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Selection of reliable reference genes for gene expression analysis in seeds at different developmental stages and across various tissues in Paeonia ostii.

Authors
  • Li, Chaoqiong1, 2
  • Hu, Lizong3
  • Wang, Xueqin3
  • Liu, Hongzhan3
  • Tian, Huihui4
  • Wang, Junsheng3
  • 1 College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, Henan, China. [email protected] , (China)
  • 2 Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, Henan, China. [email protected] , (China)
  • 3 College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, Henan, China. , (China)
  • 4 College of Journalism and Media, Zhoukou Normal University, Zhoukou, 466001, Henan, China. , (China)
Type
Published Article
Journal
Molecular Biology Reports
Publisher
Springer-Verlag
Publication Date
Dec 01, 2019
Volume
46
Issue
6
Pages
6003–6011
Identifiers
DOI: 10.1007/s11033-019-05036-7
PMID: 31446531
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Paeonia ostii seeds have recently been identified as a new source of α-linolenic acid in China. Studying the gene expression patterns of unsaturated fatty acid-related genes would be helpful for understanding the mechanism of α-linolenic acid accumulation. Quantitative real-time polymerase chain reaction (qRT-PCR) is a useful method for reliably evaluating gene expression, and it is necessary to select reliable reference genes for data normalization in qRT-PCR analysis. In this study, we evaluated the expression stability of 12 candidate reference genes using four mathematical algorithms (∆Ct, BestKeeper, NormFinder, and geNorm). The web-based tool RefFinder was used to integrate the results and to provide a comprehensive ranking order. The expression stability ranking orders of reference genes were different caculated by these four algorithms, and the ranking order analyzed by the RefFinder was UBQ > Tip41 > UCE > EF-1α > α-TUB > PP2A > ACT > GAPDH > SAM > CYP > β-TUB > 18S at the different seed development stages, and UBQ > Tip41 > EF-1α > α-TUB > PP2A > UCE > GAPDH > SAM > ACT > CYP > 18S > β-TUB in P. ostii tissues. UBQ and Tip41 are the two most stable whereas 18S and β-TUB are the two least stable reference genes for gene expression in various tissues and seeds at different developmental stages in P. ostii.

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