2016
DOI: 10.1371/journal.pone.0157370
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Validation of Reference Genes for Gene Expression by Quantitative Real-Time RT-PCR in Stem Segments Spanning Primary to Secondary Growth in Populus tomentosa

Abstract: The vertical segments of Populus stems are an ideal experimental system for analyzing the gene expression patterns involved in primary and secondary growth during wood formation. Suitable internal control genes are indispensable to quantitative real time PCR (qRT-PCR) assays of gene expression. In this study, the expression stability of eight candidate reference genes was evaluated in a series of vertical stem segments of Populus tomentosa. Analysis through software packages geNorm, NormFinder and BestKeeper s… Show more

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Cited by 22 publications
(22 citation statements)
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“…All gene‐specific primers were designed using Primer 3 Plus (Untergasser et al ., ), apart from those for amplifying EF1‐beta and eIF5A , which were adopted from Wang et al . (). The efficiency of each primer pair was evaluated with standard curve before performing qRT‐PCR (Table S1).…”
Section: Methodsmentioning
confidence: 97%
“…All gene‐specific primers were designed using Primer 3 Plus (Untergasser et al ., ), apart from those for amplifying EF1‐beta and eIF5A , which were adopted from Wang et al . (). The efficiency of each primer pair was evaluated with standard curve before performing qRT‐PCR (Table S1).…”
Section: Methodsmentioning
confidence: 97%
“…Reference genes were screened in P. euphratica leaves under six abiotic stress, the genes selected as optimal for gene expression analysis were RPL17 in ABA, EF1 a in cold, HIS in dehydration, GII in drought and short-duration salt, and TUB in long-duration salt (Wang et al, 2014). Wang et al (2016) demonstrated that the combination of three most appropriate reference genes, eukaryotic translation initiation factor 5A (eIF5A), Actin (ACT6) and elongation factor 1-beta (EF1-beta) is suitable for gene expression normalization in P. tomentosa stems for both primary and secondary growth. Gutierrez et al (2008) reported that the expression stability of several genes commonly used were unstable in various tissues of P. tremula × P. tremuloides (Gutierrez et al, 2008).…”
Section: Comprehensive Analysismentioning
confidence: 99%
“…These algorithms are widely employed to evaluate the expression stability of reference genes and identify reference genes that are consistent across various species such as Populus tomentosa Carr. [16], Bixa orellana L. [3] and Betula luminifera H.J.P. Winkl.…”
Section: Introductionmentioning
confidence: 99%