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Molecular characterization and analysis of conserved potyviral motifs in bean common mosaic virus (BCMV) for RNAi-mediated protection

  • Elizabeth A. Worrall
  • Alice C. Hayward
  • Stephen J. Fletcher
  • Neena Mitter
Original Article

Abstract

Australian bean common mosaic virus (BCMV) isolates were sequenced, and the sequences were compared to global BCMV and bean common mosaic necrosis virus (BCMNV) sequences and analysed for conserved potyviral motifs to generate in planta RNA-interference (RNAi) resistance. Thirty-nine out of 40 previously reported potyvirus motifs were conserved among all 77 BCMV/BCMNV sequences. Two RNAi target regions were selected for dsRNA construct design, covering 920 bp of the nuclease inclusion b (NIb) protein and 461 bp of the coat protein (CP). In silico prediction of the effectiveness of these constructs for broad-spectrum defence against the 77 BCMV and BCMNV sequences was done via analysis of putative 21-nucleotide (nt) and 22-nt small-interfering RNAs (siRNAs) generated from the target regions. The effectiveness of both constructs for siRNA generation and BCMV RNAi-mediated resistance was validated in Nicotiana benthamiana transient assays.

Notes

Acknowledgments

The authors would like to thank Queensland Alliance of Agriculture and Food Innovation (QAAFI), Australia and the University of Queensland, Australia, for their support.

Funding

This study was funded by the Accelerated Partnership Grant, Queensland Government (2014000652), awarded to N.M. with Nufarm Australia Limited as the industry partner. E.A.W. PhD programme with N.M. is supported by a scholarship from the University of Queensland.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Data availability

The datasets generated and/or analysed during the current study are available in the NCBI GenBank repository, https://www.ncbi.nlm.nih.gov. Only complete genomes of BCMV and BCMNV were used.

Supplementary material

705_2018_4065_MOESM1_ESM.doc (1.8 mb)
Supplementary material 1 (DOC 1792 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Elizabeth A. Worrall
    • 1
  • Alice C. Hayward
    • 1
  • Stephen J. Fletcher
    • 1
  • Neena Mitter
    • 1
  1. 1.Centre of Horticultural Science, Queensland Alliance for Agriculture and Food InnovationThe University of QueenslandBrisbaneAustralia

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