利用玻璃化超低温技术脱除草莓斑驳病毒(SMoV)的初步研究  

Preliminary study on elimination of Strawberry mottle virus(SMoV)from shoot tips by vitrification-cryopreservation treatment

作　　者：盛宏亚 万继花 徐川[1] 王红清[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193

出　　处：《中国农业大学学报》2016年第3期53-57,共5页Journal of China Agricultural University

基　　金：科技部科技支撑计划(213BAD201301);国家"863"计划(2013AA103002)

摘　　要：以携带草莓斑驳病毒（Strawberry mottle virus）的‘红颜’草莓为材料,通过对玻璃化超低温脱毒技术中各程序的优化,建立适合‘红颜’草莓的茎尖玻璃化超低温脱毒体系。结果表明：1~2mm的茎尖在0.5mol/L蔗糖的预培养基预培养3~7d,室温装载处理60min,0℃玻璃化处理180min,液氮冷冻60min,40℃解冻2min,高糖溶液卸载20min后接种到MS＋2.0mg/L 6-BA再生培养基上,成活率可达70%以上。再生培养60d后,随机选取20株再生植株,利用RT-PCR技术检测草莓斑驳病毒的脱毒效果,脱毒率为100%。利用该体系可以克服传统茎尖脱毒受茎尖大小的限制（0.1~0.3mm）,并有效脱除草莓斑驳病毒。Strawberry can be infected by virus easily,which lead to yield and quality losses.‘Benihoppe＇strawberry（Fragaria×ananassaDuch）infected with strawberry mottle virus was used as materials.By optimizing the parameters of vitrification-cryopreservation,appropriate cryopreserved detoxification of Benihoppe shoot tips was established.The results indicated：The shoot tips（1-2 mm）were pre-cultured on basic medium with 0.5 mol/L sucrosef for 3to7 days,then the pre-cultured shoot tips were loaded for 60 min at 25 ℃ and vitrified for 180 min at 0 ℃,immersed in liquid nitrogen for 1h,and rewarmed in water for 2min at 40 ℃.After uninstalling in high sugar solution for 20 min,the shoot tips were inoculated into regeneration medium.In the end,the percentage of survived shoot tips was more than70%.Twentyregeneration plants were chosen to test the effect of SMoV elimination.The result showed that elimination rate of SMoV reached 100%.

关 键 词：草莓 超低温 脱毒 草莓斑驳病毒 病毒检测 

分 类 号：S668.4[农业科学—果树学]