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Co-expression of genes ApGSMT2 and ApDMT2 for glycinebetaine synthesis in maize enhances the drought tolerance of plants

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Abstract

Glycinebetaine plays an important role in the protection mechanism of many plants under various stress conditions. In this study, genetically engineered maize plants with an enhanced ability to synthesise glycinebetaine (GB) were produced by introducing two genes, glycine sarcosine methyltransferase gene (ApGSMT2) and dimethylglycine methyltransferase gene (ApDMT2), from the bacterium Aphanothece halophytica. Southern blotting and RT-PCR analysis demonstrated that the two genes were integrated into the maize genome and expressed. The increased expression levels of ApGSMT2 and ApDMT2 under drought conditions facilitated GB accumulation in the leaves of transgenic maize plants and conferred improved drought tolerance. Under drought conditions, the transgenic plants showed an increased accumulation of sugars and free amino acids, greater chlorophyll content, a higher photosynthesis rate and biomass, and lower malondialdehyde and electrolyte leakage compared to the wild-type; these results suggest that GB provides vital protection against drought stress. Under normal conditions, the transgenic plants did not show decreased biomass and productivity, which indicated that the co-expression of ApGSMT2 and ApDMT2 in maize plays an important role in its tolerance to drought stress and does not lead to detrimental effects. It was concluded that the co-expression of ApGSMT2 and ApDMT2 in maize is an effective approach to enhancing abiotic stress tolerance in maize breeding programmes.

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Abbreviations

BADH:

Betaine-aldehyde dehydrogenase

CMO:

Choline mono-oxygenase

COD:

Choline oxidase

CTAB:

Cetyltrimethylammonium bromide

DIG:

Digoxigenin

EPSPS:

5-Enolpyruvyl-shikimate-3-phosphate synthase

GB:

Glycinebetaine

MDA:

Malondialdehyde

PEG:

Polyethylene glycol

PS II:

Photosystem II

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Acknowledgments

This research was supported by the Hi-Tech Research and Development (863) Program of China (2012AA10A306).

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Authors and Affiliations

  1. Maize Institute, Shandong Academy of Agricultural Sciences/National Maize Improvement Sub-Center, Jinan, 250100, People’s Republic of China

    Chunmei He, Qiang Liu, Tieshan Liu, Chunxiao Liu, Liming Wang & Juren Zhang

  2. School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, 250100, Shandong, China

    Ying He & Juren Zhang

Authors
  1. Chunmei He
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  2. Ying He
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  3. Qiang Liu
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  4. Tieshan Liu
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  5. Chunxiao Liu
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  6. Liming Wang
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  7. Juren Zhang
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Correspondence to Liming Wang or Juren Zhang.

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Fig. 2S

Maize seedlings growth of each line under normal and osmotic stress conditions. Maize seedlings were cultured in Hoagland solution (a1) and in Hoagland solution containing 12% (w/v) PEG-6000 (a2). Seedlings grown in Hoagland solution were transferred to Hoagland solution supplemented with 12% (w/v) PEG-6000 to be subjected to osmotic stress for 7 d. there are no significant differences between the transgenic lines and the WT seedlings under normal Hoagland solution culture, the WT plants wilted while the three transgenic lines grew much better when cultured under osmotic-stress (JPEG 1210 kb)

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He, C., He, Y., Liu, Q. et al. Co-expression of genes ApGSMT2 and ApDMT2 for glycinebetaine synthesis in maize enhances the drought tolerance of plants. Mol Breeding 31, 559–573 (2013). https://doi.org/10.1007/s11032-012-9815-7

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  • DOI: https://doi.org/10.1007/s11032-012-9815-7

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