In forest ecosystems, interactions between overstory trees and understory herbs play an important role in driving plant species diversity. However, reported links between overstory tree and understory herb species diversity have been inconsistent, due to variations in forest types and environmental conditions. Here, we measured species richness (SR) and diversity (Shannon-Wiener (H') and Simpson's (D) indices) of overstory trees and understory herbs in the protected Tianshan Wild Fruit Forest (TWFF), Northwest China, to explore their relationships along the latitudinal, longitudinal, elevational, and climatic (current climate and paleoclimate) gradients in 2018. We found that SR, and H' and D diversity indices of overstory trees and understory herbs exhibited a unimodal pattern with increasing latitude and elevation (P<0.05) and negative associations with longitude (P<0.01). Along the climatic gradients, there were U-shaped patterns in SR, and H' and D diversity indices between trees and herbs (P<0.05). SR, and H' and D diversity indices for overstory tree species were positively associated with those for understory herbs (P<0.01). These findings indicate that overstory trees and understory herbs should be protected concurrently in the TWFF to increase effectiveness of species diversity conservation programs.