- ItemRecovery ability of thermally stressed captive Coral Anthelia spp., as measured by dinoflagellate density(2022) Dunbar, Dana; Shaw, RossWarming ocean temperatures are leading to an increase in coral bleaching events. These rising temperatures are fatal to coral species as they disrupt the symbiotic relationship between corals and dinoflagellates. Among other factors, thermal stress results in dinoflagellate damage and the loss of these symbionts. The recovery ability of corals exposed to this stress is a small area of research within the larger body of coral conservation. This study aims to add to that field by examining how soft corals, specifically Anthelia spp., react to thermal stresses. Over a nine- week period, 4 different experimental tanks will be raised from 28o to 32oC before returning to 28C to observe recovery potential. Dinoflagellate density was examined twice per week using a maceration method on tissue samples, viewed under a compound microscope. These densities were used as an indication of coral health and successful recovery. Expanding the knowledge of the recovery ability of soft corals is imperative to continuing the existence of these species.
- ItemDeveloping microsatellite markers for Cypripedium passerinum (Sparrow’s egg lady’s slipper)(2022) Lim, Lina; McFadyen, DavidNatural and anthropogenic disturbances contribute to increased habitat loss and fragmentation and subsequently, species loss. Integrated conservation approaches combine both in-situ and ex-situ approaches whereby natural habitats of endangered species are conserved, and the genetic diversity of the threatened population is retained outside of their natural habitat. Therefore, an essential component of an effective conservation strategy is to assess genetic variation to ensure that the conservation approach employed is effective in preserving the diversity of the whole population. Microsatellites, highly polymorphic repetitive DNA sequences in the genome of all organisms, have proven to be a valuable tool in the assessment of genetic diversity. This project aimed to isolate microsatellite markers from Cypripedium passerinum, a native North American terrestrial orchid at risk of extinction. Fast Isolation by AFLP of Sequences Containing Repeats (FIASCO) was employed to generate a genomic DNA library enriched for AT, AC, and AAG microsatellites. Clones were selected from the libraries and bidirectionally sequenced to identify those which contain microsatellites. A total of 158 microsatellite loci were identified, of which 83% were perfect microsatellites. PCR primers were developed using the unique sequences flanking the identified microsatellites and were evaluated for their utility. Primers amplifying polymorphic loci can be used to assess the genetic diversity of C. passerinum populations both within the Wagner Natural Area, Alberta, Canada and elsewhere in its range of distribution. The project findings will contribute to the integrated conservation efforts to protect species found in Wagner Natural Area and contribute to our understanding of C. passerinum.
- ItemCloning and purification of a glycerol-specific alditol oxidase for biosensor construction(2022) Barroma, Chrissa; Kryjak, Amanda; Jung, SandyWine production is dependent on ethanol, but also on optimal glycerol concentrations, both of which are produced by S. cerevisiae fermentation. Wine characteristics like sweetness levels are influenced by glycerol concentrations. Additionally, elevated glycerol levels can be an indication of abnormal blood sugar levels. In both situations, close observations of glycerol levels are essential. One proposed method of measuring glycerol concentrations is through enzymatic oxidation with a glycerol biosensor. Alditol oxidase (AldO) is a recently discovered carbohydrate oxidase in S. coelicolor. Despite specificity for longer-chained polyols, studies have proposed that AldO can be used as a glycerol oxidase. Using random point mutations, an AldO mutant was isolated and had increased specificity for glycerol. These results suggest that potential for AldO with glycerol biosensor development. This project aimed to produce a glycerol specific alditol oxidase to be used as a biosensor. A synthetic alditol oxidase (AldOG) gene was used to produce AldOG via cloning methods. We plan to overexpress and purify the AldOG protein to use in construction of a glycerol biosensor in collaboration with Dr. Samuel Mugo (MacEwan University).
- ItemSand armour: how it provides plants with an edge up in survival(2021) Roth, JenniferPlants have evolved a dizzying array of morphological and biochemical defenses; a deceptively simple one involves sand. Some plants actively coat themselves in sand, termed psammophory, as an ingenious adaptation for survival. While the functional significance of psammophory is understudied, experimental data from Abronia latifolia and Navarretia mellita suggests that it acts as a mechanical defense against herbivory within dune habitats. This defense stems from both the damaging and non-nutritive properties of sand and the lasting detrimental effects it has on herbivore physiology. While sand armour may seem like an unusual adaptation, it certainly can deter herbivores by giving them something to chew on.
- Item9-(2-Phosphonyl-methoxyethyl)-adenine promotes erythrocytic differentiation and disrupts cell replication in chronic myelogenous leukemia K562 cells(2021) Wiseman, BrittanyDisruption during cellular differentiation can cause hematopoietic stem cells to proliferate uncontrollably, resulting in the development of cancer. Differentiation therapies are being investigated as a type of cancer treatment which involve inducing agents that promote the differentiation of cancer cells into those with similar properties to normal blood cells. These cells can then undergo apoptosis at an accelerated and controlled rate compared to cancer cells, making this a potential therapeutic technique. In this study, the ability of human chronic myelogenous leukemia K562 cells to undergo cellular differentiation in response to the inducing agent 9-(2-Phosphonyl-methoxy ethyl)-adenine (PMEA) is investigated. PMEA has previously been shown to disrupt cell replication, and promote erythrocytic differentiation in K562 cells. In order to further test the effectiveness of this inducer, cell proliferation was measured with a cell growth curve, hemoglobin presence was measured with benzidine staining, and gamma-globin expression (a protein subunit of fetal hemoglobin) was measured in both induced and uninduced K562 cell cultures via RT-qPCR and western blotting. The results indicate that PMEA slows cell replication, and promotes hemoglobin (and subsequently gamma-globin) expression in treated cells. In summary, the findings support the conclusion that PMEA is able to promote erythrocytic differentiation in K562 cells, and provides information that supports differentiation therapies as a method for cancer treatment.