Abstract: 【Objective】This research explored the tissue culture and rapid propagation of seedless Siraitia grosvenorii in order to provide technical supports on seedless Siraitia grosvenorii regenerations system establishment and factory production .【Method】The young stem of seedless Siraitia grosvenorii was used as explants to investigate the effects of different sterilization time, different hormone combinations and concentrations on the initiating, proliferation, as well as rooting cluster buds were studied.【Result】The results showed that the best sterilization method for the stem segments was agitating in 0.1% HgCl2 for 8 min. The optimum medium for initiation culture of stem segments was MS+ 1.0 mg/L 6-BA+1.5 mg/L KT , The suitable multiplication medium was MS+1.0 mg/L 6-BA+0.5 mg/L IBA, with the bud proliferation coefficient of 6.8; The suitable rooting medium was 1/2MS+1.0 mg/L IBA+0.5 mg/L NAA, in which the average root number was 7.4, the rooting rate of muhiplied buds was up to 95%.【Conclusion】It could provide theoretical basis for the large-scale production of the tissue culture and rapid propagation of seedless Siraitia grosvenorii, and the experiment provided references for the establishment of efficient Tissue culture regeneration system and the exogenous gene transduction.