Diversity of Phytophthora, Pythium, and Phytopythium Species in Recycled Irrigation Water in a Container Nursery

Recycling of irrigation water increases disease risks due to spread of waterborne oomycete plant pathogens such as Phytophthora, Pythium, and Phytopythium. A comprehensive metabarcoding study was conducted to determine spatial and temporal dynamics of oomycete communities present in irrigation water collected from a creek (main water source), a pond, retention reservoirs, a chlorinated water reservoir, and runoff channels within a commercial container nursery in Oregon over the course of 1 year. Two methods, filtration and leaf baiting, were compared for the detection of oomycete communities. Oomycete communities in recycled irrigation water were less diverse but highly enriched with biologically active plant pathogens as compared with the creek water. The filtration method captured a larger portion of oomycete diversity, while leaf baiting was more selective for plant-associated oomycete species of Phytophthora and a few Pythium and Phytopythium species. Seasonality strongly influenced oomycete diversity in irrigation water and detection with leaf baiting. Phytophthora was the major colonizer of leaf baits in winter, while all three genera were equally abundant on leaf baits in summer. The metabarcoding approach was highly effective in studying oomycete ecology, however, it failed to distinguish some closely related species. We developed a custom oomycete internal transcribed spacer (ITS)1 reference database containing shorter sequences flanked by ITS6 and ITS7 primers used in metabarcoding and used it to assemble a list of indistinguishable species complexes and clusters to improve identification. The predominant bait-colonizing species detected in recycled irrigation water were the Phytophthora citricola-complex, Phytophthora syringae, Phytophthora parsiana-cluster, Phytophthora chlamydospora, Phytophthora gonapodyides, Phytophthora irrigata, Phytophthora taxon Oaksoil-cluster, Phytophthora citrophthora-cluster, Phytophthora megasperma-cluster, Pythium chondricola-complex, Pythium dissotocum-cluster, and Phytopythium litorate.