High Throughput Localization and Characterization of Conserved Photosynthetic Proteins in Chlamydomonas Reinhardtii

Abstract

An ongoing food crisis affects humanity, and today millions face hunger worldwide. Demand for foodstuffs is projected to exceed supply within decades, yet climate change and deforestation are set to diminish the land suitable for agricultural use. One solution, already met with success, is increasing crop yield efficiency through bioengineering staple crop strains. Model green algae Chlamydomonas Reinhardtii performs a highly efficient mode of photosynthesis within unique subcellular structures. Crop strains performing photosynthesis similarly efficiently would constitute a technical achievement and resecure the agricultural sector. Chlamydomonas remains the most well understood alga, yet reverse engineering attempts are limited by a poor understanding of its chloroplast. Evolutionarily conserved genes common to land plant model Arabidopsis Thaliana offer an untapped wealth of information that informs models in both organisms. Informed by literature, we selected 289 Chlamydomonas genes whose Arabidopsis homologs are reported to the chloroplast. We then leveraged a recently certified high-throughput fluorescent tagging strategy to obtain subcellular localizations for 47 proteins in Chlamydomonas. Results yield the initial experimental report of 98% of these proteins in Chlamydomonas, including 12 chloroplast localizations. We use these to perform analysis and speculate on possible pathway roles. We propose one gene as a γ-glutyl-cyclo-transferase and offer future directions. Proposals aim to inform the work needed to understand Chlamydomonas going forward.