Project description:Proteomic optimisation of Arabidopsis thaliana leaf proteome generated using legumain (cleave C-terminal of asparagine/aspartic acid) as protease at different pH from pH 5.0 to pH 6.5.
Project description:Analysis of gene expression in Arabidopsis thaliana leaf, stem and flower tissues. Columbia (Col-0) Arabidopsis thaliana plants were grown at a density of 4 plants per 5 inch square pot either in a growth chamber or a green house set to 25*C by day, 20*C by night. Days were set to a 16hr photoperiod with 125 micro mol m-2s-1 fluorescent irradiation. Expanding leaves were harvested 15 days post germination in the middle of the photoperiod (3 replicates). Expanding upper 2" of the stem with siliques and pedicels removed were harvested 29 days post germination in the middle of the photoperiod (4 replicates). Developed flowers and unopened buds were harvested 29 days post germination in the middle of the photoperiod (4 replicates). RNA and Microarray Methods: Total RNA was extracted from the plants using the Trizol method (Invitrogen, Ramonell et al. (2002) Mol. Plant Pathol. 3: 301) and purified with a silica membrane column (Qiagen, RNeasy). Twenty micrograms biotinylated complementary RNA (cRNA) was prepared as described (Hernan et al. (2003) Cancer Res. 63, 140) from the purified total RNA. The resulting cRNA was used to hybridize ATH1 Arabidopsis GeneChips (Affymetrix) using the manufacturer's protocols. The array images were analyzed with the Affymetrix Microarray Suite 5.0 software with the target intensity set to 500. Normalized signal intensities were generated by multiplying signal intensities by ratio figure which set GapC (ID_REF = 258588_s_at) to 8500. Keywords: other
Project description:This set consists of small RNAs sequenced from two replicates of wildtype and two replicates of RDR6-15 knockout Arabidopsis thaliana Col-0 leaf samples. RDR6 is required for the production of tasRNAs (trans-acting small RNAS) and so tags associated with the tasRNA loci should be severely down-regulated or absent in the knockout compared to wildtype. The set can thus be used as containing known true positives for testing differential expression detection methods.
Project description:The douple mutant Arabidopsis thaliana soc1 ful, in contrast with WT, produces an interfascicular cambium and a large wood cylinder is the flowering stem. We present the RNAseq data for polyA mRNA of different developmental stages of cambium and wood formation in Arabidopsis thaliana. We sequenced 7 stages; 4 in the woody mutant soc1-6 ful-7 (herbaceous, cambium initiation, wood initiation and leaf) and 3 stages in the WT Col-0 (herbaceous, cambium and leaf). The corresponding stem anatomy is also presented in the manuscript indicating the stage of cambium development and the production of secondary xylem.