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Live-Cell Imaging of Rhabdovirus-Induced Morphological Changes in Plant Nuclear Membranes

July 2005 , Volume 18 , Number  7
Pages  703 - 709

Michael Goodin , 1 Sharon Yelton , 1 Debasish Ghosh , 2 Stephanie Mathews , 1 and Judith Lesnaw 2

1Department of Plant Pathology and 2Department of Biology, University of Kentucky, Lexington 40546, U.S.A.


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Accepted 9 March 2005.

Potato yellow dwarf virus (PYDV) and Sonchus yellow net virus (SYNV) belong to the genus Nucleorhabdovirus. These viruses replicate in nuclei of infected cells and mature virions accumulate in the perinuclear space after budding through the inner nuclear membrane. Infection of transgenic Nicotiana benthamiana 16c plants (which constitutively express green fluorescent protein (GFP) targeted to endomembranes) with PYDV or SYNV resulted in virusspecific patterns of accumulation of both GFP and membranes within nuclei. Using immunolocalization and a lipophilic fluorescent dye, we show that the sites of the relocalized membranes were coincident with foci of accumulation of the SYNV nucleocapsid protein. In contrast to the effects of PYDV and SYNV, inoculation of 16c plants with plusstrand RNA viruses did not result in accumulation of intranuclear GFP. Instead, such infections resulted in accumulation of GFP around nuclei, in a manner consistent with proliferation of the endoplasmic reticulum. We propose that the relocalization of GFP in 16c plants can be used to study sites of rhabdovirus accumulation in live cells. This study is the first to use live-cell imaging to characterize the effects of rhabdoviruses on plant nuclear membranes.


Additional keywords: glycoprotein , nucleus , viroplasm .

© 2005 The American Phytopathological Society