Hyoscyamine and scopolamine (HS), two valuable tropane alkaloids of significant medicinal importance, are found in multiple distantly related lineages within the Solanaceae family. Here we sequence the genomes of three representative species that produce HS from these lineages, and one species that does not produce HS. Our analysis reveals a shared biosynthetic pathway responsible for HS production in the three HS-producing species. We observe a high level of gene collinearity related to HS synthesis across the family in both types of species. By introducing gain-of-function and loss-of-function mutations at key sites, we confirm the reduced/lost or re-activated functions of critical genes involved in HS synthesis in both types of species, respectively. These findings indicate independent and repeated losses of the HS biosynthesis pathway since its origin in the ancestral lineage. Our results hold promise for potential future applications in the artificial engineering of HS biosynthesis in Solanaceae crops.

天仙子胺和东莨菪碱 (HS) 是两种具有重要药用价值的有价值的托烷生物碱，存在于茄科家族的多个远亲谱系中。在这里，我们对这些谱系中产生 HS 的三种代表性物种和一种不产生 HS 的物种的基因组进行了测序。我们的分析揭示了三种产生 H2S 的物种中负责 H2S 产生的共同生物合成途径。我们在这两种物种的家族中观察到与 HS 合成相关的高水平基因共线性。通过在关键位点引入功能获得和功能丧失突变，我们分别确认了两类物种中参与 HS 合成的关键基因的功能减少/丢失或重新激活。这些发现表明 HS 生物合成途径自其起源于祖先谱系以来就存在独立且反复的损失。我们的研究结果有望在茄科作物中 H2S 生物合成的人工工程中得到潜在的应用。