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Cyclic nucleotide gated channel 10 negatively regulates salt tolerance by mediating Na+ transport in Arabidopsis

Abstract

A number of cyclic nucleotide gated channel (CNGC) genes have been identified in plant genomes, but their functions are mainly undefined. In this study, we identified the role of CNGC10 in the response of Arabidopsis thaliana to salt stress. The cngc10 T-DNA insertion mutant showed greater tolerance to salt than wild-type A. thaliana during seed germination and seedling growth. The cngc10 mutant accumulated less Na+ and K+, but not less Ca2+, in shoots in response to salt stress. By contrast, overexpression of CNGC10 resulted in greater sensitivity to salt stress, and complementation of this gene recovered salt sensitivity. In response to salt stress, heterologous expression of CNGC10 in the Na+ sensitive yeast mutant strain B31 inhibited growth due to accumulation of Na+ at a rate greater than that of yeast transformed with an empty vector. Quantitative RT-PCR analysis demonstrated that CNGC10 was expressed mainly in roots and flowers. GUS analysis of a root cross section indicated that CNGC10 was expressed mainly in the endodermis and epidermis. Furthermore, the expression of CNGC10 in roots was dramatically inhibited by exposure to 200 mM NaCl for 6 h. These data suggest that CNGC10 negatively regulates salt tolerance in A. thaliana and may be involved in mediating Na+ transport.

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Acknowledgments

We would like to thank Dr. Alonso Rodríguez-Navarro (at Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid) to provide Na+ sensitive yeast mutant strain B31. This work is supported by grants from National Basic Research Program of China (2012CB114200), NSFC grants (31171461 and 91117003), and the Fundamental Research Funds for the Central Universities (KYTZ201402) to W Zhang, and grants from National Natural Science Foundation of China (31100194 and 31470364) and the Fundamental Research Funds for the Central Universities (KYZ201423) to Q. Zhang.

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Correspondence to Qun Zhang or Wenhua Zhang.

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10265_2014_679_MOESM1_ESM.ppt

Supplementary material 1 (PPT 396 kb). Fig. S1 Salt tolerance of seedlings. a Expression of CNGC10 as determined by RT-PCR in shoots (top) and roots (bottom) of cngc10, over-expressing lines (OE-1, OE-3), complemented lines (COM-2, COM-5), and the WT. EF1α was used as the reference gene. b Four-day-old seedlings of cngc10, OE-1, OE-3, COM-2, COM-5, and WT were transferred to 1/2 MS (control) (top) or medium containing 200 mM NaCl (middle) or 400 mM mannitol (bottom). Photographs were taken on day 4 (NaCl treatment) or 14 (mannitol treatment or control). Fig. S2 Growth curves and ion contents of yeasts grown in Na+-free medium. a Growth curves of CNGC10-, HKT1;1-, and pYES2-transformed B31 exhibited no obvious difference in Na+-free AP medium. b Na+ contents in yeasts grown for 8 h in liquid Na+-free AP medium. c K+ contents in yeasts grown for 8 h in liquid Na+-free AP medium. d Ca2+ contents in yeasts after growth for 8 h in liquid Na+-free AP medium. Values are mean ± SE (n = 5)

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Jin, Y., Jing, W., Zhang, Q. et al. Cyclic nucleotide gated channel 10 negatively regulates salt tolerance by mediating Na+ transport in Arabidopsis . J Plant Res 128, 211–220 (2015). https://doi.org/10.1007/s10265-014-0679-2

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Keywords

  • CNGC10
  • Na+ transport
  • Salt tolerance
  • Arabidopsis thaliana