Serological Specificity of the Lipopolysaccharides, the Major Antigens of Pseudomonas syringae

The nature of major antigens of Pseudomonas syringae was studied on one strain of four pathovars (pvs aptata, mors-prunorum, phaseolicola and tabaci) belonging to four separate serogroups. Bacterial antigens were prepared by 4 procedures: extraction by phenol-water (PW), by citrate-NaCl (CN), by trichloracetic acid (TCA), and precipitation of a glycoproteic extracellular complex (GP). 3-Deoxy-2-octulosonic acid (KDO) revelation in all the extracts showed that the four procedures led to antigens containing similar amounts of lipopolysaccharide (LPS). Twenty polyclonal antisera were raised in rabbits against whole bacteria and the different extracts. Serological reactions were tested by gel double diffusion (DD) and indirect immunofluorescent staining (IF). The anti-whole cell sera were shown to contain mostly anti-LPS antibodies. For each pathovar, whole bacteria used as antigens in DD gave precipitation bands identical to the bands given by the LPS extracts (PW, CN or TCA), identical to the heated bacteria (HB), and identical to LPS sidechain preparations. The GP extract itself was shown to be rich in LPS. To serotype P. syringae, it is advised to raise antisera against either whole bacteria or GP extracts; whereas the reacting antigens for DD would be heated bacteria.