Microarray analysis of Col-0 and lbd37 lb38 lbd39 triple mutant treated with Mock or ABA
Ontology highlight
ABSTRACT: In order to functionally characterise the LATERAL ORGAN BOUNDARIES DOMAIN (LBD) 37/38/39 proteins during ABA response we performed microarray analysis of Col-0 and lbd37 lb38 lbd39 treated with Mock or ABA.
Project description:ABI3 is a key regulator of seed development in Arabidopsis and other plants.To identify genes regulated by ABI3 we performed array based transcriptome analysis of Dexamethasone inducible ABI3 transgenic seedlings. ABI3 mostly in concert with abscisic acid (ABA) was found to activate genes specifically expressed during the maturation phase of seed development. Two weeks old wildtype and 35S::ABI3::GR seedlings were treated with ABA, Dexamethasone (DEX) and ABA plus Dexamethasone for 4 h. Two biological independent experiments were performed. Wildtype was induced to control for DEX-induction of genes.
Project description:Microarray experiments were performed using Arabidopsis wild type plants (Col-0) and srk2dei triple knockout mutant to investigate the functions of ABA-activated protein kinases, SRK2D/SnRK2.2, SRK2E/OST1 and SRK2I/SnRK2.3. Transcription profiles of wild type and mutants were compared under ABA treatment or dehydration stress for 0 and 90 min. The srk2dei mutant was established by crossing T-DNA insertion mutants provided from Arabidopsis Biological Resource Center.
Project description:We found that TOR kinase regulates the plant environmental stress responses through phosphorylation at a conserved serine residue of Abscisic acid receptor PYR1/PYLs/RCARs. The phosphorylation inhibits the function of PYLs and prevents the binding of PYLs to abscisic acid and its downstream substrates of PP2C, which blocks the ABA and stress signaling. Our results suggest that plant utilizes this mechanism as a hub to regulate the ABA-mediate stress responses and growth recovery
Project description:Many of duplicated genes are enriched in signaling pathways. Recently, gene duplication of kinases has been shown to provide genetic buffering and functional diversification in cellular signaling. Transcription factors (TFs) are also often duplicated. However, how duplication of TFs affects their regulatory structures and functions of target genes has not been explored at the systems level. Here, we examined regulatory and functional roles of duplication of three major ARR TFs (ARR1, 10, and 12) in Arabidopsis cytokinin signaling using wild-type and single, double, and triple deletion mutants of the TFs. Comparative analysis of gene expression profiles obtained from Arabidopsis roots in wild-type and these mutants showed that duplication of ARR TFs systematically extended their transcriptional regulatory structures, leading to enhanced robustness and diversification in functions of target genes, as well as in regulation of cellular networks of target genes. Therefore, our results suggest that duplication of TFs contributes to robustness and diversification in functions of target genes by extending transcriptional regulatory structures. Duplication of TFs can confer an extension of transcriptional regulatory structures for target genes by providing new regulatory relationships between duplicated TFs and new or old target genes. To examine the nature of the extension in the regulatory structure, we performed gene expression profiling of Arabidopsis root tissues obtained from wild-type (WT) and deletion mutants of three type-B ARR1, 10, and 12. To examine how the extended regulatory structures by the duplicated ARR TFs are utilized for the responses to external CK, we generated gene expression profiles of WT Arabidopsis roots treated with mock or exogenous CK for 1 hour. Total RNAs were isolated from two biological replicates at each condition and used to measure gene expression level.
Project description:Abscisic acid (ABA) is a plant hormone that is important in responding to various environmental stresses. Using an ABA auxotroph in Arabidopsis (aba2-2) as the plant material, we would like to identify early downstream targets of transcription in response to a small dose of ABA, 1 uM. We also added cycloheximide to preferentially obtain immediate targets of ABA addition. We believe that using a sensitized background of an ABA auxotroph would yield a set of genes that are very closely regulated by ABA at the transcriptional level. This data set will be used for network analysis of ABA signaling.
