Transcriptomic analysis of phosphatin effect-Phosphatin effect on Arabidopsis
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ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE45937 | GEO | 2013/04/11
SECONDARY ACCESSION(S): PRJNA196708
REPOSITORIES: GEO
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Project description:cea11-02_phosphatin - transcriptomic analysis of phosphatin effect - Identification of transcriptomic modifications promoted by phosphatin (AC6) (a molecule mimicking Pi addition effects on Pi starved plants). - 40 µM Phosphatin was added to MS (diluted 10 times) containing 20µM phosphate medium and controls were grown on MS (diluted 10 times) containing 20µM phosphate or supplemented with 500 µM of Pi. For each ARN extraction their was 3 plates containing each 10 seedlings grown 13 days in vertical petri dishes. 6 dye-swap - treated vs untreated comparison
2013-04-11 | E-GEOD-45937 | biostudies-arrayexpress
Project description:Rooted sugarcane plantlets, originated from in vitro meristem culture (genotype SP80-3280, CTC, Brazil), were greenhouse acclimatized by initial cultivation on 1/20th strength Hoagland and Arnon (1950) nutrient solution. Nutrient solutions were aired from an oil-less compressor and replaced every 7 days, increasing nutrient concentration to ¼ strength in 3 weeks. Plants were then individually transferred to 2.8 L pots filled with fresh ¼ strength nutrient solution. After one week, half of the plants were transferred to fresh solution containing 250 µM Pi, while the other half was transferred to nutrient solution deprived of phosphate (Pi), with H2PO4 being replaced by H2SO4 (Muchhal et al., 1996). Roots from six plants from each treatment (0 and 250 µM Pi) were harvested 6, 12, 24 and 48 h after exposure to phosphate starvation and immediately frozen in liquid nitrogen. For each time point and each treatment, root samples were aggregated in three pools of two samples each. Extraction of total RNA was performed separately on each sample pool. Keywords: time course of stress response
2007-05-20 | GSE4966 | GEO
Project description:Fourteen days plants growth under hydroponic +P condition (200 µM) were treated with +P(200µM) or –P (no phosphate) for another 7 days, shoot of plants from 3 biological repeats were sampled for Affymetrix microarray analysis. We used microarrays to detail the global programme of gene expression underlying +Pi and -Pi condition between WT and spx1spx2 double mutant. Fourteen days plants growth under hydroponic +P condition (200 µM) were treated with +P(200µM) or –P (no phosphate) for another 7 days, shoot of plants from 3 biological repeats were sampled for Affymetrix microarray analysis.
2014-10-03 | E-GEOD-60823 | biostudies-arrayexpress
Project description:Fourteen days plants growth under hydroponic +P condition (200 µM) were treated with +P(200µM) or –P (no phosphate) for another 7 days, shoot of plants from 3 biological repeats were sampled for Affymetrix microarray analysis. We used microarrays to detail the global programme of gene expression underlying +Pi and -Pi condition between WT and spx1spx2 double mutant.
2014-10-03 | GSE60823 | GEO
Project description:Diploid wild type cells in a chemostat, in different times after changing the feeding medium from low to high Pi. Diploid cells were also grown in a chemostat at a dilution rate of 0.3[1/hr] and phosphate concentration in the feeding vessel of 0.3mM Pi. Cells were allowed to reach steady state and then feeding medium was replaced to 7.3mM Pi. Samples from the chemostat were harvested in different times after the transfer (t= 0.35, 2, 5, and 10 days). Total RNA was extracted using MasterPure™ Kit (Epicentre). The samples were amplified, labeled, hybridized to yeast dual color expression microarrays and scanned, all using standard Agilent protocols, reagents, and instruments. The scanned images were analyzed using SpotReader software (Niles Scientific).
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Project description:The effects of phosphate starvation in Arabidopsis thaliana (L.) plants were compared in plants grown in liquid MS medium transferred in low or high Pi and in plants grown vertically in petri dishes during 10 days. In the transfer experiments, 2 treatments were analysed for evaluating the short-(3, 6 and 12 h pooled) and medium-(1 and 2 d pooled) term effects of Pi deficiency on the gene expression. Since some Arabidopsis genes are regulated by diurnal rhythm and circadian clocks, plantlets were harvested separately at the beginning and at the end of the photoperiod and pooled. In the long term experiment, leaves and roots were sampled separately after 10 days. Triplicates were analysed for each experiment.
2006-09-01 | E-MEXP-791 | biostudies-arrayexpress
Project description:Arabidopsis plants were grown in sand and supplied with full nutrient during 6 to 7 weeks in a growth chamber in controlled conditions (21/18°C day/night temperature, 60 % humidity, 8 h photoperiod). Then for the split-root experiments, the root system was washed in water to discard the sand and separated in two parts dipped in 2 containers (300 mL each) containing either the same medium (MS/10 supplied with 500 ?M Pi (cP500) or without Pi (cP0) or different media (500 ?M Pi and no Pi, respectively spP500 and spP0) for two days. Three plants were pooled for each condition and the experiment was triplicated.
