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    高山红景天愈伤组织的玻璃化法保存及植株再生

    Cryopreservation of calli by vitrification and plant regeneration of Rhodiola sachalinensis

    • 摘要: 采用玻璃化法对高山红景天的愈伤组织进行了超低温保存研究.高山红景天愈伤组织在8%蔗糖中暗培养5 d后,先用25℃的60%玻璃化保护剂PVS2预处理20 min,转至100%PVS2于-20℃乙醇浴中处理2 h后,投入液氮进行保存,48 h后使用40℃水浴快速化冻.冻存后的愈伤组织用液体培养基MS+6-BA 2 mg/L+NAA 0.5 mg/L+40%蔗糖洗涤3次,再转入MS+6-BA 2mg/L+NAA 0.5 mg/L固体培养基中在22℃条件下进行暗培养,2周后转入光照培养(光照强度800 lx).6周后观察到高山红景天愈伤组织呈鲜绿色,长势较为旺盛,存活率可达到78.24%.冻存后的愈伤组织可诱导成苗.

       

      Abstract: A cryopreservation procedure was presented on the callus of Rhodiola sachalinensis by vitrification.Calli were precultured in 8% sugar solution in darkness for 5 days,followed by treatment with 60% PVS2 at 25℃ for 20 min and with 100% PVS2 in an alcohol bath at-20℃ for 2 hrs.The calli were then directly immersed into liquid nitrogen for 48 hrs.After rapid thawing in a water bath at 40℃,the calli were washed three times in MS liquid medium supplemented with 6-BA 2 mg/L,NAA 0.5 mg/L and 40% sugar solution,and then moved to MS solid medium containing 6-BA 2 mg/L and NAA 0.5 mg/L.The calli were cultured at 22℃ in darkness for two weeks,then under a light of 800 lx.Six weeks later,the calli of Rh.sachalinensis were fresh and green,showing vigorous growth and the survival rate reached 78.24%.The cryopreserved calli of Rh.sachalinensis could be induced to seedlings.

       

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