Figure 4

Time course analysis of ORCA2 expression. ORCA2 transcripts generated from the endogenous ORCA2 gene and from the ORCA2 transgene were measured separately by qPCR, using primer pairs specific for each gene. The PCR primer pair used to analyze ORCA2 transgene transcripts, as expected, gives only a very low background signal for the control line (which lacks the ORCA2 transgene, data not shown). The lines analyzed are: uninduced ORCA2-OE line (□, solid line), 0.02% ethanol-induced ORCA2-OE line (■, solid line), uninduced control line (∆, dashed line) and 0.02% ethanol-induced control line (▲, dashed line). A) Relative ORCA2 transgene transcript levels are presented as ∆∆CT. Note that ORCA2 transgene transcript levels were normalized versus ORCA2 endogenous gene transcript levels in the uninduced control line at 0 h rather than against ORCA2 transgene transcript levels in the uninduced control line at 0 h because the control line does not carry the ORCA2 transgene. B) Relative ORCA2 endogenous gene transcript levels are presented as ∆∆CT. Positive ∆∆CT values indicate that ORCA2 endogenous gene transcript levels are higher in the indicated hairy root line grown for the indicated time under the indicated conditions than in the uninduced control line at 0 h. Negative ∆∆CT values indicate the reverse. Results are the average ∆∆CT value of three biological replicates, with two technical replicates for each biological replicate. Error bars indicate standard deviations. ORCA2 transcript levels in the induced versus uninduced ORCA2-OE cultures differed at the same time point with: * = p ≤ 0.05, ** = p ≤ 0.01 according to Student’s t-test. Student’s t-test results for the induced versus uninduced cultures of the control line are not depicted.