Transcription profiling by array of Arabidopsis guard cells and mesophyll cells in response to ABA treatment
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
SUBMITTER: Yingzhen Yang
PROVIDER: E-MEXP-1443 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Publications
Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool.
Yang Yingzhen Y Costa Alex A Leonhardt Nathalie N Siegel Robert S RS Schroeder Julian I JI
Plant methods 20080219
<h4>Background</h4>A common limitation in guard cell signaling research is that it is difficult to obtain consistent high expression of transgenes of interest in Arabidopsis guard cells using known guard cell promoters or the constitutive 35S cauliflower mosaic virus promoter. An additional drawback of the 35S promoter is that ectopically expressing a gene throughout the organism could cause pleiotropic effects. To improve available methods for targeted gene expression in guard cells, we isolate ...[more]
Similar Datasets
Project description:The aim of this experiment is to understand the impact of overexpression of ERD15 on the transcriptome of Arabidopsis thaliana. ERD15 was isolated from a screen for genes rapidly induced after pathogen treatment from Arabidopsis. This gene was originally found as early responsive to drought (Kiyosue et al., 1994, Plant Physiol 106, 1707). ABA is central phytohormone in drought response, but increasing information is pointing to its significant role in pathogen responses as well. We are interested to see the effect of this hormone on plants overexpressing ERD15 compared to control plants. The samples (rosette leaves)will be harvested from 3-week old soil grown plants 90 min after spraying with 100 micromolar ABA. Comparison will be made with non-treated plant samples. Experimenter name = Elina Helenius; Experimenter phone = +358 9 191 59085; Experimenter fax = +358 9 191 59079; Experimenter institute = University of Helsinki; Experimenter address = Viikinkaari 5 D; Experimenter address = P.O.Box 56; Experimenter address = Helsinki; Experimenter zip/postal_code = 00014; Experimenter country = Finland Experiment Overall Design: 4 samples were used in this experiment
2008-06-13 | E-GEOD-5521 | biostudies-arrayexpress
Project description:Heterotrimeric G proteins mediate crucial and diverse signaling pathways in eukaryotes. To gain insights into the regulatory modes of the G protein and the co-regulatory modes of the G protein and the stress hormone abscisic acid (ABA), we generated and analyzed gene expression in G protein subunit single and double mutants of the model plant Arabidopsis thaliana. Through a Boolean modeling approach, our analysis reveals novel modes of heterotrimeric G protein action. Keywords: transcriptome analysis; G protein subunit mutants; abscisic acid (ABA) Microarray data were generated from four genotypes (wild type, gpa1-4 mutant, agb1-2 mutant, agb1-2 gpa1-4 double mutant) with or without ABA treatment. Arabidopsis plants were grown in growth chambers with an 8 hr light/16hr dark. Three hundred Arabidopsis leaves excised from 60-70 five-week-old plants were used as the starting material for each guard cell microarray. Ten mature leaves taken from 3-4 plants grown side-by side with the plants for guard cell isolation were used for each leaf sample. Excised leaf and isolated guard cell samples were treated with ABA (50 μM) or EtOH (solvent control) for 3 hrs. For each type of sample (guard cells or leaves), three independent biological replicates were performed, resulting in a total of 48 microarray hybridizations (2 sample types ´ 4 genotypes ´ two treatments ´3 replicates).
2010-12-06 | E-GEOD-19520 | biostudies-arrayexpress
Project description:Common transcriptional responses of Arabidopsis thaliana protoplasts transfected with turnip crinkle virus (TCV) , hibiscus chlorotic ringspot virus (HCRSV) and their coat protein mutants.
2009-07-07 | E-MEXP-1218 | biostudies-arrayexpress
Project description:To investigate gene expression differences related to the regulation of stomatal aperture between two Arabidopsis accessions, Me-0 and tetraploid Col, we performed microarray analysis on guard cell protoplasts from these two accessions. We identified 1,777 probes as up-regulated and 1,706 probes as down-regulated differentially expressed genes (DEGs) in Me-0 compared with tetraploid Col (adjusted P-value < 0.001 and log 2 fold change of greater than 1 or less than -1). Using gene set analysis to functionally characterize DEGs, there were eight significant gene sets (at the level of P-value < 0.01) among the up-regulated genes and seven significant gene set among the down-regulated genes. Genes with the lowest P-value among the up-regulated genes were related to âplant-type cell wall looseningâ, an essential step in guard cell swelling. Most of the down-regulated genes were related to the defense responses for immunity, hypoxia, and ozone, which also influence stomatal responses or gas exchange. We propose that these gene expression differences may cause the difference in stomatal aperture between Me-0 and tetraploid Col. Four samples from Me-0 and three samples from tetraploid Col are used for a microarray analysis.
