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Sacred lotus is rich in biologically active compounds, particularly benzylisoquinoline alkaloids (BIAs). Here, we report on isolation of genes encoding ( S )-norcoclaurine synthase (NCS) in sacred lotus, which is a key entry-enzyme in BIA biosynthesis. Seven  NCS  genes, designated  NnNCS1  through  NnNCS7 , were identified in the sacred lotus genome, and five are located next to each other within a 83 kb region on scaffold 8. The  NCS  genes are divided into two subfamilies, designated  NCSI  and  NCSII . The  NCSII  genes are universal in plants, while the  NCSI  genes are only identified in a limited number of dicotyledonous taxa that produce BIAs. In sacred lotus, only  NnNCS4  belongs to the  NCSII  subfamily, whilst the rest  NCS  genes within the  NCSI  subfamily. Overall, the  NnNCS7  gene was predominantly expressed in all tested tissues, and its expression is significantly correlated with alkaloid content in leaf. In contrast, the  NnNCS4  expression shows no significant correlation with alkaloid accumulation in leaf, and its lack of expression cannot inhibit alkaloid accumulation. Taken together, these results suggest that the  NCSI  subfamily is crucial for BIA biosynthesis, and its origin may represent an important evolutionary event that allows certain plant taxa to produce BIAs.

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Evolutionary origin of the NCSI gene subfamily encoding norcoclaurine synthase is associated with the biosynthesis of benzylisoquinoline alkaloids in plants