Gene duplications generate critical components of genetic variation that can be selected upon to affect phenotypic evolution. The angiosperm GATA transcription factor family has undergone both ancient and recent gene duplications, with the HAN-like clade displaying divergent functions in organ boundary establishment and lateral organ growth. To better determine the ancestral function within core eudicots, and to investigate their potential role in floral diversification, I conducted HAN-like gene expression and partial silencing analyses in the asterid species petunia (Petunia x hybrida). My results indicate duplication of HAN-like genes at the base of Solanaceae followed by expression diversification within the flower. Although no aberrant phenotypes were apparent following single gene knockdowns, silencing of both paralogs lead to leaf senescence. Together with other functional studies, these data suggest a possible ancestral role for HAN-like genes in core eudicot shoot apical meristem development, followed by functional diversification following both speciation and duplication.

Oomycetes, like fungi, are filamentous heterotrophs, but unlike true fungi are diploid and share a photosynthetic ancestor. Many of these organisms are plant and animal pathogens, and members of the genus Phytophthora cause devastating disease on a diverse array of agricultural plant hosts. Several diverse topics in oomycete biology are investigated in this dissertation. Chapter 2 is a report on loss of heterozygosity in Phytophthora capsici in response to chemical mutagenesis.The research presented in Chapters 3 and 4 are centered on documenting biodiversity and genetic diversity of populations of Phytophthora species obtained from infected plant hosts. The final chapter (Chapter 5) involves determining genetic diversity, ecology, and enzymatic activities of Pythiaceous oomycetes from marsh wetlands of the southeastern US.

Gene duplications generate critical components of genetic variation that can be selected upon to affect phenotypic evolution. The angiosperm GATA transcription factor family has undergone both ancient and recent gene duplications, with the HAN-like clade displaying divergent functions in organ boundary establishment and lateral organ growth. To better determine the ancestral function within core eudicots, and to investigate their potential role in floral diversification, I conducted HAN-like gene expression and partial silencing analyses in the asterid species petunia (Petunia x hybrida). My results indicate duplication of HAN-like genes at the base of Solanaceae followed by expression diversification within the flower. Although no aberrant phenotypes were apparent following single gene knockdowns, silencing of both paralogs lead to leaf senescence. Together with other functional studies, these data suggest a possible ancestral role for HAN-like genes in core eudicot shoot apical meristem development, followed by functional diversification following both speciation and duplication.

Climate change has resulted in both increased mean water temperature and higher frequencies of extreme water temperatures in coastal areas. These new thermal regimes exert strong selective pressure on the thermal physiology of coastal aquatic species. Phenotypic plasticity (the ability of one genotype to display multiple phenotypes) and local adaptation (increased fitness to local environment due to natural selection) dictate both short-term (from hours to days to weeks) and long-term (from years to decades) resilience of a species. To better predict how a species will respond to the negative impacts of climate change, one first needs to know the current levels of variation in plasticity and local adaptation. Marginal populations are especially critical for the persistence of a species, as those populations can harbor unique genetic variation and the interaction between plasticity and local adaptation determines the boundaries of future distributional ranges. This dissertation focuses on the northern marginal population of spotted seatrout (Cynoscion nebulosus), an estuarine-dependent fish, and compares them with those from the core region of the distribution to elucidate the physiological, transcriptomic and genetic mechanisms of plasticity and adaptation. I discovered significant differences between fish from different areas at all three levels of biological organization: Chapter 1 shows different whole-organism metabolic physiology of fish sampled from distinct populations and the northern population is consistent with cold-adaptation, given the pressure of natural selection from more severe and frequent winter kills in the region. Chapter 2 presents functional genetic evidence that the cold-adapted northern spotted seatrout are more vulnerable to heat stress than the warm-adapted southern spotted seatrout, suggesting that differential gene expression is contributing to observed differences in thermal tolerance. A liver transcriptome is de novo assembled and serves as a valuable resource for future genetic studies of spotted seatrout. Chapter 3 discovers signatures of selection based on over 15,000 genome-wide single nucleotide polymorphism (SNP) markers. The pattern of genetic variation is consistent with thermal adaptation along the US east coast. Genes involved in metabolic pathways and transcriptional regulation are the main targets of natural selection. In summary, spotted seatrout are relatively resilient to the thermal effects of climate change due to a wide range of metabolic plasticity and adaptive potential in climate-related genetic variation. Range expansion at the leading edge, however, is largely constrained by the species’ cold tolerance limit. The northern and southern population will likely respond to climate change differently and this should be taken into consideration in future conservation management of this species.

Gibbon classification at the species and subspecies levels has been hotly debated for the last 200 years. This thesis explores the reasons for this debate. Authorities agree that siamang, concolor, kloss and hoolock are species, while there is complete lack of agreement on lar, agile, moloch, Mueller's and pileated. The disagreement results from the use and emphasis of different character traits, and from debate on the occurrence and importance of gene flow.

