Survey of oomycetes and pathogenicity of Phytophthora cinnamomi associated with root rot disease of western white pine (Pinus monticola)

Western white pine (Pinus monticola) with genetic tolerance to white pine blister rust disease caused by Cronartium ribicola constitutes an invaluable resource for the mitigation of this disease as well as the restoration of the species and its utilization in managed forests. In the last several years, typical root-rot symptoms and sudden tree death have been observed in some western white pine plantations located in British Columbia (BC) in Canada, and a few Phytophthora species were found to be associated with symptomatic trees. Our objectives were to survey one of the plantations located in south-west BC on Vancouver Island to confirm the presence of these Phytophthora species and other oomycetes and test their pathogenicity on western white pine by completing Koch's postulates. We used direct plating of root samples on PARP(H)-CMA media and by baiting for oomycetes in soil and root samples. Ten oomycetes, including four Phytophthora species (Phytophthora cinnamomi, P. cactorum-like, P. cryptogea and P. pseudocryptogea) were isolated from roots and soil samples of diseased Pi. monticola trees. The Phytophthora species were identified using ITS, beta-tubulin and TEF1 gene regions. Koch's postulates were validated with controlled inoculations of 1-year old seedlings; mortality occurred as early as 15 days after inoculation with P. cinnamomi when the seedlings were maintained in warm temperature conditions (25 degrees C). Molecular typing of two P. cinnamomi cultures using Oxford Nanopore MinION (TM) sequencing indicated that they belong to one of the two main clonal clusters of mating type A2 that have been recognized as the main drivers of the global P. cinnamomi epidemic.