Project description:au14-05_pi4kbeta1beta2 - pi4kbeta1beta2 sid2 triple mutant - What are the transcriptome changes induced in the pi4kbeta1beta2 sid2 triple mutant ? - The pi4kbeta1beta2 sid2 triple mutant plants were compared to sid2 plants or to pi4kbeta1beta2 double mutant plant. As a control the pi4kbeta1beta2 double mutant was compared to WT.
Project description:Phosphatidylinositol-4-kinases β1 and β2 (PI4Kβ1/PI4Kβ2), which are responsible for phosphorylation of phosphoinositol to phosphatidylinositol-4-phosphate, have an important role in vesicular trafficking in plants. Moreover, PI4Kβ1/PI4β2 negatively regulate biosynthesis of phytohormone salicylic acid (SA). SA plays crucial role in plant immune responses and its endogenous concentration strongly affects plant transcriptome. In this study we were focused on the effect of the PI4Kβ1/PI4Kβ2 and SA on the abundance of membrane proteins. For this purpose, we performed proteomic analysis on isolated microsomal fractions from leaves of four Arabidopsis thaliana genotypes: wild type ecotype Columbia-0; double mutant impaired in function of PI4Kβ1/PI4Kβ2 (pi4kβ1/pi4kβ2) exhibiting high SA level; sid2 mutant with impaired SA biosynthesis depending on the isochorismate synthase 1 and triple mutant sid2/pi4kβ1/pi4kβ2. In total, we identified 4534 proteins from which 1696 proteins differed in abundance between the mutants and WT. We showed that SA has a big impact on membrane proteome, because among hundreds of the identified affected proteins typical proteins associated with SA triggered pathway occurred. Our data thus point out new connections between SA pathway and clathrin independent endocytosis (flotillins) and exocytosis/protein secretion (syntaxins, tertraspanin) to be investigated in future. In contrast to SA, presence/absence of PI4Kβ1/PI4Kβ2 itself affected only 27 proteins. Nevertheless, among them we identified CERK1, plasma membrane receptor for chitin. Although PI4Kβ1/PI4Kβ2 itself did not have strong impact on A. thaliana microsomal proteome, our data clearly shows that PI4Kβ1/PI4Kβ2 enhance the proteome changes when SA pathway is modulated in parallel
Project description:Transcriptional profiling of the vegetative part of Arabidopsis comparing wild type with the shr scl23 scr triple mutant. The latter is produced by crossing the strong null alleles of shr (shr-2), scl23 (scl23-1) and scr (scr-5). The goal was to determine the effects of the GRAS transcription factors SHR, SCL23 and SCR on growth and development of the Arabidopsis shoot system by global transcriptome analysis. Two-condition experiment: Col-0 vs. shr scl23 scr triple mutant. Biological replicates: 2 WT vs. triple mutant replicates, 2 WT vs. triple mutant replicates dye-swap replicates.
Project description:Comparison of osmotic stress related gene expression in Col-0, sid2-1 single, wrky54wrky70 double and wrky54wrky70sid2-1 triple mutants.
Project description:To explore the possible cause for the enhanced tolerance to osmotic stress exhibited by wrky54wrky70 double mutant, we have used Agilent Arabidopsis (V4) Gene Expression Microarray to characterize the effect of these mutations under the osmotic stress treatment. In addition to wild type and wrky54wrky70 double mutant, sid2-1 single and wrky54wrky70sid2-1 triple mutant were also included in the microarray experiment. When comparing to untreated control samples, 58 genes were identified as representatives to show the different expression level among different mutants under the 15% PEG6000 treatment for one day. Expression of six genes (RAB18, LTI78, KIN1,NCED3,P5CS1 and PRODH) from this gene list were quantified by qRT-PCR, confirming the suppressed induction in wrky54wrky70 double mutant under the osmotic stress. Osmotic stress induced gene expression in Col-WT, sid2-1 single, wrky54wrky70 double and wrky54wrky70sid2-1 triple mutants was measured after exposure to 15% PEG6000 treatment for one day. For each array, three labeled aRNA sample were hybridized and three biological replicates for each sample with dye swaps were made.
Project description:As sessile organism, plants evolved a highly complicated signaling system to cope with unfavorable and fluctuating environmental conditions. Rapid and transient Reactive Oxygen Species (ROS) burst is a common response to both biotic and abiotic stresses. Plants exposed with O3 could trigger extracellular similar ROS production through cell wall peroxidases and NPADPH oxidases, resulting in changes in the gene expression and cell death. Whereas ROS induced cell death is not simply due to its toxicity, rather due to interplay with several other signaling pathways, such as salicylic acid (SA), jasmonic acid (JA) and ethylene signaling pathways. Furthermore, the three hormones have both synergistic and antagonistic interactions, where the suppression of JA signaling by SA is the mostly studied. In addition, ethylene promotes cell death while JA has a protective role upon O3 exposure. The role of SA is more complicated; depending on the genetic background it can have either cell death promoting or protecting roles. Hence, a clean system to deliver apoplastic ROS is required to study the role of ROS apart from con-current activation of other signaling pathways. Arabidopsis thaliana offer a convenient system to study apoplastic ROS signaling due to the availability of hormone signaling or biosynthesis mutants including the JA receptor mutant coi1-16 (CORONATINE INSENSITIVE1), the essential ethylene signaling mutant ein2 (ETHYLENE INSENSITIVE2), the SA biosynthesis mutant sid2 (SALICYLIC ACID INDUCTION DEFICIENT2 also known as ISOCHORISMATE SYNTHASE1), and essential regulators in SA/JA/ethylene-induced defense response triple mutant tga2 tga5 tga6 (Clade II TGA transcription factors). Here we used a combination of transcriptome analysis, cell death assays and mutant analysis to systematically quantified the contribution of hormone signaling in relation to apoplastic ROS signaling, identified transcription factors (TFs) involved in ROS regulation and dissected the components involved in defense hormones associated cell death. Transcriptome profiling of ozone response using two arabidopsis triple mutants coi1-16 ein2 sid2 and tga2 tga5 tga6 related to Jasmonic acid, salicylic acid and ethylene signaling to identify hormone-independant apoplastic ROS signaling