Author ORCID Identifier

https://orcid.org/0000-0001-9268-3707

Year of Publication

2020

Degree Name

Master of Science (MS)

Document Type

Master's Thesis

College

Agriculture, Food and Environment

Department

Plant Pathology

First Advisor

Dr. Michael M. Goodin

Abstract

Viruses rely on host proteins to complete their life cycles. This results in alterations to normal cell physiology to processes which benefit viral processes such as replication and movement to other cells. This may involve the relocalization of host proteins away from their original subcellular targets to sites which may benefit the virus, a process that is not as well understood as it relates to the nucleus. To identify nuclear proteins that may be involved in such processes, a library of random Nicotiana benthamiana cDNAs were expressed as GFP fusions in a transgenic marker lines expressing a histone 2B:RFP nuclear marker. Of the 1,087 proteins screened, 355 were found to be associated with the nucleus. These nuclear proteins were then expressed in plants infected with sonchus yellow net virus, a betanucleorhabdovirus which forms its replication complex in the nucleus. Of the 355 nuclear-associated proteins, 15 were found to relocalize in response to SYNV infection, and to test if this phenomenon was virus-specific, these proteins were then expressed in plants infected with tobacco etch virus (TEV), a potyvirus which replicates in the cytoplasm. Of these 16 proteins, 6 were found to show both general and virus-specific localization patterns between the two viruses. To gain insight into the cellular tropism of SYNV, a cell map was generated in order determine the effect of this virus on membranes in N. benthamiana tissues. Finally, sowthistle yellow vein virus (SYVV), another betanucleorhabdovirus and classical model virus, was identified in the wild for the first time in over 30 years. Sequence and phylogenetic analysis confirmed the identity of this virus with historical laboratory isolates. The availability of SYVV will facilitate future studies that compare the effect of rhabdovirus infection on host nuclear proteins and membranes.

Digital Object Identifier (DOI)

https://doi.org/10.13023/etd.2020.443

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