A rapid identification by TLC was performed for phytopathogenic bacteria (Burkholderia spp., Ralstonia solanacearum and Herbaspirillum rubrisubalbicans). One loopful bacterial cell was suspended in 0.2 ml of chloroform-methanol (2 : 1, v/v) in a small glass-vial and kept for 15 min in room temperature. About 10 μl of lipid extract was spotted on the origin of silica gel TLC plate and dried well. The plate was developed with chloroform-methanol-0.2% calcium chloride (55: 35: 8, v/v/v) for 1 hr at 25℃. After drying, the spots were visualized by spraying ninhydrin and successive heating at 100℃ for 10 min. The chromatograph was recorded by a photograph and/or photocopy. The lipid spots appeared on TLC plate at Rf 0.42-0.83 area were reliable benchmarks for differentiation of rRNA homology group II pseudomonads. The chromatograms of Burkholderia caryophylli, B. cepacia, B. gladioli, B. glumae, B. plantarii and B. vandii resembled each other but were distinct at species level and clearly different from R. solanacearum, B. andropogonis and H. rubrisubalbicans. The chromatographs of B. andropogonis and R. solanacearum were found roughly similar, but characteristic spots at Rf 0.42-0.52 region were absent in R. solanacearum. On the other hand, the profile of H. rubrisubalbicans was quite unique. This TLC method will be practical for rapid identification of phytopathogenic bacteria.