卵叶泥炭藓茎尖包埋玻璃化法超低温保存研究

Cryopreservation of Shoot Tips of Sphagnum ovatum Hamp. by Encapsulation-vitrification

该研究通过对脱水时间和化冻温度的探索,检验了包埋玻璃化法在超低温保存湿润生境中苔藓的可能性。结果表明:卵叶泥炭藓无菌苗在4℃条件下预培养3d后,在0℃用60% PVS_2装载30min,PVS_2脱水60min后迅速投入液氮保存,24h后用40℃水浴快速化冻2min再培养,成活率可达42.41%,且再生植株与常温状态下的植株形态指标没有显著性差异。研究认为,包埋玻璃化法超低温保存湿润环境中生长的苔藓植物是可行的。

Dehydration time and thawing temperature were explored to test the possibility of encapsulation-vitrification in the cryopreservation of bryophytes collected from moist habitats.The results showed that the regeneration rate of S.ovatum can reach 42.41%when the materials were pre-cultured at 4℃for 3 days,loaded with 60%PVS 2 for 30 minutes and dehydrated with PVS 2 for 60 minutes at 0℃,then stored in liquid nitrogen for 24 hours and thawed at 40℃for 2 minutes.There are no significant differences of morphological indicators between the pants before and after cryopreserved.The results demonstrated the feasibility of cryopreservation of bryophytes in humid environment by encapsulation-vitrification.