Nontargeted Metabolomic Analysis Of Anoectochilus Roxburghii At Different Cultivation Stages

Anoectochilus roxburghii(Wall.)Lindl is a perennial herb of anoectochilus orchid,which is a traditional Chinese herbal medicine.It has a long application history in south of China and the market value is very high.In recent years,the establishment of micropropagation system has solved the problem of shortage of resources,so that its industry is booming.However,the morphology of soil cultivated plants is indistinguishable from the tissue culture plants.It remains unclear if the soil cultivation period is essential for the accumulation of health beneficial metabolites in A.roxburghii.As a result,the necessity for soil cultivation period is still under debate,and the market is currently confusing.In this study,we performed untargeted metabolomic analysis by both GC-TOF-MS and LC-Q-TOF-MS,using the Daye and Xiye varieties of A.roxburghii plants at tissue culture stage or after 4 months of soil cultivation,optimized the extraction methods of samples in Nontargeted Metabolomic Analysis.Selected 204 and 180 metabolic markers in Daye and Xiye varieties of A.roxburghii.We identified metabolic markers of different cultivation phases and potential health beneficial components in A.roxburghii.The main results are as follows:1.The primary metabolites,such as amino acids,organic acids and oligosaccharides,were higher in the tissue culture stage.The primary metabolites are similar between different cultivars.Oxoproline was significantly higher in soil cultivation stage of both varieties,which could be referred as the metabolic marker.2.Flavonoids glycosides,esters and terpenoids were significantly increased in the soil cultivation plants.The secondary metabolites of the two varieties show significantly different.The total content of flavonoids glycosides and esters can be used as indicator to distinguish A.roxburghii under different cultivation stages.Allicin and Sterebin D,Crassostrea Secocarotenoid and Streptozocin,which could be used as metabolic markers of Daye A.roxburghii and Xiye A.roxburghii respectively.3.We identified 216 and 193 compounds in Daye and Xiye A.roxburghii respectively.Among them,polysaccharide and flavonoid glycoside are both higher in Daye A.roxburghii,so that the nutrient and medicinal value is higher.To sum up,our study using the non-targeted metabolomics analysis on A.roxburghii at different cultivation stages for the first time,made a high-throughput systemic analysis by chemometric,which find the metabolic markers of different cultivation phases,provides certain help for the further research and promote market standardization of A.roxburghii products.