果树学报2012，Vol.29Issue(6)：1069-1073,I0002,6.
刺梨茎尖的玻璃化超低温保存及植株再生
Cryopreservation of in vitro Rosa roxburghii shoot-tips by vitrification and its regeneration
摘要
Abstract
【Objective】This research aims at developing a cryopreservation system of in vitro Rosa roxburghii Tratt shoot-tips,and thereby establishing a foundation for long-term and stable preservation of Rosa roxburghii Tratt germplasm resources.【Method】Taking Rosa roxburghii Tratt as materials,factors for cryopreservation of in vitro Rosa roxburghii Tratt shoot-tips such as sucrose preculture,LS pretreatment time,and PVS2 treatment time were investigated.【Result】 The suitable medium for bud break and growth was MS + 0.5 mg·L-1 6-BA + 30 g·L-1 sucrose + 7 g·L-1 agar.A system of cryopreservation for in vitro Rosa roxburghii Tratt shoot-tips was established: the axillary shoot tips which were about 0.5 cm in length were precultured on MS medium containing 0.5 mg·L-1 6-BA,0.05 mg·L-1 IAA,0.3 mol·L-1 sucrose and 7.0 g·L-1 agar for 3 days at 4 ℃.The shoot tips which were about 2 mm in length were cut and loaded in a solution containing 2 mol·L-1 glycerol and 0.4 mol·L-1 sucrose for 40 to 50 minutes at room temperature.The shoot tips were sufficiently dehydrated with vitrification solution for 40 min at 0 ℃.Then the shoot tips were transferred into the tube with fresh PVS2 and immediately put into liquid nitrogen.24 h later,the tips were thawed in a water bath at 40 ℃,and then rinsed twice with MS medium containg 1.2 mol·L-1 sucrose,100 mg·L-1 Vc and 0.5 mg·L-1 GA3.Then the shoot tips were plated on MS medium supplemented with 0.3 mol·L-1 sucrose under darkness for 1 day and subcultured on MS medium supplemented with 0.1 mg·L-1 6-BA,0.05 mg·L-1 IAA,30 g·L-1 sucrose and 7 g·L-1 agar under darkness for one week prior to exposure to the light.The survival rate of shoot-tips after cryopreservation was 44.3%.【Conclusion】 Cryopreservation system of in Vitro Rosa roxburghii Tratt Shoot-tips was developed successfully.关键词
刺梨/茎尖/超低温保存/玻璃化法Key words
Rosa roxburghii Tratt/Shoot-tips/Cryopreservation/Vitrification分类
农业科技引用本文复制引用
陈红,程秀枝,林田..刺梨茎尖的玻璃化超低温保存及植株再生[J].果树学报,2012,29(6):1069-1073,I0002,6.基金项目
贵州省科技攻关项目 （）
