豫杂一号泡桐茎尖的玻璃化法超低温保存方法研究  

Conservation of Stem Tip Tissue of Paulownia tomentosa×P.fortunei with Vitrification Cryopreservation

作　　者：翟晓巧[1,2] 程斐[2] 张晓申[2] 

机构地区：[1]河南省林业科学研究院,河南郑州450008 [2]河南农业大学,河南郑州450002

出　　处：《西部林业科学》2011年第3期1-5,共5页Journal of West China Forestry Science

基　　金：中国博士后科学基金(20070410874)

摘　　要：以豫杂一号泡桐健壮组培苗为研究材料,从低温锻炼时间、预处理时间、装载液处理时间、玻璃化液处理时间及液氮保存时间5个方面研究其茎尖玻璃化法超低温保存方法。结果表明,豫杂一号泡桐茎尖玻璃化法超低温适宜的保存方法为,2 cm的茎尖在附加0.4 mol·L-1甘油和0.3 mol·L-1蔗糖的MS培养基上预培养2天,然后剥取2～3 mm的茎尖,用装载液(MS+2 mol·L-1甘油+0.4 mol·L-1蔗糖)装载60 min,再用玻璃化溶液(PVS2)于0℃下处理1.5 h,换1次玻璃化溶液后迅速投入液氮,保存l h后于40℃水浴中化冻70 s,再转到室温下用去装载液(MS+1.2 mol·L-1蔗糖)洗涤2次,每次10 min。以TTC法检测,豫杂一号泡桐茎尖成活率最高可达85.74%,保存效果较好。The tissue of stem tip of Paulownia tomentosa×P.fortunei was used to study its cryopreservation method in duration of exercise of it in low temperatures,duration of pre-disposal,duration of loading solution,disposal duration of vitrified solution and duration of it in liquid nitrogen.It showed that proper disposal of that would be by putting the tissue of 2cm long on the surface of MS culture with additional 0.4 mol·L-1 glycerol and 0.3 mol·L-1 sucrose,cultivated for 2 days,then taking the tissue of about 2~3 cm into loading solution（MS＋2 mol·L-1 glycerol＋0.4 mol·L-1 sucrose） for 60 minutes,then in PVS2 for 1.5 h under 0℃,after changing the solution once more and immediately put it into liquid nitrogen for 1 h then thawing in 40 ℃ water both for 70 seconds.Washing it 2 times with de-loading solution（MS＋1.2 mol·L-1 sucrose）,each lasting for 10 min.by this method,the tissue＇s survival rate reached 85.74% after detecting it with TTC method.

关 键 词：豫杂一号泡桐 茎尖 超低温保存 玻璃化法 成活率 

分 类 号：S792.43[农业科学—林木遗传育种]