2016-01-14 | E-GEOD-71914 | biostudies-arrayexpress
Project description:Transient genetic modification of plant protoplasts is a straightforward and rapid technique for the analysis of numerous aspects of plant biology. One drawback in the analysis of transformed protoplast suspensions is that they are a heterogeneous mix of cells that have and have not been successfully transfected. To overcome this problem, we have developed a system that employs a fluorescent positive selection marker in combination with flow cytometric analysis as well as fluorescence activated cell sorting (FACS) to isolate responses in the transfected protoplasts exclusively. This recombinase-compatible system enables high-throughput screening of genetic circuitry. Moreover, the use of FACS allows in depth downstream analysis. Lastly, over-expression is an effective means to dissect regulatory networks, especially where redundancy exists. Here, this system has been applied to the study of auxin signaling in order to investigate reporter gene activation and genome-wide transcriptional changes in response to manipulation of the auxin-response network. We have transiently over-expressed dominant negative mutant isoforms of Aux/IAA transcription factors (IAA7mII and IAA19mII; Tiwari et al., 2001) in Arabidopsis Pwer::GFP root protoplasts, making use of a RFP fluorescent positive selection marker and FACS to isolate the dually labeled (IAAnmII expressing and Pwer::GFP-positive) cells. We have compared the transcriptional differences between an empty vector control, IAA7mII and IAA19mII protoplasts that had either been treated with 5microM IAA or mock-treated for 3 hours. Experiment Overall Design: 18 samples with 3 replicates for each condition and transformation vector: 3x empty vector mock treated, 3x empty vector IAA treated, 3x IAA7mII over-expressor mock treated, 3x IAA7mII over-expressor IAA treated, 3x IAA19mII over-expressor mock treated and 3x IAA19mII over-expressor IAA treated.
2009-03-07 | E-GEOD-13783 | biostudies-arrayexpress
Project description:The goal of this experiment was to investigate the early transcript changes (6h) induced by hypoxia treatment in mesophyll protoplasts. A single pair (control & hypoxia) of GeneChips® was used to confirm that hypoxia treatment altered the expression of an overlapping set of genes controlled by KIN10 (At3g01090) in Arabidopsis mesophyll protoplasts. Experiment Overall Design: For the hypoxic treatment protoplasts were submerged in mannitol buffer and the hypoxia condition was confirmed by the activation of the well-established early marker genes ADH (alcohol dehydrogenase, At1g77120) and PDC1 (pyruvate decarboxylase-1, At4g33070) based on RT-PCR and GeneChip® analyses. Results show a clear overlap in genes induced under hypoxic conditions and those induced by KIN10 expression in protoplasts. The overlap of the gene expression profiles was confirmed by triplicated real-time quantitative RT-PCR analyses of selected marker genes.
2008-06-16 | E-GEOD-8248 | biostudies-arrayexpress
Project description:Dedifferentiation signifies the capacity of somatic cells to acquire stem cell-likeproperties. This process characterizes the transition of differentiated plant cells toprotoplasts (plant cells devoid of cell walls), a transition accompanied by widespreadchromatin decondensation. Transcriptome profiling of dedifferentiating cells revealedstriking similarities with senescing cells; both display a large increase in the expressionof genes of specific transcription factor (TF) families including ANAC, WRKY, bZIPand C2H2. We further showed that leaves induced to senesce by exposure to dark displaycharacteristic features of dedifferentiating cells including widespread chromatindecondensation, disruption of the nucleolus and condensation of rRNA genes.Considering that premature senescence can be induced by various stress conditions, ourresults suggest that plant cells response to certain stresses converges on cellulardedifferentiation whereby cells first acquire stem cell-like state prior to acquisition of anew cell fate (e.g., reentry into the cell cycle, or death). Experiment Overall Design: Labeled cRNA was prepared and hybridized to Affymetrix ATH1 GeneChips, 4 biological repeats (2 Ler and 2 kyp-2) for protoplasts and 3 biological repeats for leaves (2 Ler and 1 kyp-2). Experiment Overall Design: Partek data processing results linked below as supplementary file.
2009-04-11 | E-GEOD-15515 | biostudies-arrayexpress
Project description:Stomata are highly specialized organs which consist of pairs of guard cells and regulate gas and water vapor exchange in plants. While early stages of guard cell differentiation have been described and were interpreted in analogy to processes of cell type differentiation in animals, the downstream development of functional stomatal guard cells remains poorly understood. We have isolated an Arabidopsis mutant, scap1 (stomatal carpenter 1), that develops irregularly shaped guard cells and lacks the ability to control stomatal aperture, including CO2-induced stomatal closing and light-induced stomatal opening. SCAP1 was identified as a plant-specific Dof-type transcription factor expressed in maturing guard cells but not in guard mother cells. SCAP1 regulates the expression of genes encoding key elements of stomatal functioning and morphogenesis, such as a K+ channel protein, MYB60 transcription factor, and pectin methyl esterase. Consequently, ion homeostasis was disturbed in scap1 guard cells, and esterification of extracellular pectins was impaired so that the cell walls lining the pores did not mature normally. We conclude that SCAP1 regulates essential processes of stomatal guard cell maturation and functions as a key transcription factor regulating the final stages of guard cell differentiation. We isolated guard cell protoplasts from 4-week-old WT(Col-0) and scap1 mutant plants and extracted RNA independently. Four biological replicates were performed for each experiment.
