Expression profiling and functional verification of flavonoid 3'-hydroxylase gene from leaves of Euryale ferox.
10.19540/j.cnki.cjcmm.20210614.101
- Author:
Zong-Hui JING
1
;
Meng-Jiao YIN
1
;
Qian WANG
1
;
Ke BAO
1
;
Pei-Na ZHOU
1
;
Chan-Chan LIU
1
;
Qi-Nan WU
2
Author Information
1. College of Pharmacy, Nanjing University of Chinese Medicine Nanjing 210046, China.
2. College of Pharmacy, Nanjing University of Chinese Medicine Nanjing 210046, China Collaborative Innovation Center of Chinese Medicinal Resources Industrialization Nanjing 210023, China National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine Nanjing 210023, China.
- Publication Type:Journal Article
- Keywords:
Euryale ferox;
eukaryotic expression;
flavonoid 3′-hydroxylase;
qRT-PCR;
quercetin
- MeSH:
Anthocyanins;
Cytochrome P-450 Enzyme System/metabolism*;
Plant Leaves/metabolism*;
Plant Proteins/metabolism*;
Transcriptome
- From:
China Journal of Chinese Materia Medica
2021;46(18):4712-4720
- CountryChina
- Language:Chinese
-
Abstract:
Leaves of Euryale ferox are rich in anthocyanins. Anthocyanin synthesis is one of the important branches of the flavonoid synthesis pathway, in which flavonoid 3'-hydroxylase(F3'H) can participate in the formation of important intermediate products of anthocyanin synthesis. According to the data of E. ferox transcriptome, F3'H cDNA sequence was cloned in the leaves of E. ferox and named as EfF3'H. The correlation between EfF3'H gene expression and synthesis of flavonoids was analyzed by a series of bioinforma-tics tools and qRT-PCR. Moreover, the biological function of EfF3'H was verified by the heterologous expression in yeast. Our results showed that EfF3'H comprised a 1 566 bp open reading frame which encoded a hydrophilic transmembrane protein composed of 521 amino acid residues. It was predicted to be located in the plasma membrane. Combined with predictive analysis of conserved domains, this protein belongs to the cytochrome P450(CYP450) superfamily. The qRT-PCR results revealed that the expression level of EfF3'H was significantly different among different cultivars and was highly correlated with the content of related flavonoids in the leaves. Eukaryotic expression studies showed that EfF3'H protein had the biological activity of converting kaempferol to quercetin. In this study, EfF3'H cDNA was cloned from the leaves of E. ferox for the first time, and the biological function of the protein was verified. It provi-ded a scientific basis for further utilizing the leaves of E. ferox and laid a foundation for the further analysis of the biosynthesis pathway of flavonoids in medicinal plants.
