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Enhanced salinity tolerance in transgenic mungbean overexpressing Arabidopsis antiporter (NHX1) gene

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Abstract

Efficient compartmentalization of Na+ ions into the vacuole through heterologous overexpression of vacoular antiporter gene NHX1 is a promising approach to develop salt tolerance in plants. Mungbean (Vigna radiata L. Wilczek) is an important grain legume widely cultivated in Southeast Asia for its protein rich grains. Salinity affects growth and productivity of mungbean. In this paper, we report overexpression of an Arabidopsis NHX1 (AtNHX1) in transgenic mungbean plants conferred enhanced salt tolerance. Cotyledonary node explants were transformed via Agrobacterium tumefaciens mediated transformation using pCAMBIA2301 vector that harbours 35S::AtNHX1 in its T-DNA. Putative transformed plants were selected on kanamycin containing medium. Polymerase chain reaction and Southern blot analysis confirmed the presence, integration and copy number of transgenes in T1 transgenic lines. Reverse transcription-PCR analysis showed higher expression of AtNHX1 in transgenic plants as compared to wild type plants (WT). Under salt stress conditions, T2 transgenic lines displayed less damage and stronger growth phenotypes with concurrent physiological changes as compared to WT. In addition, T2 transgenic lines under salt stress accumulated higher K+/Na+ in the aerial parts and higher [Na+] in roots than WT. Moreover, the T2 transgenic lines showed under NaCl treatment reduced membrane lipid peroxidation and H2O2 and O2 − accumulation, higher levels of antioxidant enzyme activity and increased accumulation of proline and ascorbate than WT. These results indicated that the activity of heterologous AtNHX1 protein contributing enhanced salt tolerance in transgenic mungbean.

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Acknowledgments

We express our sincere thanks to Prof. Sampa Das, Bose Institute for providing Agrobacterium tumefaciens strain, EHA105 and Center for Application of Molecular Biology to International Agriculture (CAMBIA), Australia for pCAMBIA2301. The research was supported partially by various Grants from Department of Biotechnology, Government of India (BT/PR10818/AGR/02/591/2008, BT/01/NE/PS/08 and BT/PR13560/COE/34/44/2015) to LS. DPS and SM are grateful to DBT and MHRD for Research Fellowship at IIT Guwahati.

Authors contribution

LS conceived and designed the experiments. DPS and SK produced the transgenic plants. DPS, SM and YK performed molecular characterisation of transgenics. DPS performed the physiological and biochemical experiments. SKP assisted in physiological assays. LS critically analysed the data and wrote the manuscript.

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Authors and Affiliations

  1. Department of Bioscience and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, India

    Debee Prasad Sahoo, Sanjeev Kumar, Sagarika Mishra, Yasufumi Kobayashi & Lingaraj Sahoo

  2. Department of Life Science and Bioinformatics, Assam University, Silchar, 788011, India

    Debee Prasad Sahoo & Sanjib Kumar Panda

  3. Faculty of Applied Biological Sciences, Gifu University, Gifu, 501-1193, Japan

    Yasufumi Kobayashi

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  1. Debee Prasad Sahoo
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  2. Sanjeev Kumar
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Correspondence to Sanjib Kumar Panda or Lingaraj Sahoo.

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Sahoo, D.P., Kumar, S., Mishra, S. et al. Enhanced salinity tolerance in transgenic mungbean overexpressing Arabidopsis antiporter (NHX1) gene. Mol Breeding 36, 144 (2016). https://doi.org/10.1007/s11032-016-0564-x

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