Author

Roaa Alnajjar

Date of Award

12-2018

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Narayanaswamy Bharathan

Second Advisor

Seema Bharathan

Third Advisor

Robert Major

Abstract

Rhizoctonia solani, is a soil-borne plant pathogenic fungus infecting crop plant. The double-stranded (ds) RNA from these viruses have been implicated in reduced pathogenic ability. This research focuses on isolate EGR4 to develop primers for the cloned sequence and perform Real-Time PCR to test viral dsRNA. In order to find this objective, double-stranded (ds) RNA of 3.8 to 4kb of EGR4 was extracted. The dsRNA was reverse transcribed into cDNA and ligated into a p-jet clone vector. The plasmid of different clones of EGR4 was purified using a fast digest enzyme EcoRI. The largest fragment containing full length cDNA was sequenced, then the forward and reverse primers, and the probe were designed to test viral dsRNA. The primers were used to amplify dsRNA from purified EGR4. The sensitivity of detection of EGR4 dsRNA was further enhanced by di-hybrid probes.

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