Chimeric Virus Made from crTMV RNA and the Coat Protein of Potato Leafroll Virus is Targeted to the Nucleolus and Can Infect Nicotiana benthamiana Mechanically.

Konstantin O Butenko,Eugene V Skurat,Yuri F Drygin,Olga A Kondakova,Inna A Chaban

doi:10.3390/ht9020011

Abstract

A genetically engineered chimeric virus crTMV-CP-PLRV composed of the crucifer-infecting tobacco mosaic virus (crTMV) RNA and the potato leafroll virus (PLRV) coat protein (CP) was obtained by agroinfiltration of Nicotiana benthamiana with the binary vector pCambia-crTMV-CPPLRV. The significant levels of the chimeric virus enabled direct visualization of crTMV-CP-PLRV in the cell and to investigate the mechanism of the pathogenesis. Localization of the crTMV-CP-PLRV in plant cells was examined by immunoblot techniques, as well as light, and transmission electron microscopy. The chimera can transfer between vascular and nonvascular tissues. The chimeric virus inoculum is capable to infect N. benthamiana mechanically. The distinguishing feature of the chimeric virus, the RNA virus with the positive genome, was found to localize in the nucleolus. We also investigated the role of the N-terminal sequence of the PLRV P3 coat protein in the cellular localization of the virus. We believe that the gene of the PLRV CP can be substituted with genes from other challenging-to-study plant pathogens to produce other useful recombinant viruses.

Highlights

Potato leafroll virus (PLRV) is the type species of the genus Polerovirus, family Luteoviridae
Earlier [7], a vector was constructed on the base of the TMV U1, PLRV (TMV∆coat proteins (CP)-PLRV-CP), in which the CP PLRV gene replaced the normal CP gene
When the chimeric virus TMV∆CP-PLRV-CP was agroinoculated into N. benthamiana, this vector showed limited accumulation in Nicotiana

Summary

Introduction

Potato leafroll virus (PLRV) is the type species of the genus Polerovirus, family Luteoviridae. It is a highly pathogenic, phloem-limited cytoplasmic virus that accumulates in infected plants in low quantities. PLRV, like other luteoviruses, cannot be transmitted mechanically It infects the plants only when delivered into phloem tissues by aphids, grafting, or agroinoculation. It is challenging and complicated to obtain the virus antigen, the coat protein, to study its role in virus infection and to raise virus-specific antibodies for diagnostics in practice. The obtaining of virus antigen, the coat protein, for investigation of its role in viral infection and for increasing of antibodies production for diagnostics, is a challenging and complicated task

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Results

Conclusion