Overexpressing of chalcone isomerase (CHI) gene enhances flavonoid accumulation in Glycyrrhiza uralensis hairy roots
10.16438/j.0513-4870.2020-0951
- VernacularTitle:过表达CHI基因提高甘草毛状根中黄酮类化合物含量的研究
- Author:
Jia-ming HOU
1
;
Yan-chao YIN
2
;
Shao-kai TIAN
1
;
Zhi-xin ZHANG
1
;
Lin YANG
1
;
Wen-dong LI
3
;
Ying LIU
1
Author Information
1. School of Life Sciences, Beijing University of Chinese Medicine, Beijing 102488, China
2. Department of Pharmacy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China
3. Beijing Institute for Drug Control, Beijing 102206, China
- Publication Type:Research Article
- Keywords:
italic>Glycyrrhiza uralensis;
chalcone isomerase;
overexpression;
hairy root;
UPLC;
qRT-PCR
- From:
Acta Pharmaceutica Sinica
2021;56(1):319-327
- CountryChina
- Language:Chinese
-
Abstract:
Chalcone isomerase (CHI) is the second rate-limiting enzyme involved in the biosynthetic pathway of flavonoids in Glycyrrhiza uralensis. Based on our previous studies, we selected the specific CHI haplotype (GenBank Accession No. KY115232) to maximize flavonoid accumulation. We constructed a plant binary expression vector for overexpression of this CHI gene by the gene fusion method and transfected the plasmid into Agrobacterium tumefaciens ACCC10060 by electroporation. The recombinant A. tumefaciens ACCC10060 subsequently was used to infect cotyledons and hypocotyls of G. uralensis to obtain transgenic hairy roots. A qRT-PCR method was used to determine the copy number of CHI and a UPLC method was used to assay the content of four flavonoids in different hairy root lines. The qRT-PCR results showed that the copy number of CHI in hairy roots was 1 or 5. UPLC results showed that the content of total flavonoids, liquiritin, liquiritigenin, and isoliquiritigenin in transgenic hairy root samples was significantly higher than that in wild-type samples. This study demonstrates that overexpression of CHI significantly increases the content of flavonoids in hairy roots of G. uralensis. This work provides a theoretical basis for clarifying the function of CHI. Three transgenic hairy root lines of G. uralensis were isolated which can be used to increase the accumulation of licorice flavonoids in vitro.
