Thermal cycler for plant molecular genetics

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This proposal requests funds to purchase a thermal cycler to support for PCR (polymerase chain reaction) to support research on the biology and biotechnology of plant cell walls and plant biomass production. PCR and instruments of this type are essential in molecular biology and genetics labs for many methodologies and experimental approaches including gene cloning, genotype determination, non-quantitative or semi-quantitative gene expression, and DNA sequencing. All of these approaches are central to the research being carried out in the applicant's lab. The instrument will support research in three areas related to plant cell wall biosynthesis and the improvement of cell walls in woody species for practical purposes. One set of studies involves the regulation of secondary cell wall biosynthesis (which form the bulk of the woody biomass in trees and in lignocellulosic bioenergy feedstocks). We have identified a negative feedback loop in the regulatory network governing secondary wall formation in the model plant Arabidopsis, and are investigating the functions of the transcription factors involved in this regulation work using a variety of approaches that depend on PCR. In another set of studies, we are translating this and other information into poplar, a fast growing tree that is a strong candidate as a bioenergy feedstock. PCR-aided approaches will be used to investigate regulation of secondary cell wall (wood) formation in poplar, and the use natural variants in important cell wall genes to improve the feedstock. A final area of research study investigates the formation of the strongest plant cell wall material known, the outer wall (exine) of pollen grains. We are using Arabidopsis as a model system to investigate the genes encoding required for exine biosynthesis, which is poorly understood, and will rely heavily on PCR and the requested thermal cycler to carry out these studies.