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First infectious clone of the propagatively transmitted Oat blue dwarf virus

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Abstract

Oat blue dwarf virus (OBDV) is a small, phloem-limited marafivirus that replicates in its leafhopper vector. We have developed complete cDNA clones of OBDV from which infectious transcripts may be derived––the first such clones for any propagatively transmitted plant virus. Prior to clone construction, the reported sequences of the 5′ and 3′ ends were confirmed using 5′ RACE, primer extension, and ligation-anchored PCR. Using vascular puncture of maize seeds with capped transcripts, multiple clones were shown to be infectious at an average rate of 24.3% (range 14–36%). Aster leafhoppers successfully transmitted OBDV to oats and barley after feeding on detached, infected maize leaves. Proteins and RNAs consistent in size with those expected in OBDV infection were detected in young leaves via western and northern blotting, respectively. One construct, pOBDV-2r, was designated as the reference clone. An infectious clone of OBDV will be valuable in examining the interaction of this virus with both its insect and plant hosts.

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Acknowledgments

The authors wish to thank Ryne Hendrickson and Renee McClean for technical assistance. We also thank Dr. Shiaoman Chao for assistance with primer extension analysis and Dr. Margaret Redinbaugh for helpful discussion regarding the vascular puncture technique.

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Authors and Affiliations

  1. Northern Crop Science Laboratory, USDA-ARS Cereal Crops Research Unit, P.O. Box 5677, Fargo, ND, 58105-5677, USA

    Michael C. Edwards & John J. Weiland

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  1. Michael C. Edwards
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  2. John J. Weiland
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Correspondence to Michael C. Edwards.

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Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.

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Edwards, M.C., Weiland, J.J. First infectious clone of the propagatively transmitted Oat blue dwarf virus . Arch Virol 155, 463–470 (2010). https://doi.org/10.1007/s00705-010-0603-6

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  • DOI: https://doi.org/10.1007/s00705-010-0603-6

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