Identification And Metabonomics Of Flavonoids From Different Tartary Buckwheat Sprout And Seedling

Fagopyrum tataricum(L.)Gaertn,also known as Tatar buckwheat,is an annual herb of tartary buckwheat in Polygonaceae of dicotyledons and Caryophyllaceae.The seeds of fagopyrum tataricum are rich in secondary metabolites.Flavonoids,phenols,terpenes and organic acids are the main components of fagopyrum tataricum.Flavonoids mainly include flavonol,flavanol,chalcone,flavanone,procyanidins and anthocyanins.Pharmacological studies showed that the flavonoids in buckwheat seeds,such as procyanidins,rutin and quercetin,had the pharmacological effects of anti-inflammatory,anti-oxidation,hypoglycemic and anti-tumor.Tartary buckwheat seeds contain high activity of rutin degrading enzyme and trypsin in-hibitor.After germination,it can effectively overcome the degradation of rutin,reduce the activity of trypsin in-hibitor,and significantly improve the content of active nutrients.In recent years,with the tartary buckwheat seedling being listed as a new resource food,there are new sprout vegetables represented by tartary buckwheat sprout and tartary buckwheat seedling in the market.At the same time,tartary buckwheat sprout also has broad application prospects in medicine,cosmetics,health care products,food and other aspects,so more and more attention has been paid to the study of buckwheat bud and seedling.In this study,firstly,the flavonoids in tartary buckwheat seedlings were identified by LC-QTOF-MS,and the flavonoids in different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings were analyzed by LC-QQQ-MSMS,so as to study the yellow in different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings at the metabolic and molecular levels The difference of ketone metabolism and the expression of related genes in flavonoid synthesis pathway lay the foundation for screening high nutrition and high quality tartary buckwheat sprouts.Acquisition of experimental materialsThe results of this study are as follows:1.The establishment of flavonoid spectrum in tartary buckwheat seedlingsUsing LC-QTOF-MS mass spectrometry to analyze the mixed samples of tartary buckwheat seedlings under ESI~+,ESI~-ionization mode,based on the flavonoids in tartary buckwheat,and using LC-QQQ-MS/MS sub ion scanning mode to obtain the fragment ion information of the compounds,46 flavonoids were identified.2.Establishment of detection method for tartary buckwheat seedling samplesBased on UPLC-QQQ-MS/MS,the target metabonomics method of 46flavonoids in tartary buckwheat seedlings samples was established.The parameters of son ion,mother ion,voltage value and collision energy were optimized during the study,which laid a foundation for the target detection of flavonoids in the samples.3.Screening of different compounds40 different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings have differences in seed color and shape,including black(n=20),yellow brown(n=20),long(n=20),short(n=20)according to the shape.Using multivariate statistical analysis to screen the different compounds in seed color and seed shape of different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings,3 and 4 different compounds were screened out in different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings,4 and 1 different compounds were screened out in different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings,and they were screened out in total 28 different compounds were identified.Among them,the variation trend of different compounds in different seed color of tartary buckwheat sprouts and tartary buckwheat seedlings is different.The difference compounds in long seed shape of tartary buckwheat seedlings are higher than that in short seed shape of tartary buckwheat seedling,but the difference compounds in long seed shape of tartary buckwheat sprouts are lower than that in short seed shape of tartary buckwheat sprouts,and they have the opposite trend in the difference compounds;and the difference between tartary buckwheat sprouts and tartary buckwheat seedlings 25different compounds were screened out.The results showed that the flavonoids in different kinds of tartary buckwheat sprouts and tartary buckwheat seedlings were different.4.Expression of related genes in flavonoids synthesisAccording to the related metabolism data,1508-TB115 was selected as the material to detect the MRM pattern of catechin,epicatechin,rutin and quercetin in tartary buckwheat sprouts and seedlings respectively,and qRT-PCR was used to detect the related genes in the synthesis pathway of flavonoids.The results showed that the peak areas of catechin,epicatechin,rutin and quercetin in tartary buckwheat seedlings were lower than those in tartary buckwheat sprouts.and the expression levels of genes FtPAL?FtC4H?Ft4CL?FtCHS?FtCHI?FtF3H?FtF3'H?FtFLS?FtDFR?FtLAR?FtANS?FtANR in tartary buckwheat sprouts were higher than those intartary buckwheat seedlings.The related experiments laid a foundation for transcriptome analysis,molecular experiments and selection of good varieties of tartary buckwheat.