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    乙烯对春石斛类原球茎玻璃化超低温保存存活率的影响

    Effects of ethylene on the viability of cryopreserved Dendrobium protocorm-like bodies

    • 摘要:
        目的  为探究乙烯对PLBs超低温保存存活率的影响,以期为提高其存活率开拓思路。
        方法  以春石斛‘红梦幻’类原球茎(PLBs)为材料,分别在其玻璃化超低温保存的预培养和装载阶段添加外源乙烯供体乙烯利和乙烯合成抑制剂硝酸钴,测定PLBs超低温保存过程中内源乙烯前体1-氨基环丙烷羧酸(ACC)含量、ACC氧化酶(ACO)活性及PLBs液氮保存后细胞存活率。
        结果  结果显示:无论在预培养还是装载阶段,外源乙烯供体乙烯利处理后,超低温保存过程中PLBs内源ACC含量和ACO活性均显著提高,同时液氮保存后存活率也显著提高;在这两个阶段添加外源乙烯合成抑制剂硝酸钴后,超低温保存过程中PLBs内源ACC含量、ACO活性显著降低,液氮保存后存活率也显著降低。相关分析显示,PLBs液氮冻后存活率与其内源ACC含量和ACO活性呈极显著正相关。
        结论  本研究表明,外源乙烯调节剂乙烯利和硝酸钴通过调节内源乙烯含量而对玻璃化超低温保存的春石斛类原球茎存活率产生显著影响。

       

      Abstract:
        Objective  This paper aims to explore the effects of ethylene on the survival rate of PLBs under cryopreservation to develop ideas for improving the survival rate of PLBs.
        Method  The protocorm-like bodies (PLBs) of Dendrobium nobile ‘Hamana Lake Dream’ were used as materials, exogenous ethylene donor ethephon and ethylene synthesis inhibitor cobalt nitrate were added during the preculture and loading stages of its vitrification cryopreservation, respectively to study the effects of ethylene on cell survival rate, endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) content and ACC oxidase (ACO) activity of the cryopreserved PLBs.
        Result  The addition of exogenous ethylene ethephon can significantly improve the endogenous ACC content, ACO activity and cell survival rate of Dendrobium PLBs after cryopreservation by vitrification. While, the addition of exogenous ethylene inhibitor cobalt nitrate significantly reduced the endogenous ACC content, ACO activity and the cell survival rate of PLBs after cryopreservation by vitrification. In addition, the survival rate of PLBs after cryopreservation was significantly positively correlated with endogenous ACC content and was extremely significantly positively correlated with endogenous ACO activity.
        Conclusion  This study shows that the exogenous ethylene regulator ethephon and cobalt nitrate has a significant effect on the viability of Dendrobium PLBs after vitrification-cryopreservation by regulating endogenous ethylene.

       

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