Project description:We investigated the transcriptional response of hybrid poplar (Populus trichocarpa x deltoides) leaves to methyl jasmonate treatment over a 24 hour time course. Experiments were conducted using clonal trees under greenhouse conditions at the University of British Columbia. We used the 15.5K poplar cDNA microarray platform previously described by Ralph et al. (Molecular Ecology 2006, 15:1275-1297). Differentially expressed genes were determined using three criteria: fold-change between methyl jasmonate-treated and tween-treated control leaves > 1.5-fold, P value < 0.05 and Q value < 0.05. This study identified > 1,000 differentially expressed genes in response to methyl jasmonate treatment. A factorial hybridization design was chosen to assess gene expression between tween-treated control leaves, and leaves subjected to methyl jasmonate treatment. For each tree, all but the lowest five mature leaves were covered with a light-weight plastic bag and then sprayed with either methyl jasmonate or tween, the solvent control. Leaves were harvested 2, 6 or 24 hours after the initiation of each treatment and total RNA was individually isolated from each tree. For each treatment and time point, equal amounts of total RNA were combined from each of the five biological replicate trees prior to cDNA microarray analysis. A total of 18 hybridizations were performed to directly compare total RNA from methyl jasmonate-treated and tween-treated control leaves at each time point, as well as among methyl jasmonate-treated leaves across time points, with dye balance.
Project description:We investigated the transcriptional response of hybrid poplar (Populus trichocarpa x deltoides) leaves to methyl jasmonate treatment over a 24 hour time course. Experiments were conducted using clonal trees under greenhouse conditions at the University of British Columbia. We used the 15.5K poplar cDNA microarray platform previously described by Ralph et al. (Molecular Ecology 2006, 15:1275-1297). Differentially expressed genes were determined using three criteria: fold-change between methyl jasmonate-treated and tween-treated control leaves > 1.5-fold, P value < 0.05 and Q value < 0.05. This study identified > 1,000 differentially expressed genes in response to methyl jasmonate treatment.
Project description:Potato cultivar Russet Norkotah was grown in a growth chamber at 16 hour light/8 hour dark at 22°C fertilized with Osmocote. Plants were from tissue culture, transplanted to soil and used approximately 4 weeks after transplanting. RNA was extracted from leaves using Trizol. Plants were sprayed with one of four treatments, either 1mM salicylic acid, 100 µM methyl jasmonate, 1 mg/ml arachidonic acid or 100 µM methyl jasmonate plus 1 mM salicylic acid together. Samples were harvested at 3, 24 and 72 hours after spraying. Each salicylic acid or arachidonic acid treatment has 3 biological replicates done in parallel, using leaves pooled from 2 plants. The control for SA or AA treated plants were water sprayed plants, from which leaves of 3 different plants were pooled at each timepoint, done side-by-side with the treated samples. Methyl jasmonate or MJ+SA treatments had 2 biological replicates each, using leaves pooled from 2 plants. A separate set of water sprayed plants (from the SA and AA control treatments) were used as controls, and pooled leaves from 3 plants were used at each time-point, and done in parallel with the treated samples. Keywords: Direct comparison
Project description:Illumina HiSeq technology was used to generate mRNA profiles from Populus tremula x alba INRA 717-1B4 roots treated with Methyl jasmonate. Samples were harvested after two weeks either from untreated control roots or from Methyl jasmonate treated roots. Paired-end (2X100bp) reads were generated and aligned to the Populus trichocarpa (http://www.phytozome.net/poplar.php) using CLC Genomics Workbench 6. mRNA profiles from Populus tremula x alba INRA 717-1B4 roots treated with Methyl jasmonate as well as from control roots were generated by paired-end (2X100bp) Illumina HiSeq sequencing. Four samples were sequenced per lane, two biological replicates per treatment.