Quantification of Pythium populations in ginseng soils by culture dependent and real-time PCR methods §

Ginseng (Panax quinquefolius L.) is widely cultivated in North America as a medicinal herb. However, yields are often reduced by various root pathogens, including Pythium species, which cause damping-off in young plants. In order to improve the prediction of disease risk, real-time PCR assays were developed and used in conjunction with dilution plating on selective media to quantify populations of Pythium irregulare Buisman and Pythium ultimum Trow directly from soil. The assays were tested on artificially infested soils and then used on a variety of naturally infested, ginseng-cultivated soils in south-western Ontario. Data on P. ultimum DNA concentrations were positively correlated with the number of P. ultimum viable propagules on selective media. However, in the case of P. irregulare, the presence of cryptic species resulted in incongruent relationships between the dilution plate and real-time PCR data. We therefore sequenced the ITS region of a large number of P. irregulare isolates in order to determine the proportion that would be detected by the real-time PCR assay. The ability to quantify pathogen populations directly from soils using real-time PCR (calibrated with data on inoculum potential) will improve disease risk assessment and lead to a reduction in pesticide application.