Abstract: ‘Ganoderma lucidum No.1’ was taken as the test materials, a liquid nitrogen preservation orthogonal test with three factors and three levels was conducted.Four indicators, included germination rate, recovery time, mycelial growth rate, and dry weight of mycelium, were used to comprehensively evaluate the preservation effect, in order to establish the liquid nitrogen preservation method for Ganoderma lucidum strains, 10 different strains were used to verify the preservation effect of this method.The results showed that the optimal preservation scheme involved cooling with liquid nitrogen at-80 ℃,using 10% glycerol as a protective agent, and recovering at room temperature.Ten different Ganoderma lucidum strains could germinate after using this preservation method, and the recovery time was between 1-7 days.There was no significant change in mycelium growth rate before and after preservation of 80% strains, and no significant change in mycelium dry weight of 100% strains before and after preservation.A total of 611 fragments were amplified by ISSR,ranging in size from 300-3 000 bp.The amplification bands of pre-preservation and post-preservation strains had no difference with the same primers, and no variation was found.In conclusion, the liquid nitrogen preservation method in this study could guarantee the germination rate, recovery time, mycelium growth rate, mycelium dry weight and other indicators of frozen strains.At the same time, this method was suitable for the preservation of various Ganoderma lucidum strains.It had strong universality, wide application range, simple operation and good application prospect.