AUTHOR=Xu Leifeng , Xu Hua , Cao Yuwei , Yang Panpan , Feng Yayan , Tang Yuchao , Yuan Suxia , Ming Jun 

TITLE=Validation of Reference Genes for Quantitative Real-Time PCR during Bicolor Tepal Development in Asiatic Hybrid Lilies (Lilium spp.)

JOURNAL=Frontiers in Plant Science

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2017.00669

DOI=10.3389/fpls.2017.00669

ISSN=1664-462X

ABSTRACT=<p>Quantitative real-time PCR (qRT-PCR) is a reliable and high-throughput technique for gene expression studies, but its accuracy depends on the expression stability of reference genes. To date, several reliable reference gene identifications have been reported in <italic>Lilium</italic> spp., but none has been obtained for lily tepals at different developmental stages. In this study, ten candidate reference genes were selected and evaluated for their expression stability in <italic>Lilium</italic> ‘Tiny Padhye’ during the process of bicolor tepal development. The expression stability of these candidates was evaluated by three software programs (geNorm, NormFinder, and BestKeeper) and the comparative ΔCt method, and comprehensive stability rankings were generated by RefFinder. As a result, <italic>TIP41-like family gene</italic> (<italic>TIP41</italic>) and <italic>actin</italic> (<italic>ACT</italic>) were the best combination of reference genes for tepals at different developmental stages; <italic>TIP41</italic> and <italic>F-box family gene</italic> (<italic>F-box</italic>) for tepals under shading treatment; <italic>ACT, actin11 (ACT11)</italic>, and <italic>elongation factor 1-α (EF1-α)</italic> for different tissues; and <italic>ACT, TIP41</italic>, and <italic>ACT11</italic> for all samples. The selected optimal reference genes were further verified by analyzing the expression levels of <italic>flavonoid 3′-hydroxylase</italic> (<italic>LhF3′H</italic>) and <italic>anthocyanidin 3-O-glucosyltransfersae (LhUFGT</italic>) in tepals at different developmental stages. This study provides useful information for gene expression characterization in lilies under different experimental conditions, and can serve as a basis for similar research in other closely related species.</p>