| Abstract Detail
Plant-Pathogen Interactions Huang, Tyng-Shyan [1], Wang, Aiming [2]. The molecular identification and functional characterization of host proteins associated with Plum pox virus (PPV) infection. Plum pox virus (PPV) belongs to the genus Potyvirus, family Potyviridae, which is the largest and most economically important plant virus family. PPV is considered as one of the most serious threats to the stone fruits and ornamental Prunus species. Currently, there is no natural resistance to PPV that can be introduced into elite cultivars through conventional breeding. Despite extensive research and study on PPV interaction with its host, its exact replication mechanism remains unclear. PPV encodes a limited number of viral proteins in its genome; hence it has evolved to recruit host proteins to aid its translation and replication upon infection. In order to identify PPV host proteins involved in the viral replication complex (VRC), a PPV infected peach cDNA library was constructed and used in a yeast two-hybrid screen. Two PPV-encoded proteins, NIb (viral RNA polymerase) and VPg (viral genome linked protein) were cloned as baits in the yeast two-hybrid assay. The library screening was conducted at three different temperatures to include interactions that are temperature sensitive. Three rounds of screening have identified a total of 215 host candidates that interacted strongly with NIb and VPg. Based on their predicted functions, five of these candidate genes were selected for further functional analysis. Interactions between the viral proteins and these selected host proteins are being verified by a GST pull-down assay followed by in planta confirmation using the bimolecular fluorescence complementation assay (BiFC). Arabidopsis T-DNA insertion lines of corresponding homologous genes are being used for PPV infection test in order to determine if these candidate genes are required for PPV infection. Further biochemical assays and reverse genomics tools will be used to elucidate the molecular mechanisms of the identified genes in the PPV infection process.
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1 - The University of Western Ontario, Department of Biology, Biological & Geological Building, 1151 Richmond Street, London, Ontario, N6A 5B7, Canada
2 - Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research Centre
Keywords:
plant-virus interaction
protein-protein interaction
Gene Silencing
viral replication complex
Plum pox virus
virus interacting host factors
recessive resistance
RNA viruses.
Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P15019
Abstract ID:293 |
