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

• Main Authors: Lingjiang Zeng, Qiaozhuo Zhang, Chunxue Jiang, Yueyue Zheng, Youwei Zuo, Jianbo Qin, Zhihua Liao, Hongping Deng
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: International Journal of Molecular Sciences
• Subjects: <i>Atropa belladonna</i> L.; CRISPR/Cas9; hyoscyamine 6<i>β</i>-hydroxylase; tropane alkaloids; transgenic plant
• Online Access: https://www.mdpi.com/1422-0067/22/4/1731

<i>Atropa belladonna</i> L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the <i>in planta</i> hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from the tropane alkaloids in <i>A. belladonna</i>. Therefore, it is vital to develop <i>A. belladonna</i> plants with high yields of hyoscyamine, and without anisodamine and scopolamine. In this study, we generated <i>A. belladonna</i> plants without anisodamine and scopolamine, via the CRISPR/Cas9-based disruption of hyoscyamine 6<i>β</i>-hydroxylase (<i>AbH6H</i>), for the first time. Hyoscyamine production was significantly elevated, while neither anisodamine nor scopolamine were produced, in the <i>A</i>. <i>belladonna</i> plants with homozygous mutations in <i>AbH6H</i>. In summary, new varieties of <i>A. belladonna</i> with high yields of hyoscyamine and without anisodamine and scopolamine have great potential applicability in producing hyoscyamine at a low cost.