Abstract: Cryopreservation of callus of Pinus elliottii × Pinus caribaea were conducted in this research. The results showed that the best process of cryopreservation for the embryogenic callus of P. elliottii × P. caribaea was as follows: the callus were sub-cultured for 9-12 days in callus proliferation medium firstly and were pre-cultured in 0.5 mol/L sorbol for 4 days, and then pretreated 20 minutes in buffer of 0.6 mol/L sorbol + 10% DMSO, cooled to -40 ℃ at rate of -1 ℃/min, maintained at -40 ℃ for 10 min, preserved in liquid nitrogen after cooled to -90 ℃ at rate of -5 ℃/min. The resuscitate callus were kept in 37 ℃ water bath, clean three times in 1 mol/L sorbol liquid medium, passage in solid medium support by filter paper. New embryogenic callus will be produced within one month.