Objective To observe the anatomical morphology of root canal system of human mandibular first premolar based on the reconstructed images of cone-beam CT (CBCT). Methods The CBCT images of 1 102 mandibular first premolars meeting the inclusion criteria were reconstructed. We observed the root canal morphology and root canal groove (depression)of mandibular first premolars, and measured the thinnest root canal wall thickness of single premolars in different cross sections by one volume viewer software. Results Among the 1 102 mandibular first premolars, there were 958 type I root canals, 92 type V root canals, 42 type C root canals and 10 type III root canals. The detection rates of type I, type III, type V and C root canals in 424 male patients were 75. 0%, 2. 4%, 15. 6% and 7. 1%, respectively. The detection rates of type I, type III, type V and C root canals in 678 female patients were 94. 4%, 0, 3. 8%, and 1. 8%, respectively (all P<0. 05). Among the 1 102 mandibular first premolars, 524 (564) had root groove (depression), and the incidence of root groove (depression)was 47. 5%. Among the 524 teeth with root surface groove (depression), the starting positions were 4 at 1/3 of root crown, 500 at 1/3 of root middle and 20 at 1/3 of root apex. Among the 564 root surface grooves (depressions), 356 were on the mesial root surface, 194 on the distal root surface, 10 on the lingual root surface, and 4 on the buccal root surface. The starting positions were 4 at 1/3 of the root crown, 536 at 1/3 of root middle, and 24 at 1/3 of root apex. The thickness of the thinnest root canal wall at the junction of crown and root was (1. 74±0. 14)mm, and the thickness of the thinnest root canal wall at the junction of crown and root in men was greater than that in women (P< 0. 05). The thinnest root canal wall thickness at the junction of 1/3 of root neck and 1/3 of root middle was (1. 24± 0. 10)mm, with no significant difference between men and women (P>0. 05). The thinnest root canal wall thickness at the junction of 1/3 of the middle root and 1/3 of the apical root was (0. 85±0. 11)mm, with no significant difference between men and women (P>0. 05). Conclusions The root canal morphology of mandibular first premolar is complex and changeable. Type I is the most common root canal type, and there are differences between men and women. The incidence of root surface groove (depression)of mandibular first premolar is high. And it often occurs on the mesial root surface and mostly starts at 1/3 of the root. The thickness of the thinnest root canal wall of a mandibular first premolar with a single root canal in different cross sections is different, and the thickness of the thinnest root canal wall at the junction of crown and root differs in gender. [ABSTRACT FROM AUTHOR]