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  • Integrated Transcript and Metabolite Profiles Reveal That EbCHI Plays an Important Role in Scutellarin Accumulation in Erigeron breviscapus Hairy Roots.

Integrated Transcript and Metabolite Profiles Reveal That EbCHI Plays an Important Role in Scutellarin Accumulation in Erigeron breviscapus Hairy Roots.

Frontiers in plant science (2018-07-18)
Ruibing Chen, Xianghui Chen, Tingting Zhu, Jianghua Liu, Xing Xiang, Jian Yu, Hexin Tan, Shouhong Gao, Qing Li, Yichao Fang, Wansheng Chen, Lei Zhang, Baokang Huang
ABSTRACT

Scutellarin, a flavonoid 7-O-glucuronide, is an essential bioactive compound of Erigeron breviscapus (Vaniot) Hand.-Mazz. used for the treatment of cerebrovascular diseases. However, due to overexploitation and overuse, E. breviscapus is facing the problems of extinction and habitat degradation. In this study, a correlation analysis between the transcript and metabolite profiles of methyl jasmonate (MeJA)-treated E. breviscapus at different time points indicated that chalcone isomerase (EbCHI) was the primary contributor to scutellarin accumulation during flavonoid biosynthesis. EbCHI was then further characterized as a chalcone isomerase that efficiently converted chalcone to naringenin in vitro. Optimal parameters derived by comparing different culture conditions were successfully used to establish hairy root cultures of E. breviscapus with a maximum transformation rate of 60% in B5 medium. Furthermore, overexpression of EbCHI significantly enhanced scutellarin accumulation in E. breviscapus hairy roots with a maximum content of 2.21 mg g-1 (dw), 10-fold higher than that of natural roots (0.21 mg g-1 dw). This study sheds new light on a method of effective gene-based metabolic engineering by accurate and appropriate strategies and provides a protocol for hairy root cultures that accumulate high levels of scutellarin, providing a promising prospect for relieving the overexploitation and unavailability of E. breviscapus in the future.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Scutellarein, ≥98% (HPLC)
Sigma-Aldrich
Naringenin, natural (US), 98%
Supelco
Scutellarin, analytical standard
Sigma-Aldrich
Methyl jasmonate, 95%
Sigma-Aldrich
3′,5′-Dimethoxy-4′-hydroxyacetophenone, 97%