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Cloning and Characterization of MxVHA-c, a Vacuolar H+-ATPase Subunit C Gene Related to Fe Efficiency from Malus xiao**ensis

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Abstract

The vacuolar H+-ATPase plays a crucial role in secondary transport and in plant response to environmental stress. In this study, a vacuolar H+-ATPase (MxVHA-c) gene, consisting of an ORF of 498 base pairs and 165 amino acid residues, has been cloned from the iron-efficient genotype of Malus xiao**ensis. Subsequently, this gene has been targeted to the tonoplast using transient expression analysis. Quantitative real-time (qRT) PCR results reveal that the MxVHA-c gene is expressed in both roots and leaves of Fe-deficient plants; however, it is sensitive to iron stress in roots. This suggests that MxVHA-c expression in roots may mediate iron-dependent responses. MxVHA-c expression is up-regulated following exogenous treatment with abscisic acid (ABA) and down-regulated following treatment with CaCl2. Overexpression of the MxVHA-c gene in yeast strains has revealed that MxVHA-c transiently alleviated cadmium toxicity via the Cd2+/H+ antiport protein. H+-ATPase activity is slightly increased in yeast overexpressing the MxVHA-c gene compared to that in yeast transformed with an empty vector. In addition, this transgenic yeast strain can grow in a liquid medium containing 40 μM ferrozine. These findings may provide useful information in elucidating molecular mechanisms that mediate resistance to iron deficiency.

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Abbreviations

qRT-PCR:

Quantitative real-time polymerase chain reaction

VHA :

Vacuolar H+-ATPase

YPD:

Yeast growth medium

SD:

Selection medium

RFU:

Relative fluorescence unit

eGFP:

Enhanced green fluorescent protein

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Acknowledgments

This study was financially supported by the National Natural Science Foundation of China (30971982), The National Transgenic Special Project (2009ZX08009-122B), and The Key Laboratory of Bei**g Municipality of Stress Physiology and Molecular Biology for Fruit Trees.

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Authors and Affiliations

  1. Institute for Horticultural Plants, China Agricultural University, Yuanmingyuan West Road, Haidian District, Bei**g, 100193, People’s Republic of China

    Qian Zhang, Yi Wang, **n Zhong Zhang, Li Li Yin, Ting Wu, Xue Feng Xu, Wen Suo Jia & Zhen Hai Han

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  1. Qian Zhang
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  2. Yi Wang
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  3. **n Zhong Zhang
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  4. Li Li Yin
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  7. Wen Suo Jia
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  8. Zhen Hai Han
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Correspondence to Zhen Hai Han.

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Qian Zhang and Yi Wang contributed equally to this work.

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Zhang, Q., Wang, Y., Zhang, X.Z. et al. Cloning and Characterization of MxVHA-c, a Vacuolar H+-ATPase Subunit C Gene Related to Fe Efficiency from Malus xiao**ensis . Plant Mol Biol Rep 30, 1149–1157 (2012). https://doi.org/10.1007/s11105-012-0426-6

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  • DOI: https://doi.org/10.1007/s11105-012-0426-6

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