Identifying Genes Involved in Leaf Polarity in Solanum lycopersicum
Tomato leaves are dorsiventral, having distinct adaxial (upper/top) and abaxial (lower/bottom) tissue. Because these distinct plant tissues contribute to proper plant function via photosynthesis and gas exchange, understanding development mechanisms is important to the fundamental knowledge of plant systems. To investigate which genes may contribute to this natural asymmetry in leaf development, Sussex surgery—a procedure which separates the shoot apical meristem from the incipient leaf primorida, and has been known to result in radial, abaxialized leaves—was performed on young tomato plants, followed by an RNAseq analysis. Using this RNAseq data, six differentially expressed genes were identified as candidates that may contribute to leaf tissue development processes in tomato based on level of differential expression, statistical significance, and any known function related to leaf development. To further investigate these candidates, in situ hybridizations were performed using RNA probes from three of the genes using 2-week-old tomato shoot apical meristem tissue to spatially visualize each gene’s RNA expression within the meristem tissue. The results from the in situ hybridization experiments will provide a basis for selecting which of the six candidate genes will be used to create CRISPR tomato mutants. These mutants may provide further information regarding the gene’s function within the context of leaf dorsiventrality. These experiments will further elucidate the role of specific genes in leaf tissue development and the establishment of differentiated leaf tissue.
This summer has been wonderful all around. My experience in the Scanlon lab has greatly improved my molecular biology skillset, and has introduced me to new and interesting questions within plant science research related to development. I grew by working independently, as well as working with other experienced lab members all of whom supported my efforts. As of now, I plan to attend graduate school to grow my knowledge and understanding of plant systems and genetics as they relate to crop improvement and food security. I really want a career that puts me at the center of innovative agricultural technologies and applying those advances to address important issues in the realm of food security and sustainability. The BTI internship experience has been instrumental in meeting and engaging with like-minded individuals with a passion for plants, and I am grateful for this experience!