1) The research shows variation in the acylsucrose biosynthesis pathway between wild tomato species like S. pennellii and the cultivated tomato S. lycopersicum. Through enzyme assays and mutagenesis, the research manipulated acyl donors and acceptors in this secondary metabolic pathway. 2) The pathway differs between S. pennellii, which fully acylates the pyranose ring, versus S. lycopersicum which adds acyl chains to positions R2 and R3. Mutagenesis identified amino acid changes that allow enzymes to utilize different substrates. 3) Future work includes sequence alignments to understand pathway evolution, analyzing retention time shifts from mutagenesis, and developing NMR structures to identify