BRC-BIO: Coding for Stress Response: Characterization of stress-responsive peptides from sORF-containing long intergenic non-coding RNAs in Arabidopsis thaliana

Plants face increasing challenges from climate change-induced stressors, resulting in unpredictable crop yields and increasing regional famines, with the greatest implications for areas of the world already grappling with food insecurity. These changes threaten food security and at worst, have the potential to trigger regional famine and civil unrest and are thus a threat to national security. Understanding plant development and stress response processes is crucial to producing crops that are more resilient to increasingly extreme environmental conditions. Small peptides play significant roles in changes during development and stress responses in plants. This project aims to explore the functional role of 20 newly identified small peptides in plant development and response to environmental stressors to identify potential targets for crop improvement. This project will engage undergraduate students in the research process through hands-on experience in a course-based undergraduate research project in Molecular Biology and in more immersive summer research positions at Gustavus Adolphus College and the Boyce Thompson Institute at Cornell University. Additionally, undergraduate students will collaborate to develop and test next-generation science standards-aligned educational modules for K-12 educators on genetics, genetic engineering, and climate impacts on food security. Long intergenic non-coding RNAs (lincRNAs) play a significant role in gene expression changes during development and stress responses in plants. Some lincRNAs contain small open reading frames (sORFs) that can encode short peptides, known as sORF-encoded peptides or SEPs, with regulatory functions. Identification of SEPs has been challenging due to their small size and unique gene expression profiles but have recently come to light through ribosomal profiling and mass spectrometry. The current project seeks to target 20 translated SEP loci, for knock-out using CRISPR/Cas9 gene editing. Edited plants will be evaluated using low-cost, high-throughput phenotyping-based approaches to determine the consequences of loss of SEP expression. SEP mutants that display altered growth will be further investigated using genetic and molecular techniques to characterize their role in plant development and stress responses. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.