原创力文档selection of reference genes for quantitative gene expression normalization in flax (linum usitatissimum l.)选择的参考基因定量基因表达规范化亚麻(亚麻属植物usitatissimum l .).pdfVIP
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selection of reference genes for quantitative gene expression normalization in flax (linum usitatissimum l.)选择的参考基因定量基因表达规范化亚麻(亚麻属植物usitatissimum l .)
Huis et al. BMC Plant Biology 2010, 10:71
/1471-2229/10/71
R E S E A R C H A R T I C L E Open Access
Research article
Selection of reference genes for quantitative gene
expression normalization in flax (Linum
usitatissimum L.)
Rudy Huis, Simon Hawkins and Godfrey Neutelings*
Abstract
Background: Quantitative real-time PCR (qRT-PCR) is currently the most accurate method for detecting differential
gene expression. Such an approach depends on the identification of uniformly expressed housekeeping genes
(HKGs). Extensive transcriptomic data mining and experimental validation in different model plants have shown that
the reliability of these endogenous controls can be influenced by the plant species, growth conditions and organs/
tissues examined. It is therefore important to identify the best reference genes to use in each biological system before
using qRT-PCR to investigate differential gene expression. In this paper we evaluate different candidate HKGs for
developmental transcriptomic studies in the economically-important flax fiber- and oil-crop (Linum usitatissimum L).
Results: Specific primers were designed in order to quantify the expression levels of 20 different potential
housekeeping genes in flax roots, internal- and external-stem tissues, leaves and flowers at different developmental
stages. After calculations of PCR efficiencies, 13 HKGs were retained and their expression stabilities evaluated by the
computer algorithms geNorm and NormFinder. According to geNorm, 2 Transcriptional Elongation Factors (TEFs) and
1 Ubiquitin gene are necessary for normalizing gene expression when all studied samples are considered. However,
only 2 TEFs are required for normalizing expression in stem tissues. In contrast, NormFinder identified glyceraldehyde-
3-phosphate dehydrogenase (GADPH) as the most stably expressed gene when al
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