Functional study of glycosyltransferase genes CtUGT25 in the flavone biosynthesis pathway of Carthamus tinctorius L.
UDP glycosyltransferase(UGT)is a terminal modifying enzyme for the formation of flavonoid glycosides.In this study,we obtained two glycosyltransferase genes,CtUGT25 and CtUGT18,which are closely related to the synthesis of safflower flavonoids,through the Pierce correlation analysis of the expression of glycosyltransferase genes in the safflower corolla transcriptome database at different developmental stages with the contents of the major constituents of safflower metabolome database,and bioinformatically analyzed the gene and protein sequences of the two genes.Expression pattern analysis revealed that CtUGT25 was mainly expressed in the corolla,with the highest expression on day 3 of flowering stage;CtUGT18 was mainly expressed in the root,with the highest expression on day 1 of flowering stage.Functional validation was verified in safflower by Agrobacterium-mediated pollen-tube pathway transgenesis method,demonstrating that CtUGT25 promoted the accumulation of kaempferol-3-O-glucoside and hydroxysafflor yellow A(HSYA),and CtUGT18 promoted the accumulation of kaempferol-3-O-glucoside and orientin,both of which may be the glycosyl-modifying enzymes for the synthesis of safflower flavonoids.Meanwhile,in vitro experiments demonstrated the catalytic activity of CtUGT25 protein on naringenin,quercetin,apigenin,kaempferol,luteoin,2-hydroxynaringenin and galangin.This study serves as reference for future advancements in regulating the quality of safflower using molecular biotechnology,particularly focuses on the industrial production of safflower exclusive component HSYA.Additionally,it offers valuable insights for researching related genes in other plants.
safflowerflavonoidglycosyltransferasebiosynthetic pathwayhydroxysafflor yellow A
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