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Exogenous dehydrin NnRab18 improves the Arabidopsis cryopreservation by affecting ROS metabolism and protecting antioxidase activities

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Abstract

Seeds usually dehydrate and reach a vitrification state during maturation, conferring stress tolerance and storage tolerance on mature seeds. Late embryogenesis abundant (LEA) proteins accumulate and play multiple protective roles in this process. In this study, recombinant dehydrin NnRab18 of LEA family from Nelumbo nucifera seeds was added into loading solution and plant vitrification solution (PVS2) to verify the optimization effect of NnRab18 on plant cryopreservation. The NnRab18-optimized method approximately doubled the survival rate of Arabidopsis thaliana L. seedlings, and the survival rates were linearly dependent on concentrations of exogenous NnRab18 protein. Evans blue staining revealed that NnRab18 alleviated the plasma membrane damages of seedling roots. In the NnRab18-optimized group, OH· scavenging activity was increased by 50.0%, and H2O2 and malondialdehyde contents were decreased by 9.3% and 29.7%, respectively. Quantitative gene expression analysis showed that oxidative stress–related genes including oxidative signal-inducible 1 (OXI1), Cu/Zn superoxide dismutase (Cu/Zn SOD), catalase2 (CAT2), CAT3, monodehydroascorbate reductase (MDHAR), glutathione reductase 1 (GR1), and glutathione peroxidase 6 (GPX6) were significantly downregulated in the NnRab18-optimized group. Additionally, the Cu/Zn SOD, POD, and CAT activities of the NnRab18-optimized group were 25.0%, 16.2%, and 30.1% higher than those of the control group, respectively. NnRab18 addition alleviated L-Lactate dehydrogenase inactivation in the in vitro enzyme activity protection assay, demonstrating that NnRab18 can directly protect enzyme activity during cryopreservation. This study suggests that NnRab18 can improve the cryopreservation efficiency by affecting ROS metabolism and signal transduction, alleviating plasma membrane peroxidation, and protecting antioxidase activities.

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Funding

This work was sponsored by the National Natural Science Foundation of China (No. 31971705) and Natural Science Foundation of Shanghai (No. 21ZR1434200).

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Authors and Affiliations

  1. School of Design, Shanghai Jiao Tong University, Shanghai, 200240, China

    Jiangyuan Sheng, Tao Liu & Di Zhang

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  1. Jiangyuan Sheng
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  2. Tao Liu
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  3. Di Zhang
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Contributions

DZ designed the research and reviewed the manuscript. TL conducted the cryopreservation experiments. JS performed the physiological indexes detection, analyzed the data, and wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Di Zhang.

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Sheng, J., Liu, T. & Zhang, D. Exogenous dehydrin NnRab18 improves the Arabidopsis cryopreservation by affecting ROS metabolism and protecting antioxidase activities. In Vitro Cell.Dev.Biol.-Plant 58, 530–539 (2022). https://doi.org/10.1007/s11627-022-10254-z

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