Cloning,expression and activity analysises of chalcone synthase genes in Carthamus tinctorius  

作　　者：Xiaohui Tang Chaoxiang Ren Jing Hu Jiang Chen Jie Wang Rui Wang Qinghua Wu Wan Liao Jin Pei 

机构地区：[1]State Key Laboratory of Southwestern Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China [2]College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China [3]Sichuan Academy of Grassland Sciences,Chengdu 611731,China

出　　处：《Chinese Herbal Medicines》2023年第2期291-297,共7页中草药（英文版）

基　　金：supported by grants from the National Natural Science Foundation of China(No.82274039,81803669);China Postdoctoral Science Foundation(No.2018M643790XB)。

摘　　要：Objective:Flavonoids are the bioactive compounds in safflower(Carthamus tinctorius),in which chalcone synthase(CHS)is the first limiting enzyme.However,it is unclear that which chalcone synthase genes(CHSs)are participated in flavonoids biosynthesis in C.tinctorius.In this study,the CHSs in the molecular characterization and enzyme activities were investigated.Methods:Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C.tinctorius.Chalcone biosynthase genes in C.tinctorius(CtCHSs)were cloned from cDNA of flowers of C.tinctorius.The cloned gene sequences were analyzed by bioinformatics,and their expression patterns were analyzed by real-time PCR(RT-PCR).The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot.A recombinant vector of CtCHS was constructed.The CtCHS recombinant protein was induced and purified to detect the enzyme reaction(catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone).The reaction product was detected by HPLC and LC-MS.Results:Two full-length CtCHS genes were successfully cloned from the flowers of safflower(CtCHS1 and CtCHS3),with gene lengths of 1525 bp and 1358 bp,respectively.RT-PCR analysis showed that both genes were highly expressed in the flowers,but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers.WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers.HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone.Conclusion:CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.

关 键 词：Carthamus tinctorius L. chalcone synthase expression analysis FLAVONOIDS functional identification gene cloning SAFFLOWER 

分 类 号：R284[医药卫生—中药学]