Serological and conductimetric assays for the detection of Pseudomonas syringae pathovar pisi in pea seeds

Test protocols for detecting Pseudomonas syringae pv. pisi, the causal agent of bacterial blight, in pea seeds are generally based on dilution-plating assays. These assays are usually very specific and reliable, but are time-consuming and laborious. Tests suited for large scale screening of seed lots are therefore needed. Conductimetric assays, immunofluorescence microscopy (IF) and an enzyme-linked immunosorbent assay (ELISA) for detecting Ps. syr. pv. pisi in pea seed extracts were compared with dilution-plating by two extraction methods, viz. 6 h soaking of seeds and 2 h soaking of flour of ground pea seeds in water. In general, the detection of Ps. syr. pv. pisi with conductimetric, IF and dilution-plating assays in the suspension water of the ground and 2 h-soaked pea samples was less sensitive than detection in suspension water of the 6 h-soaked pea seeds. The detection threshold of these assays varied per seed lot between 0 and 4.08 log cfu ml-1 for the 6 h soaking procedure. The detection threshold of ELISA varied for both extraction methods generally between 4.08 and 6.08 log cfu ml-1. Detection times recorded in conductimetric assays correlated well (— 0.89 < r < —0.98) with the log colony-forming units of Ps. syr. pv. pisi added to seed extracts at 27 as well as 17°. However, confirmation of results by isolation on semi-selective media after conductimetry was more successful at 17° than at 27°, because of the relatively lower activity of saprophytic Pseudomonas spp. at this temperature.