Enhancing Genetic Analysis and Sequencing Facility

Requested are funds for two eight-capillary DNA sequencers and a thermocycler configured for 96- well format and dedicated to sequencing reactions. These instruments are the first and most important components of a high throughput sequencing facility that the PIs and colleagues are assembling for agriculturally significant molecular research at the University of Kentucky. The requested instrumentation will provide throughput of up to 384 samples per day at a cost of $3-$4 per sample. The facility will be staffed with technicians trained and experienced with related instrumentation, and will be sited at a dedicated location being constructed close to the great majority of college research groups. Six examples are presented of research programs that are relevant to the sustainability of agriculture, will facilitate future success in competitive grant funding, and require the requested instrumentation. Schardl and Webb study symbiotic fungi and viruses, respectively, that playa significant role in biological control of agricultural pests. Schardl focuses on grass endophytes and the basis for their protective effects on forage grasses. Webb focuses on viruses that enable wasps to parasitize lepidopteran insects. Perry and Gan focus on molecular control of seed development and leaf senescence, respectively. Both of these stages in plant development are key determinants of productivity. The Howe program on apicomplexan pathogens of animals focuses on expressed sequences to identify potential diagnostic and therapeutic targets. The Nagy program focuses on RNA recombination in plant viruses, which is a concern related to transgenic plants, but also gives rise to "defective interfering" RNAs that are potential antiviral agents. The requested instrumentation will significantly increase the productivity and performance of these research programs and will greatly facilitate development of other research programs in the College of Agriculture at the University of Kentucky using molecular genetic approaches that rely upon high throughput DNA sequencing capability.