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Identification of novel motifs in unaligned protein sequences Genetic and Biochemical analysis of enzymes that regulate plant growth and metabolism

Faculty Mentor: Leeann Thornton

Students: Joseph Montes, Dylan McDivitt, & Amanda Soler

In order grow enough food and plant products for the increasing human population, agriculture productivity must continue to improve. Currently, most plants only yield about 20 percent of their potential productivity when grown in the field. Much is unknown about how plants respond to stressful conditions. We aim to determine how a group of enzymes, called Cytochrome P450s (CYPs), influence plant metabolism in response to suboptimal growth conditions. We are studying these enzymes from Arabidopsis thaliana as a model for other plants. We took both a molecular genetics approach and a biochemical approach. For the molecular genetics approach, we isolated two plants with less enzyme activity for CYP72A11 and CYP72A13. We are examining these plants for growth deficiencies that will inform us about enzyme function. For the biochemical approach, CYP72A9 and CYP72A13 were expressed in yeast cells.  Multiple yeast growth conditions were examined to obtain the optimal amount of plant enzyme. Under one condition, CYP72A13 produces a small amount of active enzyme while no functional CYP72A9 has been obtained. We will continue to optimize expression of both enzymes and work towards purifying functional enzyme samples. The goal is to perform this experiment to determine enzyme function. Our studies on A. thaliana will contribute to the better understanding of plant response to stressful conditions to improve crop plant productivity.