Reference gene selection for quantitative real-time polymerase chain reaction in Populus.
Accurate quantification of gene expression with quantitative real-time polymerase chain reaction (qRT-PCR) relies on the choice of an appropriate reference gene. In this study, nine candidate reference genes were selected to study the expression stability for qRT-PCR normalization in adventitious rooting of Populus hardwood cuttings. geNorm, NormFinder, and BestKeeper analysis revealed that actin isoform B (ACT) was the most unstable gene across developmental stages, whereas elongation factor 1 alpha (EF1a) and 18S recombinant RNA (18S) emerged as the most appropriate reference genes for qRT-PCR analysis in this complex developmental process.
Xu
Meng
M
Jiangsu Key Laboratory for Poplar Germplasm Enhancement and Variety Improvement, Nanjing Forestry University, Nanjing 210037, China.
Zhang
Bo
B
Su
Xiaohua
X
Zhang
Shougong
S
Huang
Minren
M
D000199:Actins / Q000235:genetics / Q000592:standards; D001483:Base Sequence; D017343:Genes, Plant; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000592:standards; D032107:Populus / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics / Q000592:standards; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards
D000199:Actins; D020648:Peptide Elongation Factor 1; D012337:RNA, Ribosomal, 18S