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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 102, Issue 3, pp 337–345 | Cite as

Overexpression of a Malus vacuolar Na+/H+ antiporter gene (MdNHX1) in apple rootstock M.26 and its influence on salt tolerance

  • Yonghong Li
  • Yanzi Zhang
  • Fengjuan Feng
  • Dong Liang
  • Lailiang Cheng
  • Fengwang Ma
  • Shouguo Shi
Original Paper

Abstract

Soil salinity is a major factor limiting apple production in some areas. Tonoplast Na+/H+ antiporters play a critical role in salt tolerance. Here, we isolated MdNHX1, a vacuolar Na+/H+ antiporter from Luo-2, a salt-tolerant rootstock of apple (Malus × domestica Borkh.), and introduced it into apple rootstock M.26 by Agrobacterium-mediated transformation. PCR and DNA gel blot analyses confirmed successful integration of MdNHX1. RT-PCR analysis indicated that the gene was highly expressed in transgenic plants, but the degree of this expression varied among lines. Its overexpression conferred high tolerance to salt stress. Analysis of ion contents showed that, when exposed to salinity stress, the transgenics compartmentalized more Na+ in the roots and also maintained a relatively high K+/Na+ ratio in the leaves compared with non-transformed plants. Under normal conditions, however, amounts of potassium and sodium did not differ significantly between transgenic and control plants.

Keywords

Apple rootstock M.26 Genetic transformation Na+/H+ antiporter MdNHX1 Salt tolerance 

Abbreviations

6-BA

6-Benzyladenine

Cef

Cefotaxine

CTAB

Cetyltrimethylammonium bromide

Kan

Kanamycin

Lx

Lux

MdNHX1

Apple vacuolar Na+/H+ Exchanger

NAA

α-Naphthaleneacetic acid

Npt II

Neomycin phosphotransferase II

OD

Optical density

ORF

Opening reading frame

PCR

Polymerase chain reaction

RACE

Rapid amplification of cDNA ends

RT-PCR

Reverse transcriptase polymerase chain reaction

UTR

Untranslated region

LB

Luria-Bertani broth

Notes

Acknowledgments

This work was supported by the Hi-Tech Research and Development Program of China (2008AA10Z157). The authors would like to thank Dr. Chengchao Zheng in the Plant Biochemistry and Molecular Biology Program in the College of Life Science at Shandong Agricultural University and Dr. Cankui Zhang in the Department of Plant Biology at Cornell University for their help in cloning MdNHX1.

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Yonghong Li
    • 1
  • Yanzi Zhang
    • 1
  • Fengjuan Feng
    • 1
  • Dong Liang
    • 1
  • Lailiang Cheng
    • 2
  • Fengwang Ma
    • 1
  • Shouguo Shi
    • 1
  1. 1.College of HorticultureNorthwest A & F UniversityYanglingChina
  2. 2.Department of HorticultureCornell UniversityIthacaUSA

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