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Nuclear Import And Interactions Of Potato Yellow Dwarf Virus Nucleocapsid, Matrix, And Phosphoprotein, Gavin Lloyd Franklin Anderson Jan 2014

Nuclear Import And Interactions Of Potato Yellow Dwarf Virus Nucleocapsid, Matrix, And Phosphoprotein, Gavin Lloyd Franklin Anderson

Theses and Dissertations--Plant Pathology

Potato yellow dwarf virus (PYDV) is the type species of the genus Nucleorhabdovirus and, like all members of this genus, replication and morphogenesis occurs inside the nuclei of infected cells. Protein localization prediction algorithms failed to identify a nuclear localization signal (NLS) in PYDV nucleocapsid (N) protein, although PYDV-N has been shown to localize exclusively to the nucleus when expressed as a green fluorescent protein (GFP):N fusion in plant cells. Deletion analysis and alanine-scanning mutagenesis identified two amino acid motifs, 419QKR421 and 432KR433, that were shown to be essential for nuclear import and interaction with …


Evaluation Of Pcr-Based Methods For Rapid, Accurate Detection And Monitoring Of Verticillium Dahliae In Woody Hosts By Real-Time Polymerase Chain Reaction, Baker Diwan Getheeth Aljawasim Jan 2014

Evaluation Of Pcr-Based Methods For Rapid, Accurate Detection And Monitoring Of Verticillium Dahliae In Woody Hosts By Real-Time Polymerase Chain Reaction, Baker Diwan Getheeth Aljawasim

Theses and Dissertations--Plant Pathology

Verticillium wilt, caused by Verticillium dahliae Kleb, is one of the most economically important diseases of woody hosts such as ash (Fraxinus spp.), sugar maple (Acer saccharum), and redbud (Cercis canadensis). The causal agent has a broad host range, including not only woody hosts but also important vegetable and field crops, and it is distributed worldwide. Diagnosis of V. dahliae in infected woody hosts is often based on the occurrence of vascular discoloration and time-consuming isolation. However, not all woody hosts exhibit vascular discoloration symptoms, and not all vascular discoloration symptoms are due to infection …


Ether Bridge Formation And Chemical Diversification In Loline Alkaloid Biosynthesis, Juan Pan Jan 2014

Ether Bridge Formation And Chemical Diversification In Loline Alkaloid Biosynthesis, Juan Pan

Theses and Dissertations--Plant Pathology

Loline alkaloids, found in many grass-Epichloë symbiota, are toxic or feeding deterrent to invertebrates. The loline alkaloids all share a saturated pyrrolizidine ring with a 1-amine group and an ether bridge linking C2 and C7. The steps in biosynthesis of loline alkaloids are catalyzed by enzymes encoded by a gene cluster, designated LOL, in the Epichloë genome. This dissertation addresses the enzymatic, genetic and evolutionary basis for diversification of these alkaloids, focusing on ether bridge formation and the subsequent modifications of the 1-amine to form different loline alkaloids.

Through gene complementation of a natural lolO mutant and comparison …


Rna Sequence Determinants Of A Coupled Termination-Reinitiation Strategy For Translation Of Downstream Orf In Helminthosporium Victoriae Virus 190s And Other Victoriviruses (Family Totiviridae), Hua Li Jan 2014

Rna Sequence Determinants Of A Coupled Termination-Reinitiation Strategy For Translation Of Downstream Orf In Helminthosporium Victoriae Virus 190s And Other Victoriviruses (Family Totiviridae), Hua Li

Theses and Dissertations--Plant Pathology

Double-stranded RNA fungal virus Helminthosporium victoriae virus 190S (genus Victorivirus, family Totiviridae) contains two large open reading frames (ORFs) that overlap in the tetranucleotide AUGA. Translation of the downstream ORF, which encodes the RNA-dependent RNA polymerase (RdRp), was previously proposed to depend on ribosomal reinitiation following termination of the upstream ORF, which encodes the capsid protein. In this study, I provided evidence to confirm that coupled termination-reinitiation (stop-restart) is indeed used. A dual-fluorescence method was established to define the RNA sequence determinants for RdRp translation. Stop-restart depends on a 32-nt stretch of RNA sequence immediately upstream of the …


Key Roles Of Sub-Cellular Membranes And Co-Chaperone In Tombusvirus Replication, Kai Xu Jan 2014

Key Roles Of Sub-Cellular Membranes And Co-Chaperone In Tombusvirus Replication, Kai Xu

Theses and Dissertations--Plant Pathology

Positive strand RNA viruses, inculding tombusviruses, are known to utilize cellular membranes to assemble their replicase complexes (VRCs). Two tombusviruses , Tomato bushy stunt virus (TBSV) and Carnation Italian ringspot virus (CIRV), replicate on different organellar membranes, peroxisomes or endoplasmic reticulum (ER) for TBSV and mitochodria outer membranes in case of CIRV. I showed that both TBSV and CIRV replicase proteins could assemble VRCs and replicate viral RNA on purified microsomes (ER) and mitochondria. Different efficiencies of assembly was shown determined by multiple domains on TBSV or CIRV replication proteins.

To study why VRC assembly could occur on an alternative …