Characterizing BIO3/BIO1 in Brachypodium distachyon Timothy C. Davenport1, Shih − heng Su1, and Patrick Masson1 1Department of Genetics, University of Wisconsin, Madison, WI 53706 Abstract Future Studies Root growth stunting from exogenous cadaverine treatment is conferred within the Bio3/Bio1 gene of Arabidopsis thaliana. The knowledge of whether this response in universal to other plant models is still unknown. The purpose of my investigations has been to characterize the role of BIO3/BIO1 in Brachypodium distachyon. This task was explored • Rerun of cadaverine dosage curve with Bd21, Luc1, and Bis1 through applications of exogenous cadaverine within B.distachyon single-nucleotide • Biotin rescue-effect investigations polymorphic lines within the BIO3/BIO1 gene. Exogeneous biotin treatments in tandem with a cadaverine stress environment were also undertaken to observe possible cadaverine • Initiate four step cloning strategy of Bd-BIO3/BIO1 rescue effects as previously observed in A.thaliana. Future investigations shall employ 1) Plasmid Modification (pET His6 GFP TEV LIC ) golden gate cloning of the Brachypodium BIO3/BIO1 gene into A.thaliana to determine whether the effects of cadaverine are conferred within BIO3/BIO1 of B.distachyon. 2) Insertion of Bd-BIO3/BIO1 into Vector 3) Transformation into E.coli Cadaverine Effect in Arabidopsis thaliana 4) Golden Gate Cloning Implications (Fig. 2): BIO3-BIO1 Bifunctional Domain Complex. (2) (Fig.1): Cadaverine Metabolism synthesis pathway in plants. Cadaverine stress (Fig.3): Partial box plot of BIO3/BIO1 amino acid sequence comparison between response is partially conferred in BIO3- BIO1 activity within A.thaliana. common model organisms in plant biology. Defining BIO3/BIO1 functional characteristics in B.distachyon shall become a gateway for establishing a holistic understanding of cadaverine response pathways Brachypodium distachyon: A Novel Model System in a variety of plants as opposed to a single model organism. Understanding cadaverine response components of model organisms for crop plants can establish Presently, there is a lackluster of literature on a genetic characterization of relevance to the agriculture industry. Observation of cross-species BIO3/BIO1 cadaverine stress response in monocots. B.distachyon has been selected as a functionality will contribute to a holistic understanding of plant-stress response. candidate for understanding the presence of a cadaverine effect in monocots and if this effect is conferred within the BIO3-BIO1 gene. References Preliminary Investigations with B. distachyon (1) Jancewicz A., Gibbs N., and Masson P. (2016) Cadaverine’s Functional Role in Plant Development and Environmental Response. Frontiers in Plant Science (7). pp. 870. Selection of B.distachyon lines with (2) Cobessi D., Dumas R., Pautre V., Meinguet C., Ferrer JL. and Alban C. (2012) synonymous and nonsynonymous Biochemical and structural characterization of the Arabidopsis bifunctional enzyme single-nucleotide-polymorphisms dethiobiotin synthase-diaminonpelargonic acid aminotransferase: evidence for within the Brachypodium BIO3-BIO1 substrate channeling in biotin synthesis. Plant Cell (4) 1609-25. gene. At 400uM Cadaverine treatment, the reference line Bd21 (Bradi3g18970.2) and nonsynonymous Acknowledgements line Luc1 seem to have conflicting root growth response to exogenous I would like to thank Dr. Shih-heng Su and Dr. Patrick Masson for their continued mentorship cadaverine. and support. .