| Summary: | <p>Abstract</p> <p>Background</p> <p>The first two enzymatic steps of monoterpene indole alkaloid (MIA) biosynthetic pathway are catalysed by strictosidine synthase (STR) that condensates tryptamine and secologanin to form strictosidine and by strictosidine β-D-glucosidase (SGD) that subsequently hydrolyses the glucose moiety of strictosidine. The resulting unstable aglycon is rapidly converted into a highly reactive dialdehyde, from which more than 2,000 MIAs are derived. Many studies were conducted to elucidate the biosynthesis and regulation of pharmacologically valuable MIAs such as vinblastine and vincristine in <it>Catharanthus roseus </it>or ajmaline in <it>Rauvolfia serpentina</it>. However, very few reports focused on the MIA physiological functions.</p> <p>Results</p> <p>In this study we showed that a strictosidine pool existed <it>in planta </it>and that the strictosidine deglucosylation product(s) was (were) specifically responsible for <it>in vitro </it>protein cross-linking and precipitation suggesting a potential role for strictosidine activation in plant defence. The spatial feasibility of such an activation process was evaluated <it>in planta</it>. On the one hand, <it>in situ </it>hybridisation studies showed that CrSTR and CrSGD were coexpressed in the epidermal first barrier of <it>C. roseus </it>aerial organs. However, a combination of GFP-imaging, bimolecular fluorescence complementation and electromobility shift-zymogram experiments revealed that STR from both <it>C. roseus </it>and <it>R. serpentina </it>were localised to the vacuole whereas SGD from both species were shown to accumulate as highly stable supramolecular aggregates within the nucleus. Deletion and fusion studies allowed us to identify and to demonstrate the functionality of CrSTR and CrSGD targeting sequences.</p> <p>Conclusions</p> <p>A spatial model was drawn to explain the role of the subcellular sequestration of STR and SGD to control the MIA metabolic flux under normal physiological conditions. The model also illustrates the possible mechanism of massive activation of the strictosidine vacuolar pool upon enzyme-substrate reunion occurring during potential herbivore feeding constituting a so-called "nuclear time bomb" in reference to the "mustard oil bomb" commonly used to describe the myrosinase-glucosinolate defence system in Brassicaceae.</p>
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