In order to gain high yield strain of huperzine A (HupA) from Huperzia serrata (Thunb.) Trev. in vitro, the hydrogen peroxide (H₂O₂) induced thallus was studied. The results showed that the HupA content in induced thallus significantly increased. The strain SH42 had highest yield of HupA with relative growth rate and HupA content for 4499.28% and 261.17 μg g^-1 DW, respectively, which increased 2.35 times and 2.43 times of the initial strain, respectively. Moreover, there were significant difference spectral band in SOD isozymes and soluble protein among strains. The variation characteristics were stable after nine generations. Therefore, it was suggested that H₂O₂ had good induced effect on thallus of Huperzia serrata, which could be used for screening high-yield strain of HupA in vitro.