Abstract: Pine wilt disease is currently a serious threat to global forest ecosystems, which is highly contagious. It is a common and urgent issue to screen effective biocontrol strains all over the world. From March 2023 to February 2024, five healthy Yunnan pine trees in Cangshan National Nature Reserve were selected to monitor the nematode-trapping fungi (NTF) resources. Pine bark samples were collected, and NTF were cultured, isolated, and purified in the laboratory. Species identification was conducted using morphological and molecular techniques, and the spatio-temporal dynamics of NTF communities were analyzed across different heights of the tree trunk: upper (80 cm ＜ height ≤ 200 cm), middle (20 cm ＜ height ≤ 80 cm), and lower (0 cm ≤ height ≤ 20 cm). The results revealed the presence of NTF in pine bark, with fungi of the genus Dactylellina being the dominant group, and Dactylellina ellipspspora as the dominant species (6.07%). The highest number of species (four species) was detected in March 2023, and the largest number of strains (eleven strains) was found in February 2024. A spatio-temporal variation in the number of detected NTF species and strains was observed. Indigenous NTF in pine trees can serve as ideal candidates for screening efficient biocontrol strains against pine wilt disease.