Qingtiankui Tissue Cryopreservation of complex.doc

 PAGE \* MERGEFORMAT 10 Qingtiankui Tissue Cryopreservation of complex [Abstract] Objective method of ultra-low temperature preservation of tissue culture Qingtiankui material. Methods The single-factor comparison method, uniform design study the growth of week-old, cryoprotectant, pre-incubation time, cooling method, thaw mode and other factors on Qingtiankui Cryopreservation of tissue culture materials The influence of cell life. Results of tissue culture material Qingtiankui better cryopreservation procedures: 5-week-old tissue culture material, cold acclimation at 4 ℃ for 12 d, to 12.5% DMSO +0.25% LH as a cryoprotectant at 4 ℃ for put it aside for dealing with 30 min, and then in -18 ℃ , stay put in liquid nitrogen after 1 h preservation, material removed after the thaw at 20 ℃ , sterile washing again after inoculation, tissue culture material to survive and continue to grow. Conclusion cryopreservation method can be successfully saved Qingtiankui of tissue culture material germplasm resources. [Keywords:] Qingtiankui Cryopreservation of tissue culture material Abstract: ObjectiveTo preserve the tissue cultures of Nervilia fordii using cryopreservation method. MethodsSuitable factors in cryopreservation (-196 ℃ ) were studied by monofactorial and uniform design.ResultsThe experiments showed that five-weeked tissue cultures were the best material for cryopreservation. Tissue culures treated for 12days at 4 ℃ were the best cryotolerant. The optimal cyroprectants was 12.5% DMSO +0.25% LH. The best freezing procedure was to reduce the temperature from 0 ℃ to -18 ℃ , and stay at that temperature for 1h, then drop them in liquid nitrogen (-196 ℃ ) to be preserved. 20 ℃ was good for melting the frozen material. Unfrozen tissue cultures survived when they were recultured. ConclusionTissue cultures of Nervil ia fordii can be preserved by cryopreservation method. Keywords:: Nervilia fordii; Tissue cultures; Cryopreservation Qingtiankui alias alone Yelena