The experiment was carried out in April 2021 to clarify the effects of different application ratios（20%, 40%, 60% and 80% by volume） of mushroom cultivation residue on the quality of rice seedlings of hybrid rice combinations Jingliangyou 534 and Yongyou 1540. The results showed that, compared with control（cultivated soil for raising seedlings）, the application of a certain proportion of mushroom cultivation residue in the seedling substrate had no significant effect on the agronomic traits such as root number, stem base width and dry matter accumulation of rice seedlings. With the increase in the proportion of mushroom cultivation residues, the uniformity of the seedlings increases. At the 25-day seedling age of the two tested varieties, the uniformity in the treatments with 60% and 80% mushroom cultivation residues were 9.8%~13.1% and 12.0%~19.4% higher than the control, respectively, showing significant differences. There is no significant difference in uniformity among different treatments at 40-day seedling age. Compared with the control, the chlorophyll contents in the leaves in the treatment with mushroom residue increased, especially in the treatments with 60% and 80% of the mushroom residue at 40-day seedling age, the chlorophyll contents of the leaves of the two combinations were 13.4%~35.9% and 7.6%~17.3% higher than the control, respectively. Therefore, 60%~80% mushroom residue addition to the cultivated soil was beneficial to improve the uniformity of the seedlings and the chlorophyll contents of the leaves, thereby improving the quality of the rice seedlings.