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

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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.

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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

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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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Authors and Affiliations

  1. Institute of Plant Sciences, ETH Zurich, Universitätstrasse 2, Zürich, 8092, Switzerland

    Hervé Vanderschuren, Wilhelm Gruissem & Peng Zhang

  2. Institute of Botany, University of Basel, Schönbeinstrasse 6, Basel, 4056, Switzerland

    Rashid Akbergenov, Mikhail M. Pooggin & Thomas Hohn

  3. Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China

    Peng Zhang

Authors
  1. Hervé Vanderschuren
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  2. Rashid Akbergenov
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  3. Mikhail M. Pooggin
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  4. Thomas Hohn
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  5. Wilhelm Gruissem
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  6. Peng Zhang
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Correspondence to Peng Zhang.

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Vanderschuren, H., Akbergenov, R., Pooggin, M.M. et al. Transgenic cassava resistance to African cassava mosaic virus is enhanced by viral DNA-A bidirectional promoter-derived siRNAs. Plant Mol Biol 64, 549–557 (2007). https://doi.org/10.1007/s11103-007-9175-6

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  • DOI: https://doi.org/10.1007/s11103-007-9175-6

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