2013-02-28 | E-GEOD-43964 | biostudies-arrayexpress
Project description:UV-B (280-320 nm) exposure causes serious damage in plants, limiting their growth and survival, effects that are partly counteracted by repair mechanisms active in plants receiving accompanying visible radiation. Though no particular UV-B receptor has been identified to date, there is strong evidence to indicate that certain aspects of UV-B perception are receptor-mediated. Investigations of down-stream signalling events have thus far indicated broad similarities to pathogen-induced defence responses in plants. In order to identify genes in Arabidopsis that may be up- or down- regulated specifically in response to UV-B exposure and compare them to genes whose expression is altered in plants challenged by an avirulent isolate of Peronospora parasitica (downy mildew), we propose to analyse the transcriptional profiles for the following treatments:; 1. UV-B Responses; "A-1" Columbia (Col-0) exposed to supplementary UV-B/UV-A* with a background of low photosynthetically active radiation (PAR of 20 micromol m-2 s-1) for 1.5 photoperiods (photoperiod = 12h). [UV-B treatment]; "A-2" Col-0 exposed to supplementary UV-A and low PAR for 1.5 photoperiods [control for UV-B treatment]; "A-3" Col-0 exposed to visible light only (low PAR) (no UV) for 1.5 photoperiods [control for UV effects in general].* There are no pure sources of UV-B light available. 2. Pathogen Responses; "A-4" Col-0 spray-inoculated with P. parasitica isolate HIKS-1 (recognised by the R-gene RPP7). After spraying, plants were kept covered in plant propagators and transferred to an 18 degreeC growth chamber. !Samples for RNA extraction were taken 72h after inoculation. "A-5" The viability of spores was also checked by parallel spraying of the susceptible mutant, Col-rpp7. [pathogen treatment]; "A-6" Col-0 mock treated with water, covered and transferred to an 18 degree C growth chamber, 72h prior to sampling. [control for pathogen treatment]; In all experiments, we are using RNA from leaves taken at the same time of day (6 h into the 12 h photoperiod) from 4.5-week old plants grown under 12h photoperiod. All treatments were normalised against PR-1 expression levels to ensure comparability between UV-B and pathogen treatments. Due to the difficulty in distinguishing between local and systemic induced responses in UV-B treated plants, we are using RNA from whole rosettes for both the UV-B and pathogen treatment for better comparability among treatments. The degree of similarity between these two sets of transcriptional changes will complement and help interpret our experimental data on changes in resistance to pathogens in plants pre-treated with UV-B. Moreover, the data set obtained would allow for identification of UV-B specific changes in gene expression including cis-acting UV-B-responsive promoter elements. Experimenter name = Julia Brueggemann; Experimenter phone = 01789 470 382; Experimenter fax = 01789 470 552; Experimenter address = Horticulture Research International; Experimenter address = Wellesbourne; Experimenter address = Warwickshire; Experimenter zip/postal_code = CV35 9EF; Experimenter country = UK Experiment Overall Design: 6 samples were used in this experiment
2008-06-14 | E-GEOD-5731 | biostudies-arrayexpress
Project description:The goal of this experiment was to explore the extent of KIN10 (At3g01090) transcriptional regulation and identify its early target genes in Arabidopsis mesophyll protoplasts. Results suggest that KIN10 targets a remarkably broad array of genes that orchestrate transcription networks, promote catabolism and autophagy, and suppress anabolism and ribosome biogenesis. The transient expression condition ruled out secondary or long-term effects of metabolism and growth, and circumvented experimental limitations caused by redundancy and embryonic lethality observed in mammals and plants. Experiment Overall Design: Analysis of the transcript changes induced by transient KIN10 (At3g01090) expression (6h) in Arabidopsis mesophyll protoplasts using protoplasts similarly transfected with control plasmid DNA for comparison. To maximize the significance of the duplicated experimental data for defining differentially expressed genes, the biologically independent experiments were carefully performed by two individuals using different soil, plants, growth chambers, DNAs and microarray facilities for hybridization and scanning to cover potential technical and biological variations. For each pair (control & KIN10) of large-scale protoplast transfection experiments, about 3 million protoplasts were isolated from the fifth and sixth leaves of 36 randomly picked plants and pooled. Experiment Overall Design: RNA samples isolated from control and KIN10-transfected protoplasts were subjected to quality controls, including RNA quality and quantity inspection by gel electrophoresis and staining as well as consistent gene expression changes by duplicated real-time quantitative RT-PCR analyses of selected marker genes.
2008-06-16 | E-GEOD-8257 | biostudies-arrayexpress