Cloning and function analysis of chalcone isomerase gene and chalcone synthase gene in Lonicera macranthoides.
10.19540/j.cnki.cjcmm.20220212.101
- Author:
Juan ZENG
1
;
Yu-Qing LONG
1
;
Can LI
1
;
Mei ZENG
1
;
Min YANG
1
;
Xin-Ru ZHOU
1
;
Xiang-Dan LIU
2
;
Ri-Bao ZHOU
2
Author Information
1. School of Pharmacy, Hunan University of Chinese Medicine Changsha 410208, China Key Laboratory of Germplasm Resources and Standardized Planting of Hunan Large-scale Genuine Medicinal Materials Changsha 410208, China.
2. School of Pharmacy, Hunan University of Chinese Medicine Changsha 410208, China Key Laboratory of Germplasm Resources and Standardized Planting of Hunan Large-scale Genuine Medicinal Materials Changsha 410208, China Key Laboratory of Traditional Chinese Medicine Modernization Research in General Colleges and Universities of Hunan Province Changsha 410208, China.
- Publication Type:Journal Article
- Keywords:
Lonicera macranthoides;
chalcone isomerase;
chalcone synthase;
content determination;
expression analysis;
gene cloning
- MeSH:
Acyltransferases/metabolism*;
Chalcone;
Cloning, Molecular;
Intramolecular Lyases;
Lonicera/metabolism*;
Plant Breeding
- From:
China Journal of Chinese Materia Medica
2022;47(9):2419-2429
- CountryChina
- Language:Chinese
-
Abstract:
In order to explore the functions of genes of key rate-limiting enzymes chalcone isomerase(CHI) and chalcone synthase(CHS) in the biosynthesis of flavonoids in Lonicera macranthoides, this study screened and cloned the cDNA sequences of CHI and CHS genes from the transcriptome data of conventional variety and 'Xianglei' of L. macranthoides. Online bioinformatics analysis software was used to analyze the characteristics of the encoded proteins, and quantitative reverse-transcription polymerase chain reaction(qRT-PCR) to detect the expression of CHI and CHS in different parts of the varieties at different flowering stages. The content of luteo-loside was determined by high performance liquid chromatography(HPLC) and the correlation with the expression of the two genes was analyzed. The results showed that the CHI and CHS of the two varieties contained a 627 bp and 1170 bp open reading frame(ORF), respectively, and the CHI protein and CHS protein were stable, hydrophilic, and non-secretory. qRT-PCR results demonstrated that CHI and CHS of the two varieties were differentially expressed in stems and leaves at different flowering stages, particularly the key stages. Based on HPLC data, luteoloside content was in negative correlation with the relative expression of the genes. Thus, CHI and CHS might regulate the accumulation of flavonoids in L. macranthoides, and the specific functions should be further studied. This study cloned CHI and CHS in L. macranthoides and analyzed their expression for the first time, which laid a basis for investigating the molecular mechanism of the differences in flavonoids such as luteoloside in L. macranthoides and variety breeding.
