Chromosome-level genome assembly of Prunella vulgaris L. provides insights into pentacyclic triterpenoid biosynthesis.

Prunella vulgaris is one of the bestselling and widely used medicinal herbs. It is recorded as an ace medicine for cleansing and protecting the liver in Chinese Pharmacopoeia and has been used as the main constitutions of many herbal tea formulas in China for centuries. It is also a traditional folk medicine in Europe and other countries of Asia. Pentacyclic triterpenoids are a major class of bioactive compounds produced in P. vulgaris. However, their biosynthetic mechanism remains to be elucidated. Here, we report a chromosome-level reference genome of P. vulgaris using an approach combining Illumina, ONT, and Hi-C technologies. It is 671.95 Mb in size with a scaffold N50 of 49.10 Mb and a complete BUSCO of 98.45%. About 98.31% of the sequence was anchored into 14 pseudochromosomes. Comparative genome analysis revealed a recent WGD in P. vulgaris. Genome-wide analysis identified 35 932 protein-coding genes (PCGs), of which 59 encode enzymes involved in 2,3-oxidosqualene biosynthesis. In addition, 10 PvOSC, 358 PvCYP, and 177 PvUGT genes were identified, of which five PvOSCs, 25 PvCYPs, and 9 PvUGTs were predicted to be involved in the biosynthesis of pentacyclic triterpenoids. Biochemical activity assay of PvOSC2, PvOSC4, and PvOSC6 recombinant proteins showed that they were mixed amyrin synthase (MAS), lupeol synthase (LUS), and ß-amyrin synthase (BAS), respectively. The results provide a solid foundation for further elucidating the biosynthetic mechanism of pentacyclic triterpenoids in P. vulgaris.

Chromosome-level genome assembly of Salvia miltiorrhiza with orange roots uncovers the role of Sm2OGD3 in catalyzing 15,16-dehydrogenation of tanshinones.

Salvia miltiorrhiza is well known for its clinical practice in treating heart and cardiovascular diseases. Its roots, used for traditional Chinese medicine materials, are usually brick-red due to accumulation of red pigments, such as tanshinone IIA and tanshinone I. Here we report a S. miltiorrhiza line (shh) with orange roots. Compared with the red roots of normal S. miltiorrhiza plants, the contents of tanshinones with a single bond at C-15,16 were increased, whereas those with a double bond at C-15,16 were significantly decreased in shh. We assembled a high-quality chromosome-level genome of shh. Phylogenomic analysis showed that the relationship between two S. miltiorrhiza lines with red roots was closer than the relationship with shh. It indicates that shh could not be the mutant of an extant S. miltiorrhiza line with red roots. Comparative genomic and transcriptomic analyses showed that a 1.0 kb DNA fragment was deleted in shh Sm2OGD3m. Complementation assay showed that overexpression of intact Sm2OGD3 in shh hairy roots recovered furan D-ring tanshinone accumulation. Consistently, in vitro protein assay showed that Sm2OGD3 catalyzed the conversion of cyptotanshinone, 15,16-dihydrotanshinone I and 1,2,15,16-tetrahydrotanshinone I into tanshinone IIA, tanshinone I and 1,2-dihydrotanshinone I, respectively. Thus, Sm2OGD3 functions as tanshinone 15,16-dehydrogenase and is a key enzyme in tanshinone biosynthesis. The results provide novel insights into the metabolic network of medicinally important tanshinone compounds.

Comparative efficacy of Chinese herbal injections for angina pectoris: A Bayesian network meta-analysis of randomized controlled trials.

OBJECTIVE: The severity of angina pectoris has been recognized. It is believed that Chinese herbal injections have an outstanding clinical effect on this condition. This network meta-analysis was devised to investigate the comparative efficacy of eight Chinese herbal injections (Ciwujia injection, Dazhuhongjingtan injection, Huangqi injection, Shenfu injection, Shengmai injection, Shenmai injection, Shenqi Fuzheng injection, Yiqifumai injection) in the treatment of angina pectoris. METHODS: A literature search was performed in PubMed, Embase, and the Cochrane Library, Chinese Biological Medicine Database, China National Knowledge Infrastructure, Wanfang Database, and Chinese Scientific Journal Database from their inception to June 25, 2018. A pre-designed eligibility criterion was utilized in this network meta-analysis, and a methodological quality analysis was conducted. Data analysis was performed by WinGUGS 1.4.3, Stata 13.0 and TSA software, and the odds ratio or mean difference with the 95% credible interval was reported for symptomatic improvement, electrocardiography improvement, fibrinogen, triglyceride and cholesterol. The ranking probability of interventions in various outcomes was also utilized. RESULTS: A total of 73 randomized controlled trials with 6639 patients were identified. Integrating network meta-analysis results, Shenqi Fuzheng injection plus western medicine therapy and Shenmai injection plus western medicine therapy were shown to be more efficacious than other therapies. In addition, Huangqi injection plus western medicine therapy and Shenmai injection plus western medicine therapy performed well in improving the haemorheology index and serum lipid parameters. CONCLUSIONS: Eligible Chinese herbal injections plus western medicine therapy might have a better impact on angina pectoris patients than western medicine therapy alone. While this study had limitations, the findings should be interpreted with caution. In addition, more high-quality randomized controlled trials with a large sample must be conducted to support this study.