Project ChemicalBlooms: Collaborating with Citizen Scientists to Survey the Chemical Diversity and Phylogenetic Distribution of Plant Epicuticular Wax Blooms

Abstract

Plants use chemistry to overcome diverse challenges. A particularly striking chemical trait that some plants possess is the ability to synthesize massive amounts of epicuticular wax that accumulates on the plants surfaces as a white coating visible to the naked eye. The ability to synthesize basic wax molecules appears to be shared among virtually all land plants and our knowledge of ubiquitous wax compound synthesis is reasonably advanced. However, the ability to synthe-size thick layers of visible epicuticular crystals ("wax blooms") is restricted to specific lineages and our knowledge of how wax blooms differ from ubiquitous wax layers is less developed. Here, we recruited the help of citizen scientists and middle school students to survey the wax bloom chemistry of 78 species spanning dicot, monocot, and gymnosperm lineages. Using gas chromatography-mass spectrometry, we found that the major wax classes reported from bulk wax mixtures can be present in wax bloom crystals, with fatty acids, fatty alcohols, and alkanes being present in many species bloom crystals. In contrast, other compounds including aldehydes, ketones, secondary alcohols, and triterpenoids were present in only a few species wax bloom crystals. By mapping the 78 wax bloom chemical profiles onto a phylogeny and using phylogenetic comparative analyses, we found that secondary alcohol and triterpenoid-rich wax blooms were present in lineage-specific patterns that would not be expected to arise by chance. That finding is consistent with reports that secondary alcohol biosynthesis enzymes are found only in certain lineages, but was a surprise for triterpenoids, which are intracellular components in virtually all plant lineages. Thus, our data suggest that a lineage-specific mechanism other than biosynthesis exists that enables select species to generate triterpenoid-rich surface wax crystals. Overall, our study outlines a general mode in which research scientists can collaborate with citizen scientists as well as middle and high school classrooms not only to enhance data collection and generate testable hypotheses, but also directly involve classrooms in the scientific process and inspire future STEM workers. Significance StatementPlants coat themselves in a protective layers of waxes. Some plants produce exceptionally thick layers of wax ("wax blooms"), and these thick layers are associated with enhanced abilities to tolerate stress, including drought and insect attack. In collaboration with citizen scientists and middle school classrooms, we provide an overview of the chemistry and phylogenetic distribution of plant epicuticular wax blooms. These data constitute a foundation upon which future studies of diverse wax blooms and their functions can build.