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Overexpression of TaNHX2 enhances salt tolerance of ‘composite’ and whole transgenic soybean plants

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Abstract

Salinity is a major factor resulting in extensive loss of agricultural production. Genetic transformation has become a powerful tool for studying gene function and for improving crop salt tolerance. In this study, a TaNHX2 gene was transformed into a plant cloning vector under the control of cauliflower mosaic virus 35S promoter, and then introduced into Agrobacterium rhizogenes strain K599. Explants of soybean were transformed with A. rhizogenes and ‘composite’ plants consisting of wild-type shoots and transgenic hairy roots overexpressing TaNHX2 were produced. When exposed to salt stress, ‘composite’ plants displayed high salinity tolerance at 171 mM NaCl in vermiculite and in solid medium supplemented with up to 200 mM NaCl, whereas control plants displayed chlorosis and died within 15 days under above treatment conditions. We subsequently obtained soybean plants overexpressing TaNHX2 through A. tumefaciens-mediated transformation and studied four homozygous lines of TaNHX2. Transgenic lines displayed an enhanced salt tolerance in plant biomass and flower number per plant, compared with wild type plants grown on sand culture containing 150 mM NaCl. Furthermore, transgenic plants of line C12-11 showed longer survival, less growth inhibition and greater number of flowers than wild type plants. Taken together, these results indicated that TaNHX2 gene could enhance salt tolerance of soybean, and A. rhizogenes-mediated transformation system could be used as a complementary tool of A. tumerfaciens-mediated transformation to rapidly investigate candidate gene function in soybean.

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Abbreviations

CaMV:

Cauliflower mosaic virus

CCM:

Co-cultivation medium

BA:

6-Benzylaminopurine

MSB:

Murashige and Skoog basal nutrient salts with B5 vitamins

RT-PCR:

Reverse transcription-PCR

SD(I%):

Salt damage index (%)

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Acknowledgments

We are grateful to Professor Guihua Shao for her valuable advice in designing salt treatment and data analysis, Drs. Shikui Song, Hongbo Sun and Chunhui Chen for their suggestions, Miss Jun Zhang for her technical assistance, Professor Shouyi Chen for providing the plasmid pBin438-TaNHX2, Professor Peter Gresshoff for providing A. rhizogenes strain K599 and the binary vector pGFPGUSPlus+ and Professor Kan Wang for providing vector pTF101.1. We especially thank for Dr. Richard Ann for improving the language of the manuscript. This work was supported by Natural Science Foundation of China (30771358), the State Key Basic Research and Development Plan of China (2009CB11840), State Grand Project for Transgenic Organism Breeding (2008ZX08010-004) and Special Fund for Establishment of China Agricultural Research System (CARS-04).

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Authors and Affiliations

  1. Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin, 150081, Heilongjiang, People’s Republic of China

    Dong Cao & Xiaobing Liu

  2. The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, The Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Dong Cao, Wensheng Hou, Wei Liu, Weiwei Yao, Cunxiang Wu & Tianfu Han

  3. The Graduate University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Dong Cao

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  1. Dong Cao
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  2. Wensheng Hou
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  3. Wei Liu
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  4. Weiwei Yao
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  5. Cunxiang Wu
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  6. Xiaobing Liu
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Correspondence to Xiaobing Liu or Tianfu Han.

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Dong Cao and Wensheng Hou have equally contributed to this work.

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Cao, D., Hou, W., Liu, W. et al. Overexpression of TaNHX2 enhances salt tolerance of ‘composite’ and whole transgenic soybean plants. Plant Cell Tiss Organ Cult 107, 541–552 (2011). https://doi.org/10.1007/s11240-011-0005-9

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