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Rice choline monooxygenase (OsCMO) protein functions in enhancing glycine betaine biosynthesis in transgenic tobacco but does not accumulate in rice (Oryza sativa L. ssp. japonica)

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Abstract

Glycine betaine (GB) is a compatible quaternary amine that enables plants to tolerate abiotic stresses, including salt, drought and cold. In plants, GB is synthesized through two-step of successive oxidations from choline, catalyzed by choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH), respectively. Rice is considered as a typical non-GB accumulating species, although the entire genome sequencing revealed rice contains orthologs of both CMO and BADH. Several studies unraveled that rice has a functional BADH gene, but whether rice CMO gene (OsCMO) is functional or a pseudogene remains to be elucidated. In the present study, we report the functional characterization of rice CMO gene. The OsCMO gene was isolated from rice cv. Nipponbare (Oryza sativa L. ssp. japonica) using RT-PCR. Northern blot demonstrated the transcription of OsCMO is enhanced by salt stress. Transgenic tobacco plants overexpressing OsCMO results in increased GB content and elevated tolerance to salt stress. Immunoblotting analysis demonstrates that a functional OsCMO protein with correct size was present in transgenic tobacco but rarely accumulated in wild-type rice plants. Surprisingly, a large amount of truncated proteins derived from OsCMO was induced in the rice seedlings in response to salt stresses. This suggests that it is the lack of a functional OsCMO protein that presumably results in non-GB accumulation in the tested rice plant.

Key message Expression and transgenic studies demonstrate OsCMO is transcriptionally induced in response to salt stress and functions in increasing glycinebetaine accumulation and enhancing tolerance to salt stress. Immunoblotting analysis suggests that no accumulation of glycinebetaine in the Japonica rice plant presumably results from lack of a functional OsCMO protein.

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Acknowledgments

This work was Supported by the 973 Program from Chinese Ministry of Science and Technology (2011CB100401), the National Science Fund of China for Distinguished Young Scientists (30825030), the Key Project from Chongqing Local Government (2010AA1019), National Natural Science Foundation of China (30970260, 30770466 and 30971752) and the National Science and Technology Key Project of China on GMO Cultivation for New Varieties (Nos. 2009ZX08001-011B, 2009ZX08009-072B).

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Authors and Affiliations

  1. Ministry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Science and State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu, 610064, China

    Di Luo, Jinde Yu & Yongsheng Liu

  2. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, 230009, China

    Xiangli Niu & Yongsheng Liu

  3. Laboratory for Chemistry of Traditional Chinese medicine, Chengdu University, Chengdu, 610106, China

    Jun Yan & Xiaojun Gou

  4. Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, Fudan University, Shanghai, 200433, China

    Bao-Rong Lu

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  1. Di Luo
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  3. Jinde Yu
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Correspondence to Bao-Rong Lu or Yongsheng Liu.

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Communicated by S. Schillberg.

D. Luo and X. Niu contributed equally to the work.

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Luo, D., Niu, X., Yu, J. et al. Rice choline monooxygenase (OsCMO) protein functions in enhancing glycine betaine biosynthesis in transgenic tobacco but does not accumulate in rice (Oryza sativa L. ssp. japonica). Plant Cell Rep 31, 1625–1635 (2012). https://doi.org/10.1007/s00299-012-1276-2

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