【摘 要】
：
超低温保存(-196℃)是植物种质资源保存最安全经济的技术,本研究初步建立了崇明水仙的超低温保存流程:(1)取崇明水仙的3～4年生鳞茎,用70％酒精消毒1min,0.1％的升汞消毒15 min,无菌水冲洗若干次后,将带3～4层鳞片及部分茎盘的鳞茎切块接种于含BA及NAA的MS培养基上诱导出小鳞茎;(2)将继代3个月崇明水仙的鳞茎诱导后,切取2～3 mm大小的茎尖,(3)接种到含0.3 mol·L-
【机 构】
：
上海市农业生物基因中心,上海闵行区北翟路2901号,201106 上海市农业科学院林木果树研究所,
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超低温保存(-196℃)是植物种质资源保存最安全经济的技术,本研究初步建立了崇明水仙的超低温保存流程:(1)取崇明水仙的3～4年生鳞茎,用70％酒精消毒1min,0.1％的升汞消毒15 min,无菌水冲洗若干次后,将带3～4层鳞片及部分茎盘的鳞茎切块接种于含BA及NAA的MS培养基上诱导出小鳞茎;(2)将继代3个月崇明水仙的鳞茎诱导后,切取2～3 mm大小的茎尖,(3)接种到含0.3 mol·L-1蔗糖的MS培养基内,25℃下培养3～5 d,(4)放于预处理溶液中于室温下处理30min后,(5)转入玻璃化保护剂中进行70～90min的冰浴处理,(6)再将茎尖转移至装有预冷过的新鲜玻璃化保护剂的冷冻管中,8～10个茎尖/管,迅速投入液氮中保存;(7)液氮保存24h后,从液氮中取出冻存的冷冻管,立即放入40℃水浴中快速解冻2min,(8)用MS+1.2 mol·L-1蔗糖液体培养基洗涤20min,(9)滤纸吸干后接种到恢复培养基中,(10)在25℃下暗培养7d后转入正常光照培养,30d后再生率可达50％以上.本研究建立崇明水仙的超低温保存方法对有效保护崇明水仙种质资源,探索快速、有效、稳定的崇明水仙离体脱毒方法,提纯复壮、加速产业化进程具有现实意义,对其他鳞茎和球宿根类植物遗传资源的超低温保存来说,具有较好的参考价值.
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