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当前位置: 首页 > 文章 > 莪术茎尖玻璃化法超低温保存技术 热带农业科学 2018,38 (4) 52-57
Position: Home > Articles > Vitrification Cryopreservation of Shoot-tips of Curcuma zedoaria Chinese Journal of Tropical Agriculture 2018,38 (4) 52-57

莪术茎尖玻璃化法超低温保存技术

作  者：
吴怡;顾雅坤;符丽;曾琳
关键词：
莪术;茎尖;种质资源;玻璃化法;超低温保存
摘  要：
为莪术种质资源稳定保存提供一条新途径.以莪术组培苗茎尖为试材,用玻璃化法进行超低温保存后,于25℃用0.1% TTC溶液培养24 h后检测生活力.结果表明,切取2~3 mm长、 继代培养60 d的莪术组培苗茎尖,避光条件下,用含0.3 mol/L蔗糖的MS固体培养基预培养7 d,室温下,茎尖用60%的PVS2装载液装载10 min, 0℃用100%的PVS2玻璃化保护液对茎尖脱水处理30 min,快速投入液氮中进行超低温保存16 h;取出茎尖于40℃水浴中解冻2 min,室温下用US溶液洗涤20 min,立即用0.1% TTC溶液于25℃人工气候培养箱中培养24 h,莪术茎尖的生活力最高可达100%.表明该方法可用于莪术种质资源的保存.
译  名：
Vitrification Cryopreservation of Shoot-tips of Curcuma zedoaria
关键词：
Curcuma zedoaria%shoot-tips%germplasm resources%vitrification%cryopreservation
摘  要：
Shoot tips of tissue cultured Curcuma zedoaria were subcultured,vitrified and cryopreserved to get a new approach for stable conservation of C. zedoaria. The test-tube plantlets of C. zedoaria were subcultured for 60 d, and their shoot tips were cut 2~3 mm long and cultured in the MS solid medium supplemented with 0.3 mol/L sucrose for 7 days in the dark. Then the shoot tips were loaded with 60% PVS2 for 10 min at 25℃, dehydrated for 30 min at 0℃ with 100% PVS2 as vitrification protection solution and placed immediately into liquid nitrogen for cryopreservation for 16 h. The cryopreserved shoot tips were thawed in aqueous bath for 2 min at 40℃,and rinsed with US solution for 20 min at the room temperature. The survival rate of the shoot tips was measured by using 0.1% TTC solution and the highest survival rate was up to 100.00%.This indicates that the vitrified cryopreservation may be used for preservation of germplasm resources of C.zedoaria.

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