Global gene expression analysis of Arabidopsis Col-0 under normal and low oxygen conditions
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ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
SUBMITTER: Danny Llewellyn
PROVIDER: E-GEOD-21504 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Global gene expression was compared between Arabidopsis lines with altered expression of ANAC102 (over-expressed and knocked-out) and wild-type. ANAC102 is a putative NAC domain transcription factor. Gene expression was compared between an ANAC102 over-expressing line and parental ecotype C24 under ambient atmosphere to determine which genes ANAC102 is capable of regulating. Gene expression was also compared between three week old plants of an ANAC102 knock-out line and parental ecotype Col-0 under 0.1% Oxygen and ambient atmosphere conditions to determine which genes may require ANAC102 for appropriate expression under these conditions. Gene expression was also compared between imbibed seeds of an ANAC102 knock-out line and parental ecotype Col-0 following a 0.1% Oxygen treatment. Experiment Overall Design: Three related experiments were performed. In the first two, three week old Arabidopsis seedlings (4-6 leaves) were used. In the first experiment one line of ANAC102 over-expressing plants were compared to ecotype C24 (the parental line used in the creation of the ANAC102 over-expressing line). Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from whole plants. In the second experiment, one line of plants bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to ecotype Col-0 (the parental line used for the T-DNA mutagenesis) both without treatment and following treatment with 0.1% Oxygen for four hours. Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from root tissue only. In the third experiment imbibed seeds of one Arabiopsis line bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to seeds of ecotype Col-0 (the parental line used for the T-DNA mutagenesis) following treatment with 0.1% Oxygen for six days. Three biological replicates were used for each line and for each biological replicate, ~5mg (pre-imbibition) of seed were bulked. RNA was extracted from whole seed.
2009-01-25 | E-GEOD-14420 | biostudies-arrayexpress
Project description:Non-symbiotic hemoglobins are ubiquitously expressed proteins known to interact with nitric oxide, an inhibitor of mitochondrial respiration and an important signalling component. We evaluated the underlying molecular mechanisms of AtHb1 (also referred as AtGLB1 or AHb1) function, its effects on stress response and the interplay with nitric oxide. For this purpose, AtHb1 was overexpressed in Arabidopsis thaliana under control of the seed-specific promoter LeB4. We performed comparative transcriptome analysis of developing siliques from wild type (WT, Col-0) and transgenic plants subjected to control and moderate hypoxic conditions. The experimental design was used to analyze the underlying molecular mechanisms of AtHb1 function and to assess differences in the hypoxic response between WT and AtHb1-overexpressing seeds/siliques. Hypoxic and control (normoxic) treatment was carried out with complete transgenic (T3) and WT plants 45 days after germination (DPG). Plants were aerated with a gas mixture containing 10.5% oxygen or ambient gas containing 21% oxygen for control samples in darkness. After one hour, plants were decapitated, immediately frozen in liquid nitrogen, and siliques were dissected in liquid nitrogen for RNA extraction and hybridization to Affymetrix microarrays. Hypoxic and control treatment runs with WT and AtHb1-overexpressing plants were repeated twice to provide 3 biological replicates (total number of samples = 12).
2011-02-15 | E-GEOD-23846 | biostudies-arrayexpress
Project description:Reference: De Vos and Jander, 2009 - Plant, Cell, and Environment Aim: Identification of genes responding to aphid saliva. Background: During feeding on phloem sap aphids repeatedly salivate into the sieve element. It is thought that compounds in aphid saliva play a role in sustainable feeding. These compounds may include proteins and small molecules, which can function as virulence factors. Growth conditions Plants: Seeds of wild-type Arabidopsis thaliana (Col-0) were obtained from the were kept in 0.1% Phytagar (Invitrogen, Carlsbad, CA) for 24 h at 4°C prior to planting on Cornell mix with Osmocoat fertilizer. Plants were grown in Conviron growth chambers in 20- x 40-cm nursery flats at a photosynthetic photon flux density of 200 mmol m-2 s-1 and a 16-h photoperiod. The temperature in the chambers was 23°C and the relative humidity was 50%. Plants were grown for 3 weeks and used in experiments before flowering. Aphids: All experiments were conducted with a tobacco-adapted red lineage of M. persicae. Aphids were raised on cabbage (Brassica oleracea) with a 16-h day (150 mmol m-2 s-1 at 24°C) and an 8-h night (19°C) at 50% relative humidity. Experimental set-up/treatment: Fifty aphids were allowed to feed from 50 µL artificial diet, containing sucrose and amino acids (Kim and Jander, 2007) between two layers of Parafilm. After 24 h, artificial diet from 20 aphids and control (0 aphids) diet cups was collected and infiltrated into leaves of intact Arabidopsis plants using a 1-mL syringe without the needle. Plants for control diet and aphid saliva containing diet were grown in the same pot to allow for a paired comparison. Eighteen leaves (3 leaves from 6 plants) treated with control and aphid saliva containing diet were harvested and immediately frozen in liquid nitrogen. This experiment was repeated 3 times to function as independent biological replicates. RNA extraction + processing: RNA was extracted using the Qiagen Plant RNeasy kit. RNA quality and quantity was assessed with an Agilent BioAnalyser 2100. Samples were processed by the Cornell Microarray facility. Whole genome gene expression profiling was done using Affymetrix ATH1 GeneChips. Data analysis: Raw data from the microarrays was normalized at probe-level using gcRMA algorithm. The detection calls (present, marginal, absent) for each probe set was obtained using the GCOS system. Significance of gene expression was determined using the LIMMA (Smyth, 2004) program and raw p values of multiple tests were corrected using False Discovery Rate (FDR). 6 samples were used in this experiment
