STRATA

: Genetic and Morphological Analysis of Quercus welshii: A Test of Hypothesized Hybrid Origin; Quercus welshii is geographically restricted to the Navajo Basin area of NE Arizona and SE Utah. Two hypotheses have been formed in order to explain how Q. welshii evolved. One states that Q. welshii is a relic population left over from when its ancestor, Q. havardii, populated the region. The other states that Q. welshii is stabilized hybrid of Q. gambelii and Q. turbinella, the two dominate species in the area. In order to test if Q. welshii is a hybrid or relic population, both genetic and morphological data was gathered from sample populations. Analysis showed that Q. welshii possesses its own genetic signature, lending evidence in support of the theory that Q. welshii is a relic population. The data also showed that Q. welshii is not morphologically a hybrid of Q. turbinella and Q. gambelii, but has its own distinct phenotype.

: Genetic and Morphological Evidence of Gene Flow Among Oaks (Quercus) in the Four Corners Region; Gene flow is a common occurrence among plant taxa with low barriers to reproduction. Oaks (Quercus) are known for their low levels of reproductive isolation and production of fertile hybrids. In the Four Corners Region a number of Oak taxa occur sympatrically and give rise to various hybrid forms which are generally well segregated based on morphology. To understand the extent with which gene exchange is occurring among the oak species of the Four Corners Region I evaluated a set of five taxa in nine populations for both phenotypic and genotypic divergence. Genetic measures showed that gene flow was extensive among all the taxa while phenotypic identity continued to be largely preserved. This indicates that even under conditions of high gene exchange unique morphological forms can be maintained, likely the result of secondary interactions between genetic structure and ecological factors.

: Genius: Individual Phenomenon or Social Phenomenon?; Genius is not an absolute concept and seems to vary in its considerations from era to era, discipline to discipline, culture to culture, and person to person. Accepted formal definitions of genius are used to indicate a person of exceptional intellectual or extraordinary creative ability, to the inclusion of strong inclination or natural talent. Yet if this is the case, why is such a humanly diverse description limited by accepted example to predominantly men of academia or formal training active between the renaissance and 18th centuries? Einstein died in 1955, and presumably, so too did our living model of genius. This research was conducted to explore how we define and distinguish genius today. Is genius externally defined, or internally refined? Participants were 50 college students who voluntarily filled out a 5-section Genius Perception Survey/ Questionnaire created by the researchers. The questionnaire assessed overall perception of genius, whether the concept was perceived as an internal/ individual event or an external/ societal circumstance, some of the factors associated with that perception, and whether genius is considered natured or nurtured. Results indicated a significant means difference between selected statements representing either very internal (3.74) or external (2.66) perceptions of genius, with a larger mean of respondent's perceptions reflecting genius as a very internal/ individual anomaly, t (40) = 2.88, p = .006. Participants indicated that a genius is an "introspective individual" who is relatively socially extroverted, and "able to participate in and excel in communicating their ideas." Additionally, there were significant paradoxes between definitions given from differing academic disciplines. In short, it appears that genius and society opportunistically collide. One cannot exist and evolve without the benefits of the other - genius needs a society, through comparison and acknowledgement, to exist, and in turn, a society needs a genius, through innovation and increased awareness, to evolve - this cycle must repeat itself indefinitely for humanity to move forward.

: Genomic Analysis of Excreted Enzymes in Acidobaterium Capsulatum; Acidobacterium capsulatum (A. capsulatum) is a gram-negative, acidophilic, heterotrophic, rod-shaped, bacterium belonging to the phylum Acidobacteria, originally isolated out of acid drainage from the Yanahara pyrite mine in Japan. A. capsulatum is thought to play an important role in carbon and nutrient cycling in soil and aquatic environments. A. capsulatum has a genome size of approximately 4.1 Mbp, 88.3% of the genome codes for RNA or proteins, but only 68% of the protein genes encode for proteins of known function. Two genes encoding excreted cellulase enzymes, endo-1,4-D-glucanase and β-glucosidase, were selected for further characterization, cloning and expression. These genes are located at nucleotide positions 109,786-110,952 and 22,187-24,517. BLAST searches were done on the nucleic and amino acid sequences to determine the organism with the highest homology to the gene and protein sequences. Structural homologs in the Protein Data Bank were used to identify conserved catalytic residues in the active sites of these enzymes. Primary sources were identified describing the enzyme assays that will be used to test for enzyme activity in cell-free lysates and to test the activity of the proteins after cloning and expression. Forward and reverse primers of these genes were designed such that the gene can be amplified and inserted directionally into a TOPO TA cloning and expression plasmid. Polymerase chain reaction (PCR) was optimized and utilized to amplify the genes and characterize the inserts in the plasmid. Agarose gel electrophoresis was used to determine the size and purity of the amplicons produced by PCR. The amplicons were quantified by absorbance at 260 nm and cleaned up using a PCR cleanup kit prior to cloning. Directionality of the inserts was confirmed by sequencing the insert using manufacturer's provided primers. These results suggest that these genes do encode the putative enzymes, but that they might have unique characteristics. Future work will include optimizing A. capsulatum growth conditions that can result in expression of these active enzymes.

