Cryopreservation of spores of Haplocladium microphyllum (Hedw.) Broth
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摘要: 通过对干燥时间及预处理温度等超低温保存条件的实验探索,筛选出细叶小羽藓(Haplocladium micr ophyllum)孢子超低温最适保存条件,利用最适保存条件对细叶小羽藓孢子进行不同时间(1d、15d、30d、90d、180d)的超低温保存研究,并与未经任何处理直接投入液氮的孢子进行比较. 结果表明:(1)细叶小羽藓孢子在硅胶干燥5h、--20oC~低温预处理和室温自来水化冻的条件下的平均萌发率最高(88.26%),干燥20 h、常温处理的孢子与干燥5h、--20oC~低温预处理的孢子平均萌发率差异不显著;(2)液氮保存1d后的孢子平均萌发率(88.26%)高于未用液氮保存的孢子平均萌发率(77.90%),保存30 d后的孢子平均萌发率降为6.03%, 而保存90d后平均萌发率又出现上升的趋势, 达到88.47%, 保存180d后平均萌发率仍维持在49.46%, 相对保持率为63.49%. 因此,使用液氮超低温长期保存细叶小羽藓孢子是可行的.Abstract: Drying time and pretreatment temperature were explored to find out the optimum conditions of spores of Haplocladium microphyllum storaged in Liquid nitrogen. Different time (1 d, 15 d, 30 d, 90 d, 180 d) of cryopreservation were studied using the optimum storage conditions and without any treatment. The spore average germination rate and protonema growth were compared before and after cryopreservation. The results showed that (1) After drying 5 h, --20oC low temperature pretreatment and thawing at room temperature conditions, the spore average germination rate was highest (88.26%), there were no significant difference between the average germination of spores of drying 5 h, 15--18oCand 20 h, --20oC low temperature pretreatment; (2) The average germination rate (88.26%) of spores stored in liquid nitrogen for 1 d was comparatively higher than that of the control (77.90%), the average germination rate of spores stored in liquid nitrogen for 30 d dropped to 6.03%, the spore average germination rate stored for 90 d appeared to rise and reaching 88.47%, the spore average germination rate stored for 180 d remained at 49.46% and the relative retention rate reached 63.49%. The results demonstrated the feasibility of cryopreservation for the long-term storage of spores of Haplocladium microphyllum.
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Key words:
- bryophyte /
- Haplocladium microphyllum /
- spore /
- cryopreservation
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