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Foxtail Millet CBL4 (SiCBL4) Interacts with SiCIPK24, Modulates Plant Salt Stress Tolerance

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Abstract

The calcineurin B-like (CBL) protein and the CBL-interacting protein kinase (CIPK) signaling pathway play important roles in plant abiotic stress tolerance. To investigate the molecular mechanism of salt stress tolerance of foxtail millet, SiCBL4 and SiCIPK24 were identified and functionally characterized. Both SiCBL4 and SiCIPK24 were induced by salt, abscisic acid (ABA), methyl viologen (MV), and heat shock stress in foxtail millet seedlings. Yeast two-hybrid and bimolecular fluorescence complementation assay showed that SiCBL4 interacted with SiCIPK24. The mutation of the N-myristoylation site of SiCBL4 changed the sub-cellular localization of SiCBL4 and directed the SiCBL4-SiCIPK24 protein complex from plasma membrane to cytoplasm, and disrupted its function in plant salt stress tolerance. Overexpression of SiCBL4 or SiCIPK24 in Arabidopsis sos3-1 or sos2-1 mutant plants rescued the mutant salt hypersensitivity phenotype. In addition, overexpression of SiCIPK24 also enhanced the salt stress tolerance of Arabidopsis wild-type plants. This work helps to understand the structure and function of the foxtail millet CBL and CIPK genes and confirmed that the foxtail millet CBL-CIPK pathway can be manipulated to enhance the plant salt stress tolerance.

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Acknowledgements

We thank Dr. Yuanqing Jiang for providing us the Atsos3-1 and Atsos2-1 mutants. This research was supported by the special fund for transgenic research from the Ministry of Agriculture in China 2014ZX0800920B, the National Natural Science Foundation of Shaanxi Province 2014JM3073 (to T.Z), and the National Natural Science Foundation of Shanxi Province 2015011071 (to J.Z).

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  1. Yumin Zhang and Jingjing Linghu contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China

    Yumin Zhang, Jingjing Linghu, Dong Wang, Xi Liu, Fengting Li & Tianyong Zhao

  2. The Key Laboratory of Biology and Genetics Improvement of Maize in Arid Area of Northwest Region, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, China

    Yumin Zhang, Jingjing Linghu, Dong Wang, Xi Liu, Fengting Li & Tianyong Zhao

  3. Millet Research Institute, Shanxi Academy of Agricultural Sciences/Shanxi Key Laboratory of Genetic Resources and Breeding in Minor Crops, Changzhi, 046011, China

    Aili Yu & Jinfeng Zhao

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  1. Yumin Zhang
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Correspondence to Jinfeng Zhao or Tianyong Zhao.

Electronic supplementary material

Supplemental Figure 1

Phylogenetic relationship of CBL proteins from maize, rice, Arabidopsis and foxtail millet. Organism and accession numbers are listed in supplemental data set1. (JPEG 1861 kb)

Supplemental Figure 2. Phylogenetic relationship of CIPK proteins from maize, rice, Arabidopsis and foxtail millet. Organism and accession numbers are listed in are listed in supplemental data set1

(JPEG 3355 kb)

ESM 1

The List of CBL and CIPK proteins from Arabidopsis, rice, mazie and foxtail millet genome. (XLS 40 kb)

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Zhang, Y., Linghu, J., Wang, D. et al. Foxtail Millet CBL4 (SiCBL4) Interacts with SiCIPK24, Modulates Plant Salt Stress Tolerance. Plant Mol Biol Rep 35, 634–646 (2017). https://doi.org/10.1007/s11105-017-1051-1

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