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Characterization and expression analyses of the H+-pyrophosphatase gene in rye

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Abstract

The H+-pyrophosphatase (H +-PPase) gene plays an important role in maintaining intracellular proton gradients. Here, we characterized the full-length complementary DNA (cDNA) and DNA of the H +-PPase gene ScHP1 in rye (Secale cereale L. ‘Qinling’). We determined the subcellular localization of this gene and predicted the corresponding protein structure. We analysed the evolutionary relationship between ScHP1 and H +−PPase genes in other species, and did real-time quantitative polymerase chain reaction to explore the expression patterns of ScHP1 in rye plants subjected to N, P and K deprivation and to cold, high-salt and drought stresses. ScHP1 cDNA included a 2289 bp open reading frame (ORF) encoding 762 amino acid residues with 14 transmembrane domains. The genomic ScHP1 DNA was 4354 bp and contained eight exons and seven introns. ScHP1 was highly homologous with other members of the H +-PPase gene family. When the full-length ORF was inserted into the expression vector pA7-YFP, the fluorescent microscopy revealed that ScHP1-YFP fusion protein was located in the plasma membrane. Rye plants that were subjected to N deprivation, cold and high-salt stresses, ScHP1 expression was higher in the leaves than roots. Conversely, plants subjected to P and K deprivation and drought stress, ScHP1 expression was higher in the roots than leaves. Under all the investigated stress conditions, expression of ScHP1 was lower in the stem than in the leaves and roots. Our results imply that ScHP1 functions under abiotic stress response.

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Acknowledgements

This work was supported by the China Transgenic Research Programme (2013ZX08002-005) and the International Science and Technology Cooperation Programme of China (no. 2015DFA 30600).

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    Authors and Affiliations

    1. Triticeae Research Institute, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China

      CHANG-SHUI WANG, QIAN-TAO JIANG, JIAN MA, XIU-YING WANG, JI-RUI WANG, GUO-YUE CHEN, PENG-FEI QI, YUAN-YING PENG, XIU-JIN LAN & YU-MING WEI

    2. Key Laboratory of South-Western Crop Germplasm Utilization, Ministry of Agriculture, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

      YOU-LIANG ZHENG

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    Correspondence to YU-MING WEI.

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    [Wang C.-S., Jiang Q.-T., Ma J., Wang X.-Y., Wang J.-R., Chen G.-Y., Qi P.-F., Peng Y.-Y., Lan X.-J., Zheng Y.-L. and Wei Y.-M. 2016 Characterization and expression analyses of the H+-pyrophosphatase gene in rye. J. Genet. 95, xx–xx]

    Chang-Shui Wang and Qian-Tao Jiang contributed equally to this work.

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    WANG, CS., JIANG, QT., MA, J. et al. Characterization and expression analyses of the H+-pyrophosphatase gene in rye. J Genet 95, 565–572 (2016). https://doi.org/10.1007/s12041-016-0664-8

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