ARS | Publication request: REAL-TIME PCR IDENTIFICATION OF PYTHIUM SPP. FROM CEREALS IN EASTERN WASHINGTON

Submitted to: APS Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: May 20, 2004
Publication Date: July 20, 2004
Citation: Schroeder, K.L., Paulitz, T.C. 2004. Real-time pcr identification of pythium spp. from cereals in eastern Washington. APS Annual Meeting. Phytopathology 94: S94.

Technical Abstract: A number of Pythium spp. cause root rot and yield loss in wheat and barley. These species possess varying degrees of virulence and many different species can be found in a single soil sample. Determining the species composition within a soil sample using classical methods is time consuming and difficult. Divergent portions of the ITS rDNA were used to design species-specific primers for P. abappressorium, P. debaryanum, P. aff. echinulatum, P. heterothallicum, P. intermedium, P. irregulare, P. paroecandrum, P. rostratum, and P. ultimum. Five to 11 isolates per species were screened using each primer pair. The SYBR Green dye was used in conjunction with capillary PCR to develop standard curves for each primer pair and to demonstrate that each pair was specific for isolates from the same species. Soils were inoculated with each Pythium spp. to examine amplification directly from soils and correlate DNA amplification with pathogen populations. This technique will prove invaluable to study the population dynamics of these pathogens in soil and to determine how Pythium spp. respond to