Cloning of casbene and neocembrene synthases from Euphorbiaceae plants and expression in Saccharomycescerevisiae

A large number of diterpenes have been isolated from Euphorbiaceae plants, many of which are of interest due to toxicity or potential therapeutic activity. Specific Euphorbiaceae diterpenes of medical interest include the latent HIV-1 activator prostratin (and related 12-deoxyphorbol esters), the analgesic resiniferatoxin, and the anticancer drug candidate ingenol 3-angelate. In spite of the large number of diterpenes isolated from these plants and the similarity of their core structures, there is little known about their biosynthetic pathways. Other than the enzymes involved in gibberellin biosynthesis, the only diterpene synthase isolated to date from the Euphorbiaceae has been casbene synthase, responsible for biosynthesis of a macrocyclic diterpene in the castor bean (Ricinus communis). Here, we have selected five Euphorbiaceae species in which to investigate terpene biosynthesis and report on the distribution of diterpene synthases within this family. We have discovered genes encoding putative casbene synthases in all of our selected Euphorbiaceae species and have demonstrated high-level casbene production through expression of four of these genes in a metabolically engineered strain of Saccharomyces cerevisiae. The only other diterpene synthase found among the five plants was a neocembrene synthase from R. communis (this being the first report of a neocembrene synthase gene). Based on the prevalence of casbene synthases, the lack of other candidates, and the structure of the casbene skeleton, we consider it likely that casbene is the precursor to a large number of Euphorbiaceae diterpenes. Casbene production levels of 31 mg/L were achieved in S. cerevisiae and we discuss strategies to further increase production by maximizing flux through the mevalonate pathway.     

A geospatial assessment on the distribution,condition, and vulnerability of Wyoming's wetlands

Wetlands serve critical functions including natural flood control and providing wildlife habitat, yet despite these values they remain highly threatened systems. Here we present a landscape-scale geospatial assessment of wetlands in Wyoming. Areas containing high densities of wetlands were identified and mapped, and wetland complexes were quantified as a function of their biological diversity, protection status, susceptibility to climate change, and proximity to sources of impairment. Our results indicate there are 280591 wetlands in Wyoming, totaling 371758 surface hectares, and 222 wetland complexes. The majority (67%) of wetlands are classified as temporary. Low elevation wetland complexes are the least protected, in the poorest current condition, and the most vulnerable to future land use changes. This fundamental information will provide a tool decision-makers can use to more effectively allocate limited resources to conserve, manage, and restore Wyoming's wetlands.     

Modeling Tick-Borne Disease: A Metapopulation Model

Recent increases in reported outbreaks of tick-borne diseases have led to increased interest in understanding and controlling epidemics involving these transmission vectors. Mathematical disease models typically assume constant population size and spatial homogeneity. For tick-borne diseases, these assumptions are not always valid. The disease model presented here incorporates non-constant population sizes and spatial heterogeneity utilizing a system of differential equations that may be applied to a variety of spatial patches. We present analytical results for the one patch version and find parameter restrictions under which the populations and infected densities reach equilibrium. We then numerically explore disease dynamics when parameters are allowed to vary spatially and temporally and consider the effectiveness of various tick-control strategies.