DEVELOPMENT AND APPLICATIONS OF GENOMIC RESOURCES FOR TWO NORTH AMERICAN HARDWOOD TAXA

Hardwoods comprise about half of the biomass of forestlands in North America and present many uses including economic, ecological and aesthetic functions. Forest trees rely on the genetic variation within tree populations to overcome the many biotic, abiotic, anthropogenic factors which are further worsened by climate change, that threaten their continued survival and functionality. To harness these inherent genetic variations of tree populations, informed knowledge of the genomic resources and techniques, which are currently lacking or very limited, are imperative for forest managers. The current study therefore aimed to develop genomic microsatellite markers for the leguminous tree species, honey locust, Gleditsia triacanthos L. and test their applicability in assessing genetic variation, estimation of gene flow patterns and identification of a full-sib mapping population. We also aimed to test the usefulness of already developed nuclear and gene-based microsatellite markers in delineation of species and taxonomic relationships between four of the taxonomically difficult Section Lobatae species (Quercus coccinea, Q. ellipsoidalis, Q. rubra and Q. velutina. We recorded 100% amplification of G. triacanthos genomic microsatellites developed using Illumina sequencing techniques in a panel of seven unrelated individuals with 14 of these showing high polymorphism and reproducibility. When characterized in 36 natural population samples, we recorded 20 alleles per locus with no indication for null alleles at 13 of the 14 microsatellites. This is the first report of genomic microsatellites for this species. Honey locust trees occur in fragmented populations of abandoned farmlands and pastures and is described as essentially dioecious. Pollen dispersal if the main source of gene flow within and between populations with the ability to offset the effects of random genetic drift. Factors known to influence gene include fragmentation and degree of isolation, which make the patterns gene flow in fragmented populations of honey locust a necessity for their sustainable management. In this follow-up study, we used a subset of nine of the 14 developed gSSRs to estimate gene flow and identify a full-sib mapping population in two isolated fragments of honey locust. Our analyses indicated that the majority of the seedlings (65-100% - at both strict and relaxed assignment thresholds) were sired by pollen from outside the two fragment populations. Only one selfing event was recorded confirming the functional dioeciousness of honey locust and that the seed parents are almost completely outcrossed. From the Butternut Valley, TN population, pollen donor genotypes were reconstructed and used in paternity assignment analyses to identify a relatively large full-sib family comprised of 149 individuals, proving the usefulness of isolated forest fragments in identification of full-sib families. In the Ames Plantation stand, contemporary pollen dispersal followed a fat-tailed exponential-power distribution, an indication of effective gene flow. Our estimate of δ was 4,282.28 m, suggesting that insect pollinators of honey locust disperse pollen over very long distances. The high proportion of pollen influx into our sampled population implies that our fragment population forms part of a large effectively reproducing population. The high tendency of oak species to hybridize while still maintaining their species identity make it difficult to resolve their taxonomic relationships. Oaks of the section Lobatae are famous in this regard and remain unresolved at both morphological and genetic markers. We applied 28 microsatellite markers including outlier loci with potential roles in reproductive isolation and adaptive divergence between species to natural populations of four known interfertile red oaks, Q. coccinea, Q. ellpsoidalis, Q. rubra and Q. velutina. To better resolve the taxonomic relationships in this difficult clade, we assigned individual samples to species, identified hybrids and introgressive forms and reconstructed phylogenetic relationships among the four species after exclusion of genetically intermediate individuals. Genetic assignment analyses identified four distinct species clusters, with Q. rubra most differentiated from the three other species, but also with a comparatively large number of misclassified individuals (7.14%), hybrids (7.14%) and introgressive forms (18.83%) between Q. ellipsoidalis and Q. velutina. After the exclusion of genetically intermediate individuals, Q. ellipsoidalis grouped as sister species to the largely parapatric Q. coccinea with high bootstrap support (91 %). Genetically intermediate forms in a mixed species stand were located proximate to both potential parental species, which supports recent hybridization of Q. velutina with both Q. ellipsoidalis and Q. rubra. Analyses of genome-wide patterns of interspecific differentiation can provide a better understanding of speciation processes and taxonomic relationships in this taxonomically difficult group of red oak species.

WATER RESOURCES, HEALTH, AND THE SUSTAINABILITY OF INTERVENTIONS TO ACHIEVE WATER AND SANITATION TARGETS OF THE MILLENNIUM DEVELOPMENT GOALS IN A CHANGING WORLD

This dissertation addresses sustainability of rapid provision of safe water and sanitation required to meet the Millennium Development Goals. Review of health-related literature and global statistics demonstrates engineers' role in achieving the MDGs. This review is followed by analyses relating to social, environmental, and health aspects of meeting MDG targets. Analysis of national indicators showed that inadequate investment, poor or nonexistent policies and governance are challenges to global sanitation coverage in addition to lack of financial resources and gender disparity. Although water availability was not found to be a challenge globally, geospatial analysis demonstrated that water availability is a potentially significant barrier for up to 46 million people living in urban areas and relying on already degraded water resources for environmental income. A daily water balance model incorporating the National Resources Conservation Services curve number method in Bolivian watersheds showed that local water stress is linked to climate change because of reduced recharge. Agricultural expansion in the region slightly exacerbates recharge reductions. Although runoff changes will range from -17% to 14%, recharge rates will decrease under all climate scenarios evaluated (-14% to -27%). Increasing sewer coverage may place stress on the readily accessible natural springs, but increased demand can be sustained if other sources of water supply are developed. This analysis provides a method for hydrological analysis in data scarce regions. Data required for the model were either obtained from publicly available data products or by conducting field work using low-cost methods feasible for local participants. Lastly, a methodology was developed to evaluate public health impacts of increased household water access resulting from domestic rainwater harvesting, incorporating knowledge of water requirements of sanitation and hygiene technologies. In 37 West African cities, domestic rainwater harvesting has the potential to reduce diarrheal disease burden by 9%, if implemented alone with 400 L storage. If implemented in conjunction with point of use treatment, this reduction could increase to 16%. The methodology will contribute to cost-effectiveness evaluations of interventions as well as evaluations of potential disease burden resulting from reduced water supply, such as reductions observed in the Bolivian communities.