Date of Award

5-2015

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Narayanaswamy Bharathan, Ph.D.

Second Advisor

Seema Bharathan, Ph.D.

Third Advisor

Carl Luciano, Ph.D.

Abstract

Rhizoctonia solani is a soil-borne pathogenic fungus with several distinct isolates that have been classified based on their anastomosis groups. Many isolates of this fungus contain double-stranded (ds) viral RNA that are cytoplasmic and viral in nature. In the present study, the dsRNA from isolates EGR4, Tom7 and a hyphal anastomosis reaction of EGR4/Tom7 was extracted and analyzed by gel-electrophoresis and micro-chip analysis. Interaction between the isolates showed effects on ribosomal RNA which may be due to the presence of the viral dsRNA. Real-time PCR was optimized with primers and probes, specific to the dsRNA sequence, via plasmid cultures that had previously been transformed using the extracted dsRNA from the isolate EGR4. The ideal primer/probe concentration was determined through comparing the lowest real-time PCR cycle threshold values supporting that a specific isolate can be selectively identified by means of an optimized real-time PCR protocol.

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