Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum.
10.19540/j.cnki.cjcmm.20201024.103
- Author:
Hao WANG
1
;
Qi-Zhong CAI
1
;
Lu LIU
1
;
Quan YANG
1
;
Liang-Yun ZHOU
1
Author Information
1. Guangdong Engineering Research Center of Good Agricultural Practice, Comprehensive Development for Cantonese Medicinal Materials,Guangzhou Comprehensive Experimental Station of National Industrial Technology System for Chinese Materia Medica,Key Laboratory of State Administration of Traditional Chinese Medicine for Production, Development of Cantonese Medicinal Materials, School of Traditional Chinese Medicine, Guangdong Pharmaceutical University Guangzhou 510006, China.
- Publication Type:Journal Article
- Keywords:
Polygonum multiflorum;
Real-time quantitative PCR;
reference gene
- MeSH:
Fallopia multiflora;
Gene Expression Profiling;
Gene Expression Regulation, Plant;
Genes, Plant/genetics*;
Real-Time Polymerase Chain Reaction;
Reference Standards;
Reproducibility of Results
- From:
China Journal of Chinese Materia Medica
2021;46(1):80-85
- CountryChina
- Language:Chinese
-
Abstract:
To select suitable references gene of Polygonum multiflorum for gene expression analysis in different tissues, five candidate reference genes like Actin,GAPDH,SAND,PP2A,TIP41 were selected from the transcriptome data of P. multiflorum, then the specific primers were designed. The expression stability of the five reference genes in different tissues of P. multiflorum was analyzed by Real-time quantitative PCR through avilable analysis methods such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, to ensure the reliability of the analysis results. The results showed that there were significant differences in the expression levels and stability of candidate genes in different tissues of P. multiflorum. Ct distribution analysis of the expression levels of candidate genes showed that the expression levels of Actin and GAPDH genes were relatively high in different tissues, while the expression levels of SAND, PP2A and TIP41 were lower. The stability of each candidate gene was analyzed by different methods. The results of geNorm analysis showed that the expression of PP2A and GAPDH was the most stable, the expression stability of SAND was the worst, the stability of PP2A was the highest in both NormFinder and Delta CT, the stability of SAND was the lowest, and the stability of Actin was the most stable in BestKeeper analysis. Through the comprehensive evaluation and analysis of the stability of candidate genes by RefFinder, it is concluded that the stability of PP2A gene is the highest, followed by GAPDH, Actin, TIP41, SAND, and SAND gene is the worst. Therefore, the PP2A gene is an ideal reference gene for the analysis of gene expression in different tissues of P. multiflorum.
