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Appropriate Reference Genes for RT-qPCR Normalization in Various Organs of Anemone flaccida Fr. Schmidt at Different Growing Stages
- Source :
- Genes, Vol 12, Iss 459, p 459 (2021), Genes, Volume 12, Issue 3
- Publication Year :
- 2021
- Publisher :
- MDPI AG, 2021.
-
Abstract
- Anemone flaccida Fr. Schmidt is a traditional medicinal herb in southwestern China and has multiple pharmacological effects on bruise injuries and rheumatoid arthritis (RA). A new drug with a good curative effect on RA has recently been developed from the extract of A. flaccida rhizomes, of which the main medicinal ingredients are triterpenoid saponins. Due to excessive exploitation, the wild population has been scarce and endangered in a few of its natural habitats and research on the cultivation of the plant commenced. Studies on the gene expressions related to the biosynthesis of triterpenoid saponins are not only helpful for understanding the effects of environmental factors on the medicinal ingredient accumulations but also necessary for monitoring the herb quality of the cultivated plants. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) as a sensitive and powerful technique has been widely used to detect gene expression across tissues in plants at different stages<br />however, its accuracy and reliability depend largely on the reference gene selection. In this study, the expressions of 10 candidate reference genes were evaluated in various organs of the wild and cultivated plants at different stages, using the algorithms of geNorm, NormFinder and BestKeeper, respectively. The purpose of this study was to identify the suitable reference genes for RT-qPCR detection in A. flaccida. The results showed that two reference genes were sufficient for RT-qPCR data normalization in A. flaccida. PUBQ and ETIF1a can be used as suitable reference genes in most organs at various stages because of their expression stabilitywhereas the PUBQ and EF1Α genes were desirable in the rhizomes of the plant at the vegetative stage.
- Subjects :
- Medicinal herb
Genetics
Normalization (statistics)
education.field_of_study
food.ingredient
lcsh:QH426-470
Anemone flaccida Fr. Schmidt
Data normalization
RT-qPCR
Population
Reference gene
Biology
Rhizome
lcsh:Genetics
food
Real-time polymerase chain reaction
Herb
Reference genes
Gene expression
education
Gene
Genetics (clinical)
Subjects
Details
- ISSN :
- 20734425
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Genes
- Accession number :
- edsair.doi.dedup.....e10219c2890ddff875deb7df44f621fd