Caffeoyl-coenzyme A O-methyltransferase mediates regulation of carbon flux fluctuations during phenylpropenes and lignin biosynthesis in the vegetative organ roots of Asarum sieboldii Miq

Pingping Ji; Maoyi Lin; Mengying Chen; Muhammad Haneef Kashif; Yuling Fan; Tahir Ali; Ruixian Dai; Chongsheng Peng; Zhiqing Wang; Zhong Liu

doi:10.1016/j.plaphy.2023.107855

Caffeoyl-coenzyme A O-methyltransferase mediates regulation of carbon flux fluctuations during phenylpropenes and lignin biosynthesis in the vegetative organ roots of Asarum sieboldii Miq

Plant Physiol Biochem. 2023 Aug:201:107855. doi: 10.1016/j.plaphy.2023.107855. Epub 2023 Jul 5.

Authors

Affiliations

¹ School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
² School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; School of Life Sciences, Beijing University of Chinese Medicine, Beijing 102488, China.
³ School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China.
⁴ School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: cspeng@sjtu.edu.cn.
⁵ School of Chinese Medicinal Materials, Jilin Agricultural University, Changchun 130118, China. Electronic address: wangzhiqing@jlau.edu.cn.
⁶ School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: liuzhong@sjtu.edu.cn.

PMID: 37433236
DOI: 10.1016/j.plaphy.2023.107855

Abstract

Asarum sieboldii Miq. possesses remarkable medicinal value due to its essential oil enriched with phenylpropenes (e.g., methyleugenol and safrole). Although the biosynthesis of phenylpropenes shares a common pathway with lignin, the regulation mechanisms in carbon flux allocation between them are unclear. This study is the first to genetically verify the carbon flux regulation mechanism in A. sieboldii roots. We regulated the expression of Caffeoyl-coenzyme A O-methyltransferase (CCoAOMT), an essential enzyme in the common pathway, to investigate carbon flux allocation in vegetative organs. Here, the lignin and phenylpropene content fluctuation was analyzed by wet chemistry and GC-MS methods. A bona fide CCoAOMT gene from A. sieboldii was firstly cloned and verified. Preliminary heterologous expression validation in transgenic Arabidopsis thaliana showed that RNAi-induced CCoAOMT down-regulation significantly decreased lignin content by 24% and increased the S/G ratio by 30%; however, AsCCoAOMT over-expression in A. thaliana resulted in a 40% increase in lignin content and a 20% decrease in the S/G ratio when compared to the wild type. Similar trends were noted in homologous transformation in A. sieboldii, although the variations were not conspicuous. Nevertheless, the transgenic A. sieboldii plants displayed substantial differences in the level of phenylpropene compounds methyleugenol and safrole leading to a 168% increase in the methyleugenol/safrole ratio in the over-expression line and a 73% reduction in RNAi-suppression line. These findings suggest that the biosynthesis of phenylpropene constituents methyleugenol and safrole seems to be prioritized over lignin. Furthermore, this study indicated that suppression of AsCCoAOMT resulted in marked root susceptibility to pathogenic fungal disease, implying a significant additional role of CCoAOMT in protecting plant vegetative parts from diseases. Overall, the present study provides important references and suggests that future research should be aimed at elucidating the detailed mechanisms of the carbon flux allocation between phenylpropenes and lignin biosynthesis, as well as the disease resistance competency.

Keywords: Asarum sieboldii Miq.; Caffeoyl-coenzyme A O-Methyltransferase; Carbon flux allocation; Chinese traditional medicine; Lignin biosynthesis; Phenylpropenes.