A Rapid Extraction - TLC Method for Differentiation of Burkholderia spp., Ralstonia solanacearum, Herbaspirillum rubrisubalbicans and Pseudomonas syringae Pathovars

A rapid extraction -TLC method was conducted for differentiation of Burkholderia spp., Ralstonia solanacearum, Herbaspirillum rubrisubalbicans and Pseudomonas syringae pathovars. Lipids were extracted from bacterial cells in a small glass-vial using 0.2 ml of chloroform - methanol - 0.3% NaCl solution (2 : 1 0.4, v/v/v). The development of TLC plate was performed with chloroform - methanol- 0.2% CaCl_2・2H_2O solution (55 : 35 : 8, v/v/v) for 1 h at 25℃ and the amino-lipid spots were visualized by spraying ninhydrin and successive heating at 100℃ for 10 min. The chromatograms were characteristic at both the generic and species level. This method will be practical for rapid differentiation as well as identification of Burkholderia spp., R. solanacearum, H. rubrisubalbicans and P. syringae pathovars