A Genetic Approach to the Identification of Plant Genes Involved in Viral Movement
Abstract
Plant viruses encode a movement protein that is required for movement and systemic infection of host plants. We propose a genetic approach to identifying host proteins involved in plant viral movement. We are using as a model red clover necrotic mosaic virus (RCNMV) which infects, but does not kill Arabidopsis.. We plan to create two transgenic plant lines of Arabidopsis, one that expresses the firefly luciferase (Luc) gene, and one that expresses the codA (cytosine deaminase, CD) gene. Both genes will be under the transcriptional control of the RCNMV subgenomic promoter. We will identify transgenic plant lines that express Luc or CD upon infection with RCNMV. Phenotypically these lines will either be luminescent (Luc) or sensitive to 5-fluorouracil (5FU) (CD) when infected with RCNMV. A library of EMS mutants will be generated for Luc- and CD-expressing plant lines. Plantlets from the mutagenized Luc-expressing lines will be infected with RCNMV and screened for the loss of luminescence. Plantlets from the mutagenized CD-expressing lines will be infected with RCNMV and selected for resistance to 5FU. Surviving mutants will be examined for the presence of RCNMV in the inoculated and upper leaves. Mutants that are unable to support RCNMV infection will be selfed and their progeny tested as described above. The mutated gene in each of these lines will then be cloned and sequenced.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 30, 1999
- Accession Number
- ADA388438
Entities
People
- Donna M. Cookmeyer
- Steven A. Lommel
Organizations
- North Carolina State University