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Functional characterization of a Glycine soja Ca2+ATPase in salt–alkaline stress responses

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Abstract

It is widely accepted that Ca2+ATPase family proteins play important roles in plant environmental stress responses. However, up to now, most researches are limited in the reference plants Arabidopsis and rice. The function of Ca2+ATPases from non-reference plants was rarely reported, especially its regulatory role in carbonate alkaline stress responses. Hence, in this study, we identified the P-type II Ca2+ATPase family genes in soybean genome, determined their chromosomal location and gene architecture, and analyzed their amino acid sequence and evolutionary relationship. Based on above results, we pointed out the existence of gene duplication for soybean Ca2+ATPases. Then, we investigated the expression profiles of the ACA subfamily genes in wild soybean (Glycine soja) under carbonate alkaline stress, and functionally characterized one representative gene GsACA1 by using transgenic alfalfa. Our results suggested that GsACA1 overexpression in alfalfa obviously increased plant tolerance to both carbonate alkaline and neutral salt stresses, as evidenced by lower levels of membrane permeability and MDA content, but higher levels of SOD activity, proline concentration and chlorophyll content under stress conditions. Taken together, for the first time, we reported a P-type II Ca2+ATPase from wild soybean, GsACA1, which could positively regulate plant tolerance to both carbonate alkaline and neutral salt stresses.

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Acknowledgments

We thank members of the lab for discussions and comments on this manuscript. This work was supported by the National Natural Science Foundation of China (31171578), National basic scientific talent training fund projects (J1210069), the National Natural Science Foundation of China (31500204), the Natural Science Foundation of Heilongjiang Province (C2015035), and Graduate student innovation research project of Northeast Agricultural University (yjscx14056).

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Authors and Affiliations

  1. Key Laboratory of Agricultural Biological Functional Gene, Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Mingzhe Sun, Bowei Jia, Na Cui, Yidong Wen, Huizi Duanmu, Qingyue Yu, Jialei Xiao & Yanming Zhu

  2. Crop Stress Molecular Biology Laboratory, Heilongjiang Bayi Agricultural University, Daqing, 163319, People’s Republic of China

    Mingzhe Sun, Bowei Jia & Xiaoli Sun

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  1. Mingzhe Sun
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  2. Bowei Jia
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  3. Na Cui
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Correspondence to Xiaoli Sun or Yanming Zhu.

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11103_2015_426_MOESM1_ESM.tif

Supplementary Fig. S1 Gene architecture and phylogenetic analyses of soybean ECAs. a Phylogenetic relationship among ECAs in soybean. b Representation of intron-exon and domain structures in ECAs proteins. The intron-exon structures were from Phytozome website. The domains found in the ECA subfamily contained cation transporter/ATPase N-terminus (red), E1-E2 ATPase (orange), haloacid dehalogenase-like hydrolase (purple) and cation transporter/ATPase C-terminus (blue). (TIFF 84 kb)

11103_2015_426_MOESM2_ESM.tif

Supplementary Fig. S2 Multiple amino acid sequence alignment of ECAs from soybean and Arabidopsis. The figure showed the conserved motifs (red boxes) and the predicted transmembrane domains (designated as M1-M10) (marked by red lines on the top). The amino acid sequences were aligned by the ClustalX software. The existence and position of transmembrane domains were verified and marked according to previous study (Chung et al. 2000). (TIFF 1817 kb)

11103_2015_426_MOESM3_ESM.tif

Supplementary Fig. S3 Expression patterns of ACA subfamily genes in Glycine soja under 50 mM NaHCO3 (pH 8.5) treatment based on the microarray data. a Expression patterns of ACAs in Glycine soja roots. b Expression patterns of ACAs in Glycine soja leaves. The color saturation reflected the log2 fold change (LogFC) to 0 h for each gene. Green color indicated higher transcript levels than 0 h, whereas green color meant lower transcript levels. (TIFF 154 kb)

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Sun, M., Jia, B., Cui, N. et al. Functional characterization of a Glycine soja Ca2+ATPase in salt–alkaline stress responses. Plant Mol Biol 90, 419–434 (2016). https://doi.org/10.1007/s11103-015-0426-7

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