Project description:To investigate the response of Arabidopsis thaliana plants to non-freezing, cool temperatures, we subjected four week old plants to various chilling temperatures at defined times during the diurnal cycle to control for diurnal effects on transcription. From the same plants, metabolites and enzyme activities were measured as well. Interestingly a gradual change could be observed over a wide range of temperatures. Some of which could be attributed to the CBF program. Keywords: time course, different temperatures
Project description:For the detailed and sensitive detection of the responses to a specific treatment it is important to perform tissue or cell type specific analyses, but this is not easily achievable for the different leaf tissues. Here we developed a method termed MeSelect to effectively separate leaf epidermis, vascular and mesophyll cells basically without contamination from other tissues. The high yield of the MeSelect method allowed for tissue specific high-throughput proteome analyses after inhibition of the proteasome with Syringolin A and the affinity enrichment of polyubiquitylated proteins in epidermis, mesophyll and vasculature.
Project description:For the detailed and sensitive detection of the responses to a specific treatment it is important to perform tissue or cell type specific analyses, but this is not easily achievable for the different leaf tissues. Here we developed a method termed MeSelect to effectively separate leaf epidermis, vascular and mesophyll cells basically without contamination from other tissues. The high yield of the MeSelect method allowed for tissue specific high-throughput proteome analyses after inhibition of the proteasome with Syringolin A and the affinity enrichment of polyubiquitylated proteins in epidermis, mesophyll and vasculature.
Project description:Plant growth and survival depends to a large extent on the diurnal regulation of cellular processes. Although extensively studied at the transcript level, notably less is known about diurnal fluctuations at the protein level. Here, we report a high-resolution quantitative time-course of the Arabidopsis rosette proteome and phosphoproteome over a 12 h light:12 h dark diel cycle. We monitored the proteome every 2 h and the phosphoproteome immediately before and after the light-to-dark and dark-to-light transitions. Notably, we quantified nearly 5000 proteins and 1800 phosphopeptides, of which 288 and 225, respectively, were found to fluctuate over the time-course. Diurnal proteome and phosphoproteome changes were related to diverse biological processes, including protein translation, light detection, photosynthesis, metabolism and transport. Together, these datasets represent the most comprehensive proteomic analysis of Arabidopsis rosettes to date, allowing us to make multi-level inferences about the diurnal regulation of key cellular plant processes plants.
Project description:Diurnal time-course transcriptional profiling of rice leaf in the field comparing a circadian clock related mutant, osgi, with the wild-type (WT).
Project description:How do the transcript levels of leaf-expressed genes change in a normal day-night cycle? The interest is in genes that are regulated by the circadian clock and the diurnal component (i.e. light, metabolite changes). Plants were grown on soil in a 12/12 h light/dark rythm at 20°C day and night. 5 weeks after germination the rosettes of the non-flowering plants were harvested, 15 plants per sample. Plants were harvested at 6 timepoints every 4 hours beginning with the end of the night (still in darkness). Keywords: repeat
