Fungi, serving as real-time bioindicators to environmental changes and stressors, are crucial for effective forest conservation and management practices under ongoing global change. However, the large-scale assessment of soil fungi still encounters challenges in striking a balance between the extensive sampling costs and the limited accuracy of minimal sampling. In this study, we analyzed 1,606 soil samples collected from 625 quadrats (20 m × 20 m) within a 25-ha subtropical forest dynamic plot in East China. Our primary objective was to explore the impact of different sampling schemes, in conjunction with remote sensing (RS) technologies, on the interpolation of soil fungal diversity using Ordinary Kriging (OK) and Co-kriging (CoK) models. Our findings suggested that a sampling scheme including points at 0 m (the base points) and 8 m within each quadrat, totaling to 26 points/ha, would be a sufficient scheme. This scheme with OK model yielded comparable results to those of more intensive schemes (at 0, 2, 5 and 8 m), but required the fewest sampling points. Upon incorporating each RS variable separately into the CoK models, including two vegetation indices (normalized difference vegetation index and transformed chlorophyll absorption ratio index 2), three terrain attributes (Elevation, Aspect and Slope), and the synthesis of these RS variables, the accuracy of the predicted results was further improved for each sampling scheme. By leveraging high-precision soil DNA sequencing in conjunction with cost-effective RS technologies, this study proposes a rapid and affordable approach for monitoring soil fungal diversity on a large scale. This will facilitate data collection for understanding responses of forest soil fungi to ongoing global change.