This study used environmentally friendly nano-xylan to enhance the drug loading and preservative performance (especially against white-rot fungi) of pine wood (Pinus massoniana Lamb), determine the best pretreatment, nano-xylan modification process, and analyze the antibacterial mechanism of nano-xylan. High-temperature, high-pressure steam pretreatment-assisted vacuum impregnation was applied to enhance the nano-xylan loading. The nano-xylan loading generally increased upon increasing the steam pressure and temperature, heat-treatment time, vacuum degree, and vacuum time. The optimal loading of 14.83% was achieved at a steam pressure and temperature of 0.8 MPa and 170 °C, heat treatment time of 50 min, vacuum degree of 0.08 MPa, and vacuum impregnation time of 50 min. Modification with nano-xylan prohibited the formation of hyphae clusters inside the wood cells. The degradation of integrity and mechanical performance were improved. Compared with the untreated sample, the mass loss rate of the sample treated with 10% nano-xylan decreased from 38 to 22%. The treatment with high-temperature, high-pressure steam significantly enhanced the crystallinity of wood.