Date of Award

2017

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Clore, Amy

Area of Concentration

Biology

Abstract

Endophytes consist of either bacterial or fungal populations that exist in the tissues of a plant, and have been discovered in all plants that have been tested. They are microorganisms that live inside the host tissue and can contribute to the growth and survivability of plant species by providing certain hormones or resources. Research has shown that different tissues of plants contain unique endophytic bacterial populations. In the present study, meristematic (unspecialized) tissue was taken from store-bought cauliflower and allowed to differentiate (or specialize) in culture. The explants produced several different organs without the need for supplemental hormones. Samples from the resulting roots, buds, stems, leaves and flowers were then harvested and surface sterilized to remove external microorganisms, then metagenomic DNA was extracted. This DNA was initially sent to Molecular Research Laboratory LP (MrDNA) to have 16S rRNA gene sequenced. After failure of the lab to amplify DNA due to reported low quality, in-house PCR reactions were carried out with more universal primers. These PCR reactions were successful and the amplified DNA was isolated using a PCR clean up kit, at which point the amplicons were sent back to MrDNA for a second attempt. More PCR reactions were run using fluorescently tagged primers. These samples were then digested using RsaI and HsaI and will be sent to the University of Illinois for terminal restriction fragment length polymorphism analysis (TRFLP), another but less specific method for characterizing endophyte populations, as a backup. Several gels were run and bands corresponding to 16S rRNA DNA were present in all samples. However, analysis from MrDNA and University of Illinois were not complete at the time of this writing.

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