2009-11-11 | E-GEOD-18960 | biostudies-arrayexpress
Project description:Anoxia induces several heat shock proteins and a heat pre-treatment can acclimatize Arabidopsis seedlings to a subsequent anoxic treatment. In this work we analyzed the response of Arabidopsis seedlings to anoxia, heat and a combined heat+anoxia stress. A significant overlapping between the anoxic and heat shock responses has been observed by whole-genome microarray analysis. Experiment Overall Design: We treated Arabidopsis seedling, 4-days old, dark germinated with: Experiment Overall Design: -Control (23°C, dark, liquid Murashige-Skoog medium containing 30mM sucrose). Experiment Overall Design: -Heat-treated (38°C for 90 minutes, dark, liquid Murashige-Skoog medium containing 30mM sucrose). Experiment Overall Design: -Anoxia-treated (23°C, under anoxia for 6h, dark, liquid Murashige-Skoog medium containing 30mM sucrose). Experiment Overall Design: -combined heat+Anoxia-treatment (23°C, treated at 38°C for 90 min and thereafter under anoxia for 6h, dark, liquid Murashige-Skoog medium containing 30mM sucrose). Experiment Overall Design: Two biological replicates for each condition.
2009-06-08 | E-GEOD-16222 | biostudies-arrayexpress
Project description:Global gene expression was compared between root RNA samples from three-week-old Arabidopsis Col-0 plants subjected to 0.1% oxygen (balance nitrogen) or ambient atmospheric conditions.
2010-04-27 | GSE21504 | GEO
Project description:- Background and Aims: Oxygen can fall to low concentrations within plant tissues, either because of environmental factors that decrease the external oxygen concentration or because the movement of oxygen through the plant tissues cannot keep pace with the rate of oxygen consumption. Recent studies document that plants can decrease their oxygen consumption in response to relative small changes in oxygen concentrations to avoid internal anoxia. The molecular mechanisms underlying this response have not been identified yet. The aim of this study was to use transcript and metabolite profiling to investigate the genomic response of Arabidopsis roots to a mild decrease in oxygen concentrations. - Methods: Arabidopsis seedlings were grown on vertical agar plates at 21, 8, 4 and 1% (v/v) external oxygen for 0.5, 2 and 48h. Roots were analyzed for changes in transcript levels using Affymetrix whole genome DNA microarrays, and for changes in metabolite levels using routine GC-MS based metabolite profiling. Root extension rates were monitored in parallel to investigate adaptive changes in growth. - Key results: Results show that root growth was inhibited and transcript and metabolite profiles were significantly altered in response to a moderate decrease in oxygen concentrations. Low oxygen leads to a preferential up-regulation of genes that might be important to trigger adaptive responses in the plant. A small but highly specific set of genes is induced very early in response to a moderate decrease in oxygen concentrations. Genes that were down-regulated mainly encoded proteins involved in energy-consuming processes. In line with this, root extension growth was significantly decreased which will ultimately save ATP and decrease oxygen consumption. This was accompanied by a differential regulation of metabolite levels at short and long term incubation at low oxygen. - Conclusions: Results show that there are adaptive changes in root extension involving large-scale reprogramming of gene expression and metabolism when oxygen concentration is decreased in a very narrow range. Experiment Overall Design: Arabidopsis seedlings were grown on vertical agar plates and treated with various oxygen concentrations (1%, 4%, 8%, and 21% as a control), 350ppm CO2 and N2 (rest) for different time periods (0.5 hours, 2 hours and 2 days. At the end of the experiment, the roots of the seedlings were immediately frozen in liquid nitrogen and used for gene expression analysis.
2008-10-25 | E-GEOD-11558 | biostudies-arrayexpress
Project description:We used microarrays to detail Arabidopsis gene expression in response to paraquat, a herbicide that acts as a terminal oxidant of photosystem I that in the light leads to the enhanced generation of superoxide and hydrogen peroxide inside plastids. Within a few hours after paraquat treatment changes in nuclear gene expression occur. Distinct sets of genes were activated that were different from those induced by another reactive oxygen species, singlet oxygen. Experiment Overall Design: Arabidopsis thaliana rosette leaves were harvested 1, 2, and 4 h after spraying either with a solution of 20 microM paraquat (methyl viologen, Sigma) in 0.1% Tween or with Tween alone for RNA extraction and hybridization on Affymetrix ATH1 microarrays. Plants were grown on soil for 3 weeks under continuous light at 90 mmol. m-2 . s-1. For each sample, the rosette leaves of five to six 3-week-old plants (before they start bolting) were collected for RNA extraction. Total RNAs from two separate biological experiments were pooled for the preparation of cDNA and the subsequent synthesis of biotin-labeled complementary RNA as recommended by Affymetrix.