Clostridium scatologenes is an anaerobic bacterium that demonstrates some unusual metabolic traits such as the production of 3-methyl indole. The availability of genome level sequencing has lent itself to the exploration and elucidation of unique metabolic pathways in other organisms such as Clostridium botulinum. The Clostridium scatologenes genome, with an estimated length 4.2 million bp, was sequenced by the Applied Biosystems Solid method and the Roche 454 pyrosequencing method. The resulting DNA sequences were combined and assembled into 8267 contigs with an average length of 1250 bp with the Newbler Assembler program. Comparision of published subunits of csd gene and assembled contigs identified that one contig contained all three subunits. In addition a gene with similarity to clostridium carboxidivorans butyrate kinase was found lined next to csd gene. An alignment of the contig and csdgene sequences identified three deletions in the contig within the 4066 bases of the alignment. This implies that there is about 0.07% error rate in the sequencing itself requiring more finishing. Even without finishing the genome assembly into single contig, contigs were annotated in RAST pipeline predicting 2521 protein encoding genes (PEGs). The PEGs were classified by their metabolic function and compared to classified PEGs found in the closely related clostridium species, Clostridium carboxidivorans and Clostridium. ljungdahlii, which have similarly sized genomes. According to the RAST analysis, Clostridium scatologenes had 35% subsystem coverage of all known metabolic processes with its 2521 PEGs. This compares to 41% for Clostridium carboxidivorans with 4174 PEGs (29) and 42% for Clostridium ljungdahlii with 4184 PEGs (30), indicating that Clostridium scatologenesmay still have more genes to be identified. Comparison of the percent genes found in the metabolic subsystems was similar except in motility and chemotaxis. The contigs, on which the csd gene and tryptophan metabolizing genes lay, were examined to see if additional genes might support these metabolic pathways. Butyrate kinase was associated with the csd genes but no other associations were found for the two tryptophan metabolizing genes. The tryptophan biosynthesis operon genes were all found on one contig (contig 6771) and were syntenic with other bacterial species.

Cardiovascular disease (CVD) is a major cause of morbidity and mortality in the U.S. and worldwide. Atherosclerosis, the buildup of plaque in the arteries, is a common cause of CVD. For many years, research in atherosclerosis was focused on lipid metabolism and the accumulation of low-density lipoprotein in the arteries. While this research set public health guidelines for lipid management, lipid concentration was not the only factor influencing atherosclerosis and CVD events. Many scientists, as far back as the 1850’s recognized the role of inflammation in the progression of atherosclerotic disease. The continuous low levels of immune activation in the body contribute to atherosclerosis. Research in animal models and epidemiologic studies have shown the involvement of both the innate and the adaptive immune systems in plaque development and to elucidate the roles of monocytes and T cells. In addition to animal studies and epidemiologic research, CVD and atherosclerotic research has extended to genetic analysis in the search for associations with risk factors and outcomes. The first chapter is a review of the literature studying the immune system’s involvement in atherosclerosis. Beginning with an examination of the impact of CVD and atherosclerosis, the basic pathophysiology, and the involvement of the innate and adaptive immune systems through animal models and epidemiology. Some of the significant cohort studies in CVD and genome wide association studies are also discussed. Chapter 2 examines the associations of soluble interleukin 2 receptor alpha (sIL-2Rα) with clinical events in the Cardiovascular Health Study and genetic variants. Interleukin 2 (IL-2) and its receptor regulate both tolerance and immunity, IL-2 induces the proliferation and differentiation of T cells, part of the adaptive immune system. The results showed an association between sIL-2Rα and CVD events. The genome-wide association study found 52 variants to be significantly associated with sIL-2Rα in European Americans. Chapter 3 assesses the involvement of the innate immune system in atherosclerosis through the associations of soluble CD163 (sCD163). CD163 is a marker of macrophage activation, specifically associated with M2 macrophages. In CHS, sCD163 levels were analyzed for associations with cardiovascular events and genetic variants. sCD163 was found to be associated with CVD risk factors and with cardiovascular events. In a genome-wide association study six variants in European Americans and three variants in African Americans were found to be significant. Chapter 4 summarizes the results and discusses some bench to bedside translational science already seen in atherosclerosis treatment and prevention. Continued investigation of markers of T-cell and monocyte differentiation in animal models and cohort studies may lead to opportunities for the prevention of atherosclerosis and/or treatment through an increased understanding of the biology and genetics of the innate and adaptive immune.

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Down Syndrome is a human condition caused by an extra copy of a #21 chromosome. At least one to two percent of free (not translocated) trisomy 21 cases are mosaics, i.e., they have two or more distinct cell lines. Usually, one cell line is 47 ,XX or XY ,+21 while the other cell line is normal 46,XX or 46,XY. The purpose of the study was to establish the etiologies of the separate cell lines by determining whether the zygote was trisomic or normal. Meiotic non-disjunction in the formation of a gamete could lead to a trisomic zygote; loss of a #21 chromosome during a later mitotic division could then lead to a chromosomally normal cell line. Alternatively, a mitotic error in a normal embryo can produce a trisomy 21 cell line.

This thesis proposal shows the efficiency of different methods of Myostatin inhibition by using CRISPR/CAS9. With the data cataloged by this thesis, researchers will have a better understanding of what methods are better to achieve their goals. The data was collected by reading multiple scientific articles involving the usage of CRISPR/CAS9 to inhibit the Myostatin protein. The data collected from all the different studies were analyzed in the same categories. The experiment that used CRIPSR/CAS9 on in vitro specimens had a superior Myostatin inhibition overall, therefore presenting higher muscle mass. The method using CRISPR/CAS9 to inhibit the Myostatin in vivo and in vitro depends on what the researcher is trying to accomplish. By reading this thesis researchers can understand that if they choose to use an in vitro treatment the results would be way more severe than using an in vivo application treatment.