Cloning and expression analysis of PsAP3 gene in tree peony
Superior flower type breeding is one of the bottleneck problems encountered by the tree peony(Paeonia suffruticosa)industry.In this study,the single flower and stamen petalody varieties were used as plant materials.As the entry point,the key gene PsAP3 regulating peony stamen development,was char-acterized by gene clone,subcellular localization,expression analysis and yeast two-hybrid.PsAP3 silenc-ing vector was constructed and successfully transformed into Agrobacterium tumefaciens for subsequent transient silencing assays in peony.The results showed that the full length of PsAP3 gene is 612 bp(Gen-Bank accession number:MT225537),encoding 203 amino acids,with a molecular weight of 23 891.18.It is a hydrophilic protein and contains a MADS domain and a K domain.Phylogenetic analysis showed that PsAP3 was most closely related to Quercus variabilis.Subcellular localization results showed that PsAP3 protein was localized in the nucleus.Expression analysis showed that PsAP3 expression was significantly higher in the inner petals of stamen petalized varieties compared to the stamens of single petal varieties.These indicated that the expansion of PsAP3 gene may be expanded in the floral tube and significant high expression in stamens might lead to stamen petalody in tree peony.Yeast two-hybrid results showed that PsAP3 protein interacted with PsAP1-1,PsPI,PsSEP and PsAG protein.This study provides a preliminary analysis of the function of PsAP3 in peony,provided a theoretical basis for revealing the molecular mecha-nism of stamen petalody,and laid a theoretical basis for further fine-controling the flower type in tree peony.
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