Figure 1
Silencing Na α-dox1 results in lines with two different growth phenotypes correlating with the suppression level of α-DOX activity. The growth phenotype of (A) 35-day-old and (B) 60-day-old WT and T3 homozygous Nicotiana attenuata plants transformed with an inverted repeat (IR) construct to silence Naα-dox1. More than half of 18 independently transformed lines are severely stunted (IR α-dox S) whereas the other lines (IR α-dox M) are morphologically indistinguishable from wild-type plants (WT). (C) α-DOX activity (mean ± SE of 4 biological replicates) of protein extracts from leaves by in vitro measurements of the formation of 8,11,14-heptadecatrienal (HDT). Leaves had been wounded and treated with 2.5 μL of OSWT every 30 min for four times. Leaves were harvested 30 min after the last treatment. (D) Leaf levels of 2-hydroxyoctadetrienoic acid (2-HOT; mean ± SE of 4 biological replicates), the main α-DOX product in planta. Asterisks signify significant differences between IR α-dox and WT plants (unpaired t-test WT vs. lines P < 0.05). nd: not detected.