Our overall goal is to use genomic approaches to drive an increased understanding of the network of enzymatic and other processes involved in the production of seed TAGs, and in particular how these processes have coevolved to facilitate the synthesis and accumulation of unusual fatty acids. 

Our specific objectives under this grant are:

• To identify components of the mFA-producing networks that evolved to allow for efficient accumulation of mFA. We will conduct comparative transcriptomics between species of mFA accumulators and closely-related non-accumulators. Identification of regulatory factors will be complemented by yeast-one-hybrid and co-immunoprecipitation assays.


• To assemble the mFA biosynthetic pathways. We will investigate and optimize the activities of FAH, CONJ and CPS using protein engineering and directed evolution. Roles of candidate genes/enzymes involved in the mFA fluxes through PC will be assessed through expression in transgenic plants. Stable isotope labeling experiments and rigorous mathematical modeling will be used to answer specific questions about the metabolic bottlenecks of mFA during oil synthesis.


• To optimize the accumulation of high levels of mFA in transgenic seeds. We will evaluate plant responses to mFA accumulation by gene expression changes in transgenic plants using deep sequencing of transcripts. Endogenous, competing enzymes will be suppressed through gene silencing. In addition, we will evaluate the feedback inhibition of fatty acid synthesis in transgenic lines using directed proteomics approaches and use the information gained to mitigate the inhibition.


• To broaden public understanding and excite young students about the potential of plants to serve as chemical factories of the future.  We will train current and future plant scientists to apply genomics in a focused way to solve long-standing questions in complex biosynthetic pathways and to appreciate how the answers to these questions can aid society.