Influence of Isolation Method on Recovery of Pythium Species from Forest Nursery Soils in Oregon and Washington.

Pythium species are common damping-off pathogens that can cause stunting, chlorosis, and death of conifer seedlings in the Pacific Northwest (PNW) region of the United States. Despite the prevalence and importance of these pathogens in forest nurseries, relatively little is known about the identity of Pythium species associated with forest nursery soils in Washington and Oregon. A limited number of studies have reported P. aphanidermatum, P. irregulare, P. mamillatum, and P. ultimum as the predominant species in the PNW, but most studies of this genus in forest nurseries have not reported Pythium species identity. In an attempt to identify Pythium species associated with forest nursery soils, field surveys were conducted at three forest nurseries (two in Oregon and one in Washington) in 2008 using three isolation methods. Pythium species were isolated by plating soil onto a semiselective medium or by baiting soil with rhododendron leaf disks and Douglas-fir needle segments. One hundred isolates were randomly selected from each isolation method at each nursery (900 isolates total) and identified on the basis of the internal transcribed spacer (ITS) sequence. Nineteen Pythium species were identified during the survey. Species richness and abundance were strongly influenced by both nursery and isolation method. Of the 300 isolates obtained from each nursery, P. irregulare was the most commonly isolated species from nursery A in Washington (65% incidence). P. 'vipa' and P. dissotocum were the most commonly isolated species from nurseries B and C in Oregon, respectively (53 and 47% incidence, respectively).