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Selectable marker-free transgenic barley producing a high level of cellulase (1,4-β-glucanase) in developing grains

  • Genetic Transformation and Hybridization
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Abstract

The use of barley grains as bioreactors for high-level production of cellulase (1,4-β-glucanase) was investigated. A hybrid cellulase gene, cel-hyb1, driven by the rice GluB-1 promoter was expressed specifically in developing endosperm. Codon usage optimisation of cel-hyb1 increased its expression in barley grains 527-fold and led to cellulase production of up to 1.5% of total grain protein. CEL-HYB1 enzyme in barley grains was highly stable during post-harvest storage. Selectable marker gene (hph) was subsequently eliminated from transgenic lines through segregation of hph from synthetic cel-hyb1 (syn.cel-hyb1) in T1 progeny, using a binary plasmid containing hph and syn.cel-hyb1 in separate T-DNAs. These data suggest that barley grains can potentially be used for the commercial production of cellulase.

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Fig. 1A, B.
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Abbreviations

AZCL:

Azurine-crosslinked

CM:

Carboxymethyl

HE:

Hydroxyethyl

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Acknowledgement

This work was partly supported by Australian Grains Research and Development Corporation.

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Authors and Affiliations

  1. CSIRO Plant Industry, 120 Meiers Road, Indooroopilly, Brisbane, Australia

    G. P. Xue, M. Patel, J. S. Johnson, D. J. Smyth & C. E. Vickers

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  1. G. P. Xue
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  2. M. Patel
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  3. J. S. Johnson
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  4. D. J. Smyth
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  5. C. E. Vickers
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Corresponding author

Correspondence to G. P. Xue.

Additional information

Communicated by P. Lakshmanan

G. P. Xue and M. Patel made an equal contribution

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Xue, G.P., Patel, M., Johnson, J.S. et al. Selectable marker-free transgenic barley producing a high level of cellulase (1,4-β-glucanase) in developing grains. Plant Cell Rep 21, 1088–1094 (2003). https://doi.org/10.1007/s00299-003-0627-4

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  • DOI: https://doi.org/10.1007/s00299-003-0627-4

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