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Dephosphorylation of Nitrate Reductase Protein Regulates Growth of Rice and Adaptability to Low Temperature

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Abstract

Nitrate reductase (NR) is an important enzyme for nitrate assimilation in plants, and its activity is regulated by post-translational phosphorylation. The change of nitrogen uptake affects the response of rice to low temperature and its growth. To investigate the effect of NIA1 protein dephosphorylation on the growth of rice and its adaptability to low temperature, we analyzed phenotype, chlorophyll content, nitrogen utilization, and antioxidant capacity at low temperature in lines with a mutated NIA1 phosphorylation site (S532D and S532A), an OsNia1 over-expression line (OE), and wild-type Kitaake rice (WT). Plant height, dry matter weight, and chlorophyll content of S532D and S532A were lower than those of WT and OE under normal growth conditions but were higher than those of WT and OE at low temperature. Compared with WT and OE, the nitrite, H2O2, and MDA contents of S532D and S532A leaves were higher under normal growth conditions. The difference in leaf nitrite content between transgenic lines and WT was narrower at low temperature, especially in S532D and S532A, while H2O2 and MDA contents of S532D and S532A leaves were lower than those in WT and OE leaves. The NH4+-N and amino acid contents of S532D and S532A leaves were higher than those of WT and OE leaves under normal or low temperature. qRT-PCR results revealed that transcription levels of OsNrt2.4, OsNia2, and OsNADH-GOGAT were positively correlated with those of OsNia1, and the transcription levels of OsNrt2.4, OsNia2, and OsNADH-GOGAT were significantly higher in transgenic lines than in WT under both normal and low temperature. Phosphorylation of NR is a steady-state regulatory mechanism of nitrogen metabolism, and dephosphorylation of NIA1 protein improved NR activity and nitrogen utilization efficiency in rice. Excessive accumulation of nitrite under normal growth conditions inhibits the growth of rice; however, accumulation of nitrite is reduced at low temperature, enhancing the cold tolerance of rice. These results provide a new insight for improving cold tolerance of rice.

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Data Availability

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request. Sequences used in this study are available from the National Center for Biotechnology Information (NCBI).

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Acknowledgements

We would like to thank the research group of Wan Jianmin, academician of the Institute of Crop Science of the Chinese Academy of Agricultural Sciences, for their platform and technical support in vector construction and genetic transformation.

Funding

The research was supported by the National Natural Science Foundation of China (31760350, 31560350), and the Modern Agricultural Scientific Research Collaborative Innovation Special Project of Jiangxi Province (JXXTCXBSJJ202010).

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Author notes

  1. RuiCai Han, YuPeng Wang, ChenYan Li and ZiMing Wu have contributed equally to this work and share first authorship.

Authors and Affiliations

  1. Key Laboratory of Crop Physiology, Ecology and Genetic Breeding, Ministry of Education, College of Agronomy, Jiangxi Agricultural University, Nanchang, 330045, China

    RuiCai Han, YuPeng Wang, ChenYan Li & ZiMing Wu

  2. Key Laboratory of Rice Physiology and Genetics of Jiangxi Province, Rice Research Institute, Nanchang Subcenter of Rice National Engineering Laboratory, Jiangxi Academy of Agricultural Sciences, Nanchang, 330200, China

    RuiCai Han

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Contributions

RH designed and participated in all experimental procedures, performed data analysis, and drafted the manuscript. CL, YW, QS, XP, HL, HJ, and ZW supervised the study and critically revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to ZiMing Wu.

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Han, R., Wang, Y., Li, C. et al. Dephosphorylation of Nitrate Reductase Protein Regulates Growth of Rice and Adaptability to Low Temperature. J Plant Growth Regul 42, 6920–6932 (2023). https://doi.org/10.1007/s00344-023-10985-2

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