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Isolation, functional characterization and stress responses of raffinose synthase genes in sugar beet

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Abstract

Raffinose (sucrosylgalactoside oligosaccharide) is a water soluble carbohydrate and accumulates in response to abiotic stresses in plants. Plant raffinose synthases are poorly characterized, and the genes involved in raffinose biosynthesis are unknown in sugar beet. Here, we report the isolation of two genes encoding raffinose synthase (BvRS1 and BvRS2) as well as a gene encoding galactinol synthase (BvGolS1) from sugar beet. BvRS1 and BvRS2 show high homologies to Arabidopsis raffinose synthase AtRS5. BvRS1 and BvGolS1 were expressed in Escherichia coli. Crude extracts showed the activities of raffinose synthase and galactinol synthase. The K m values of BvRS1 for galactinol and sucrose and the K m values of BvGolS1 for UDP-galactose and myo-inositol were determined. The expression levels of BvRS1 were significantly higher than that of BvRS2. The mRNA for BvRS1 was rapidly induced by cold stress whereas the mRNA for BvRS2 was slowly induced by cold and salt stresses. These data suggest that BvRS1 and BvRS2 encode raffinose synthase genes responsible to cold and salt stress, respectively.

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Abbreviations

Bv:

Beta vulgaris

DTT:

Dithiothreitol

Gol:

Galactinol

GolS:

Galactinol synthase

Raf:

Raffinose

RafS:

Raffinose synthase

RFO:

Raffinose family of oligosaccharides

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Acknowledgements

This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan and the International Center for Green Biotechnology of Meijo University to T.T.

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Author notes

  1. Koji Yamane

    Present address: Graduate School of Agricultural Sciences, Kinki University, Nara, 631-8505, Japan

Authors and Affiliations

  1. Graduate School of Environmental and Human Sciences, Meijo University, Nagoya, 468-8502, Japan

    Kunihide Kito, Takahiro Yamamori, Yoshito Tanaka & Teruhiro Takabe

  2. Research Institute, Meijo University, Nagoya, Aichi, 468-8502, Japan

    Koji Yamane & Teruhiro Takabe

  3. Hokkaido Agricultural Research Center, National Agriculture and Food Organization, Hokkaido, 082-0081, Japan

    Hiroaki Matsuhira

Authors
  1. Kunihide Kito
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  2. Koji Yamane
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  5. Yoshito Tanaka
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Corresponding author

Correspondence to Teruhiro Takabe.

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The authors declare that they have no conflict of interest.

Additional information

Nucleotide sequence data for raffinose synthase genes, BvRS1 and BvRS2, and galactinol synthase gene, BvGolS1, from sugar beet are available in the DDJB database under the accession numbers LC179838, LC179839 and LC179840, respectively.

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Kito, K., Yamane, K., Yamamori, T. et al. Isolation, functional characterization and stress responses of raffinose synthase genes in sugar beet. J. Plant Biochem. Biotechnol. 27, 36–45 (2018). https://doi.org/10.1007/s13562-017-0413-y

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  • DOI: https://doi.org/10.1007/s13562-017-0413-y

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