2010-11-01 | E-MEXP-2601 | biostudies-arrayexpress
Project description:Pi availability is a significant limiting factor for plant growth in both natural and agricultural systems. To cope with such limiting conditions, plants have adapted developmental and biochemical strategies to enhance Pi acquisition and to avoid starvation. A myriad of genes that are involved in the regulation and display of these strategies have been identified. However, the possible epigenetic components regulating the phosphate starvation responses have not been thoroughly investigated. DNA methylation is a major epigenetic mark involved in diverse biological processes and it may play a critical role in Pi starvation stress adaptation, also changes in DNA methylation can lead to a unique gene expression pattern in response to specific developmental and environmental conditions. Here in we demonstrate that non-CpG DNA methylation is required for proper expression of a number of Pi-limitation responsive genes in Arabidopsis thaliana and results in altered morphologic and physiologic phosphate starvation responses.Our data suggest that DNA methylation is involved in the modulation of Pi starvation responses via the transcriptional regulation of a set of phosphate-starvation responsive genes. Analysis of 8 different treatments, 2 different Organs (Root and Shoot), 2 different Phosphate treatments (High Pi, Low Pi), 2 different Times (Short Term, Long Term), 2 biological replicates for treatment
2015-12-31 | E-GEOD-72770 | biostudies-arrayexpress
Project description:To explore the transcriptional regulations in lpcat1/2 mutant and wild type plants grown on 1/2 MS media (control), or 1/2 MS media containing 20ug/ml lyso-PAF or 20ug/ml lyso-PAF plus 50 µM ONO
2023-03-30 | GSE128799 | GEO
Project description:Proteomic analysis of pervanadate-induced tyrosine-phosphorylated proteins in hepatocellular carcinoma WRL 68 cells Protein tyrosine phosphorylation plays critical roles in modulating biological processes such as cellular proliferation, differentiation, migration, apoptosis and metabolism. To profile tyrosine phosphorylated (pTyr) proteins as well as search novel pTyr proteins as cross-talk points among different cellular pathways, we developed a rapid and efficient approach to identify cellular pTyr proteins and their complexes by a combination of subcellular proteomics approach with signal transduction strategies. Human hepatocytic cells from WRL68 cell line were treated with pervanadate (POV), subfractionated into four fractions and then subjected to immunoaffinity purification with anti-pTyr antibody. The eluted mixtures of the anti-pTyr purification were identified by LC-MS/MS. Subcellular fractionation and affinity purification of tyrosine-phosphorylated proteins: WRL68 cells were first grown to 80% confluence in MEM complete medium and then the medium replaced with serum free media. After 15 h, the cells were either untreated or stimulated with 0.1 mM pervanadate (1 mM sodium orthovanadate, 3 mM H2O2) for 10 min. 150-mm cultures of WRL 68 cells were rinsed twice with 4? PBS and then scraped from the dish in 750?l of hypotonic buffer (10 mM Tris, 1 mM NaF, 10 mM IAA, pH 7.5) containing a cocktail of protein inhibitors. After a 20-min incubation on ice, the cells were passed about five times through a 25-g needle. The resulting lysate was subjected to a 15min centrifugation at 1000 rpm at 4?, after which the pellet was resuspended in 250?l of hypotonic buffer and re-extracted by a second round of the trituration and centrifugation. The supernatants of the first and second spins were combined, adjusted to 0.25 M NaCl, and separated into cytosolic (supernatant) and membrane (pellet) fractions bycentrifugation at 19,000 rpm (43,000 g) for 90 min at 4?. All the pellets and total cell lysate were resuspended in RIPA buffer (50 mM Tris-HCl, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 1% deoxycholic acid sodium) containing 1mM pervanadate with a cocktail of protein inhibitors with sonication aid. Cleared cell lysates were incubated overnight at 4? with 30?l monoclonal anti-phosphotyrosine-agrose (Sigma). Precipitated immune complexes were washed three times with 1×HNTG (20 mM HEPES, 150 mM NaCl, 0.1% Triton X-100, 10% Glycerol, pH 7.5) and then eluted with 100 mM phenyl phosphate (Sigma) in lysis buffer at 4?. Enzyme digestion, mass spectrometry and protein identification: The sample was digested according to the published method. Chromatography was performed using a surveyor LC system (Thermo Finnigan,SanJose,CA) on C18 reverse phase column(RP, 180 µm x 150 mm, BioBasic® C18, 5 µm, Thermo Hypersil-Keystone). The pump flow rate is split 1:100 for a colum flow rate of 1.5 µL/min.The column effluent is directly electrosprayed using the orthogonal metal needle source without further splitting.Mobile phase A is 0.1% formic acid in water,and the B mobile phase is 0.1% formic acid in acetonitrile. The separation of peptides obtained by enzymatic digest of bile sample was achieved with a gradient of 2-80% B over 360 min.The column effluent from the reverse phase column was analyzed by LCQ Deca XP ion-trap mass spectrometer.The micro-electrospray interface uses a 30 µm metal needle that is orthogonal to the inlet of the LCQ.The mass spectrometer was set that one full MS scan was followed by three MS/MS scans on the three most intense ion from the MS spectrum with the following Dynamic Exclusion™ settings: repeat count, 1; repeat duration, 0.5 min; exclusion duration, 3.0 min. The acquired MS/MS spectra were automatically searched against protein database for human proteins (SWISS-PROT/TrEMBL) using the TurboSEQUEST program in the BioWorks™ 3.0 software suite. An accepted SEQUEST result had to have a ?Cn score of at least 0.1 (regardless of charge state) and Xcorr (one charge?1.5, two charges?2.0, three charges?2.5). Single peptides that alone identify a protein were manually validated after meeting the above criteria. Bioinformatics analysis: The pI and MW of the proteins were analyzed using ExPASy proteomics tools accessed from http://cn.expasy.org/tools/#proteome. The grand average hydropathicity (GRAVY) values were determined according to Kyte-Doolittle. The protein subcellular location annotation was from SwissProt &TrEMBL protein database (http://us.expasy.org/sprot/). The protein function family was categorized according to Gene Ontology (GO) annotation terms extracted by InterPro (http://www.ebi.ac.uk./interpro/). The annotation of protein phosphorylation was from SwissProt &TrEMBL protein database (http://us.expasy.org/sprot/) and PhosphoSite (http://www.phosphosite.org). The kinases were annotated according to human kinome. Keywords: other
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