Original Paper

Plant Molecular Biology Reporter

, Volume 30, Issue 5, pp 1149-1157

First online:

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

  • Qian ZhangAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Yi WangAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Xin Zhong ZhangAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Li Li YinAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Ting WuAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Xue Feng XuAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Wen Suo JiaAffiliated withInstitute for Horticultural Plants, China Agricultural University
  • , Zhen Hai HanAffiliated withInstitute for Horticultural Plants, China Agricultural University Email author 

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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 xiaojinensis. 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.

Keywords

H+-ATPase activity Iron deficiency Malus xiaojinensis Vacuolar H+-ATPase