LE3 .A278 2023

2023

Stewart, Don

Acadia University

Bachelor of Science

Honours

Sociology

Sociology

Rare and elusive species often lack sufficient monitoring to efficiently carry out conservation efforts, and this is the case for the aquatic lycophyte Isoetes prototypus in Nova Scotia. This species is known from only 20 locations globally, 10 of which are in Nova Scotia. Due to the difficulties of accessing, identifying, and surveying this species, no monitoring has been conducted since 2004. However, eDNA analysis provides a solution to these challenges as it presents a simple, cost effective, and reliable alternative to visual surveys and physical monitoring techniques. The technique of eDNA also reduces uncertainty related to species identification as it is based on genetic sequences unique to that species, also known as DNA barcoding. This project involved conducting a series of surveys for I. prototypus in Nova Scotia to establish a baseline of population changes since 2004, developing eDNA primers, and conducting a pilot study to test efficacy of this technique. The field surveys were carried out to the extent of the required scope of this project, and samples were collected for the remainder of the research. Methods were optimized to extract DNA from plant tissue, and primers were designed and tested for PCR amplification. Genetic sequencing revealed that the primers were amplifying the correct region and that they could reliably distinguish I. prototypus from other native Isoetes species. Extractions for eDNA analysis from positive control samples were analysed and the process is ready to be applied to samples collected from field sites. The eDNA protocols for this species have not been finalized, however, results suggest that with further optimization, eDNA will be a valuable additional tool for ongoing monitoring of I. prototypus.

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https://scholar.acadiau.ca/islandora/object/theses:4004