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A synthetic xylanase as a novel reporter in plants

  • Genetic Transformation and Hybridization
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Abstract

Transient gene expression assays are often used to screen promoters before stable transformation. Current transient quantification methods have several problems, including a lack of reporter gene stability and expense. Here we report a synthetic, codon-optimised xylanase gene (sXynA) as a reporter gene for quantitative transient analyses in plants. Azurine-crosslinked xylan (AZCL-xylan) was used as a substrate for assaying xylanase activity. The enzymatic nature of the protein allows for sensitive assays at the low levels of transgene protein found in transiently transformed tissue extracts. The xylanase (XYN) protein is stable, activity slopes are linear over long time periods and assays are cost-effective. Coupled with the GUSPlus reporter gene, the XYN reporter allows sensitive and accurate quantification of gene control sequences in transient expression systems.

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Abbreviations

Act1 :

Rice actin promoter

AZCL-xylan:

Azurine cross-linked xylan

AU:

absorbance units

Blt4.9 :

Barley lipid transfer protein promoter

GEB :

GUS extraction buffer

GFP :

Green fluorescent protein

GluB-1 :

Rice glutelin B-1 promoter

GUS :

β-Glucuronidase

LUC :

Luciferase

sXynA :

Synthetic xylanase A gene

Ubi-1 :

Maize ubiquitin promoter

XAB :

Xylanase assay buffer

XYN :

Xylanase

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Acknowledgements

This work was supported by the Grains Research and Development Corporation grant GRS19.

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Authors and Affiliations

  1. CSIRO Plant Industry, 306 Carmody Rd, St Lucia, 4067, Brisbane, QLD, Australia

    C. E. Vickers & G. P. Xue

  2. School of Life Sciences, The University of Queensland, CSIRO Plant Industry, St Lucia, 4072, Brisbane, QLD, Australia

    C. E. Vickers & P. M. Gresshoff

  3. ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, St Lucia, 4072, Brisbane, QLD, Australia

    P. M. Gresshoff

Authors
  1. C. E. Vickers
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  2. G. P. Xue
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  3. P. M. Gresshoff
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Correspondence to C. E. Vickers.

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Communicated by P. Lakshmanan

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Vickers, C.E., Xue, G.P. & Gresshoff, P.M. A synthetic xylanase as a novel reporter in plants. Plant Cell Rep 22, 135–140 (2003). https://doi.org/10.1007/s00299-003-0667-9

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  • DOI: https://doi.org/10.1007/s00299-003-0667-9

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