Abstract: 【Objective】To establish a technological system of tissue culture and rapid propagation of hybrid germplasm of Camellia nitidissima C.W. Chi that would provide technical support for the preservation and rapid propagation of elite hybrid germplasm of C. nitidissima C.W. Chi.【Method】With sterile seedlings from hybrid seeds of C. impressinervis Chang et S.Y. Liang♀× C. japonica ‘Dormi'♂(S1), C. chrysanthoides ‘Huangzun'♀ × C. chrysantha var. longistyla ♂(S2), and C. chrysantha var. longistyla ♀ × C. amplexicaulis ♂(S3)as material, we determined the difference in induction rate from different explants, including the apical bud, stem section, and radicle of a sterile seedling, effects of 3 hybrids, effects of 4 basic media including MS, B5, Hyponex No. 1, and 1/2MS+Hyponex No. 1, effects of concentration combinations of different plant growth regulators(PGRs), including 6-BA, IBA, and NAA on the proliferation and rooting of shoots, effects of sucrose concentration on rooting rate of shoots, and the difference in transplantation survival rate of 3 hybrids on 4 substrates, including pure yellow soil, 3V_{pure yellow soil}+1V_vermiculite, 3V_{pure yellow soil}+1V_perlite , and 3V_{pure yellow soil}+1V_coconut .【Result】The average induction rate of apical bud and stem section of the 3 hybridized combinations was 12% and 100%, respectively. The difference in proliferation times of shoots was significant for the 3 hybridized combinations, the 4 basic media, and the different concentrations of 6-BA. The proliferation times of S1,S2 and S3 were higher at 7.4, 6.6 and 7.5, respectively, on the subculture medium B5+6-BA 2.5 mg/L+IBA 0.05 mg/L+30 g/L sucrose for 40 d. The shoots grew poorly with falling leaves on the basic medium MS for several generations of multiplication. The difference in the rooting rate of the 3 hybrids was significant on concentration combinations of different PGRs and sucrose concentration, and the higher the sucrose concentration, above 10 g/L, the lower the rooting rate. A higher rooting rate for S1,S2 and S3 was obtained on the medium 1/2 B5+ABT 1.0 mg/L+sucrose 5 g/L, at 92%, 85% and 89%, respectively. The difference in the transplantation survival rate of shoots was significant on the 4 substrates, with the average survival rate of S1, S2 and S3 reaching 94.0% on substrote of 3V_{yellow soil}+1V_vermiculite after hardening for 30 d.【Conclusion】Stem sections of sterile seedlings were the best explants for tissue cultures of the 3 hybridized combinations of C. nitidissima C.W. Chi. The best subculture and rooting medium was B5+6-BA 2.5 mg/L+IBA 0.05 mg/L+30 g/L sucrose and 1/2 B5+ABT 1.0 mg/L+sucrose 5 g/L, respectively. The optimum transplant mixture was 3V_{yellow soil}+1V_vermiculite. Tissue culture technology was the best preservation method and method for plantlet regeneration for the elite hybrid germplasm of C. nitidissima C.W. Chi.