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Cloning, characterization and expression of OsGLN2, a rice endo-1,3-β-glucanase gene regulated developmentally in flowers and hormonally in germinating seeds

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Abstract

We report here the isolation and characterization of a new endo-1,3-β-glucanase (1,3-β-GLU) cDNA, OsGLN2, that is expressed both in flowers and in germinating seeds of rice (Oryza sativa L.). The isolated OsGLN2 gene encoded a protein which displayed 72%, 93% and 92% identity at the amino acid level with those encoded by barley GII, rice Gns4 and glu1 1,3-β-GLU genes, respectively. A GST-OsGLN2 recombinant protein expressed in Escherichia coli preferentially hydrolyzed Laminaria digitata 1,3;1,6-β-glucan and liberated only oligosaccharides, suggesting that the enzyme can be classified as a 1,3-β-GLU. Northern analysis with a 3′-UTR gene-specific probe revealed that OsGLN2 is expressed exclusively in the paleae and lemmas during flowering, and no expression of OsGLN2 was detected in other tissues such as leaf blades, leaf sheaths, stems, nodes and roots in mature rice plants. The OsGLN2 gene is also expressed in germinating seeds, where its expression is predominant in endosperms rather than embryos. In de-embryonated rice half-seeds, addition of gibberellin A3 (GA) greatly enhanced expression of the OsGLN2 gene, while the GA-induced gene expression was suppressed strongly by abscisic acid (ABA). This is the first report, to our knowledge, that OsGLN2 encodes a 1,3-β-GLU and is expressed specifically in paleae and lemmas during flowering and in germinating seeds, where its expression is enhanced by GA and suppressed by ABA.

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Abbreviations

ABA :

Abscisic acid

GA :

Gibberellin A3

1,3-β-GLU :

Endo-1,3-β-glucanase

GST :

Glutathione S-transferase

IPTG :

Isopropyl-1-thio-β-galactopyranoside

3′-UTR :

3′-Untranslated region

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Acknowledgements

We thank Dr. James R. Ketudat Cairns for critical reading of the manuscript. This work was supported in parts by a grant from the Science and Technology Agency of Japan to M.A.P and by a grant (Japan Rice Genome Project Grant no. PR-1103) from the Ministry of Agriculture Forestry and Fisheries of Japan to T.A.

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  1. Department of Low-Temperature Science, National Agricultural Research Center for Hokkaido Region, 1 Hitsujigaoka, Toyohira-ku, Sapporo, 062-8555, Hokkaido, Japan

    Takashi Akiyama, M. Arumugam Pillai & Naoki Sentoku

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  1. Takashi Akiyama
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  2. M. Arumugam Pillai
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  3. Naoki Sentoku
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Correspondence to Takashi Akiyama.

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Akiyama, T., Pillai, M.A. & Sentoku, N. Cloning, characterization and expression of OsGLN2, a rice endo-1,3-β-glucanase gene regulated developmentally in flowers and hormonally in germinating seeds. Planta 220, 129–139 (2004). https://doi.org/10.1007/s00425-004-1312-8

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