| The plant pigment anthocyanin is not only a sign of fruit ripening,but also an important secondary metabolite in the process of fruit maturation.It has extremely important significance in physiological function activity.The Lonicera edulis is rich in anthocyanins compared to other plants,and is therefore highly valued by scholars from various countries.In recent years,scholars from all over the world have focused on the development and utilization of biological functional activities of the Lonicera edulis anthocyanins.In terms of molecular metabolism,the space-time synthesis mechanism has been studied in depth,but the molecular regulation of Lonicera edulis research on several key areas such as mechanisms is not yet in depth,and related genetic information is rarely reported.To study the mechanism of anthocyanin biosynthesis and accumulation at different growth stages of Lonicera edulis,and to understand the mechanism of anthocyanin synthesis at the molecular level,it is of great significance to screen and cultivate Lonicera japonica varieties with high content of anthocyanins.In this study,we used the solvent extraction method combined with p H for differe nt varieties(wild type “Shangganling”,Canada imported varieties “Blue Sky”,“Blue Forest”,“Blue Pacific”,and “Blue Velvet”).Differential scanning method for total cyanine content,Filin reagent method for total soluble sugar content,ELISA for detection of key enzymes in the anthocyanin synthesis pathway,and LC-ESI-MS/MS analysis for anthocyanin intermediate metabolism The content of the product and the relative expression of the structural gene were detected by fluorescence quantitative PCR.The following results were obtained.(1)The total anthocyanin contents of the five Lonicera edulis varieties at maturit y were: 75.24 mg/100 g,263.00 mg/100 g,253.73 mg/100 g,127.28 mg/100 g and 242.22 mg/100 g;(2)The total soluble sugar content of five Lonicera edulis varieties at maturity were 2.91%,3.11%,1.98%,3.55%,2.31%,indicating that Lonicera edulis was a low-glucose fruit and the accumulation of soluble sugars promoted the synthesis of anthocyanins;(3)The LC-ESI-MS/MS method focuses on the content of nine metabolites in the anthocyanin biosynthetic pathway during Canada’s introduction of the “Blue Sky” No.6 and Canada’s No.9 “Blue Forest” fruits.The results showed that L-phenylalanine,cinnamic acid,4-coumaric acid,dihydroquercetin,cyanidin and cyanidin 3-O-glucoside were the main intermediates in the ripening of Lonicera edulis.And the conversion of cyanidin and sugar to the blue cyanidin 3-O-glucoside was started from the third half of the color change period,and the process is very rapid;(4)The relative expression levels of most of the nine structural genes in the nine structural genes of the anthocyanin synthesis pathway in Lonicera edulis showed a normal distribution that increased first and then decreased.The decrease indicates that most of the structural genes are highly expressed at the beginning of the fruit coloring of Lonicera edulis fruits;(5)The three enzymes PAL,F3 H,and DFR played a key role in the biosynthesis of Lonicera edulis.PAL showed high activity in all stages of fruit ripening and showed a similar trend with PAL gene expression,but it was not an absolute positive correlation.F3 H and DFR may be the key structural genes that cause the anthocyanin biosynthesis to be blocked,and the role of F3 H and DFR was mainly in the later stage of fruit ripening. |
