Reference gene selection for miRNA and mRNA normalization in potato in response to potato virus Y. (February 2021)

• Record Type: Journal Article
• Authors: Yin, Zhimin; Xie, Fuliang; Michalak, Krystyna; Zhang, Baohong; Zimnoch-Guzowska, Ewa
• Is Part Of: Molecular and cellular probes. Volume 55(2021)
• Publication Date: 2021-02
• Subjects: Hypersensitive resistance -- microRNA -- Protein coding reference gene -- Solanum tuberosum -- Stem-loop RT-qPCR -- PVY; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2020.101691
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: This was the first report on evaluating candidate reference genes for quantifying the expression profiles of both coding (e.g., mRNA) and non-coding (e.g., miRNA) genes in potato response to potato virus Y (PVY) inoculation. The reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) method was employed to quantify the expression profiles of eight selected candidate reference genes; their expression stability was analyzed by four statistical algorithms, i.e., geNorm, BestKeeper, NormFinder and RefFinder. The most stable reference genes were sEF1a, sTUBb and seIF5 with a high stability. The least stable ones were sPP2A, sSUI1 and sGAPDH . The same reference gene allows for normalization of both miRNA and mRNA levels from a single RNA sample using cDNAs synthesized in a single RT reaction, in which a stem-loop primer was used for miRNAs and the oligo (dT) for mRNAs. Highlights: The same protein-coding reference gene for miRNA and mRNA normalization. cDNA synthesis in a single RT reaction using a single RNA sample. Stem-loop primer for miRNA and oligo (dT) for mRNA target and the reference gene.