Project description:To investigate differences in plant responses to salt and ABA stimulus, differences in gene expression in Arabidopsis in response to salt and ABA were compared using an Agilent oligo microarray. Four-week-old Arabidopsis thaliana ecotype Columbia (Col-0) seedlings were treated with either 150 mM NaCl or 10 M-NM-<M ABA for 6 hours; unstressed seedlings (control sample) were collected in parallel to avoid the possible effects of circadian rhythms. The results revealed that 31 genes were up regulated by both NaCl and ABA stress, and 23 genes were down-regulated by these stressors. To provide further validation of our microarray experiment data, ten genes from this signature were quantified in the same RNA samples by quantitative real-time PCR. Differentially expression genes of Arabidopsis thaliana were measured under salt stressed, ABA stressed and normal condition for 6 hours, respectively. Three independent experiments were performed at each treatment using different plants for each experiment.
Project description:The experiment was designed to enable comparison between Columbia and ahk2/ahk3, ahk3/ahk4 double and ahk2/ahk3/ahk4 triple mutants Arabidopsis seedlings
Project description:ZFP3, a nuclear C2H2 zinc finger protein acts as a negative regulator of ABA- suppressed germination. Regulated over-expression of ZFP3 and closely related ZFP1, ZFP4, ZFP6 and ZFP7 confers ABA insensitivity to seed germination while the zfp3,zfp4 double mutant displays enhanced ABA susceptibility. Reduced expression of a large set of ABA-induced genes such as RAB18 and transcription factors ABI3, ABI4, ABI5 and RGL2 in ZFP3ox seedling suggests that ZFP3 indeed negatively regulates ABA signaling. Analysis of ZFP3ox plants revealed multiple phenotypic alterations such as semidwarf growth habit, defects in fertility and enhanced sensitivity of hypocotyl elongation to red but not to far-red or blue light. Analysis of genetic interactions with phytochrome and abi mutants suggested that ZFP3 amplifies red light signals perceived by photoreceptors other than phyB, and controlled by ABI5 downstream of ZFP3. Comparison of ZFP3 overexpressing and wild type Arabidopsis seedlings
Project description:4 weeks old rooted plantlets of P. × canescens (Clone INRA717 1-B4) were cultivated in hydroponics in 2 l pots in Long Ashton nutrient solution in a culture room for 8 weeks before treatments started. Three treatments were applied to the plants: control treatment (-ABA), continuous 100 µM ABA treatment (+ABA) and discontinuous 100 µM ABA treatment (±ABA). ABA was fed to +ABA plants during the whole treatment period of 30 days. ABA was fed to ±ABA plants for three days in two weeks. Developing xylem and mature xylem were collected separately during the harvest and shortly frozen in liquid nitrogen. RNA was extracted from these samples and followed by RNA-sequencing.
Project description:In order to identify new regulators of the phosphate (Pi) starvation signaling pathway in plants, we analyzed variation in the abundance of nuclear-enriched nuclear proteins isolated from Arabidopsis roots that depends on Pi supply. We used 2-D Fluorescence Difference Gel Electrophoresis and MALDI-TOF/TOF techniques for nuclear proteome separation, visualization and relative protein abundance quantification and identification. Pi-controlled proteins identified in our analysis included components of the chromatin remodeling, DNA replication, and mRNA splicing machineries. In addition, by combining Pi starvation conditions with proteasome inhibitor treatments, we characterized the role of the ubiquitin- (Ub)-proteasome system (UPS), a major mechanism for targeted protein degradation in eukaryotes, in the control of the stability of Pi-responsive proteins. Among Pi-responsive proteins, the histone chaperone NAP1;2 was selected for further characterization, and was shown to display differential nucleo-cytoplasmic accumulation in response to Pi deprivation. We also found that mutants affecting three members of the NAP1 family accumulate lower Pi levels and display reduced expression of Pi starvation-inducible genes, reflecting a regulatory role for these chromatin-remodelling proteins in Pi homeostasis.