杂交兰种苗超低温脱毒技术研究

以携带建兰花叶病毒(Cy MV)和齿兰环斑病毒(ORSV)的杂交兰根状茎为试材,采用包埋玻璃化超低温保存法对杂交兰病毒脱除进行研究。结果表明:低温炼苗21 d的茎尖,在0.5 mol/L的蔗糖浓度预培养基中培养3 d,然后浸入0.4 mol/L高糖液体培养基中加载90 min,之后转入玻璃化溶液PVS2(protect vitrification solution 2)中冰上处理6 h,再液氮保存1 h,最后于37℃水浴中解冻2~3 min,1.2 mol/L高糖液体培养基中卸载20 min,待恢复培养后,杂交兰茎尖成活率为64%以上,随机检测30个样品,两种病毒脱毒率均为100%。研究为杂交兰种苗脱毒奠定了一定的理论基础。

Study of Hybrid Orchid Seedlings on Virus Elimination Using Cryopreservation Technology

Taking rhizomes of hybrid Cymbidium with CyMV and ORSV as test material, the paper studied virus elimination of hybrid orchid by encapsulation vitrification cryopreservation method. The results showed that treating shoot tips under low temperature for 21 d then cultured in 0.5 mol/L sucrose of preculture medium for 3 d，and immersed them in the liquid culture medium with 0.4 mol/L high sucrose for 90 min, and put into PVS2 for 6 h on ice, then preserved in liquid nitrogen for 1 h and cultured in liquid medium with 1.2 mol/L high sucrose for 20 min afterwards treated at 37℃ for 2~3 min. Their survival rate was over 64% after being cultured for recovery. 30 samples were selected randomly for virus detection, 2 of them were virus-free seedlings. Their virus detoxification rates were 100%. This study laid certain theoretical basis for virus detoxification of hybrid orchid seedlings.