Project description:<p><em>Tripterygium wilfordii</em> is a vine used in Traditional Chinese Medicine (TCM) from the family Celastraceae. The active ingredient celastrol is a friedelane-type pentacyclic triterpenoid, with a putative role as an anti-tumor, immunosuppression, and obesity agent. Here we reported a reference genome assembly of <em>T. wilfordii</em> with high-quality annotation by using a hybrid sequencing strategy, which obtained a 340.12 Mb total genome size, a contig N50 reaching 3.09 Mb, 31593 structure genes, and the repeat percentage was 44.31%. Comparative evolutional analyses showed that <em>T. wilfordii</em> diverged from species of Malpighiales about 102.4 million years ago. 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 of genome, transcriptome and metabolite analyses, as well as in vivo and in vitro enzyme assays of the two CYP450 genes, <em>TwCYP712K1</em> and <em>TwCYP712K2</em> the second biosynthesis step of celastrol was investigated and elucidated. Syntenic analysis revealed that <em>TwCYP712K1</em> and <em>TwCYP712K2</em> derived from a common ancestor. These results have provided insights into further investigating pathways for celastrol and valuable information to aid the conservation of resources and helped us reveal the evolution of Celastrales.</p>
Project description:Triptolide or triptonide, major bioactive components isolated from the traditional Chinese herb Tripterygium wilfordii Hook f. (TWHF), have been shown to exert various pharmacological effects on systemic autoimmune disease. However, the severe toxicity, especially reproduction toxicity, of triptolide or triptonide prevents wide clinical use for those people with fertility needs. Long noncoding (lnc) RNAs are critical for spermatogenesis, and their dysregulation might lead to male infertility. However, whether they are involved in triptolide or triptonide -induced reproduction toxicity is unknown. Here we found that exposure of spermatocyte cell line (GC2) to triptolide or triptonide led to reduction in cell viability and induced cell apoptosis and autophagy. Then the total RNAs of triptolide or triptonide -treated GC2 cells were used to investigate lncRNA expression profiles by strand-specific RNA sequencing at the transcriptome level to help determine any RNA-related mechanisms in triptolide or triptonide-induced toxicity. Gene ontology and pathway analyses showed that the functions of differentially expressed lncRNA targets and mRNAs were closely linked with many processes involved in spermatogenesis.