| Description: | Simple RNA viruses such as tobacco mosaic virus (TMV)are known to alter the cellular machinery of their hosts in order 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 hosts' genomes. Our ealier microarray experiments (Proposal numbers 945-CJC-890-HFG-1 and 958-BDG-334-IBH-7) were aimed at identifying host genes that are differentially regulated during TMV infection. The Arabidopsis ecotype, Shahdara, was used for these experiments since it allows rapid systemic movement of TMV as well as displays marked disease symptoms such as stunting, necrosis, leaf curling and late bolting. Gene expression was monitored in infected whole tissues of Shahdara at 4 days (inoculated tissue) and 14 days (systemic tissue) after inoculation with TMV. Expression profiles obtained from these two time points i.e. 4dpi and 14dpi, should reflect host gene expression in response to the presence of a virulent viral pathogen and gene expression in symptom production, respectively. We now propose to verify the expression profiles of the previous two experiments using biological replicates from similarly obtained poly A+ samples. For the proposed experiments, four microarray slides will be used - the first two slides to monitor expression of host genes at 4 dpi and the second set of two slides to similarly monitor gene expression at 14 dpi. Mature Shahdara leaves will be inoculated with 10 ug of purified TMV. At 4 dpi and 14 dpi, total RNA will be extracted in duplicate sets from inoculated and systemic leaves, respectively. Control RNA will be similarly obtained from mock-inoculated Shahdara plants. Poly A+ RNA isolated from the test and control total RNA samples will be used for microarray analysis. Results thus obtained from the use of biological and technical replicates should help verify the accuracy of expression profiles from the previous two experiments. Together this information should provide a better understanding of the physiological changes occurring within the host in response to the establishment of a virulent viral pathogen. |
NASCArrays Information at The BAR
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