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Overexpression of CsCLCc, a Chloride Channel Gene from Poncirus trifoliata, Enhances Salt Tolerance in Arabidopsis

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Abstract

Chloride channels (CLCs) have been reported to be involved in plant adaptation to salt stress by regulating Cl− homeostasis. In this study, a putative CLC-encoding gene, CsCLCc, was isolated from trifoliate orange [Poncirus trifoliata (L.) Raf]. The deduced amino acids of CsCLCc share high identity with other CLC-like sequences, which also contain domains common in CLC-like transmembrane segments and cystathionine β-synthase domains. Quantitative reverse transcription PCR analysis revealed that the CsCLCc gene expressed in the leaves and roots of trifoliate orange was upregulated by ABA, 4 °C, and NaCl. Transformation of Arabidopsis AtCLCc mutant clcc with 35S::CsCLCc improved seed germination in transgenic plants under salinity. In addition, under 200 mM NaCl treatment, the reduction in fresh weight, electrolyte leakage, and chlorophyll content was lower in the transgenic seedlings than that in mutant or wild type. This was further supported by the observation that the total Cl− accumulated in the roots and shoots was lower in transgenic plants than that in mutant or wild type. Our results indicate that CsCLCc could play an important role in Cl− homeostasis and could be an important gene target for improving salt tolerance in plants.

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Abbreviations

CBS:

Cystathionine beta-synthase

CLCs:

Chloride channels

EL:

Electrolyte leakage

TMs:

Transmembrane segments

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Acknowledgments

This work was supported by the National Modern Agriculture (Citrus) Industry System of Special Funds (CARS-27) and Special Fund for Agro-scientific Research in the Public Interest (201203075). The authors thank Dr. Zhiyong Pan for critical reading of this manuscript and Prof. Yanxiu Liu for her help in improving the English.

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Authors and Affiliations

  1. College of Horticulture and Forestry Science, Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University, Wuhan, 430070, China

    Qingjiang Wei, Yongzhong Liu, Gaofeng Zhou, Qiaohong Li, Chengquan Yang & Shu-ang Peng

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  1. Qingjiang Wei
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  2. Yongzhong Liu
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  3. Gaofeng Zhou
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  4. Qiaohong Li
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Corresponding author

Correspondence to Shu-ang Peng.

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ESM 1

Alignment of the amino acid sequences of CsCLCc and CLCs from other plants. The accession numbers of the corresponding proteins are PtrCLC-7 (XP_002331119), VvCLC (XP_002268066), AtCLCc (NP_199800), CLC-Nt1 (CAA64829.1), GmCLC (AAY43007.1), AtCLCa (NP_198905.1), StCLC1 (CAA71369.1), and ZmCLC (NP_001183936.1). Black and gray shading indicate sequence identity and similarity, respectively. The conserved motifs GxGIPE, GKxGPxxH, and PxxGxLF are indicated by a box, and the transmembrane segments (TM1-10) and cystathionine β-synthase domains (CBS1-2) are indicated by the line above the sequences (TIFF 3423 kb)

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Wei, Q., Liu, Y., Zhou, G. et al. Overexpression of CsCLCc, a Chloride Channel Gene from Poncirus trifoliata, Enhances Salt Tolerance in Arabidopsis . Plant Mol Biol Rep 31, 1548–1557 (2013). https://doi.org/10.1007/s11105-013-0592-1

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