Selection of optimal q RT-PCR reference genes for gene expression studies in foxtail millet

Zhiyong Li; Yanbin Zhu; Nan Wang; Hui Bai; Li Dong; Jianzhang Quan; Zhiping Dong

Zhiyong Li Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Yanbin Zhu Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Nan Wang Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Hui Bai Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Li Dong Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Jianzhang Quan Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China
Zhiping Dong Millet Institute, Hebei Academy of Agricultural and Forestry Sciences; National Foxtail Millet Improvement Center; Minor Cereal Crops Laboratory of Hebei Province, 126 Road hengshan, 050035 Shijiazhuang, China

Keywords: Foxtail millet, reference genes, geNorm, normFinder, bestKeeper

Abstract

Appropriate reference gene is very important for gene expression analysis by the quantitative RT-PCR (qRT-PCR), but no systematics research have been carried out in foxtail millet. In this study, the mRNA expression were investigated by qRT-PCR for six candidate reference genes, 25S rRNA, 18S rRNA, 17S rRNA, GAPDH, ACTB and UBQ under three different experimental conditions including different stages of leaf rust, five plant organs and seven different foxtail millet cultivars. The results obtained with GeNorm, NormFinder and BestKeeper software tools showed stable and uniform expression of 25S rRNA among different cultivars, 18S rRNA under the different conditions of leaf rust infection and 17S rRNA in different foxtail millet organs The list of reference genes confirmed in present study will be useful for genes expression analysis in foxtail millet under different conditions.