| Description: | Simple RNA plant viruses such as tobacco mosaic virus (TMV) are known to alter the cellular machinery of their hosts in order to replicate their genomes and spread both locally and systemically. In addition, plants actively respond to viral infections in a variety of ways that are aimed at limiting the infection process, including host derived hypersensitive responses and gene silencing. Thus, viruses have a significant impact on the expression patterns of their host's genome. The goal of this proposal is to identify plant genes that are differentially regulated during viral infections that result in disease responses. Specifically, the interaction between TMV, the type member of the tobamovirus group, and the Arabidopsis ecotype Shahdara will be examined. This ecotype allows rapid systemic movement of TMV into upper non-inoculated leaves within five days post-inoculation. This rapid movement is accompanied by severe disease symptoms that include stunting, leaf curling, and necrosis. In comparison, ecotypes such as Columbia permit only limited systemic TMV movement after 15 days post-inoculation and display no symptoms. In addition, Northern blot analysis indicates that the host pathogen related proteins, PR-1 and PR-5, are induced in Shadhara upon infection with TMV but not in ecotype Columbia. The observed responses of Shahdara to TMV infection are similar to other virus - host interactions previously studied in tobacco, tomato, and pepper. This makes the TMV - Shahdara system ideally suited for the investigation of fundamental plant ? virus interactions on a genomic level. For the proposed array experiments, mature Shahdara leaves will be inoculated with 10 micrograms of purified TMV. RNA will be extracted from these leaves at three days post-inoculation. This is the earliest time point in which reliable PR-1 and PR-5 host gene induction can be observed. Both PR-1 and PR-5 will also serve as internal controls to determine the quality of isolated mRNA. Control RNA will be acquired in a similar fashion from mock inoculated leaves of Shahdara. Duplicate sets of RNA pooled from multiple TMV and mock inoculated leaves will be provided for the analysis. It is anticipated that information obtained from these experiments should provide new insights into how viruses regulate the gene expression of their hosts and how plants respond to the presence of a virulent viral pathogen. |
NASCArrays Information at The BAR
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