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

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:1~2 mm的茎尖在0.5 mol/L蔗糖的预培养基预培养3~7 d,室温装载处理60 min,0 ℃玻璃化处理180 min,液氮冷冻60 min,40 ℃解冻2 min,高糖溶液卸载20 min后接种到MS+2.0 mg/L 6-BA再生培养基上,成活率可达70%以上。再生培养60 d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(0.1~0.3 mm),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 3 to 7 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 2 min 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 than 70%.Twentyregeneration plants were chosen to test the effect of SMoV elimination.The result showed that elimination rate of SMoV reached 100%.