Comparison of five endogenous reference genes for specific PCR detection and quantification of Brassica napus.
Five previously reported Brassica napus endogenous reference genes, including acetyl-CoA carboxylase gene (BnACCg8), phosphoenolpyruvate carboxylase (PEP), oleoyl hydrolase gene (FatA), high-mobility-group protein I/Y gene (HMG-I/Y) and cruciferin A gene (CruA), were analyzed for their PCR specificity between B. napus and other species and the quantification stability among different B. napus cultivars. PCR and sequencing results indicated that none of these systems was species-specific as required by the genetically modified organism labeling policy. When these genes were employed in real-time PCR, BnACCg8 and HMG-I/Y systems showed relatively greater heterogeneity among 10 different cultivars. The sequencing results showed that the single nucleotide polymorphism in the primer binding sites was the potential source of the instability in the HMG-I/Y system. The bias of BnACCg8 was thought to be associated with the inconsistent copy number of this gene.
Wu
Gang
G
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, No. 2 Xudong Second Road, Wuhan, 430062, China.
Zhang
Li
L
Wu
Yuhua
Y
Cao
Yinglong
Y
Lu
Changming
C
D001483:Base Sequence; D001937:Brassica / Q000235:genetics*; D017931:DNA Primers; D017343:Genes, Plant; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D017931:DNA Primers