RUI: Cloning and Characterization of OPENED IMMATURE FLOWER

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Intellectual Merit: Many studies of gene expression have focused on transcription levels, leaving alternative splicing, a conserved mechanism that increases transcript diversity, relatively unexplored. This project seeks to deepen understanding of alternative splicing by identifying new factors within the splicing network of plants. In Arabidopsis thaliana, SR45 belongs to a family of factors that play important roles in splicing events. The gene OPENED IMMATURE FLOWER (OIF) was recently identified as a genetic enhancer of SR45. To understand how OIF modulates the splicing regulatory function of SR45, the OIF gene will be isolated by map-based cloning and functional complementation by transformation of candidate OIF genes into plants that are mutant for SR45 and OIF. New factors in the splicing network will be sought by screening for additional enhancers that produce an opened immature flower phenotype in the sr45 mutant background. Mutant phenotypes will be further characterized by scanning electron microscopy, and the splicing patterns of potential gene targets will be analyzed using RT-PCR. Together, these approaches will provide new insights into the role of alternative splicing in plant development. Broader Impact: Undergraduate students will be engaged in a unique research experience that can be replicated at other undergraduate institutions. First, through a partnership between St. Bonaventure University and the University of Maryland, students will be exposed to active research by gaining first-hand laboratory experience at both institutions and by attending national and international conferences. Second, a new integrative course, Plant Development and Physiology, including research-based lectures, open-ended research lab sessions, and interactive discussions, will be designed and offered to undergraduates at the sophomore/junior/senior level. Prior to each lab session, a student will be designated and trained to lead the lab with help from the instructor. Students from the entire class will participate in generating a poster describing their combined results from all lab sessions and share co-authorship. One student will be selected to present the poster at a conference. This innovative design will engage students in critical thinking, active research and creative discussion. All the opportunities provided to the students in this project are aimed at stimulating enthusiasm and building research experience in preparation for future careers in scientific research.