Department of Chemistry, University of New Brunswick, Fredericton, NB, Canada.
Department of Chemical Engineering, University of New Brunswick, Fredericton, NB, Canada.
Methods Mol Biol. 2022;2505:69-77. doi: 10.1007/978-1-0716-2349-7_5.
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.
植物长春花以其在细胞内和细胞间水平上对裂环烯醚萜和单萜吲哚生物碱（MIA）生物合成的空间分离而闻名，这一观点综合了使用叶片表皮进行的 RNA 原位杂交、酶学和转录组学研究以及荧光蛋白标记研究的结果。尽管在其他植物物种中有相关记载，但直到最近的一项研究表明，长春花中的裂环烯醚萜糖苷，如獐牙菜苦苷，是从根部运输到地上部分的，通过将低含量的裂环烯醚萜/MIA 突变接穗嫁接到野生型母株上，这种远距离运输才为人所知。本章介绍了长春花植物的离体培养和嫁接技术，以实现关于器官间裂环烯醚萜/MIA 运输的研究。同时还提供了裂环烯醚萜和 MIA 的分析方法。
