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- 2018
羽状鸡冠花无菌幼苗的超低温保存研究
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Abstract:
以羽状鸡冠花无菌幼苗为材料，研究建立了其玻璃化超低温保存技术程序.具体为：取胚根长2~3 mm的无菌幼苗，25 ℃装载40 min，0 ℃下PVS2脱水60 min，更新PVS2溶液并迅速投入液氮，冻存24 h以上.取出玻璃化幼苗，用40 ℃水浴化冻60 s，再用Unloading溶液处理20 min，然后将幼苗在恢复培养基上暗培养7 d后，转入光照培养(光照强度约40 μmol/(m2·s)，14 h/d).在此条件下，幼苗再生率可达60%以上.装载时导入0.1 mmol/L ASA，卸载时导入400 U/mL CAT可提高无菌幼苗存活率.
A procedure has been developed for vitrification cryopreservation of plumed cockscomb (Celosia plumosa 'Fresh Look') aseptic seedlings. First, aseptic seedlings with a radicle 2-3 mm in length were loaded at 25 ℃ for 40 min and then dehydrated in a vitrification solution (PVS2) at 0 ℃ for 60 min. Next, the PVS2 solution was replaced by a fresh PVS2 solution and the seedlings were directly immersed into LN2 and frozen for more than 24 hours. Then the vitrified seedlings were taken out and thawed in a water bath at 40 ℃ for 60 s, and washed twice (10 min each time) with an unloading solution. Finally, the vitrified seedlings were recultured on MS medium in darkness for 7 days prior to exposure to the light. Under the above conditions, the regeneration rate was as high as 60% or more. Addition of 0.1 mmol/L ASA in the loading solution or 400 U/mL CAT in the unloading solution helped to increase the regeneration rates of the aseptic seedlings
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