Development of Atropa belladonna L. Plants with High-Yield Hyoscyamine and without Its Derivatives Using the CRISPR/Cas9 System

L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from the tropane alkaloids in . Therefore, it is vital to develop plants with high yields of hyoscyamine, and without anisodamine and scopolamine. In this study, we generated plants without anisodamine and scopolamine, via the CRISPR/Cas9-based disruption of hyoscyamine 6 -hydroxylase ( ), for the first time. Hyoscyamine production was significantly elevated, while neither anisodamine nor scopolamine were produced, in the . plants with homozygous mutations in . In summary, new varieties of with high yields of hyoscyamine and without anisodamine and scopolamine have great potential applicability in producing hyoscyamine at a low cost.