Scientia Silvae Sinicae ›› 2009, Vol. 12 ›› Issue (10): 49-54.doi: 10.11707/j.1001-7488.20091009
Previous Articles Next Articles
Zhai Xiaoqiao1,2,Cheng Fei1,Zhu Yanlin2
Received:
Revised:
Online:
Published:
Supported by:
Abstract:
Robinia bella-rosea calli were initiated from the leaves on MS medium supplemented with 5 mg·L-1 6-BA. The calli were cooled down at 4 ℃ for 2 h in a cryoprotectant (MS+10%DMSO+0.5 mol·L-1 Sucrose), and further refrigerated to -7 ℃ at a rate of 1 ℃·min-1 by using a programmed temperature-changing freezer. After one hour at -7 ℃ for 1 h, the calli in the cryoprotectant were further refrigerated to -20 ℃ at a rate of 0.1 ℃·min-1. Being incubated for another hour at -20 ℃, the calli were cooled down to -40 ℃ at a rate of 0.3 ℃·min-1 and then the calli were transferred and preserved in liquid nitrogen for a day. The low temperature treated calli were thawed at 38 ℃ water bath, and the thawed calli were washed with liquid MS medium with 6-BA 5 mg·L-1 and 30 g·L-1 sucrose, then transferred to solid medium and cultured in darkness. Two weeks later, the calli were cultured in light, and showed able to produce fresh and green new callus after 3 d, and the survival rate of the low temperature treated calli reached to 52%. New-produced calli were able to regenerate plantlets.
Key words: Robinia bella-rosea, callus, cryopreservation, freezing method
CLC Number:
 
Zhai Xiaoqiao;Cheng Fei;Zhu Yanlin. Methods of Lowering Temperature for Cryopreservation of Calli of Robinia bella-rosea[J]. Scientia Silvae Sinicae, 2009, 12(10): 49-54.
0 / / Recommend
Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks
URL: http://www.linyekexue.net/EN/10.11707/j.1001-7488.20091009
http://www.linyekexue.net/EN/Y2009/V12/I10/49