In greenhouses, high humidity, low light, and inadequate ventilation conditions, along with continuous and high-density planting, promote the proliferation of soilborne pathogens. Among these pathogens, Fusarium oxysporum Schltdl (F. oxysporum) is a notably challenging one, causing root rot of tomato plants in greenhouse cultivation. To address this issue, this study applied a pulsed electric field (PEF) to target the elimination of F. oxysporum in suspension and soil media. Initially, PEF parameters were systematically explored in suspensions to determine the effective ranges for the elimination of F. oxysporum. The results revealed that the effective ranges for achieving the desired microbial reduction were an electric field strength (EFS) between 5–15 kV·cm−1, a pulse number within the range of 100–500, and a pulse width of 10–20 µs. Subsequently, the impact of soil moisture content, soil bulk density, and soil type on soil dielectric breakdown field strength was analyzed within the range from previous results. Based on these findings, the soil experiments were conducted with parameters designed to prevent dielectric breakdown. Specifically, for sampling soil with a moisture content of 16.2% and a bulk density of 1.31 g·cm−3, the maximum effective application of electric field strength was 9.5 kV·cm−1, accompanied by 1000 pulses and a pulse width of 20 µs. Finally, building on these results, soil samples were sterilized within a parameter range that spanned an electric field strength of 5–9.5 kV·cm−1, a pulse number between 100–500, and a pulse width of 10–20 µs. Response surface methodology (RSM) analysis further identified the optimal parameter combination: an electric field strength of 8.2 kV·cm−1, 306 pulses, and a pulse width of 15 µs, resulting in an average lethal rate of 76.16% for F. oxysporum sterilization in soil. These findings suggest the potential use of PEF against F. oxysporum and other pathogens in greenhouse soils, and provide theoretical foundations for further experiments, thereby contributing to the sustainable advancement of greenhouse agriculture.
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