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Cloning and characterization of MxHA7, a plasma membrane H+-ATPase gene related to high tolerance of Malus xiaojinensis to iron deficiency

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Abstract

Malus xiaojinensis, an iron-efficient apple rootstock, was used to study the molecular mechanisms of iron uptake. Increased H+ extrusion under iron-deficient conditions has been related with H+-ATPases. Thus, a 2,901-bp plasma membrane H+-ATPase gene, MxHA7, encoding 966 amino acids was isolated. Quantitative real-time PCR showed that MxHA7 was specifically induced in the roots of M. xiaojinensis during iron-deficient conditions, not in M. baccata. A functional complementation assay indicated that the high tolerance of MxHA7-transgenic aha7 Arabidopsis thaliana (HA7) plants to iron deficiency was significantly enhanced. Under iron-deficient conditions, Fe2+ contents in the roots and chlorophyll concentrations in the leaves of HA7 plants were increased up to about 2 to 3 times compared to Col-0, aha7 and empty vector (EV) (aha7 transformed with an empty vector) plants. The zinc and manganese contents in the roots of HA7 plants were also higher significantly than in aha7 and EV plants under iron-deficient conditions. Meantime, the HA7 plants have less increasing for iron uptake-related genes than those Col-0, aha7 and EV other plants after iron deficiency, which means MxHA7 gene apparently contributed to help Arabidopsis tolerance to iron deficiency.

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Abbreviations

CTAB:

Cetyltrimethylammonium-bromide

DW:

Dry weight

MS:

Murashige and Skoog

NCBI:

National Center for Biotechnology Information

qRT-PCR:

Quantitative real-time PCR

IBA:

Indole-3-butytric acid

PM:

Plasma membrane

6-BA:

6-Benzylaminopurine

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Acknowledgments

This study was financially supported by the National Special Funds for Scientific Research on Public Causes (Agriculture) (201203075), the National High Technology Research and Development Program of China (2011AA001204), the Modern Agricultural Industry Technology System (Apple) (CARS-28), Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Nutrition and Physiology), Ministry of Agriculture.

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Authors and Affiliations

  1. Institute of Horticultural Plants, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China

    Qian Zha, Qian Zhang, Xinzhong Zhang, Zhenhai Han & Yi Wang

Authors
  1. Qian Zha
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  2. Qian Zhang
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  3. Xinzhong Zhang
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  4. Zhenhai Han
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  5. Yi Wang
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Corresponding author

Correspondence to Yi Wang.

Additional information

Communicated by J.-H. Liu.

Electronic supplementary material

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Supplementary material 1 (TIFF 334 kb) The amino acids of MxHA7

11738_2013_1474_MOESM2_ESM.tif

Supplementary material 2 (TIFF 8773 kb) The standard curve and melt curve of MxHA7 in M. xiaojinensis (A and B) and M. baccata (C and D)

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Zha, Q., Zhang, Q., Zhang, X. et al. Cloning and characterization of MxHA7, a plasma membrane H+-ATPase gene related to high tolerance of Malus xiaojinensis to iron deficiency. Acta Physiol Plant 36, 955–962 (2014). https://doi.org/10.1007/s11738-013-1474-8

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