Cre 2. 2. 1, was selected from which loss of the 35S ABF3 nos transgene was confirmed by PCR. F2 progeny from the selected F1 plants were then backcrossed selleck screening library to wild type plants to eliminate Cre recombinase. The F1 plants from this cross were again PCR genotyped to identify those from which the Cre gene was lost, using primers CRE. F and CRE. R which are both specific for the Cre recombinase gene. One plant from each cross, Control 48 1. 1. 2. 5, Control 57 1. 2. 2. 5 and Control 59 2. 2. 1. 4, was selected from which loss of the Cre recombinase gene was confirmed by PCR. The selected F1 plants represent the control plant lines and the F2 progeny of these plants were used in subsequent experiments. Growth of Arabidopsis plants Seeds were surface sterilized by soaking for 20 min in 25% commercial Clorox and 0.

05% Triton X 100, and then Inhibitors,Modulators,Libraries rinsing four times with distilled water. Seeds were ger minated on MS media and grown at 22 C with a 16 h photoperiod. To determine the growth rate of plants, three day old seedlings were transplanted onto fresh MS plates and after one week or four weeks of growth the fresh weight of Inhibitors,Modulators,Libraries the seedlings Inhibitors,Modulators,Libraries was measured. To measure transpiration rate, leaves of a similar developmental stage were excised from four week old plants and the loss of weight over a 24 h period was measured. For microarray analysis, seeds were germinated on MS media and one week old Inhibitors,Modulators,Libraries seedlings were collected. For the drought stress, seedlings were transferred onto paper towels and harvested after 2 and 24 h. Microarray Analysis For each sample, three biological replicates were pre pared.

RNA Inhibitors,Modulators,Libraries was extracted from seedlings using the RNeasy Plant Mini Kit, following the manufacturers protocol. A 2100 Bioanalyser was used to determine the quality of the RNA. Microarray analysis was performed using the GeneChip Arabidopsis ATH1 Genome Array. Standard RNA processing, hybridization, and scanning protocols were followed as recommended by the GeneChip Expression Analysis Technical Manual. Hybridization, and scanning were performed at Agriculture and Agri Food Canada by Mark Jordan. RMA procedure was performed to normalize data using the AffylmGUI R software package from Bioconductor. Analysis of differential expression was done using a moderated t test with empirical Bayes smoothing. Microarray data from this study have been deposited at ArrayExpress.

Reverse transcription PCR DNaseI treated RNA was used for first strand cDNA synthesis using Superscript III reverse transcriptase and oligo 18 primers according to the manufacturers protocol. Between 24 and 30 cycles of PCR amplification was performed using gene specific primers. As an internal directly control, elongation factor 1 a was amplified. Sequences of all primers used in RT PCR analysis can be found in Additional file 4.