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Plant Molecular Biology

, Volume 64, Issue 5, pp 549–557 | Cite as

Transgenic cassava resistance to African cassava mosaic virus is enhanced by viral DNA-A bidirectional promoter-derived siRNAs

  • Hervé Vanderschuren
  • Rashid Akbergenov
  • Mikhail M. Pooggin
  • Thomas Hohn
  • Wilhelm Gruissem
  • Peng ZhangEmail author
Article

Abstract

Expression of double-stranded RNA (dsRNA) homologous to virus sequences can effectively interfere with RNA virus infection in plant cells by triggering RNA silencing. Here we applied this approach against a DNA virus, African cassava mosaic virus (ACMV), in its natural host cassava. Transgenic cassava plants were developed to express small interfering RNAs (siRNA) from a CaMV 35S promoter-controlled, intron-containing dsRNA cognate to the common region-containing bidirectional promoter of ACMV DNA-A. In two of three independent transgenic lines, accelerated plant recovery from ACMV-NOg infection was observed, which correlates with the presence of transgene-derived siRNAs 21–24 nt in length. Overall, cassava mosaic disease symptoms were dramatically attenuated in these two lines and less viral DNA accumulation was detected in their leaves than in those of wild-type plants. In a transient replication assay using leaf disks from the two transgenic lines, strongly reduced accumulation of viral single-stranded DNA was observed. Our study suggests that a natural RNA silencing mechanism targeting DNA viruses through production of virus-derived siRNAs is turned on earlier and more efficiently in transgenic plants expressing dsRNA cognate to the viral promoter and common region.

Keywords

African cassava mosaic virus Cassava RNA interference Common region Transcriptional gene silence siRNA Plant recovery 

Abbreviations

ACMV

African cassava mosaic virus

AV1

coat protein

BC1

movement protein

CMD

cassava mosaic disease

CR

common region

dsDNA

double-stranded DNA

dsRNA

double-stranded RNA

Pro

bidirectional promoter of ACMV

PTGS

post-transcriptional gene silencing

RdRP

RNA-dependent RNA polymerase

REn/AC3

replication enhancer protein

Rep/AC1

replication associated protein

siRNA

small interfering RNAs

ssDNA

single-stranded DNA

TrAP/AC2

transcriptional activator protein

TGS

transcriptional gene silencing

MYMV

Mungbean yellow mosaic virus

Wt

wild type

Notes

Acknowledgements

We thank Dr. John Stanley (John Innes Centre) for the ACMV clones, Dr. Johannes Fütterer (ETH Zurich) for helpful discussions. This work was supported by grants from the Swiss Centre for International Agriculture (ZIL), the Eiselen-Foundation-Ulm, the Bill & Melinda Gates Foundation through the Grand Challenges in Global Health initiative and the Indo-Swiss Collaboration in Biotechnology.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Hervé Vanderschuren
    • 1
  • Rashid Akbergenov
    • 2
  • Mikhail M. Pooggin
    • 2
  • Thomas Hohn
    • 2
  • Wilhelm Gruissem
    • 1
  • Peng Zhang
    • 1
    • 3
    Email author
  1. 1.Institute of Plant SciencesETH ZurichZürichSwitzerland
  2. 2.Institute of BotanyUniversity of BaselBaselSwitzerland
  3. 3.Institute of Plant Physiology and EcologyShanghai Institutes for Biological Sciences, Chinese Academy of SciencesShanghaiChina

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