软枣猕猴桃休眠芽超低温保存技术研究  

Study on the cryopreservation of dormant buds by vitrification in Actinidia arguta ‘Kuilü’

作　　者：白晓雪 秦红艳[1] 韩先焱 杨义明[1] 范书田[1] 路文鹏[1] 李昌禹[1] BAI Xiaoxue;QIN Hongyan;HAN Xianyan;YANG Yiming;FAN Shutian;LUWenpeng;LI Changyu(Institute of Special Wild Economic Animals and Plants,Chinese Academy of Agricultural Sciences,Changchun 130112,Jilin,China)

机构地区：[1]中国农业科学院特产研究所,长春130112

出　　处：《果树学报》2020年第8期1247-1255,共9页Journal of Fruit Science

基　　金：吉林省科技发展计划项目(20170203006NY);中国农业科学院科技创新工程(201409)。

摘　　要：【目的】探讨软枣猕猴桃种质资源的长期保存技术体系。【方法】以软枣猕猴桃‘魁绿’休眠芽为试材,研究了预培养液种类、预培养时间、装载时间、玻璃化保护液、恢复培养基等因素对玻璃化法超低温保存后休眠芽成活率的影响。【结果】休眠芽在0.3 mol·L^-1蔗糖+1 mol·L^-1甘油的预培养液中震荡培养2 d,常温下用装载液(2 mol·L^-1甘油+0.4mol·L^-1蔗糖+MS)处理20 min,0℃条件下用PVS2(30%甘油+15%乙二醇+15%二甲基亚砜+0.4 mol·L^-1蔗糖+MS)脱水120 min,换新鲜PVS2后迅速投入液氮中冻存24 h以上。取出后立即用38℃水浴解冻2 min,用含1.2 mol·L^-1蔗糖的MS卸载液洗涤30 min,无菌滤纸吸干后接种到MS+2 mg·L^-16-BA+0.02 mg·L^-1NAA恢复培养基上,休眠芽成活率达86.30%。用流式细胞仪鉴定再生植株倍性,没有发现明显变化。【结论】建立了高效的软枣猕猴桃‘魁绿’休眠芽玻璃化超低温保存体系。【Objective】Actinidia arguta is a precious resource of cold-resistant fruit tree in China.It is extremely resistant,and rich in nutrientsand vitamin C,so it is of important nutritional and economic values.A.arguta is one of the important utilization values in the Actinidia and an important germplasm resource for variety improvement.Due to the increasing commercialization and large-scale production of A.arguta,resulting in a single variety and the disappearance of many excellent genetic resources,the research on ultra-low temperature preservation technology for kiwifruit germplasm resources is of great significance.The tara vine’Kuilü’is an excellent variety that has been bred by the Institute of Special Wild Economic Animals and Plants of the Chinese Academy of Agricultural Sciences for more than 10 years.Using’Kuilü’as a test material,the objective of this project is to study the ultra-low temperature preservation method,so as to provide a reference for the long-term preservation of kiwifruit germplasm resources.【Methods】The’Kuilü’dormant branches were cut into single-bud stem sections with a length of about 2 cm.Firstly,the single-bud stem sections were peeled,then soaked in 75%alcohol for30 s,and finally,sterilized by shaking with 0.1%HgCl2 for 30 min.The dormant buds were peeled for ultra-low temperature preservation by vitrification.Basic procedure of dormant bud vitrification cryopreservation was as follows:the dormant buds soaked in preculture fluid containing 0.3 mol·L^-1 sucrose+1 mol·L^-1 glycerol+MS were shaken and cultured for 2 d,and thereafter they were transferred into 2 mL cryopreservation tubes under sterile conditions,with 10 buds per tube,and repeated 3 times.Loading was made at room temperature for 20 min with loading solution(2 mol·L^-1 glycerol+0.4 mol·L^-1 sucrose+MS),dehydration was conducted with PVS2 for 120 min at 0℃,and then immediately put into liquid nitrogen to freeze for more than 24 h after they were changed to fresh PVS2.The tubs were taken out,and tha

关 键 词：软枣猕猴桃 ‘魁绿’ 休眠芽 玻璃化法 超低温保存 

分 类 号：S663.4[农业科学—果树学]