The genus Burkholderia is a heterogeneous group with extraordinary nutritional versatility and which occupies a diversity of niches. In recent decades, members of Burkholderia have been shown to be active participants in plant-microbe interactions, imparting beneficial effects as plant-growth-promoting bacteria (PGPB) or as pathogens. The study of quorum sensing and cell-density-dependent gene regulation, which play an important role in host colonization and pathogenesis, is extremely important in such a versatile organism. We report the identification and characterization by thin-layer chromatography (TLC) of N-acyl homoserine lactone (HSL) quorum sensing signal molecules by PGP Burkholderia. The Burkholderia spp. strains CBMB40, CBPB-HOD and CBPB-HIM investigated in this study were isolated from rice and possessed one or more PGP characteristics. Culture extracts of these strains contained detectable levels of hexanoyl (C(6)-), heptanoyl (C(7)-) and octanoyl (C(8)-) HSLs. Burkholderia sp. strain CBMB40 produced an additional molecule that migrated along decanoyl (C(10)-) HSL. Inoculation of HSL-producing Burkholderia strains through seed bacterization to canola stimulated root elongation. Signal molecules produced by Burkholderia strains could also be detected in planta, as determined by plate assays and TLC analysis of plant extracts. This study advances the hypothesis that signaling molecules by PGPB in planta might play a substantial role in increasing the pathogen resistance of plants.