Abstract
Atropa belladonna L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the in planta hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from the tropane alkaloids in A. belladonna. Therefore, it is vital to develop A. belladonna plants with high yields of hyoscyamine, and without anisodamine and scopolamine. In this study, we generated A. belladonna plants without anisodamine and scopolamine, via the CRISPR/Cas9-based disruption of hyoscyamine 6β-hydroxylase (AbH6H), for the first time. Hyoscyamine production was significantly elevated, while neither anisodamine nor scopolamine were produced, in the A. belladonna plants with homozygous mutations in AbH6H. In summary, new varieties of A. belladonna with high yields of hyoscyamine and without anisodamine and scopolamine have great potential applicability in producing hyoscyamine at a low cost.
Highlights
Atropa belladonna L. is an important herbal plant used by human beings
Modern pharmaceutical science has revealed that A. belladonna plants produce anticholinergic tropane alkaloids (TAs), including hyoscyamine, anisodamine, and scopolamine [1]
Hyoscyamine is useful for the treatment of arrhythmias and organophosphate poisoning; anisodamine has been applied to treat infective shock, gastrointestinal colic and vascular spasm; scopolamine is well-known for curing motion sickness [5,6,7]
Summary
Atropa belladonna L. is an important herbal plant used by human beings. Modern pharmaceutical science has revealed that A. belladonna plants produce anticholinergic tropane alkaloids (TAs), including hyoscyamine, anisodamine, and scopolamine [1]. A. belladonna plants produce hyoscyamine as a major compound of TAs, and produce the derivatives of hyoscyamine, including anisodamine and scopolamine, as minor compounds [6] It is necessary for the pharmaceutical industry to separate one compound from the others. NHEJ usually results in nucleotide insertion or deletion (indels), leading to a loss of or change in gene function Of these genome-editing technologies, the CRISPR/Cas system is the most popular because of its advantages, such as its ease of use, its high efficiency, and its capacity for adaptation to diverse organisms [17]. In order to establish germplasm of A. belladonna without anisodamine and scopolamine, we used the CRISPR/Cas system to genetically edit the H6H gene, and analyzed the production of tropane alkaloids in planta
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