Abstract: The long-term use of chemical fertilizers and pesticides significantly threatens soil biodiversity, and biological control can effectively reduce their impact on farmland biota and the environment. Yunnan Province has established a green control strategy of ‘Four Pests and One Agent’, combining Aphidius gifuensis, Telenomus remus, predatory mites, Cantheconidea furcellata and endogenous antiviral agents. Soil nematodes, as one of indicators for evaluating soil health, can provide reference for guiding pest control strategies in tobacco fields. In order to detect the effects of different control measures on the nematode community in tobacco field ecosystems and enhance the implementation of green control of tobacco pests and diseases, this study employed a ‘Four Pests and One Agent’ green control strategy and a traditional control strategy primarily using the endogenous antiviral agents in tobacco fields in Yuxi City, Yunnan Province. Conventional pesticide and fertilizer management was used as a control. Continuous monitoring of soil nematode communities and soil factors was conducted from July to September 2023. The results revealed that: (1) 23,742 nematodes belonging to 22 families and 35 genera were identified, of which bacterivorous were the most prevalent taxa, accounting for 46.19% of the total. Significant differences in the numbers of plant-parasitic and omnivore-predator nematodes were observed under different pest control measures, which is correlated with the growth phases of crops. (2) Compared with traditional control treatments, conventional pesticide and fertilizer applications significantly decreased the total abundance, number of taxa, and Shannon-Wiener index of soil nematodes, indicating the negative impact of conventional management practices on soil nematode diversity. (3) The nematode community composition of the green control and traditional control treatment groups were similar, but the former showed a significantly higher Plant Parasitic Index (PPI) than the latter. The Wasilewska Index (WI) of the green control group was significantly lower than that of the conventional treatment group. (4) Soil pH, total nitrogen, and total phosphorus were the main factors affecting the composition of the nematode community′s trophic groups. In summary, green control positively affects the diversity of soil nematodes in tobacco cultivation, but it might increase the number of phytophagous nematodes (especially Meloidogyne), which will impede crop growth at later stages. Therefore, it is recommended that future tobacco cultivation management fully utilize green control techniques, reasonably control the amount of pesticide used, and strengthen the monitoring of harmful nematodes such as root-knot nematodes.