Tripterygium wilfordii is a Traditional Chinese Medicine (TCM) from family Celastraceae and celastrol is one of the strongest active ingredients belonging to friedelane-type pentacyclic triterpenoid, which has a large clinical application value of anti-tumor, immunosuppression, and obesity treatment. The first committed biosynthesis step of celastrol is the cyclization of 2, 3-oxidosqualene to friedelin, catalyzed by the oxidosqualene cyclase, while the rest of this pathway is still unclear. In this study, we reported a reference genome assembly of T. wilfordii with high-quality annotation by using a hybrid sequencing strategy (Nanopore, Bionano, Illumina HiSeq, and Pacbio), which obtained a 340.12 Mb total size and contig N50 reached 3.09 Mb. In addition, we successfully anchored 91.02% sequences into 23 pseudochromosomes using Hi-C technology and the super-scaffold N50 reached 13.03 Mb. Based on integration genome, transcriptom and metabolite analyses, as well as in vivo and in vitro enzyme assays, two CYP450 genes, TwCYP712K1 and TwCYP712K2 have been proven for C-29 position oxidation of friedelin to produce polpunonic acid, which clarifies the second biosynthesis step of celastrol. Syntenic analysis revealed that TwCYP712K1 and TwCYP712K2 derived from the common ancestor. These results have provided insight into illustrating pathways for both celastrol and other bioactive compounds found in this plant.

中文翻译：

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雷公藤的基因组分析显示，TwCYP712K1和TwCYP712K2负责Celastrol生物合成途径中Friedelin的氧化

雷公藤（雷公藤）是Celestraceae家族的传统中药（TCM），celastrol是属于Friedelane型五环三萜的最强活性成分之一，具有抗肿瘤，免疫抑制和肥胖症治疗的巨大临床应用价值。celastrol的第一个重要生物合成步骤是由氧化角鲨烯环化酶催化将2，3-氧化角鲨烯环化为Friedelin，而该途径的其余部分仍不清楚。在这项研究中，我们报告了野生拟南芥的参考基因组装配通过使用杂交测序策略（Nanopore，Bionano，Illumina HiSeq和Pacbio）进行高质量注释，获得总大小340.12 Mb，重叠群N50达到3.09 Mb。此外，我们使用Hi-C技术成功地将91.02％的序列锚定到23个假染色体中，超级支架N50达到13.03 Mb。根据整合基因组，转录组和代谢产物分析以及体内和体外酶分析，两个CYP450基因TwCYP712K1和TwCYP712K2已被证明可用于Friedelin的C-29位置氧化以产生polpunonic酸，从而阐明了第二个生物合成步骤celastrol。语法分析显示TwCYP712K1和TwCYP712K2来自共同祖先。这些结果提供了洞察力，以说明该植物中发现的天青素和其他生物活性化合物的途径。