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Highly Efficient, 5′-Sequence-Specific Transgene Silencing in a Complex Polyploid

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Abstract

Sugarcane, a complex polyploid, shows highly efficient and rapidly imposed silencing of diverse transgene constructs. Silencing is 5′-sequence-specific, copy-number independent, developmentally regulated and post-transcriptional in the plants first regenerated from transgenic callus. The results challenge some early generalizations from studies in model dicotyledonous plants, about causes of transgene silencing and approaches to avoid the problem. We reason that patchy and progressive transgene silencing, seen in most other plant species, may be associated with increasing endopolyploidy during maturation of differentiated tissues. The experimentally tractable sugarcane system allows features likely to trigger or protect from silencing to be tested, and ultimately controlled to improve the efficiency of plant improvement by genetic transformation.

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Abbreviations

cDNA:

complementary DNA

DNA:

deoxyribonucleic acid

dsRNA:

double-stranded RNA

GUS:

glucuronidase (encoded by the uidA gene)

HDGS:

homology-dependent gene silencing

LUC:

luciferase (encoded by the luc gene)

NPTII:

neomycin phosphotransferase (encoded by the aphA gene)

PTGS:

post-transcriptional gene silencing

RNA:

ribonucleic acid

RNAi:

RNA interference (RNA-mediated gene silencing)

siRNA:

small interfering RNA (intermediate in RNAi)

TGS:

transcriptional gene silencing

Ubi:

ubiquitin

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Acknowledgements

We thank Tony Cavallaro, Janette McDouall Stuart, and Simon Hansom for research assistance, and BSES Limited for assistance with sugarcane. This work was supported by the Australian Research Council through the industry collaborative grant scheme, and by the Sugar Research and Development Corporation.

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Authors and Affiliations

  1. Department of Botany / BIOL, The University of Queensland, Brisbane, 4072, Australia

    Robert G. Birch, Robert S. Bower & Adrian R. Elliott

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  1. Robert G. Birch
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  2. Robert S. Bower
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  3. Adrian R. Elliott
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Correspondence to Robert G. Birch.

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Communicated by: Ray Ming

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Birch, R.G., Bower, R.S. & Elliott, A.R. Highly Efficient, 5′-Sequence-Specific Transgene Silencing in a Complex Polyploid. Tropical Plant Biol. 3, 88–97 (2010). https://doi.org/10.1007/s12042-010-9047-0

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