Abstract
Transgenic sweetpotato (cv. Lizixiang) plants exhibiting enhanced salt tolerance were developed using LOW OSMOTIC STRESS 5 (LOS5) with Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harbors the pCAMBIA1300 binary vector with the LOS5 and hygromycin phosphotransferase II (hptII) genes. Selection culture was conducted using 25 mg l−1 hygromycin. A total of 26 plants were produced from the inoculated 200 cell aggregates of Lizixiang via somatic embryogenesis. PCR analysis showed that 23 of the 26 regenerated plants were transgenic plants. All of the transgenic plants exhibited higher salt tolerance compared to the untransformed control plants by in vitro assay for salt tolerance with 86 mM NaCl. When plants were exposed to 86 mM NaCl, 16 transgenic plants had significantly higher levels of superoxide dismutase (SOD), proline, and abscisic acid (ABA) and significantly lower malonaldehyde (MDA) contents than those in untransformed control plants. Salt tolerance of these 16 plants was further evaluated with Hoagland solution containing 86 mM NaCl in a greenhouse. Four of the sixteen had significantly better growth and rooting ability than the remaining 12 plants and control plants. Stable integration of the LOS5 gene into the genome of the 4 salt-tolerant transgenic plants was confirmed by Southern blot analysis, and the copy number of integrated LOS5 gene ranged from 1 to 3. High level of LOS5 gene expression in the 4 salt-tolerant transgenic plants was demonstrated by real-time quantitative PCR analysis. This study provides an important approach for improving salt tolerance of sweetpotato.
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Abbreviations
- ABA:
-
Abscisic acid
- AS:
-
Acetosyringone
- Carb:
-
Carbenicillin
- CTAB:
-
Cetyltrimethylammonium bromide
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- EDTA:
-
Ethylenediaminetetraacetic acid
- ELISA:
-
Enzyme-linked immuno sorbent assay
- hptII:
-
Hygromycin phosphotransferase II gene
- Hyg:
-
Hygromycin
- LB:
-
Luria-Bertani
- MDA:
-
Malonaldehyde
- MS:
-
Murashige and Skoog
- SOD:
-
Superoxide dismutase
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Acknowledgments
We thank Prof. Gong ZZ, College of Biological Sciences, China Agricultural University, Beijing, China, and Prof. Wang T, State Key Laboratory of Agrobiotechnology, Beijing, China, for providing LOS5 gene and Strain EHA 105, respectively. We also thank Dr. Michael Portereiko, Ceres, Inc. USA, for English improvement. This work was supported by China Agro-industry Research System (Sweetpotato), the National High-Tech Research and Development Project of China (no. 2009AA10Z102) and the National Transgenic Plants Project of China (no. 2009ZX08009-064B).
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Shang Gao and Li Yuan contributed equally to this work.
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Gao, S., Yuan, L., Zhai, H. et al. Transgenic sweetpotato plants expressing an LOS5 gene are tolerant to salt stress. Plant Cell Tiss Organ Cult 107, 205–213 (2011). https://doi.org/10.1007/s11240-011-9971-1
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DOI: https://doi.org/10.1007/s11240-011-9971-1