欢迎访问林业科学,今天是

林业科学 ›› 2007, Vol. 43 ›› Issue (06): 57-60.doi: 10.11707/j.1001-7488.20070610

• 论文及研究报告 • 上一篇    下一篇

高山红景天愈伤组织的超低温保存

刘剑锋1 阎秀峰2 程云清1 李霞1   

  1. 1.吉林师范大学生命科学学院,四平136000;2.东北林业大学生命科学学院,哈尔滨150040
  • 收稿日期:2006-04-20 修回日期:1900-01-01 出版日期:2007-06-25 发布日期:2007-06-25

Cryopreservation of Callus of Rhodiola sachalinensis

Liu Jianfeng1,Yan Xiufeng2,Cheng Yunqing1,Li Xia1   

  1. 1. College of Life Sciences, Jilin Normal University Siping 136000; 2. College of Life Sciences, Northeast Forestry University Harbin 150040
  • Received:2006-04-20 Revised:1900-01-01 Online:2007-06-25 Published:2007-06-25

摘要:

高山红景天叶片在附加6-BA 2mg·L-1和NAA 0.5mg·L-1的改良MS培养基中诱导出愈伤组织,将愈伤组织放入10% DMSO+0.5 mol·L-1山梨醇的冰冻保护剂中,4℃冰箱中放置8h,然后以1℃.min-1的降温速度降至-40℃,停留2h后投入液氮保存,在40℃水浴中化冻。冻存后的愈伤组织用液体培养基MS+6-BA 2mg·L-1+NAA 0.5mg·L-1反复冲洗后,转入固体培养基中进行暗培养,2周后转入光照(2000 lx)培养。5周后观察到高山红景天愈伤组织呈鲜绿色,长势较为旺盛,愈伤组织存活率达到75.64%。冻存后的愈伤组织可诱导成苗。

关键词: 高山红景天, 愈伤组织, 植株再生

Abstract:

Rhodiola sachalinensis calli were initiated from leaves on modified MS medium supplemented with 2 mg·L-1 6-BA and 0.5 mg·L-1 NAA. The calli were cooled at 4 ℃ for 8 h after treated with cryoprotectants (10%DMSO+0.5 mol·L-1 sorbitol), and further refrigerated at a rate of 1 ℃·min-1 to-40 ℃, and maintained at-40 ℃ for at least 2 h. The calli then were preserved in liquid nitrogen and thawed at 40 ℃ water bath. Calli were washed with liquid MS medium with 6-BA 2 mg·L-1 and NAA 0.5 mg·L-1 for several times, transferred to solid medium and cultured in darkness. Two weeks later, the calli were cultured in light (2 000 lx). The calli of R. sachalinensis showing vigorous growth were fresh and green after 5 weeks, and the survival rate reached 75.64%. Calli of R. sachalinensis could regenerate to plantlets.

Key words: Rhodiola sachalinensis, callus, plantlet regeneration