摘要
超低温疗法是近年发展起来的一种新型高效的脱毒技术,其脱毒率显著高于传统的脱毒方法。本研究以感染草莓斑驳病毒、草莓皱缩病毒、草莓镶脉病毒和草莓轻型黄边病毒的红颜草莓茎尖为试材,对超低温疗法中茎尖长度以及关键程序(预培养和预处理)进行筛选和优化,以期建立起完善的草莓超低温疗法脱毒技术体系。研究结果表明,在病毒脱除过程中,茎尖长度对脱毒率没有影响,然而却显著影响成活率。优化的超低温保存体系为:预培养蔗糖浓度为0.3 mol/L,暗培养7 d;室温60%PVS2装载60 min;0℃100%PVS2玻璃化处理60 min;液氮处理1d后,于38℃~40℃水浴2 min。在此体系下,草莓的茎尖存活率为68.89%,但草莓皱缩病毒、草莓斑驳病毒、草莓镶脉病毒和草莓轻型黄边病毒被同时去除,脱除率为100%,上述研究结果将为草莓脱毒苗的培育提供重要参考。
Cryotherapy is a new and efficient technology for virus elimination in recent years, which detoxification rate is higher than traditional method. In order to establish the cryotherapy technical system of strawberry, the length of shoot tips and key procedure(pre-culture and pre-treatment) of cryotherapy were selected and optimized by using the shoot tips of Fragariaananassa cv. Benihoppe infected with strawberry mottle virus,strawberry crinkle virus,strawberry vein banding virus and strawberry mild yellow edge virus as the experiment material. The results showed that the length of shoot tip have no effect on virus elimination rate, but have significant effect on survival rate. The optimized cryotherapy system was that the shoot tips were pre-cultured on basic medium with 0.3 mol/L sucrose for 7 days in darkness, then loaded for 60 min at room temperature, vitrified with 100% PVS2 for 60 min at 0觷C, immersed directly in liquid nitrogen for 1 day and rewarmed in 38℃~40觷C for2 min. After the above procedure, the main four virus of strawberry were totally eliminated, and the survival rate was 68.89%. These findings will provide important reference for breeding of virus-free strawberry seedings.
出处
《分子植物育种》
CAS
CSCD
北大核心
2016年第9期2488-2494,共7页
Molecular Plant Breeding
基金
教育部留学回国人员科研基金
四川农业大学双支计划(03570109)共同资助
关键词
草莓
病毒
超低温冷冻保存
脱毒
Strawberry
Virus
Cryotherapy
Virus elimination
