Project description:We investigated genome-wide changes in mRNA translation in Arabidopsis thaliana T87 suspension cell cultures which thought to be one of the host materials for bioreactor. Global translational repression was observed in cells of 8 day after inoculation that is thought to be stressful condition by the nutrient deficiency and hypoxia. This suggested the negative effect of the global translational repression on transgene expression. On the other hand, previous study using heat stress showed that some mRNAs were actively translated under such stressful condition, suggesting the existence of mRNA that were actively translated in cells of 8 day after inoculations. To identify mRNAs that escape global translational repression on 8 day and its cis-elements would be the 1st step to make the system for higher transgene expression by the escaping global translational repression. To this end, we subjected polysomal RNA and non-polysomal RNA from sucrose gradient fractionated cell lysates to the co-hybridization on Agilent Arabidopsis 4 Oligo Microarrays. The ratio of signal intensities (polysomal RNA: total RNA) was used as an indicator of the translation state for each transcript.
Project description:The goal of this study was to determine how the exometabolome of defense-elicited Arabidopsis suspension cells inhibits virulence gene expression and growth of a plant pathogenic bacterium Pseudomonas syringae. Arabidopsis T87 suspension cells were treated with the pathogen-associated molecular pattern elf26 or a DMSO-control treatment for six hours, then incubated in fresh media for one hour. The conditioned medium (exudate) was collected from each culture by centrifugation and 0.22 um filter to remove plant cells. These samples are designated T=6 mock and T=6 elf26 in our experimental design. We also prepared samples in the same manner from control-treated cells except without any pre-treatment time prior to one hour exudate production. These samples are labeled T=0 mock. A total of seven biological replicates of each treatment condition were analyzed, with each replicate prepared from cells grown in separate flasks. The exudates were prepared in four independent experiments performed on separate days (1 biological replicate from first experiment, 2 biological replicates from each of the 3 remaining experiments). Four samples of the culture medium, one from each of the four independent experiments, are included.
Project description:Transcriptional profiling after inhibition of cellulose synthesis by thaxtomin A and isoxaben in Arabidopsis thaliana suspension cells Perturbations in the cellulose content of the plant cell wall lead to global modifications in cellular homeostasis, as seen in cellulose synthase mutants or after inhibiting cellulose synthesis. In particular, application of inhibitors of cellulose synthesis such as thaxtomin A (TA) and isoxaben (IXB) initiates a programmed cell death (PCD) in Arabidopsis thaliana suspension cells that is dependent on de novo gene transcription. To further understand how TA and IXB activate PCD, a whole genome microarray analysis was performed on mRNA isolated from Arabidopsis suspension cells exposed to TA and IXB. More than 75% of the genes upregulated by TA were also upregulated by IXB, including genes encoding cell wall-related and calcium-binding proteins, defence/stress-related transcription factors, signalling components and cell death-related proteins. Comparisons with published transcriptional analyses revealed an important subset of genes generally induced in response to various biotic and abiotic stress.