: Geometric Constructions; Ancient Greeks were fascinated with what could be constructed only using a compass and a straightedge without markings. They were able to construct many things like bisecting an angle, but others like doubling the cube baffled them. Legend has it that ancient Athenians were told by the oracle at Delos that a plague would end if they could double the volume of the cube at the altar to Apollo with a compass and a straightedge. The ability to show that this and other problems were or were not possible did not come until 2000 years later with the discovery of algebra in the nineteenth century. We will take a look at what it means to be constructable and how one can put these geometric constructions into a plane. I will then touch on the ability to show if there exist any type of relationship between two constructions.

: Glucose Metabolism in Acidobacterium Capsulatum Studied by 1H-NMR and In Vitro Studies of Cell Lysates and Cloning of a Putative Transaldolase Gene; In 1991 the acidophilic bacteria, Acidobacterium capsulatum was cultured from acid mine drainage in Japan. This microorganism is of interest because there is little known about its metabolic capabilities. Ward et al. (2009) showed in their annotated genome of A. capuslatum the absence of a key enzyme, aldolase, involved in the breakdown of glucose in glycolysis. If this is true, then glucose has to be shunted between use for energy production and biomass through alternative pathways, most likely the pentose phosphate shunt (PPS) which could use transaldolase as a key controlling enzyme. A recent paper by Thompson et al. (2011) suggests that transaldolase is a key enzyme involved in shunting glucose from the Calvin cycle to the PPS when marine cyanobacteria are infected with cyanobacteria phage. To test our hypothesis that transaldolase is a key enzyme in controlling carbon flux in A. capsulatum, we have used a multipronged approach to study glucose metabolism in A. capsulatum. These experiments include 1H-NMR analysis of water soluble metabolites from cell lysates, enzyme activity assays from cell lysates and cloning a gene for a putative transaldolase. Our experiments demonstrate that A. capsulatum does indeed consume all glucose provided in growth media under aerobic conditions. Under these conditions, the cells reach log phase in a 70 to 90 hour period after inoculation. 1H-NMR of water soluble metabolites extracted from cells in log phase versus cells in stationary show distinct differences in metabolite profiles. Malate dehydrogenase assays show that we can lyse the cells and preserve enzyme activity and furthermore show that malate dehydrogenase is active in log phase. If enzyme activity is preserved during cell lysis, we plan to do more complex assays for other key enzyme activities such as aldolase and transaldolase. To further understand the role of transaldolase in A. capsulatum we are cloning the putative transaldolase gene into E. coli for expression and characterization. As a whole, we have been able to characterize A. capsulatum in aerobic conditions with glucose as the sole carbon source, as well as develop a lysis procedure that maintains enzyme activities in vitro. These methods will allow for future studies that will provide more information about glucose metabolism in A. capsulatum.

: Glucose Metabolism, Cloning and Expression of Phosphofructokinase-1 from Acidobacterium Capsulatum; Acidobacteria capsulatum was first isolated from acid mine drainage in 1987. Phenotypic and chemotaxonomic studies indicate that this organism does not fit known gram-negative taxa, and as a result, the phylum Acidobacteria was created. An annotated genome of this acidophilic bacterium, published in 2009, shows no aldolase sequence, however a sequence for phosphofructokinase (PFK1) has been found. A. capsulatum has the ability to utilize glucose as a sole carbon source. In other organisms, there is little evidence of any enzyme, aside from aldolase, using the PFK1 metabolite, fructose-1,6-bisphosphate. Our hypothesis is that glucose is metabolized through a modified glycolytic pathway involving a transaldolase that has been identified in the genome. This hypothesis explains how A. capsulatum can utilize glucose as a sole carbon source for both energy production and building biomass. Experiments involving 13C-NMR, GC-MS, and cloning and expression of PFK-1 have been initiated to investigate how this organism gains energy from metabolizing glucose. Cultures were incubated in media containing 13C labeled glucose. 13C -NMR of the cell supernatant and lysate was analyzed to characterize metabolites. The gene coding for A. capsulatum PFK1 was cloned into E. coli. This strain will be used to express PFK1 to study its enzymatic and allosteric regulatory characteristics. Future work includes developing qRT-PCR assays for measuring transcriptional control of the PFK1 gene and more 13C and 1H-NMR to better understand glucose metabolism.

: Glycosylated Monoterpenoids: Miticides to Protect Honey Bee Colonies; Worldwide honey bee (Apis milifera) populations are in a state of flux, and the corresponding beekeeping profession is in economic distress. An established, primary contributor to honey bee population decline is the aptly-named, ectoparasitic mite: Varroa destructor. These mites both weaken bees' immune systems by feeding on hemolymph fluid (akin to blood), and are attributed as a vector by which viruses are transmitted between beehives. Botanically derived, naturally occurring monoterpenoids such as thymol and carvacrol (constituents of oil of thyme and oil of origanum) have recently shown promise as volatile-based acaracides against the Varroa mite. Nevertheless, efficient and selective delivery of these molecules remains a challenge. This study investigated whether glycosylated monoterpenoids could be utilized as pro-drug acaracides in honey bee hives. To this end, thymol and carvacrol were synthetically glycolsylated under phase-transfer catalysis reaction conditions with protected glucose derivatives. Bulk quantities of both molecules were produced after selective deprotection. Liberation of the pro-drug was accomplished utilizing the honey bee beta-glycosidase enzyme assay. Current efforts are focused on evaluating the acaracidal efficacy of these glycosylated monoterpenoids in bee colonies.