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Journal of Virology, September 2007, p. 9339-9345, Vol. 81, No. 17
0022-538X/07/$08.00+0     doi:10.1128/JVI.00417-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Baculovirus-Expressed Dicistrovirus That Is Infectious to Aphids

Narinder Pal,¹ Sandhya Boyapalle,^1, Randy Beckett,² W. Allen Miller,² and Bryony C. Bonning^1*

Departments of Entomology,¹ Plant Pathology, Iowa State University, Ames, Iowa 50011²

Received 27 February 2007/ Accepted 17 June 2007

Detailed investigation of virus replication is facilitated by the construction of a full-length infectious clone of the viral genome. To date, this has not been achieved for members of the family Dicistroviridae. Here we demonstrate the construction of a baculovirus that expresses a dicistrovirus that is infectious in its natural host. We inserted a full-length cDNA clone of the genomic RNA of the dicistrovirus Rhopalosiphum padi virus (RhPV) into a baculovirus expression vector. Virus particles containing RhPV RNA accumulated in the nuclei of baculovirus-infected Sf21 cells expressing the recombinant RhPV clone. These virus particles were infectious in R. padi, a ubiquitous aphid vector of major cereal viruses. The recombinant virus was transmitted efficiently between aphids, despite the presence of 119 and 210 vector-derived bases that were stably maintained at the 5' and 3' ends, respectively, of the RhPV genome. The maintenance of such a nonviral sequence was surprising considering that most RNA viruses tolerate few nonviral bases beyond their natural termini. The use of a baculovirus to express a small RNA virus opens avenues for investigating replication of dicistroviruses and may allow large-scale production of these viruses for use as biopesticides.

Published ahead of print on 27 June 2007.

Present address: H. Lee Moffitt Cancer Center & Research Institute, University of South Florida, Tampa, FL 33612.