Cloning and Characterization of a Plasma Membrane H+-ATPase (PMA) Gene from a Salt-tolerant Plant Chloris virgata  

Xinxin Zhang1 , Hui Wang1 , Shenkui Liu1 , Tetsuo Takano2
1. Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular Biology Laboratory, Northeast Forestry University, Harbin, 150040, P. R. China
2. Asian Natural Environment Science Center (ANESC), the University of Tokyo, Midori Cho 1-1-1, Nishitokyo City, Tokyo 188-0002, Japan
Author    Correspondence author
Molecular Soil Biology, 2014, Vol. 5, No. 3   doi: 10.5376/msb.2014.05.0003
Received: 16 Feb., 2014    Accepted: 20 Mar., 2014    Published: 28 Apr., 2014
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Abstract

A plasma membrane H+-ATPase (PMA) gene (ChvPMA) was isolated from a wild salt-tolerant plant Chloris virgata. The expression of ChvPMA gene in leaves and roots of Chloris virgata seedlings under salt stress (NaCl, NaHCO3) was examined. The results showed the ChvPMA gene expression was induced by salt stress. The ChvPMA gene was fused to the N-terminus of green fluorescence protein (GFP) gene, and transferred into onion epidermal cells for analyses of intracellular localization. The result showed that the ChvPMA protein was found to be in the plasma membrane of ion epiderm. Because the H+-ATPase activity is regulated by a C-terminal auto-inhibitory domain that can be displaced by phosphorylation, we analyzed transgenic yeast expressing either wild-type PMA (ChvPMA) or truncated ChvPMA lacking the C-terminal auto-inhibitory domain (ChvPMAΔC) under high salt and pH conditions. The results showed that over-expression of ChvPMA and ChvPMAΔC in transgenic yeasts increased the resistance to salt and lower pH conditions, especially, the yeast over-expressing ChvPMAΔC showed better growth than ChvPMA at an external pH 4.0 in the presence NaCl. Transgenic Arabidopsis over-expressing ChvPMAΔC also showed the better root growth than that of ChvPMA at an external pH 4.0 in the presence NaCl.

Keywords
Chloris virgata; ChvPMA; Plasma membrane H+-ATPase; Salt and pH tolerance
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