The soil bacterium Alcaligenes eutrophus is able to degrade phenoxy herbicides such as 2,4-D [(2,4-dichlorophenoxy)acetic acid]. It has been shown that several degradative genes are located on a large plasmid harbored by this organism. We have demonstrated by HPLC and gas chromatography that 2,4-dichlorophenol is a major metabolite resulting from 2,4-D degradation by A. eutrophus. A portion of the large plasmid carrying the 2,4-D catabolic genes has been cloned and mapped by restriction endonuclease digestion. Overlapping subclones have been produced and were used in conjugation experiments in order to identify the first gene involved in 2,4-D detoxification. Once sequenced, this gene will be transferred to dicotyledonous plant cells using available vector systems.