摘要
研究月季茎尖玻璃化法超低温保存技术，以期为月季茎尖超低温保存提供理论和实践依据。试验选用金奖章、粉玛雅、韩红、博共4个月季品种进行简单玻璃化法和玻璃化超低温保存试验：结果表明：简单玻璃化法保存时，不同品种的茎尖存活率差异显著，金奖章的茎尖存活率最高（55．3％）；通过研究预培养、装载处理和植物玻璃化溶液PVS，脱水时间对粉玛雅茎尖存活率的影响，初步构建了月季茎尖玻璃化法超低温保存体系，即将长1．0cm的茎尖在含0．3mol·L^-1蔗糖的培养基上预培养5～6d后．取1．5mm茎尖，室温下A液（MS＋2mol·L^-1甘油＋0．4mol·L^-1蔗糖）装载40min或B液[60％PVS2＋40％（MS＋0．15mol·L^-1蔗糖）]装载30min，用PVS2（300g·L^-1甘油＋150g·L^-1乙二醇＋150g·L^-1二甲基亚砜＋0．4mol·L^-1蔗糖十MS）于0℃下处理40min，然后液氮保存24h．在40℃水浴中化冻90s，用1．2mol·L^-1蔗糖培养液洗涤，最后转入MS＋6-BA1．0mol·L-^-1＋IBA0．1mol·L^-1。上再培养，存活率高于53.3％。
The cryopreservation technique of rose shoot tips by vitrification was studied to provide theoretical and practical basis for the cryopreservation of rose shoot tips.Four rose cultivars, Gold Medal, Pink Maya,Han Hong and Bo were selected for cryopreservation by the simple vitrification and vitrification.The simple vitrification method made significant difference in vitro shoot tips of different rose cultivars.The survival rate of Golden Medal＇s shoot tips was the highest, 55.3%. The effects of preculture, loading and plant vitrification solution 2 treatment time on survival rate were tested on Pink Maya＇s shoot tips. A method of the vitrification cryopreservation procedure of rose shoot tips was constructed. Shoot tips about lcm in length were precultured for 5-6d on MS medium supplemented with 0.3mol·L^-1 sucrose. Dissected shoot apices about 1.5mm in length were loaded with solution A（MS＋2mol·L^-1 glycerol＋0.4mol·L^-1 sucrose） for 30min or solution B[60%PVS2＋40%（MS＋0.15 mol· L^-1sucrose）] for 40min at room temperature.These excised shoot tips were sufficiently dehydrated in a highly concentrated plant vitrification solution （PVS.〈300g·L^-1 glycerol＋150g·L^-1 ethylene glycol＋150g·L^-1dimethyl sulfoxide＋0.4mol· L^-1 sucrose＋MS） for 40rain at 0℃ and then plunged into liquid nitrogen and conserved for 24h. After rapid thawed in water at 40℃ for 90s, the tips were rinsed with 1.2mol·L^-1 sucrose solution and then plated on MS medium supplemented with 1.0mol·L^-1 6-BA and 0.1mol·L^-1 IBA. The survival rate of shoot tips reached 53.3%.
出处
《沈阳农业大学学报》
CAS
CSCD
北大核心
2009年第2期156-159,共4页
Journal of Shenyang Agricultural University
关键词
月季
茎尖
超低温保存
玻璃化法
存活率
rose
shoot tips： cryopreservation
vitrification
survival rate
