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Wolbachia is a genus of intracellular bacterium that is known to infect a wide range of arthropods. Species within this clade have the ability to modify the reproductive success of their hosts to promote their own distribution throughout host populations. Wolbachia cannot be cultured outside of the host and characterizing infection of gonadal cells might be time consuming. Thus, polymerase chain reaction (PCR) assays are a common method of Wolbachia DNA amplification from arthropods. I assessed the utility of 3 DNA sequence markers (16S, ITS, and 28S) paired with the Wolbachia surface protein (wsp) gene in various PCR protocols. The developed protocol was used on four species within the Cynipini tribe of gall wasps from central Illinois to detect Wolbachia infection. A PCR protocol was optimized using wsp gene and 28S primers to confirm DNA extraction, as well as the presence of Wolbachia strains in the homed oak gall wasp, Callirhytis cornigera. My results provided evidence for the potential of the combined use of 28S nuclear DNA gene and the wsp gene of Wolbachia for detection of Wolbachia infection in cynipid wasps, but further trials are needed to fine tune PCR protocol to yield consistent results, which is necessary for Wolbachia detection in oak gall wasp communities.
Kim, Jennifer, "Optimization of Protocols for Wolbachia Detection Using PCR Methods" (2013). Undergraduate Honors Theses. 64.