Cloning and Functional Analysis of PqDFR and PqANS Genes and Its Promoters from Paeonia qiui
To explore the causes of the spring leaves of Paeonia qiui with purple-red color,PqDFR and PqANS genes were selected as key genes for anthocyanin synthesis based on the leaf transcriptional data.The two genes were cloned,analyzed for expression patterns,functional characterization and analyzed for promoter activity.The open reading frame of PqDFR was 1 095 bp,encoding 364 amino acids,containing the NADPH conserved domain and a substract-binding binding site.The open reading frame of PqANS was 1 065 bp,encoding 354 amino acids,including 2OG-FeⅡ_Oxy and conserved amino acids binding ferrous ion.The results of real-time quantitative PCR showed that the expression of PqDFR and PqANS genes showed a trend of increasing first,then decreasing during leaf development,which was consistent with the accumulation of anthocyanin.The anthocyanin content of leaf disks was decreased after silencing of PqDFR and PqANS genes in Paeonia qiui.Stable overexpression assays showed that transgenic Arabidopsis seedlings all showed significant anthocyanin accumulation compared to wild-type.Both PqDFR and PqANS gene promoters contain MYB binding sequence,light response elements(G-box),abscisic acid response element(ABRE)and growth factor response element(TGA-element).GUS staining and quantitative results showed that both PqDFR and PqANS gene promoters had good activity.This research suggests PqDFR and PqANS can promote anthocyanin synthesis in Paeonia qiui leaves.
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