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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (11): 1682-1691.doi: 10.3724/SP.J.1006.2015.01682

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Overexpression of Vacuole H+-ATPase E Subunit Gene SiVHA-E from Foxtail Millet Enhances Salt Resistance in Transgenic Arabidopsis thaliana

FENG Lu1,2,ZHONG Li2,3,CHEN Dan-Dan4,MA You-Zhi2,XU Zhao-Shi2,LI Lian-Cheng2,ZHOU Yong-Bin2,CHEN Ming2,*,ZHANG Xiao-Hong1,*   

  1. 1 College of Life Science, Northwest A&F University, Yangling 712100, China; 2 Institute of Crop Science, Chinese Academy of Agricultural Science / National Key Facility for Crop Gene Response and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crop, Ministry of Agriculture, Beijing 100081, China; 3 Guizhou Provincial Institute of Grass Industry, Guiyang 550006, China; 4 College of Agronomy, Northwest A&F University, Yangling 712100, China
  • Received:2015-02-11 Revised:2015-06-01 Online:2015-11-12 Published:2015-06-29

Abstract:

The V-H+-ATPase plays an important role in processes of plant growth, development and response to stresses. In this research, SiVHA-E, a V-H+-ATPase E subunit gene, was cloned from millet by the Blast analysis against GenBank database. Phylogenetic tree showed that the gene belongs to E1/E3 subgroup and is close with ZmVHA-EL, a V-H+-ATPase E subunit from maize. The quantitative Real-time PCR (qRT-PCR) analysis revealed that the expression levels of SiVHA-E were up-regulated under treatments of high-salt, exogenous MeJA, SA, and ABA hormones, while down-regulated under stresses of cold and low nitrogen. Protein subcellular localization analysis using protoplast showed that SiVHA-E is located on tonoplast. The results of salt tolerance assay showed that the germination rate of SiVHA-E transgenic lines was significantly higher than that of wild type plant under salt stress. During seedlings period, the root lengths were significantly longer as well as fresh weight and survival rate were significantly higher in transgenic lines than in wild type plant under salt treatment. Compared with wild type plant, transgenic plant reduced the content of Na+ and increased the relative water content inside cells. In addition, the results of germination experiment used ABA showed that SiVHA-E transgenic Arabidopsis was more sensitive to ABA than wild type plant during post-germination. In short, overexpressing SiVHA-E in transgenic Arabidopsis lines enhances salt tolerance, which might be relates to positive regulation of ABA signaling pathway or reduction of Na+ accumulation and water loss in transgenic plants.

Key words: Foxtail millet, V-type H+-ATPase E subunit gene, Salt stress tolerance, Mechanism

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