In-situ remediation of contaminated farmland by horizontal transfer of degradative plasmids among rhizosphere bacteria



Horizontal transfer of catabolic genes for pollutant degradation among rhizobacteria plays an important role in environmental bioremediation, but lacks support from field trial data. To address this problem, Pseudomonas fluorescens strain TP13 was inoculated into the soils of a phenol-contaminated farmland on which tomato seedlings were growing in April 2009, 2010, 2011 and 2012. Results consistently showed that introduction of TP13 strain significantly reduced phenol content and increased plant biomass after 20 days, compared with controls. Strain TP13 was able to colonize the plant rhizosphere and the number of rhizosphere bacteria which were grown on phenol and contained the plasmids containing the gene encoding for catechol 2, 3-dioxygenase (C23O) increased gradually in the later stages of the experiment. The increase in magnitude of the plasmid-containing rhizosphere bacteria correlated well with plant biomass, while the number of plasmid-containing rhizosphere bacteria and phenol content was strongly negatively correlated. Furthermore, six strains (T1-T6) of rhizosphere bacteria were isolated and found to possess large plasmids containing identical C23O genes and similar HindIII restriction patterns. Sequence alignment showed that the C23O genes from strains T1-T6 contained almost identical sequences and the sequence of the C23O of strain T1 was the same as that of strain TP13. These data indicated that the plasmids were transferred from strain TP13 to these rhizosphere bacteria and that horizontal gene transfer stimulated phenol degradation and plant growth in the contaminated farmland. This is an important finding for in situ remediation of contaminated farmland.