扁桃小滴玻璃化超低温保存研究A study on the cryopreservation of almond buds by droplet-vitrification
姜喜;赵书珍;党艳青;张琦;焦培培;庞新安;陈家力;
摘要(Abstract):
【目的】以扁桃休眠茎尖为试材,探讨小滴玻璃化法对扁桃离体超低温保存的影响,为扁桃种质资源的长期保存提供有效途径。【方法】采用小滴玻璃化法,通过正交设计试验和单因素试验对预培养蔗糖浓度、预培养时间、装载时间和PVS2[30%(ω,后同)甘油+15%二甲基亚砜+15%乙二醇+0.4 mol·L(-1)蔗糖]脱水处理时间这4个因素影响扁桃休眠茎尖超低温保存后成活率的情况进行分析。【结果】预培养时间对休眠茎尖超低温保存后成活率的影响极显著(P<0.01),而预培养蔗糖浓度、装载时间和PVS2处理时间的影响不显著。建立了以‘莎车14号’为代表的扁桃茎尖小滴玻璃化法超低温保存体系,最佳处理方案是:用0.5 mol·L(-1)蔗糖]脱水处理时间这4个因素影响扁桃休眠茎尖超低温保存后成活率的情况进行分析。【结果】预培养时间对休眠茎尖超低温保存后成活率的影响极显著(P<0.01),而预培养蔗糖浓度、装载时间和PVS2处理时间的影响不显著。建立了以‘莎车14号’为代表的扁桃茎尖小滴玻璃化法超低温保存体系,最佳处理方案是:用0.5 mol·L(-1)预培养蔗糖液处理2 d,装载液处理20 min,PVS2脱水处理120min,放入含新鲜PVS2小滴的铝箔纸上投入液氮24 h后,浸入用40℃温水预热过的洗涤液(1.2 mol·L(-1)预培养蔗糖液处理2 d,装载液处理20 min,PVS2脱水处理120min,放入含新鲜PVS2小滴的铝箔纸上投入液氮24 h后,浸入用40℃温水预热过的洗涤液(1.2 mol·L(-1)蔗糖的MS)中30 s,然后再将休眠茎尖转入新鲜的洗涤液中洗涤30 min。恢复后接种在MS+0.3 mg·L(-1)蔗糖的MS)中30 s,然后再将休眠茎尖转入新鲜的洗涤液中洗涤30 min。恢复后接种在MS+0.3 mg·L(-1)6-BA+0.5 mg·L(-1)6-BA+0.5 mg·L(-1)IAA+0.3 mg·L(-1)IAA+0.3 mg·L(-1)GA3培养基上,扁桃成活率达80.44%。运用优化的方案对‘莎车1号’‘莎车11号’和‘莎车18号’3个扁桃品种进行超低温保存,平均成活率分别为87.41%、85.44%和83.02%。【结论】利用小滴玻璃化法可提高扁桃超低温保存成活率,建立了适合扁桃休眠芽超低温保存技术方法,为扁桃种质资源的长期保存提供理论和实践借鉴。
关键词(KeyWords): 扁桃;超低温保存;小滴玻璃化;休眠茎尖
基金项目(Foundation): 新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室开放项目(BRYB1504)
作者(Authors): 姜喜;赵书珍;党艳青;张琦;焦培培;庞新安;陈家力;
DOI: 10.13925/j.cnki.gsxb.20170345
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