The study carry out comprehensive transcriptome analysis of C. deltoidea and exploration of BIAs biosynthesis and accumulation based on UHPLC-MS/MS and combined sequencing platforms. Coptis deltoidea is an important medicinal plant with a long history of medicinal use, which is rich in benzylisoquinoline alkaloids (BIAs). In this study, Ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) and combined sequencing platforms were performed for exploration of BIAs biosynthesis, accumulation and comprehensive transcriptome analysis of C. deltoidea. By metabolism profiling, the accumulation of ten BIAs was analyzed using UHPLC-MS/MS and different contents were observed in different organs. From transcriptome sequencing result, we applied single-molecule real-time (SMRT) sequencing to C. deltoidea and generated a total of 75,438 full-length transcripts. We proposed the candidate biosynthetic pathway of tyrosine, precursor of BIAs, and identified 64 full length-transcripts encoding enzymes putatively involved in BIAs biosynthesis. RNA-Seq data indicated that the majority of genes exhibited relatively high expression level in roots. Transport of BIAs was also important for their accumulation. Here, 9 ABC transporters and 2 MATE transporters highly homologous to known alkaloid transporters related with BIAs transport in roots and rhizomes were identified. These findings based on the combined sequencing platforms provide valuable genetic information for C. deltoidea and the results of transcriptome combined with metabolome analysis can help us better understand BIAs biosynthesis and transport in this medicinal plant. The information will be critical for further characterization of C. deltoidea transcriptome and molecular-assisted breeding for this medicinal plant with scarce resources.

中文翻译：

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基于组合测序平台的黄连全长转录组分析及参与苄基异喹啉生物碱生物合成的推定基因鉴定。

本研究基于UHPLC-MS/MS和组合测序平台，对三角花进行了全面的转录组分析，并探索了BIAs的生物合成和积累。黄连是重要的药用植物，药用历史悠久，富含苄基异喹啉生物碱（BIAs）。本研究采用超高效液相色谱-电喷雾电离串联质谱（UHPLC-ESI-MS/MS）和组合测序平台，探索三角锥虫BIAs的生物合成、积累和转录组综合分析。通过代谢谱分析，使用 UHPLC-MS/MS 分析了十种 BIA 的积累，并在不同器官中观察到不同的含量。根据转录组测序结果，我们将单分子实时 (SMRT) 测序应用于 C. deltoidea 共生成了 75,438 个全长转录本。我们提出了 BIA 前体酪氨酸的候选生物合成途径，并确定了 64 个全长转录本，编码假定参与 BIA 生物合成的酶。RNA-Seq 数据表明，大多数基因在根中表现出相对较高的表达水平。BIA 的运输对于它们的积累也很重要。在这里，鉴定了 9 个 ABC 转运蛋白和 2 个 MATE 转运蛋白，与已知的与 BIA 在根和根茎中转运相关的生物碱转运蛋白高度同源。这些基于组合测序平台的发现为三角锥虫提供了有价值的遗传信息，转录组与代谢组分析相结合的结果可以帮助我们更好地了解这种药用植物中 BIA 的生物合成和转运。