Natural Infestation of Onion Seed by Pantoea ananatis, Causal Agent of Center Rot

An immunomagnetic separation and polymerase chain reaction (IMS-PCR) assay was used to detect Pantoea ananatis in naturally infested onion seeds. Using species-specific PCR primers and polyclonal antibodies, IMS-PCR consistently demonstrated detection thresholds of 10¹ to 10³ CFU/ml. There was no significant difference between the numbers of CFU recovered from onion seed wash by IMS (after repeated rinses) and by direct plating, indicating that IMS effectively captured P. ananatis cells from heterogeneous bacterial populations. Using IMS-PCR and IMS followed by plating on nutrient agar, P. ananatis was detected in 19.7% of onion seed samples harvested from two onion fields in which center rot developed naturally in 2000. When planted in germination boxes, 53% of the seed samples that tested positive for P. ananatis produced seedlings with symptoms of center rot. There was no significant difference in germination between infested and noninfested seed samples. This is the first report of natural infestation and transmission of P. ananatis in onion seed.