First report of Phytophthora ramorum on Cotoneaster sp. in the USA.

Cotoneaster (Rosaceae) is a genus of woody plants native to the Palearctic region which includes popular ornamental plants; some are invasive in parts of the USA. In May 2022 symptomatic leaves were detected on Cotoneaster pannosus (Silverleaf Cotoneaster) in Marin County, California (37.89165, -122.56755 ), an area infested heavily by Phytophthora ramorum, causal agent of Sudden Oak Death. Symptoms consisted of dark brown necrotic spots mostly near the tips and sometimes on the margin of the leaves, covering less than half of the leaf surface; no die-back or symptoms on twigs were detected. Diseased leaves were surface-sterilized with 70% ethanol, washed twice with de-ionized water, and placed on PARPH(V8) media. Two Phytophthora ramorum like isolates (NORS058 and NORS059) were obtained from different leaf samples from the same tree and the internal transcribed spacer (ITS) region was sequenced. Both sequences were deposited in GenBank (OR224345 and OR224346). NORS058 and NORS059 showed 99.88% and 99.75% sequence identity to P. ramorum strain Ex-type CPHST BL 55G (MG865581.1). Detached leaves of C. pannosus and C. lacteus (Milkflower Cotoneaster) were inoculated with mycelial plugs of P. ramorum NORS058, and incubated at 20°C. Both species developed necrotic leaf spots seven days post inoculation (dpi). Sporulation of the pathogen was observed on symptomatic leaves of C. lacteus. P. ramorum was reisolated from the symptomatic leaf tissue from both Cotoneaster species. Pathogenicity tests were also performed on whole plants of C. dammeri (Bearberry Cotoneaster) using the strain NORS058. Five plants each were inoculated using three different methods: 1) a zoospore solution (concentration 2.5 x10E5 spores/mL) were sprayed on the plant surface until run off. Ten leaves per plant were wounded with a needle, the remaining leaves were not wounded; 2) 200 µL of the zoospore solution in a PCR tube were attached to 5 leaves of each plant; and 3) 10 mL of the zoospore solution was drenched into the potting mix of the five plants. Control plants were treated as above but with water instead of the zoospore solution. Leaf spots developed 7 dpi on plants sprayed with zoospores on wounded leaves; and 10 dpi on plants treated with zoospores in the tube. P. ramorum was reisolated from symptomatic leaves treated with the first two methods mentioned above. Plants treated with a soil drench did not develop symptoms on the aerial parts or on roots that were sampled 50 dpi. Tests using AGDIA- immunostrips of the roots were negative. Control plants showed no aerial or root symptoms. To our knowledge, this is the first report of P. ramorum occurring on Cotoneaster in the USA. Previously, inoculation of detached leaves of C. dammeri and C. horizontalis with P. ramorum in Serbia resulted in symptom expression (Bulajić et al. 2010). P. ramorum was reported from a Cotoneaster sp. in the UK in 2010, but no further information were presented (FERA 2015). The tree sampled in 2022 showed symptoms again in spring 2023 and official regulatory samples were taken by the CDFA (California Department of Food and Agriculture) and confirmed by the USDA. During a survey in 2023, more symptomatic Cotoneaster plants were detected in Marin County, California, indicating Cotoneaster might play a role in the epidemiology of the disease. References: FERA 2015. https://planthealthportal.defra.gov.uk/pests-and-diseases/high-profile-pests-and-diseases/phytophthora/ Bulajić et al. 2010. Plant Dis. 94(6): 703.