Project Number: 5335-21000-030-00
Project Type: Appropriated

Start Date: Mar 17, 2006
End Date: Mar 16, 2011

Objective:
The long-term goal of this research is to develop the ability to regulate pollen development and pollen-pistil interactions. Using yeast two hybrid screens, as well as genomic and proteomic tools, we have identified candidate proteins that play roles during these processes, and have developed tools (such as cell-specific promoters) that can facilitate functional studies of these proteins. The specific objectives of this project plan are: 1. To elucidate the molecular interactions that occur during pollen development, pollen-pistil interactions, and sperm-egg interactions, using both model (e.g., Arabidopsis) and crop (e.g., tomato, corn) species. 2. To define the mechanisms underlying hybridization barriers between species and determine if manipulating gene expression levels of key genes can overcome such barriers.

Approach:
1) Use yeast 2-hybrid screens and biochemical techniques to characterize proteins expressed in female tissues and/or in pollen that are required for signaling through receptor kinases. Use similar techniques to identify sperm- and egg-expressed proteins that might mediate egg-sperm interactions. Develop sperm- and egg-specific promoter elements for gamete-specific gene manipulations. Use comparative genomics to identify protein domains important for specificity. Use gene disruptions to test interactions genetically. 2) Use pollination assays and microscopic imaging to determine where hybridization barriers occur between selected plant species pairs. Identify target genes that might act at these steps. Generate gene disruptions for selected candidates and assay for pollen tube growth/fertilization phenotypes. Introduce DNA constructs into transgenic plants. Use pollination assays to test for expanded hybridization potential in plants that overexpress or repress the expression of selected target genes. REPLACES 5335-21000-020-00D (4/06). BSL 1; 2/19/08.