ABSTRACT
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.
ABSTRACT
OBJECTIVE：To investigate the correlation of storage life and effective composition content with color value of Carthamus tinctorius ，and to provide reference for the quality evaluation of C. tinctorius with different years of storage. METHODS：Using 24 batches of C. tinctorius from same place of production with different years of storage （0，1，2 years，8 batches each type ）as samples ，the contents of hydroxysafflor yellow A （HSYA）and kaempferol were determined by HPLC. Color value [lightness value （L*），red-green value （a*），yellow-blue value （b*）] were determined by spectrophotometer. SPSS 19.0 statistical software was used to analyze the correlation of storage life and effective composition content with color value. RESULTS：Kaempferol content was still high after 1 year or 2 years of storage （0.161％，0.061％，respectively）. However ，the content of HSYA decreased with the prolongation of the storage life （the average content of HSYA were 2.46％，1.58％，and 1.51％ after storage 0，1 and 2 years，respectively），and the color of the drug became darker （a* value decreased ）. Results of correlation analysis showed that the content of HSYA was positively associated with color value L*，a*（r＝0.430，0.781，P＜0.05 or P＜0.01）；the content of HSAY was negatively associated with storage life （r＝－0.777，P＜0.01）. There was no correlation between the remaining variables （P＞0.05）. CONCLUSIONS ：The longer the storage life ，the darker the color and the lower the content of HSYA ，so it is not suitable for over year and multiyear preservation.