NDLI: Reference Gene Selection for Gene Expression Studies Using Quantitative Real-Time PCR Normalization in Atropa belladonna

Content Provider	Springer Nature Link
Author	Li, Jindi Chen, Min Qiu, Fei Qin, Baifu Liu, Wanhong Wu, Nengbiao Lan, Xiaozhong Wang, Qiang Liao, Zhihua Tang, Kexuan
Copyright Year	2014
Abstract	Quantitative PCR (qPCR) is a powerful tool for measuring gene expression levels. Accurate and reproducible results are dependent on the correct choice of reference genes for data normalization. Atropa belladonna is a commercial plant species from which pharmaceutical tropane alkaloids are extracted. In this study, eight candidate reference genes, namely 18S ribosomal RNA (18S), actin (ACT), cyclophilin (CYC), elongation factor 1α (EF-1α), β-fructosidase (FRU), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), and beta-tubulin (TUB), were selected and their expression stabilities studied to determine their suitability for normalizing gene expression in A. belladonna. The expression stabilities of these genes were analyzed in the root, stem, and leaf under cold, heat, NaCl, UV-B, methyl jasmonate, salicylic acid, and abscisic acid treatments using geNorm, NormFinder, and BestKeeper. The statistical algorithms indicated that PGK was a reliable gene for normalizing gene expression under most of the experimental conditions. The pairwise value analysis showed that two genes were sufficient for proper expression normalization, except when analyzing gene expression in heat-treated roots. However, the choice of the second reference gene depended on specific conditions. Finally, the relative expression level of the PMT gene of A. belladonna was detected to validate the selection of PGK a reliable reference gene. In summary, our results should guide the selection of appropriate reference genes for gene expression studies in A. belladonna under different organs and abiotic stress conditions.
Starting Page	1002
Ending Page	1014
Page Count	13
File Format	PDF
ISSN	07359640
Journal	Plant Molecular Biology Reporter
Volume Number	32
Issue Number	5
e-ISSN	15729818
Language	English
Publisher	Springer US
Publisher Date	2014-02-27
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Atropa belladonna Gene expression Normalization qPCR Reference gene Plant Sciences Plant Breeding/Biotechnology Proteomics Metabolomics Bioinformatics
Content Type	Text
Resource Type	Article
Subject	Plant Science Molecular Biology

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