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Isolation, Functional Characterization, and Expression Pattern of a Vacuolar Na+/H+ Antiporter Gene TrNHX1 from Trifolium repens L.

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Abstract

Plant vacuolar Na+/H+ antiporter plays an important role in salt tolerance. A vacuolar Na+/H+ antiporter gene TrNHX1 was cloned from Trifolium repens L., a forage legume, by RT-PCR and RACE methods using degenerate oligonucleotide primers. The TrNHX1 sequence contains 2,394 nucleotides and an open-reading frame of 1,626 nucleotides that encodes a protein of 541 amino acids with a deduced molecular mass of 59.5 kDa. The deduced amino acid sequence of TrNHX1 is 78% identical to that of a vacuolar Na+/H+ antiporter of Arabidopsis thaliana, AtNHX1, and contains the consensus amiloride-binding domain. TrNHX1 could partially complement the NaCl-sensitive phenotypes of yeast mutants Δnhx1 and Δena1-4Δnhx1, and a similar complementation was also observed in the presence of LiCl and KCl. In addition, it was found that TrNHX1 suppressed the hygromycin-sensitive phenotype of yeast mutant Δena1-4Δnhx1. The expression of TrNHX1 in T. repens increased in the presence of 150 mM NaCl, and this result accords with that of Na+ contents determination under the same treatment. These results suggest that TrNHX1 functions as a vacuolar Na+/H+ antiporter and plays an important role in salt tolerance and ion homeostasis in T. repens.

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Abbreviations

NHX:

Vacuolar Na+/H+ antiporter

RACE:

Rapid amplification of cDNA ends

RT:

Reverse transcription

UPM:

Universal primer mixture

ORF:

Open-reading frame

TM:

Transmembrane domain

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Acknowledgments

We thank Prof. Blumwald Eduardo (Department of Plant Sciences, UC Davis) for kindly providing all the yeast strains. This work was financially supported by National Natural Science Foundation of China.

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  1. School of Life Science, East China Normal University, Shanghai, 200062, People’s Republic of China

    Rui Tang, Cui Li, Kai Xu, Yanhua Du & Tao Xia

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  1. Rui Tang
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  2. Cui Li
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Correspondence to Tao Xia.

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Tang, R., Li, C., Xu, K. et al. Isolation, Functional Characterization, and Expression Pattern of a Vacuolar Na+/H+ Antiporter Gene TrNHX1 from Trifolium repens L.. Plant Mol Biol Rep 28, 102–111 (2010). https://doi.org/10.1007/s11105-009-0135-y

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