Identification of a central regulator of ginkgolide biosynthesis in <i>Ginkgo biloba</i> that integrates jasmonate and light signaling

J Jinfa Du (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) Z Zhen Zhao (Institute of Catalysis for Energy and Environment) L Lingqi Jin (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) L Lijin Huang (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) D Dian Jin (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) X Xiaoyan Zheng Q Qiaolei Wang (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) W Wenbo Xu (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) H Huijun Guo (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) X Xinyue Xing (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) R Raphael N. Alolga (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) L Lam-Son Phan Tran (Department of Plant and Soil Science, Institute of Genomics for Crop Abiotic Stress Tolerance, Texas Tech University) L Luis Rafael Herrera-Estrella (State Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization, Zhongshan Biological Breeding Laboratory, Department of Plant Nutrition, College of Resources and Environmental Sciences, Nanjing Agricultural University) P Ping Li X Xiaojian Yin (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University) X Xu Lu (Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University)

Abstract

Ginkgolides are secondary metabolites unique to Ginkgo biloba with the potential to prevent and treat cardiovascular and cerebrovascular diseases. Although the biosynthetic pathways of ginkgolides have been partly uncovered, the mechanism regulating their biosynthesis is still largely unknown. Here, using multiomic and genetic analyses, we report the identification of a transcription factor, named ETHYLENE RESPONSE FACTOR ASSOCIATED WITH GINKGOLIDE BIOSYNTHESIS (GbEAG), as a critical regulator of ginkgolide biosynthesis. GbEAG is highly expressed in the roots of G. biloba, and its expression is significantly induced by methyl jasmonate (MeJA). Ginkgolide content was significantly increased in roots by overexpressing GbEAG using a “cut-dip-regeneration” system. GbEAG positively regulates ginkgolide biosynthesis by directly binding to the GCC-boxes in the promoter regions of genes involved in the biosynthesis of ginkgolides, such as ISOPENTENYL DIPHOSPHATE ISOMERASE ( GbIDI ) and CYTOCHROME P450 7005C3 ( GbCYP7005C3 ). GbEAG mediates the jasmonic acid (JA)-activated ginkgolide synthesis through its direct interaction with the JASMONATE ZINC-FINGER INFLORESCENCE MERISTEM DOMAIN 3 (GbJAZ3) repressor. Importantly, we also found that the central light-response regulator ELONGATED HYPOCOTYL 5 (GbHY5) mediates light induction of ginkgolide biosynthesis by binding to the G-box in the GbEAG promoter. Our findings provide mechanistic insights into the coordinated regulation of ginkgolide biosynthesis via JA and light signals, with GbEAG as a central regulator in G. biloba, and shed light on the potential to develop ginkgolide-rich varieties through molecular breeding and gene editing.

Article Details

Volume / Issue Vol. 122, Issue 6
Published February 11, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (16)

J

Jinfa Du

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

Z

Zhen Zhao

Institute of Catalysis for Energy and Environment

L

Lingqi Jin

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

L

Lijin Huang

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

D

Dian Jin

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

X

Xiaoyan Zheng

Q

Qiaolei Wang

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

W

Wenbo Xu

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

H

Huijun Guo

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

X

Xinyue Xing

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

R

Raphael N. Alolga

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

L

Lam-Son Phan Tran

Department of Plant and Soil Science, Institute of Genomics for Crop Abiotic Stress Tolerance, Texas Tech University

L

Luis Rafael Herrera-Estrella

State Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization, Zhongshan Biological Breeding Laboratory, Department of Plant Nutrition, College of Resources and Environmental Sciences, Nanjing Agricultural University

P

Ping Li

X

Xiaojian Yin

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University

X

Xu Lu

Department of Pharmacognosy, State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University