Abstract
Aromatic compounds play essential roles in plant physiology and various industries because of their unique fragrances and beneficial properties. In this study, we investigated the transport and biosynthesis of eugenol, a prominent aromatic compound, within the Ocimum genus, using grafting experiments. Grafting sweet basil (Ocimum basilicum) scions onto diverse rootstocks, including tobacco (Nicotiana benthamiana) and thyme (Thymus vulgaris), revealed that eugenol is transported from the shoot to the root across distinct plant species. Furthermore, grafting within the Ocimum genus, which includes O. basilicum, O. tenuiflorum, and O. americanum, resulted in variations in eugenol transport and accumulation. The eugenol content in the shoots remained constant across all combinations, whereas the root eugenol levels varied depending on the scion-rootstock pair. To elucidate the biosynthetic capabilities of eugenol in Ocimum roots, we performed in vitro enzyme assays using crude protein extracts from roots, which revealed that eugenol can be synthesized in roots in addition to being transported. Expression analysis of eugenol synthase (EGSs) genes showed that EGS4 expression was influenced by grafting in O. basilicum roots, suggesting compensation by other EGSs. Our results suggest that eugenol transport and biosynthesis are multifaceted processes influenced by the interactions between different species and tissues. The potential to engineer eugenol levels in rootstocks lacking biosynthetic capacity has potential applications in agriculture and industry. This study reveals the dynamic interplay between eugenol transport and biosynthesis in the Ocimum genus, providing insights into the manipulation of aromatic compound production in plants.