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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 type of highly effective detoxification technology developed in recent years. Its detoxification rate is significantly higher than the traditional detoxification methods. In this study, the stem tip length and the key processes (pre-culture and pre-treatment) of the super-hypothermia therapy were studied with strawberry stem virus infected strawberry strawberry virus, strawberry crimple virus, strawberry mosaic virus and Strawberry light yellow edge virus Screening and optimization, in order to establish a perfect strawberry cryogenic therapy detoxification technology system. The results show that in the virus removal process, the tip length has no effect on the virus-free rate, but significantly affect the survival rate. The optimized cryopreservation system was as follows: pre-cultured sucrose at 0.3 mol / L and dark culture at 7 days; 60% PVS2 at room temperature for 60 min; 100% PVS2 vitrification at 0 ℃ for 60 min; ~ 40 ℃ water bath for 2 min. Under this system, the survival rate of strawberry tip was 68.89%. However, the strawberry shrunken virus, strawberry mottle virus, strawberry vein vein virus and strawberry light yellow edge virus were simultaneously removed and the removal rate was 100% Strawberry virus-free seedling cultivation provides an important reference.