The mechanical properties of as-quenched and tempered steels are affected by austenitizing temperature. The present work has investigated the effect of austenitizing temperature on martensitic microstructure, carbide precipitates and mechanical properties of 30NiCrMoV12 alloy steel for the axle of high-speed train. The martensitic microstructure and carbide precipitates were studied using OM, FE-SEM, TEM, EBSD and EDS. Thermodynamic calculation of equilibrium precipitation were carried out by Thermo-Calc software. The results showed that the prior austenite grains, martensitic packets, blocks and laths were coarsening with increasing austenitizing temperature. Besides, with increasing austenitizing temperature, after tempering the amount of large size carbides precipitated at martensitic lath boundaries decreased while the amount of small size carbides precipitated in matrix increased. Meanwhile, phase transformation from M 23 C 6 to M 7 C 3 during tempering was enhanced with increasing austenitizing temperature. Coarse grains and wide martensitic laths were beneficial to reducing the amount of strip-like M 23 C 6 carbides precipitated at martensitic lath boundaries due to the reduction of boundary area and thereby obtaining more fine precipitates in matrix. The strength and impact toughness could be improved to a certain extent by refining carbides in tempered steel with higher austenitizing temperature. However, the degree of favorable influence on impact toughness resulting from refining carbides was lower than the negative effect from coarse martensitic structures. Therefore, the toughness is deteriorated and the strength is improved with increasing austenitizing temperature.