In order to construct the callus induction system of Lonicera hypoglauca ‘Jincuilei’ and analyze the effect of plant growth regulator 2,4-dichlorophenoxyacetic acid(2,4-D) on the mass fraction of chlorogenic acid in callus, the young leaves of Lonicera hypoglauca ‘Jincuilei’ were used as explants to induce callus. The effects of different mass concentrations of growth regulator combinations on callus growth were compared, and the effects of different mass concentrations of 2,4-D on the mass fraction of chlorogenic acid in callus and the expression of key genes involved in chlorogenic acid synthesis were analyzed. The results showed that with the combination of 0.6 mg/L kinetin + 1.0 mg/L 2.4-dichlorophenoxyacetic acid + 0.6 mg/L 6-benzyladenine, the callus growth of ‘Jincuilei’ was the highest at 30 d, which was 5.505 g. The effect of 2,4-D on the mass fraction of chlorogenic acid in callus was promoted at low concentration and inhibited at high concentration. When the mass concentration of 2,4-D was 1 mg/L, the mass fraction of chlorogenic acid was the highest, which was 26.224 mg/g. 2,4-D treatment could promote the expression of key genes HQT and CCoAOMT in chlorogenic acid synthesis. It shows that the application of appropriate amount of plant growth regulator 2,4-D can increase the chlorogenic acid mass fraction of Lonicera hypoglauca callus.