PCR to Distinguish Diploid and Triploid Forms of Butomus umbellatus

Butomus umbellatus, also known as flowering rush, is an invasive species that was first documented in the Flathead Lake, located in northwest Montana, in 1964. There are currently no treatment methods against the invasion of flowering rush. In order to provide the correct treatment, the exact form of the species needs to be determined. There are two forms of flowering rush: diploid form (26 chromosomes) and triploid form (39 chromosomes). The chromosome squash technique to count the number of chromosomes is cumbersome and depends on obtaining rapidly dividing cells in root tips. This research is aimed at establishing an alternative method utilizing PCR to distinguish diploid or triploid forms of flowering rush. PCR is a simple, quick method that does not require any living plant tissue, only plant DNA, which can be easily isolated and stored in a freezer to be readily available. The method utilizes a single nucleotide polymorphism (SNP).  In order for a SNP to be located, regions of the plant genome need to be sequenced. Since there is virtually no sequence data on flowering rush, primers were designed within structural genes inside of exon boundaries in organisms related to flowering rush. The primers were tested, and so far an amplicon was obtained from a zinc transporter gene found in Allium cepa (onion). The PCR product was purified and sequence data has been acquired. Primer design will continue until a SNP has been located. After verifying diploid or triploid precise control treatments can be developed.