Studying Iridoid Transport in Catharanthus roseus by Grafting

The plant Catharanthus roseus is well known for its spatial separation of iridoid and monoterpenoid indole alkaloid (MIA) biosynthesis at both intracellular and intercellular levels, collectively suggested by RNA in situ hybridization, enzymatic and transcriptomic studies using leaf epidermis, and fluorescent protein tagging studies. Although documented in other plant species, the long-distance transport of iridoid glycosides, such as secologanin, has not been known in C. roseus until a recent study suggested that secologanin is transported from root to shoot, by grafting low iridoid/MIA mutant scions onto wild-type stock plants. This chapter describes the in vitro cultivation of C. roseus plants and grafting techniques to enable studies concerning iridoid/MIA transport between organs. The iridoid and MIA analysis methods are also provided.