松材线虫的低温冷冻保存

Cryopreservation of Pinewood Nematode (Bursaphelenchus xylophilus)

研究-80℃下松材线虫的冷冻保存方法,比较不同冷冻保护剂对松材线虫低温保存的保护效果,以及解冻后松材线虫的繁殖情况。结果表明:15%甘油对松材线虫保护效果最好,解冻后存活率达36.5%,其次为10%甘油、20%甘油、25%甘油,保护效果最差的是1%二甲基亚砜(DMSO);解冻后线虫能够正常生长繁殖,冻存3天和冻存30天的线虫存活率和繁殖无显著性差异。

In this study, we tested the cryopreservation of Bursaphelenchus xylophilus at - 80 ℃ , and investigated effects of different cryoprotectants on the survival and propagation. Cryopreservation with 15% （V/V） glycerol was found as the best protectant for cryopreservation, followed by 10% （ V/V）, 20% （V/V） and 25% （V/V） glycerol. The poorest protecting effect was with 1% dimethylsulfoxide. With 15% （V/V） glycerol, the survival rate reached to 36.5% after defrosting, and the nematodes retained normal propagation. There was no significant difference in the survival and propagation after cryopreservation between 3-days and 30-days cryopreservation which indicated that cryopreservation with 15% （V/V） glycerol was suitable for B. xylophilus at - 80℃.