With a conjugative plasmid pJP4 carrying strain as the donor, two bioaugmentation experiments were conducted in a microcosm biofilm reactor with 2,4-D as the sole carbon source operated in fed-batch mode, and an enlarged lab-scale sequence batch biofilm reactor with mixed carbon sources of 2.4-D and other easily biodegradable compounds, respectively. In the microcosm study under sole carbon source condition, bioaugmentation led to a persistently increased 2,4-D degradation rate in the five operation cycles with enhancement of 13-64%. For the enlarged lab-scale bioaugmentation experiment under mixed carbon source conditions, no enhancement in 2,4-D removal could be observed during start-up period. After a period of operation, biofilm samples from the bioaugmented reactor demonstrated a stronger degradation capacity than the control and showed the presence of a large number of transconjugants. This study indicates that bioaugmentation based on plasmid horizontal transfer is a feasible strategy to establish functional microbial community in a biofilm reactor, and the strong selective pressure of 2,4-D existing alone and persistently was more favorable for the success of gene augmentation. (C) 2010 Elsevier B.V. All rights reserved.