Cloning and Expression Analysis of ANS Gene in Three Flower Colors of Scutellaria baicalensis

Yin Xin; Guan Renwei; Zhang Cuicui; Zhao Yiwu; Ran Zhifang; Zhao Qiuchen; Sun Xinru; Wang Shu; Lin Huibin; Guo Fengdan

doi:10.14083/j.issn.1001-4942.2025.03.003

Abstract：Anthocyanidin synthase(ANS)is a key enzyme which catalyzes the conversion of leucoantho-cyanidin to anthocyanidin in the anthocyanin biosynthesis pathway.The full-length cDNA sequence of ANS genes were cloned from Scutellaria baicalensis with bluish-purple,rose-red and white flowers using RT-PCR method,and were named as SbANS-b,SbANS-r and SbANS-w,respectively.Sequence alignment showed that SbANS-b,SbFANS-r and SbANS-w had differences at positions 16,194,242,331,875,945 and 947 bp,and there were three amino acid different in SbANS-B,SbANS-R and SbANS-W proteins.Bioinformatics anal-ysis revealed that all the three SbANS proteins belonged to leucoanthocyanidin dioxygenase like protein super-family and were located in cytoplasm.The SbANS protein had a typical 2OG-Fe Ⅱ_Oxy conserved domain and other sequences involved in binding DHQ-2,MES/ascorbate,iron ion,2OG and DHQ-1.Phylogenetic evolu-tion based on ANS protein sequence indicated that S.baicalensis was closely related to Salvia hispanica,which were clustered in one branch,followed by Glycine max and Arabidopsis thaliana.Expression analysis showed that the expression level of SbANS gene was the highest in bluish-purple flowers.With the flower development,the expression level of SbANS gene in bluish-purple flowers showed the trend of decreasing firstly and then in-creased,while that showed the trend of increasing gradually in rose-red flowers and increasing firstly and then decreasing in white flowers.There were extremely significant differences among the different stages of flower development in bluish-purple and rose-red flowers.These results would provide important guidance for further research on the functional differences of SbANS gene in S.baicalensis with different flower colors and revealing the molecular mechanism of color variation.