The chemical biogeography of a widespread aromatic plant species shows both spatial and temporal variation

Plants produce a wide variety of secondary metabolites, but intraspecific variation in space and time can alter the ecological interactions these compounds mediate. The aim of this work was to document the spatial and temporal chemical biogeography of Monarda fistulosa. I collected leaves from 1587 M. fistulosa individuals from 86 populations from Colorado to Manitoba, extracted and analyzed their terpenes with gas chromatography, mapped monoterpene chemotypes, and analyzed chemical variation with principal component analysis. I also measured the amounts of terpenes in different plant tissues to examine intraplant variation and monitored leaf terpene chemistry over a single growing season to examine temporal patterns. Finally, I extracted terpenes from herbarium samples up to 125 years old and compared the chemotypes with recent samples from the same sites. M. fistulosa populations consisted mostly of thymol (T) and carvacrol (C) chemotypes, but geraniol (G) and (R)-(-)-linalool (L), a chemotype new to this species, were also present. A principal component analysis showed that most of the chemical variation across populations was due to the amounts of the dominant terpene in plants. Intraplant tissue chemistry revealed that leaves mostly had the greatest amounts of terpenes, followed by floral structures, stems, and roots. Short-term temporal variation in leaf chemistry of T and C plants over a growing season showed that plants produced the highest levels of biosynthetic precursors early in the season and their dominant monoterpenes peaked in mid-summer. Plant chemotype was discernable in the oldest herbarium samples, and 15 of 18 historic samples matched the majority chemotype currently at the site, indicating that population chemotype ratios may remain stable over longer time scales. Overall, the results show that plant species' secondary chemistry can vary both spatially and temporally, which may alter the biological interactions that these compounds mediate over space and time.