红掌小滴玻璃化法超低温保存研究  

Cryopreservation of Anthurium andraeanum by Droplet Vitrification

作　　者：高洁 李志英 张玄兵[1] 谢龙海 陈莹 朱振芬 符运柳 徐立 GAO Jie;LI Zhiying;ZHANG Xuanbing;XIE Longhai;CHEN Ying;ZHU Zhenfen;FU Yunliu;XU Li(Hainan University,Haikou,Hainan 570228,China;Institute of Tropical Crop Genetic Resources,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan 571101,China;Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China,Beijing 100000,China;Key Laboratory of Tropical Crops Germplasm Resources Genetic Improvement and In-novation of Hainan Province,Danzhou,Hainan 571737,China;National Gene Bank of Tropical Crops,Danzhou,Hainan 571737,China)

机构地区：[1]海南大学,海南海口570228 [2]中国热带农业科学院热带作物品种资源研究所,海南海口571101 [3]华南作物基因资源与种质创制重点实验室,北京100000 [4]海南省热带作物资源遗传改良与创新重点实验室,海南儋州571737 [5]国家热带作物中期库,海南儋州571737

出　　处：《热带作物学报》2023年第9期1909-1916,共8页Chinese Journal of Tropical Crops

基　　金：中央级科研院所基本科研业务费专项(No.1630032019039,No.1630032020026)。

摘　　要：以红掌腋芽为试材,探讨小滴玻璃化法对红掌超低温保存的影响因素,并对再生植株进行遗传稳定性检测。结果表明,红掌腋芽置于含0.4 mmol/L蔗糖和2 mmol/L甘油的MS固体培养基中预培养4 d后,在0℃下80%PVS_(2)处理50 min,转到铝箔条上PVS_(2)液滴中,将铝箔条迅速浸入液氮中,2 s后直接转入装满液氮的冻存管中,投入液氮至少保持30 min;室温下用含有1.2 mmol/L蔗糖的MS液体培养基复温并卸载20 min后置于恢复培养基上,腋芽存活率高达63.70%。通过ISSR和SSR分子标记检测,再生植株的遗传稳定性未发生改变。该研究结果为红掌种质资源的长期保存提供有效途径。Using the in vitro axillary bud of Anthurium(Anthurium andraeanum Lind.)as the materials,this article re-ports the influence of the droplet vitrification on Anthurium after cryopreservation,and plants regenerated tissues ge-netic stability were assessment.The results showed the highest survival rate of Anthurium(63.70%)was achieved after preculture for 4 days on MS solid medium containing 0.4 mmol/L sucrose and 2 mmol/L glycerol,dehydration with 80%PVS_(2)at 0℃for 50 min,cooling in droplets of PVS_(2)placed on aluminum foil strips,after immersing in liquid nitrogen for 2 s,directly transferred it to a cryotube filled with liquid nitrogen,and kept it in liquid nitrogen for at least 30 min after rewarming them with MS liquid medium containing 1.2 mmol/L sucrose at room temperature for 20 minutes,transferred to recovery medium.Through ISSR and SSR molecular marker detection,the genetic stability of regenerated plants did not changed.The results would provide an effective way for long-term preservation of Anthurium an-draeanum germplasm resources.

关 键 词：红掌 小滴玻璃化法 超低温保存 腋芽 遗传稳定性 

分 类 号：S682.14[农业科学—观赏园艺]