Sequential induction of the ethylene biosynthetic enzymes by indole-3-acetic acid in etiolated peas

Ethylene induced an increase in the accumulation of 1-aminocyclopropane-1-carboxylate (ACC) oxidase transcript level and enzyme activity in the first internode of 5- to 6-day-old etiolated pea (Pisum sativum L.) seedlings. Indole-3-acetic acid (IAA), which stimulates ethylene production by enhancing ACC synthase activity, also caused an increase in ACC oxidase transcript and activity levels. The IAA-induced increase in ACC oxidase mRNA level and enzyme activity was blocked by 2,5-norbornadiene (NBD), a competitive inhibitor of ethylene action. This indicates that IAA induced ACC oxidase through the action of ethylene. The level of ACC synthase mRNA and enzyme activity started to increase less than 1 h after the start of IAA treatment, whereas ACC oxidase activity and transcript levels began to rise after 2 h of IAA treatment. These results indicate that the enzymes of ethylene biosynthesis are sequentially induced after treatment of intact pea seedlings with IAA. The increase in ACC synthase activity leads to the production of ACC, which is converted by the low constitutive level of ACC oxidase activity to ethylene. Through a positive feedback loop, ethylene promotes the accumulation of ACC oxidase mRNA and the increase in ACC oxidase activity.