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    任瑞芬, 李泽迪, 朱梦婷, 刘燕, 张孔英. ROS诱导的氧化应激和细胞程序性死亡对超低温保存后花粉生活力的影响[J]. 北京林业大学学报, 2022, 44(2): 82-90. DOI: 10.12171/j.1000-1522.20210027
    引用本文: 任瑞芬, 李泽迪, 朱梦婷, 刘燕, 张孔英. ROS诱导的氧化应激和细胞程序性死亡对超低温保存后花粉生活力的影响[J]. 北京林业大学学报, 2022, 44(2): 82-90. DOI: 10.12171/j.1000-1522.20210027
    Ren Ruifen, Li Zedi, Zhu Mengting, Liu Yan, Zhang Kongying. Effects of ROS-induced oxidative stress and programmed cell death on pollen viability after cryopreservation[J]. Journal of Beijing Forestry University, 2022, 44(2): 82-90. DOI: 10.12171/j.1000-1522.20210027
    Citation: Ren Ruifen, Li Zedi, Zhu Mengting, Liu Yan, Zhang Kongying. Effects of ROS-induced oxidative stress and programmed cell death on pollen viability after cryopreservation[J]. Journal of Beijing Forestry University, 2022, 44(2): 82-90. DOI: 10.12171/j.1000-1522.20210027

    ROS诱导的氧化应激和细胞程序性死亡对超低温保存后花粉生活力的影响

    Effects of ROS-induced oxidative stress and programmed cell death on pollen viability after cryopreservation

    • 摘要:
        目的  超低温保存后花粉生活力呈多种变化情况,已有研究表明活性氧(ROS)是超低温保存后花粉生活力变化的主要原因之一。因此,本研究探讨液氮冻存后花粉生活力与ROS诱导的氧化应激和细胞程序性死亡间的关系,以进一步揭示ROS在超低温保存后花粉生活力变化中的作用机制。
        方法  以芍药‘粉玉奴’的花粉为材料,对比分析超低温保存不同时间长度后花粉生活力、ROS生成量、氧化应激以及细胞程序性死亡指标的变化情况。
        结果  花粉超低温保存过程中,ROS成分超氧阴离子自由基(O2•−)含量在液氮冻存1个和3个月后相对较高,过氧化氢(H2O2)含量随着保存时间的延长显著增加,羟自由基(•OH)含量在保存3个月后开始下降,且H2O2和•OH含量均与花粉生活力显著相关。其次,氧化损伤指标丙二醛(MDA)含量在保存3个月后显著升高,与花粉生活力、H2O2和•OH含量显著相关;细胞程序性死亡(PCD)指标caspase-3-like蛋白酶活性在液氮冻存1个月和3个月后均高于对照,细胞凋亡率在保存5个月和8个月后显著升高,且细胞凋亡率均与花粉生活力、H2O2和•OH含量均显著相关。此外,适宜浓度的外源抗氧化剂(抗坏血酸、谷胱甘肽)和类凋亡抑制剂(caspase-3抑制剂)显著提高了液氮冻存8个月后的花粉生活力。
        结论  芍药‘粉玉奴’花粉超低温保存过程中,ROS对超低温保存后的花粉生活力产生了重要的影响,其成分H2O2和•OH的作用尤为突出,其诱发的氧化应激反应和细胞程序性死亡是液氮冻存后花粉生活力变化的重要原因。

       

      Abstract:
        Objective  After cryopreservation, pollen viability shows various changes, studies show that ROS is one of the main reasons for the changes of pollen viability after cryopreservation. Therefore, this study investigated the relationship between pollen viability and ROS induced oxidative stress and programmed cell death after cryopreservation, to further reveal the role of ROS in pollen viability changes after cryopreservation.
        Method  The pollen of P. lactiflora ‘Fen Yu Nu’ was used as material, the changes of pollen viability, ROS production, oxidative stress and programmed cell death after cryopreservation for different lengths of time were compared and analyzed.
        Result  During the cryopreservation of pollen, the O2•− content of ROS components was relatively high after 1 and 3 months of cryopreservation, and the H2O2 content significantly increased with the extension of storage time, while •OH content began to decrease after 3 months, both H2O2 and •OH contents were significantly correlated with pollen viability. Secondly, the content of malondialdehyde (MDA), an index of oxidative damage, was significantly increased after 3 months of liquid nitrogen (LN) storage, which was significantly correlated with pollen viability, H2O2 and •OH contents. The activity of caspase-3-like protease, an indicator of programmed cell death (PCD), was higher than that of the fresh pollen after 1 and 3 months of liquid nitrogen freezing, and the apoptosis rate was significantly increased after 5 and 8 months of LN stored, and the apoptosis rate was significantly correlated with pollen viability, H2O2 and •OH contents. In addition, appropriate concentration of exogenous oxidative damage inhibitors (AsA, GSH) and apoptosis-like inhibitors (caspase-3 inhibitors) significantly improved pollen viability after 8 months of cryopreservation.
        Conclusion  During the cryopreservation of P. lactiflora ‘Fen Yu Nu’ pollen, ROS components have an important effect on pollen viability after cryopreservation, the effects of H2O2 and •OH are particularly prominent. The oxidative stress response and programmed cell death induced by H2O2 and •OH are important reasons for the changes of pollen viability after liquid nitrogen cryopreservation.

       

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