摘要
使用包埋-玻璃化法对两种来源的高山红景天茎尖进行超低温保存研究，探讨蔗糖预处理、包埋过程中渗透处理和不同温度下玻璃化液PVS2处理的时间对超低温保存后茎尖存活率的影响．结果表明，茎尖依次在0．3、0．5、0．7、1．0mol·L^-1的蔗糖溶液中各预培养1d后，转入1．0mol·L^-1的蔗糖溶液中继续培养4d，然后使用附加2mol·L^-1甘油和1mol·L^-1蔗糖的包埋液渗透处理60min，包埋珠在0℃处理180～210min后投入液氮，48h后用40℃水浴快速化冻3min，用含有1mol·L。蔗糖的改良MS培养液洗涤20min，转入MS＋6-BA1mg·L^-1＋NAAO．1mg·L^-1固体培养基中在22℃条件下进行暗培养，2d后转移入光照培养，最高成活率接近100％，再生植株生长和分化正常．
Shoot tips excised from two resources of Rhodiola sachalinensis A. Bor were cryopreserved by encapsulation-vitrification, and effects of sucrose preculture duration, osmoprotection time during encapsulation and exposure time to PVS2 on survival rate of cryopreserved shoot tips of Rhodiola sachalinensis A. Bor were discussed. Excised shoot tips were precultured with increasing sucrose concentrations of 0. 3, 0. 5, 0.7 and 1.0 mol. L 1 for 1 day respectively, then transferred to 1 mol. L 1 sucrose for 4 days, and precuhured shoot tips were encapsulated and simultaneously osmoprotected with a loading solution of 2 mol. L 1 glycerol and 1 mol·L^-1 sucrose for 60 min. Osmoprotected and encapsulated shoot tips were dehydrated with a highly concentrated vitrification solution for 180 to 210 min at 0 ℃ prior to direct immersion in liquid nitrogen for 48 h. After rapid warming in water for 3 min at 40℃, the shoot tips were rinsed with the modified MS medium containing 1 mol·L^-1 sucrose for 20 min, and then cultured on the MS medium containing 6-BA 1 mg·L^-1 and NAA 0.1 mg·L^-1 in dark prior to exposure to the light at 22℃. The best survival rate of encapsulation vitrified shoot tips amounted to nearly 100% and no abnormality was observed.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2007年第3期265-270,共6页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
黑龙江省政府博士后基金资助项目(LBH-Z05014)
关键词
高山红景天
茎尖
包埋-玻璃化法
超低温保存
Rhodiola sachalinensis A. Bor
shoot tips
encapsulation-vitrification
cryopreservation
