FIGURE 4.

qRT‐PCR analysis of PmLEC1 expression in Douglas‐fir embryonic and vegetative tissues. Total RNA was isolated from 5 biological replicates at each developmental stage. DNase I treatment and reverse transcription were carried out with 5 samples of 1 μg from each stage. The qRT‐PCR reactions were performed in quadruplicate with Douglas‐fir ribosomal L8 transcripts as internal controls. Relative gene expression was calculated according to the 2^‐ΔΔCt method (Dorak, 2006) by comparison to transcript levels in pollen cones, which showed the lowest expression. Statistical analysis was performed on the pooled groups by the Mann‐Whitney U test. Results are shown as means ± SE (n = 5). Asterisks indicate PmLEC1 transcript levels that are significantly different (p < .05) from pollen cones. DAEG: days after exposure to germination conditions