identify key players and processes mediating ethylene enhanced tolerance, the transcriptomic response of root tips was investigated directly after ethylene or control pretreatment, 2 and 4 hours of subsequent hypoxia, and 1 hour of reoxygenation. Seedlings were 4 days or 7 days old and grown on MS plates without added sugar. Only the root tips were harvested

2022-02-01 | E-MTAB-11231 | biostudies-arrayexpress

Gene expression profiles of the aerenchy maformation in maize using laser microdissection

Project description:To adapt to waterlogging in soil, some gramineous plants, such as maize (Zea mays), form lysigenous aerenchyma in the root cortex. Ethylene, which is accumulated during waterlogging, promotes aerenchyma formation. Aerencyma is formedonly in cortex in maise root. Therevore, aerenchyma is one of the model of programmed cell death. However, the molecular mechanism of aerenchyma formation is not understood. Therefore we isolated only cortex cells in maize primary root using laser microdissection. And microarray analysis was perforemed to identify the arechyma formation related genes. For microarray analysis, we designed 4 different experiments. Basal part and apical part of root were used for 1st experiment because arenchyma was formed in basal part of root, but not in apex. In second experiment, basal part of root with 6 hours waterlogged treatment and without treatment were used. And, as aerenchyma was induced by ethylene, ethylene and 1-MCP (inhibitor of ethylen perception) were used for experiment3 and 4. Gene expression analysis in 4 kinds of samples (cortex in basal reagion of maize primary root which were treated with waterlogged condition, aerobic condition, ethylene and 1-MCP and cortex in apical reagion of maize primary root which were treated with waterlogged conditio)

2011-01-28 | E-GEOD-26897 | biostudies-arrayexpress

Gene expression profiles of the aerenchy maformation in maize using laser microdissection

2011-01-28 | GSE26897 | GEO

Expression data from the Arabidopsis root epidermis

Project description:The root epidermis of Arabidopsis provides a simple and experimentally useful model for studying the molecular basis of cell fate and differentiation. The goal of this study was to define the larger gene regulatory network that governs the differentiation of the root hair and non-hair cell types of the Arabidopsis root epidermis. Transcript levels in the root epidermis of wild-type and mutant lines were assessed by purifying populations of root epidermal cells using fluorescence-based cell-sorting. Further, the role of the plant hormones auxin and ethylene on root epidermis development was assessed by defining transcript levels in the root epidermis of plants grown on media containing IAA or ACC. These microarray results were used to construct a comprehensive gene regulatory network that depicts the transcriptional control of root epidermal cell fate and differentiation in Arabidopsis.

2011-12-01 | GSE30547 | GEO

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