Undergraduate Researcher Purdue University Maple Valley, Washington
Body of Abstract: Fertilization is a critical biological process for species survival and genetic diversity. In plants, seeds are the direct products of double fertilization and are the basis of nearly all our food. Despite its importance in agriculture, little is known about the processes that lead to fertilization success and the communication between plant male and female gametes. Because fertilization occurs deep into the flower tissues and involves just a few cells in the plant, studying sperm-egg interactions is technically challenging. To overcome these difficulties, we established an estradiol-inducible system in Arabidopsis thaliana to test the potential to reprogram somatic leaf cells into gametes. Inducible UBQ10p:RKD2-mCherry and UBQ10p:DUO1-GFP, master regulators of sperm-egg cell identity, were introduced into the background of several transgenic lines that express egg and sperm cell-specific markers. Protoplasts isolated from four to six-week-old rosette leaves were monitored using fluorescence microscopy for nuclear expression of RKD2-mCherry and DUO1-GFP following induction by β-estradiol. In addition, we verified the expression of plasma membrane gamete-specific markers to validate a shift from somatic to sperm or egg-like state. We will report our preliminary findings on the ability of this system to reprogram somatic cells into sperm and egg cell fate. If successful, this novel in vitro "artificial gamete" platform can facilitate the study of protein-protein interactions occurring at the cell surface of gametes and ultimately lead to a better understanding of how fertilization and seed development are regulated in plants.