Phytophthora capsici causes root, crown, and fruit rot of vegetable and tropical hosts. Cucumber, zucchini, tomato, and pepper fruits were inoculated using 6-mm-diameter agar plugs of P. capsici, incubated in clear plastic boxes at room temperature (25±2°C and 100% relative humidity), and virulence was estimated by measuring the lesion diameter, pathogen growth diameter, and pathogen sporulation density three (cucumber, zucchini) or four (tomato, pepper) days later. When isolates were grouped by genetic cluster, significant differences in virulence were observed on cucumber and zucchini, with isolates belonging to genetic cluster five causing larger lesions than isolates from genetic cluster six. On tomato, no significant differences were observed for isolates grouped by genetic cluster, but isolates from vegetable crops were generally more virulent than isolates from tropical hosts. Isolates from fabaceous hosts sporulated better on cucumber fruits than isolates from solanaceous hosts. Isolates from vegetable hosts sporulated better on zucchini than isolates from tropical hosts. No significant differences in lesion diameter were noted on pepper when isolates were grouped by host family of origin or genetic cluster, but differences in pathogen sporulation were apparent by host family. Our findings suggest that isolate characteristics such as host family of origin and genetic cluster membership may be used to guide initial isolate selection for cucurbit fruit resistance screening. Final isolate selection should incorporate the phenotypic and genetic diversity of P. capsici, including isolates with differing virulence to the host organ of interest.