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采用包埋-玻璃化法对高山红景天茎尖进行了超低温保存研究,研究了蔗糖预处理、包埋过程中渗透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,茎尖在0.3,0.5,0.7,1.0mol.L-1的蔗糖溶液中各预培养1 d后,转入1.0 mol.L-1的蔗糖溶液中再培养4 d,使用含2 mol.L-1甘油和1mol.L-1蔗糖的包埋液渗透处理60 min,包埋珠在0℃处理180至210分钟后投入液氮,投入48 h后用40℃水浴快速化冻3 min,用含有1 mol.L-1蔗糖的改良MS培养液洗涤20 min,转入MS+6-BA1mg.L-1+NAA0.1 mg.L-1固体培养基中在22℃条件下进行暗培养,2 d后转入光照培养,最高成活率接近100%,再生植株生长和分化正常.
Cryopreservation of Rhodiola sachalinensis was carried out by embedding-vitrification method. The effect of PVS2 treatment time on vitrification solution infiltration and different temperature during sucrose pretreatment and embedding process was studied. The survival rate of shoot tip after cryopreservation Impact. The results showed that the stem tips were preincubated for 1 day in 0.3,0.5,0.7,1.0mol.L-1 sucrose solution, then transferred into 1.0 mol.L-1 sucrose solution for further 4 days, L-1 glycerol and 1mol.L-1 sucrose for 60 min. The embedded beads were treated with liquid nitrogen for 180-210 minutes at 0 ℃. After 48 hours, the embedded beads were rapidly frozen in a 40 ℃ water bath for 3 min, Washed with modified MS medium containing 1 mol.L-1 sucrose for 20 min and transferred to MS + 6-BA1mg.L-1 + NAA0.1mg.L-1 solid medium for dark culture at 22 ℃ After 2 d, the cells were transferred to light and the highest survival rate was close to 100%. The regenerated plants grew normally and their differentiation was normal.