Screening of reference genes for quantitative real-time PCR in Artemisia argyi.
10.19540/j.cnki.cjcmm.20210919.101
- Author:
Xiao-Zhe YI
1
;
Lan WU
2
;
Li XIANG
2
;
Meng-Yue WANG
2
;
Shi-Lin CHEN
3
;
Yu-Hua SHI
2
;
Xia LIU
1
Author Information
1. School of Chemistry, Chemical Engineering and Life Sciences, Wuhan University of Technology Wuhan 430070, China.
2. Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Meteria Medica, China Academy of Chinese Medical Sciences Beijing 100700, China Artemisinin Research Center, Institute of Chinese Meteria Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
3. School of Chemistry, Chemical Engineering and Life Sciences, Wuhan University of Technology Wuhan 430070, China Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Meteria Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Artemisia Argyi Folium;
Artemisia argyi;
quantitative real-time PCR;
reference gene
- MeSH:
Artemisia/genetics*;
Gene Expression Profiling;
Genes, Plant/genetics*;
Plant Leaves/genetics*;
Real-Time Polymerase Chain Reaction;
Reference Standards;
Transcriptome
- From:
China Journal of Chinese Materia Medica
2022;47(3):659-667
- CountryChina
- Language:Chinese
-
Abstract:
Artemisia Argyi Folium, a traditional Chinese medicine of important medicinal and economic value, sees increasing demand in medicinal and moxibustion product market. Screening stable and reliable reference genes for quantitative real-time PCR(qRT-PCR) is a prerequisite for the analysis of gene expression in Artemisia argyi. In this study, eight commonly used reference genes, Actin, 18s, EF-1α, GAPDH, SAND, PAL, TUA, and TUB, from the transcriptome of A. argyi, were selected as candidate genes. The expression of each gene in different tissues(roots, stems, and leaves) of A. argyi and in leaves of A. argyi after treatment with methyl jasmonate(MeJA) for different time(0, 4, 8, 12 h) was detected by qRT-PCR. Then, geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder were employed to evaluate their expression stability. The results demonstrated that Actin was the most stable reference gene in different tissues and in leaves treated with MeJA, and coming in the second was SAND. Furthermore, the expression of DXS and MCT which are involved in terpenoid backbone biosynthesis was detected in different tissues and after MeJA treatment. The results showed that the expression patterns of DXS and MCT in different tissues and under MeJA treatment calculated with Actin and SAND as internal reference genes were consistent, which validated the screening results. In conclusion, Actin is the most suitable reference gene for the analysis of gene expression in different tissues of A. argyi and after MeJA treatment. This study provides valuable information for gene expression analysis in A. argyi and lays a foundation for further research on molecular mechanism of quality formation of Artemisia Argyi Folium.
