Promoter Functional Analysis of the Key Gene PsDFR Involved in Paeonia suffruticosa Anthocyanin Biosynthesis
[Objective]To analyze the cis-acting element and activity of the tree peony anthocyanin biosyn-thetic key gene PsDFR promoter for further research on its function and regulation mechanism involved in tree peony flower coloration.[Method]The genomic DNA extracted from the petals of tree peony'Hei Hua Kui'was used as a template.PsDFR promoter was isolated by genomic walking method.The cis-acting elements of promoter were analyzed and predicted through Bioinformatics online software.Five different length of deletion fragments were fused with GUS gene to construct promoter-reporter vectors,and then were transiently expressed in tobacco leaves.The activities of deletion promoters and their response to dif-ferent stress treatments such as Abscisic acid(ABA),Methyl jasmonate(MeJA)and light were analyzed by GUS staining and GUS activity assay.[Result]A 1 687 bp sequence of PsDFR promoter was isolated.The results of bioinformatics analysis showed that the promoter contains multiple cis-acting elements asso-ciated with light signals,hormone response,stress response,and tissue-specific expression,indicating that the expression of PsDFR may be regulated by various signals such as light signaling,plant hormone and stress.GUS staining and quantitative analysis of GUS activity showed that GUS activities decreased gradually with decrease of the length of PsDFR promoters,and the region of-1 623 bp to-916 bp played an important role on the activity of the PsDFR promoter.The GUS activities were inhibited significantly by MeJA or dark treatment,and were induced obviously after light restoration.And core regulation region in-volved in ABA-response might be located between-443 and-76 bp.[Conclusion]PsDFR promoter con-tains multiple cis-acting elements associated with light signals,hormone response,stress response,and tissue-specific expression.Its activity is positively regulated by light and negatively regulated by MeJA.The region of-1 623 bp to-916 bp is important for the activity of the PsDFR promoter,and-443 and-76 bp is the core region in response to ABA treatment.This study provides a reference for further revealing the reg-ulatory mechanism of PsDFR response to environmental signals involved in tree peony flower coloration.
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