Project description:Photosynthesis in plant cells would not be possible without the supportive role of mitochondria. However, isolating mitochondria for physiological and biochemical analyses from plant cells is a lengthy and tedious process. Established isolation protocols require multiple centrifugation steps and substantial amounts of starting material. To overcome these limitations, we tagged mitochondria in plant cells with a triple haemagglutinin-tag for rapid purification by a single affinity purification step. This protocol yields highly purified mitochondria from 1 g of Arabidopsis seedlings that are suitable for enzyme activity measurements and delivers sufficient amounts of mitochondrial proteins for deep proteomic profiling. We demonstrate that the method can be applied to proteomic analysis of an Arabidopsis mutant deficient in the mitochondrial glutamate transporter À bout de souffle (BOU) and we identify 27 differentially expressed proteins in the bou mutant. Our work also sets the stage for the development of advanced isolation protocols for mitochondria from distinct cell types.
Project description:Transcriptional profiling in young flowers (stage 8) of Arabidopsis wild type control plants and sdg2-1 mutant is performed using Aligent’s Whole Arabidopsis Gene Expression Microarray (G2519F, V4, 4x44K). A significant number of genes involved in gametophyte development are found differentially regulated in the sdg2-1 mutant.
Project description:To investigate the role of this enzyme in different plant tissues, we characterized a mutant from Arabidopsis thaliana having an insertion at the GAPC gene locus. The homozygous mutant was determined to be null respect to GAPC, as it exhibited complete absence of both expression of GAPC mRNA and enzyme activity. Transcriptome analysis demonstrated that the insertion mutant plant shows altered expression of several enzymes involved in carbohydrate metabolism. Keywords: 2 channel experiment in triplicate including a dye swap comparison
Project description:Transcriptional profiling of Arabidopsis thaliana seedlings treated with safranal, highlighting to the physiological function of plant volatile chemicals by observing early response of gene expressions in Arabidopsis seedlings.
