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Article|17 Nov 2023|OPEN
ERF5.1 modulates carotenoid accumulation by interacting with CCD4.1 in Lycium
Jianhua Zhao1,4 , , Yuhui Xu1,4 , Haoxia Li2,4 , Xinlei Zhu1 , Yue Yin1 , Xiyan Zhang1 , Xiaoya Qin1 , Jun Zhou3 , Linyuan Duan1 , Xiaojie Liang1 , Ting Huang1 , Bo Zhang1 and Ru Wan1 , Zhigang Shi1 , Youlong Cao1 , Wei An,1 ,
1National Wolfberry Engineering Research Center/Wolfberry Science Research Institute, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan, 750002, China
2Institute of Forestry and Grassland Ecology, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan, 750002, China
3College of Biological Science & Engineering, North Minzu University, Yinchuan 750021, China
4These authors contributed equally: Jianhua Zhao, Yuhui Xu, Haoxia Li.
*Corresponding author. E-mail:,

Horticulture Research 11,
Article number: uhad230 (2024)
Views: 121

Received: 13 Jun 2023
Accepted: 01 Nov 2023
Published online: 17 Nov 2023


Carotenoids are important natural pigments and have medical and health functions for humans. Carotenoid cleavage dioxygenase 4 (CCD4) and ethylene responsive factor (ERF) participate in carotenoid metabolism, but their roles in Lycium have not been discovered. Here, we annotated LbCCDs from the Lycium reference genome and found that LbCCD4.1 expression was significantly correlated with the carotenoid metabolites during Lycium five fruit developmental stages. Over-expression of LbCCD4.1 in NQ’s leaves resulted in a series of significantly lower contents of carotenoid metabolites, including β-carotene and β-cryptoxanthin. Moreover, LbERF5.1, a transcription factor belonging to the ERF family that was located in the nucleus, was isolated. Significant reductions in the carotenoids, especially lutein, violaxanthin and their derivatives, were observed in over-expressing ERF5.1 transgenic NQ’s leaves. Over-expression or virus-induced gene silencing of LbERF5.1 in NQ’s leaves induced a consistent up- or down-expression, respectively, of LbCCD4.1. Furthermore, yeast one-hybrid and dual-luciferase reporter assays showed that ERF5.1 interacted with the promoter of CCD4.1 to increase its expression, and LbERF5.1 could bind to any one of the three predicted binding sites in the promoter of LbCCD4.1. A transcriptome analysis of LbERF5.1 and LbCCD4.1 over-expressed lines showed similar global transcript expression, and geranylgeranyl diphosphate synthase, phytoene synthase, lycopene δ-cyclase cytochrome, cytochrome P450-type monooxygenase 97A, cytochrome P450-type monooxygenase 97C, and zeaxanthin epoxidase in the carotenoid biosynthesis pathway were differentially expressed. In summary, we uncovered a novel molecular mechanism of carotenoid accumulation that involved an interaction between ERF5.1 and CCD4.1, which may be used to enhance carotenoid in Lycium.