Macadamia nut plantings in China are expanding year by year. In order to further promote the superior varieties, this study analyzed the effects of different varieties of rootstock and scion on the survival rate as well as the effects of 18 different ratios of substrate composition on seedling growth were analyzed. The results showed that: The survival rate of HAES788 varieties as rootstock and Guire No.1 as scion was the highest, reaching 96%. The optimal grafting time in December was better than that in March. Substrate formulations 12, 13, 15 and 16 had agglomerated soil and well-developed root systems compared to the controls. Plant height, stem diameter, leaf length, leaf width, and dry weight of aboveground and underground parts were significantly higher than those of the control. In addition, the substrate formulations 12, 13, 15 and 16 significantly improved the organic matter, total nitrogen, and total potassium content of the substrate soils, but little on total phosphorus content. All in all, macadamia grafting times are best in December, with HAES788 and Guire No.1 being the best rootstock and scion. Substrate formulations 12, 13, 15 and 16 are the optimal formula. This study provides a solid foundation to produce high-quality macadamia seedlings.