2008-04-08 | E-GEOD-10464 | biostudies-arrayexpress
Project description:Complete submergence represses photosynthesis and aerobic respiration causing rapid mortality in most terrestrial plants, but some species have evolved traits allowing them to survive prolonged flooding. Here, we studied the response to submergence of two species and their F1 hybrid in the genus Rorippa, which is related to the model Arabidopsis. We showed that these species have high tolerance to complete, deep submergence, but R. sylvestris survived longer than R. amphibia and the F1 hybrid. While the former restricted growth upon submergence, the latter two genotypes showed induced stem and petiole elongation and had higher aerenchyma contents, indicative of a low oxygen escape strategy. Arabidopsis GeneChip microarrays were used for whole-genome transcript profiling of roots of young plants exposed to air or a 24-h submergence treatment, using a probe mask based on hybridisation of genomic DNA of both species to the arrays. The induction by the submergence treatment of genes involved in glycolysis and fermentation and repression of many energy consuming pathways was similar to the response to low oxygen of Arabidopsis and rice. Notably, sucrose synthases, glycolysis and fermentation genes were more strongly induced in the less tolerant R. amphibia than in R. sylvestris, which might indicate faster carbohydrate consumption of the former, while some genes involved in hydrogen peroxide scavenging were strongly and specifically induced in the latter. F1 hybrids showed a generally weaker response to submergence and an additive mode of gene action, which did not change by the submergence treatment. Experiment Overall Design: We used replicated clones of Rorippa amphibia, Rorippa sylvestris and their F1 hybrid. Plants were grown on sand in pots for three weeks, completely submerged in rain water for 24 h, or kept in air as a control. Three biological controls were collected per genotype, each consisting of a pool of three individuals. Roots were separated from the shoots, quickly rinsed and immediately frozen and used for RNA extraction and hybridization to Arabidopsis Ath1 GeneChips. Genomic DNA of Rorippa amphibia and Rorippa sylvestris was also hybridized to the GeneChips in order to filter out badly performing probes due to sequence divergences from Arabidopsis.
2009-12-04 | E-GEOD-13641 | biostudies-arrayexpress
Project description:Plastid gene expression (PGE) acts as a signal that regulates the expression of photosynthesis-associated nuclear genes (PhANGs) via GENOMES UNCOUPLED1 (GUN1)-dependent retrograde signaling. We recently isolated Arabidopsis sugar-inducible cotyledon yellow-192 (sicy-192), a gain-of-function mutant of plastidic invertase (INV-E), and showed that following the treatment of this mutant with sucrose, the expression of PhANGs decreased while that of nitrate reductase 1 (NIA1) increased. Plastid-encoded RNA polymerase (PEP)-dependent PGE was markedly suppressed in the sicy-192 mutant by the sucrose treatment. A double mutant of sicy-192 and gun1-101, a null mutant of GUN1, revealed that metabolic perturbation in the sucrose-treated sicy-192 mutant was, at least in part, dependent on GUN1. To confirm whether there is a relationship between plastid sugar metabolism and nuclear gene expression, we performed a microarray analysis using Suc-treated 3 genotypes consisting of wild-type, sicy-192, and sicy-192 gun1-101 plants. Wild-type, sicy-192, and sicy-192 gun1-101 plants were grown on MS medium containing 100 mM sucrose in a growth chamber kept at 25°C during 16 h of light (100 µmol/m2/s) and at 22°C during 8 h of darkness for 4 days. At 4 days of age, cotyledons were collected as samples for microarray analysis.
2013-09-14 | E-GEOD-50857 | biostudies-arrayexpress
Project description:Global gene expression was compared between Arabidopsis lines with altered expression of ANAC102 (over-expressed and knocked-out) and wild-type. ANAC102 is a putative NAC domain transcription factor. Gene expression was compared between an ANAC102 over-expressing line and parental ecotype C24 under ambient atmosphere to determine which genes ANAC102 is capable of regulating. Gene expression was also compared between three week old plants of an ANAC102 knock-out line and parental ecotype Col-0 under 0.1% Oxygen and ambient atmosphere conditions to determine which genes may require ANAC102 for appropriate expression under these conditions. Gene expression was also compared between imbibed seeds of an ANAC102 knock-out line and parental ecotype Col-0 following a 0.1% Oxygen treatment. Keywords: Genetic modification
2009-01-16 | GSE14420 | GEO