Defining reference genes for quantitative real-time PCR analysis of anther development in rice.
Quantitative real-time polymerase chain reaction (qPCR) is one of the most accurate and widely used methods for gene expression analysis. However, the choice of reference genes for normalization is critical for accurate quantification of gene expression. As development of genomics, mining large-scale datasets such as microarray and RNA-sequencing data becomes a new approach for exploitation of new reference genes. In this study, we analyzed an RNA-sequencing dataset of rice anther and 167 microarray datasets involving different tissues and developing stages of rice anthers and pollens. We selected 12 candidate genes and other 5 reference genes, including ACT1, eEF-1α, GAPDH, Exp2, and CCDC72 used in previous studies, and evaluated their expression in eight tissues and different developmental stages of anthers in rice variety 9311 and Yuetai. UPF3, eIF4A-3, GAPDH, and PPP6 were identified as the most suitable reference genes for qPCR analysis of anther development in rice. The new candidate reference genes showed more stable expression than the traditionally used reference genes. These results provide a set of reliable reference genes for studies in rice anther developmental process.
State Key Laboratory of Hybrid Rice, Key Laboratory for Research and Utilization of Heterosis in Indica Rice of Ministry of Agriculture, Engineering Research Center for Plant Biotechnology and Germplasm Utilization of Ministry of Education, College of Life Science, Wuhan University, Wuhan 430072, China.
Yanxiao
Y
Ji
Ping
P
Tu
Kun
K
Wang
Feng
F
Gao
Weilong
W
Yang
Yingguo
Y
Zhu
Shaoqing
S
Li
D001483:Base Sequence; D017931:DNA Primers; D035264:Flowers / Q000254:growth & development*; D017343:Genes, Plant; D012275:Oryza / Q000235:genetics* / Q000254:growth & development; D060888:Real-Time Polymerase Chain Reaction
D017931:DNA Primers
anther; qPCR; reference gene; rice