Skip to main content

ORIGINAL RESEARCH article

Front. Plant Sci.
Sec. Plant Metabolism and Chemodiversity
Volume 15 - 2024 | doi: 10.3389/fpls.2024.1374912
This article is part of the Research Topic Plant Secondary Metabolite Biosynthesis View all 10 articles

Systematic characterization of gene families and functional analysis of PvRAS3 and PvRAS4 involved in rosmarinic acid biosynthesis in Prunella vulgaris

Provisionally accepted
Chao Yan Chao Yan 1,2Caili Li Caili Li 1Maochang Jiang Maochang Jiang 1Yayun Xu Yayun Xu 1Sixuan Zhang Sixuan Zhang 1Xiangling Hu Xiangling Hu 1,2Yuhang Chen Yuhang Chen 2*Shanfa Lu Shanfa Lu 1*
  • 1 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, Beijing Municipality, China
  • 2 College of Pharmaceutical Sciences, Chengdu Medical College, Chengdu 610500, China, Chengdu, Sichuan Province, China

The final, formatted version of the article will be published soon.

    Prunella vulgaris is an important material for Chinese medicines with rosmarinic acid (RA) as its index component. Based on the chromosome-level genome assembly we obtained recently, 51 RA biosynthesis-related genes were identified. Sequence feature, gene expression pattern and phylogenetic relationship analyses showed that 17 of them could be involved in RA biosynthesis. In vitro enzymatic assay showed that PvRAS3 catalyzed the condensation of p-coumaroyl-CoA and caffeoyl-CoA with pHPL and DHPL. Its affinity toward p-coumaroyl-CoA was higher than caffeoyl-CoA. PvRAS4 catalyzed the condensation of p-coumaroyl-CoA with pHPL and DHPL. Its affinity towards p-coumaroyl-CoA was lower than PvRAS3. UPLC and LC-MS/MS analyses showed the existence of RA, 4-coumaroyl-3',4'-dihydroxyphenyllactic acid, 4-coumaroyl-4'-hydroxyphenyllactic acid and caffeoyl-4'-hydroxyphenyllactic acid in P. vulgaris. Generation and analysis of pvras3 homozygous mutants showed significant decrease of RA, 4-coumaroyl-3',4'-dihydroxyphenyllactic acid, 4-coumaroyl-4'-hydroxyphenyllactic acid and caffeoyl-4'-hydroxyphenyllactic acid and significant increase of DHPL and pHPL. It suggests that PvRAS3 is the main enzyme catalyzing the condensation of acyl donors and acceptors during RA biosynthesis. The role of PvRAS4 appears minor. The results provide significant information for quality control of P. vulgaris medicinal materials.

    Keywords: biosynthetic pathway, CRISPR/Cas9, in vitro enzymatic activity assay, Prunella vulgaris, PvRAS3, PvRAS4, Rosmarinic acid, Rosmarinic acid synthase

    Received: 23 Jan 2024; Accepted: 15 Apr 2024.

    Copyright: © 2024 Yan, Li, Jiang, Xu, Zhang, Hu, Chen and Lu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Yuhang Chen, College of Pharmaceutical Sciences, Chengdu Medical College, Chengdu 610500, China, Chengdu, Sichuan Province, China
    Shanfa Lu, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193, Beijing Municipality, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.