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The ROS-associated programmed cell death causes the decline of pollen viability recovered from cryopreservation in Paeonia lactiflora

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After cryopreservation, the occurrence of apoptosis-like programmed cell death events induced by the accumulation of ROS reduces pollen viability.

Cryopreservation, as a biotechnological means for long-term preservation of pollen, has been applied to many species. However, after cryopreservation, the viability of pollen significantly decreases via a mechanism that is not completely clear. In this study, the pollen of Paeonia lactiflora ‘Zi Feng Chao Yang’, which exhibits significantly reduced viability after liquid nitrogen (LN2) storage, was used to study the relationship among pollen viability, programmed cell death (PCD) and reactive oxygen species (ROS). The apoptosis rate was increased significantly in pollen with decreased viability after cryopreservation, and the changes in ROS generation and hydrogen peroxide (H2O2) were consistent with the apoptosis rate. Correlation analysis results showed that the apoptosis rate is positively correlated with ROS generation and H2O2 content. In addition, ascorbic acid (AsA), glutathione (GSH) and ascorbic acid reductase (APX) levels were significantly correlated with ROS and H2O2. After LN2 preservation for 8 months, the exogenous antioxidants AsA and GSH at appropriate concentrations significantly decreased H2O2 content, inhibited PCD indicator levels, and increased cryopreserved pollen viability. These observations suggest that PCD occurred in pollen during LN2 preservation for 1–8 months and was induced by the accumulation of ROS in pollen after cryopreservation, thus explaining the main reasons for the reduction in pollen viability after cryopreservation in LN2.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (No. 31370693 and No. 31770741). In addition, we thank Prof. Barbara M. Reed for editing the manuscript. We thank Jiao Pengcheng and Ji Jiaojiao (Core Facility, Center of Biomedical Analysis, Tsinghua University) for technical support with flow cytometry analysis.

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Authors and Affiliations

  1. Beijing Laboratory of Urban and Rural Ecological Environment, Beijing Municipal Education Commission, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, National Engineering Research Center for Floriculture, College of Landscape Architecture, Beijing Forestry University, Beijing, 100083, China

    Ruifen Ren, Zedi Li, Xueru Jiang & Yan Liu

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  1. Ruifen Ren
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  2. Zedi Li
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  3. Xueru Jiang
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  4. Yan Liu
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Contributions

RR designed the research, completed part of experiment, analyzed the data and drafted the manuscript. ZL designed the research and completed part of experiment. XJ offered some help on the research designed. YL conceived the project, supervised the analysis and critically revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Yan Liu.

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Communicated by Neal Stewart.

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Ren, R., Li, Z., Jiang, X. et al. The ROS-associated programmed cell death causes the decline of pollen viability recovered from cryopreservation in Paeonia lactiflora. Plant Cell Rep 39, 941–952 (2020). https://doi.org/10.1007/s00299-020-02540-0

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