Selection and validation of reference genes for quantitative real-time PCR analysis in Paeonia veitchii.
10.19540/j.cnki.cjcmm.20230726.101
- Author:
Meng-Ting LUO
1
;
Jun-Zhang QUBIE
1
;
Ming-Kang FENG
1
;
A-Xiang QUBIE
1
;
Bin HE
1
;
Yue-Bu HAILAI
1
;
Wen-Bing LI
2
;
Zheng-Ming YANG
2
;
Ying LI
1
;
Xin-Jia YAN
2
;
Yuan LIU
2
;
Shao-Shan ZHANG
2
Author Information
1. College of Pharmacy,Southwest Minzu University Chengdu 610041,China Sichuan Provincial Qiang-Yi Medicinal Resources Protection and Utilization Technology and Engineering Laboratory Chengdu 610225,China Tibetan Plateau Ethnic Medicinal Resources Protection and Utilization Key Laboratory of National Ethnic Affairs Commission of the People's Republic of China Chengdu 610225,China.
2. Sichuan Provincial Qiang-Yi Medicinal Resources Protection and Utilization Technology and Engineering Laboratory Chengdu 610225,China Tibetan Plateau Ethnic Medicinal Resources Protection and Utilization Key Laboratory of National Ethnic Affairs Commission of the People's Republic of China Chengdu 610225,China.
- Publication Type:Journal Article
- Keywords:
Actin;
GAPDH;
Paeonia veitchii;
real-time fluorescence quantitative PCR;
reference gene
- MeSH:
Real-Time Polymerase Chain Reaction/methods*;
Paeonia/genetics*;
Actins/genetics*;
Reproducibility of Results;
Transcriptome;
Glyceraldehyde-3-Phosphate Dehydrogenases/genetics*;
Reference Standards;
Gene Expression Profiling/methods*
- From:
China Journal of Chinese Materia Medica
2023;48(21):5759-5766
- CountryChina
- Language:Chinese
-
Abstract:
Paeonia veitchii and P. lactiflora are both original plants of the famous Chinese medicinal drug Paeoniae Radix Rubra in the Chinese Pharmacopoeia. They have important medicinal value and great potential in the flower market. The selection of stable and reliable reference genes is a necessary prerequisite for molecular research on P. veitchii. In this study, two reference genes, Actin and GAPDH, were selected as candidate genes from the transcriptome data of P. veitchii. The expression levels of the two candidate genes in different tissues(phloem, xylem, stem, leaf, petiole, and ovary) and different growth stages(bud stage, flowering stage, and dormant stage) of P. veitchii were detected using real-time fluorescence quantitative technology(qRT-PCR). Then, the stability of the expression of the two reference genes was comprehensively analyzed using geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that the expression patterns of Actin and GAPDH were stable in different tissues and growth stages of P. veitchii. Furthermore, the expression levels of eight genes(Pv-TPS01, Pv-TPS02, Pv-CYP01, Pv-CYP02, Pv-CYP03, Pv-BAHD01, Pv-UGT01, and Pv-UGT02) in different tissues were further detected based on the transcriptome data of P. veitchii. The results showed that when Actin and GAPDH were used as reference genes, the expression trends of the eight genes in different tissues of P. veitchii were consistent, validating the reliability of Actin and GAPDH as reference genes for P. veitchii. In conclusion, this study finds that Actin and GAPDH can be used as reference genes for studying gene expression levels in different tissues and growth stages of P. veitchii.
