Effect Of Methyl Jasmonate On Secondary Metabolites Of Panax Notoginseng Adventitious Roots Based On Metabolomics

Radix notoginseng, a rare and specific Chinese material medica in China, has function of promoting blood circulation to remove blood stasis, calming, relieving pain, and nourishing body. The output of Radix notoginseng can't meet the increasing requirement of the market because of its slower growing speed and specific living conditions. Therefore, it is urgent to investigate an alternative approach to solve the problem. To cultivate adventitious root of Panax notoginseng in vitro is established in our lab. The next step is to discover a highly effective way to produce plenty of active secondary metabolites in vitro.In this paper, mass-spectrometry based metabolomics analysis method was established to investigate the adventitious roots. The active secondary metabolites of Methyl Jasmonate influenced adventitious roots, adventitious roots, flower buds and native roots were compared using rapid ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) and multivariate statistical analysis technique. Methyl Jasmonate was proved to be an effective elicitor. And the time course effect of Methyl Jasmonate on the adventitious roots was investigate in detail using the established method.A rapid analytical method for metabolite profiling and quantifying 15 ginsenosides in P. notoginseng by ultra performance liquid chromatography (UPLC) coupled to electrospray ionization mass spectrometry (ESI–MS) is described. ESI–MS was optimized for different cone voltages at positive ionization mode to allow simultaneous analysis of all analytes in a relatively short time. Qualitative methodological considerations, including the linear range, precision, limit of quantification, limit of detection, recovery and sensitivity are also provided. UPLC-ESI-MS coupled with multivariate statistical analysis technique was powerful to discriminate the differences of native roots and adventitious roots.The active secondary metabolites of Methyl Jasmonate influenced adventitious roots, adventitious roots, flower buds and native roots were compared using rapid ultra-performance liquid chromatography- electrospray ionization mass spectrometry (UPLC-ESI-MS) and multivariate statistical analysis technique. Principle component analysis (PCA) of the UPLC-ESI-MS data showed a clear separation of compositions among the flower bud, roots, methyl jasmonate influenced adventitious roots and adventitious roots of P. notoginseng. The critical saponins accountable for such variations were identified through the corresponding loading weights and were further verified by mass and retention times of available standard saponins.The adding time and amount of methyl jasmonate was optimized. The effect of Methyl Jasmonate on the secondary metabolites of adventitious roots was investigated by the established method. The trajectory on the PCA score plot showed that methyl jasmonate has an effective influence on synthesis of secondary metabolites at the seventh day, especially saponins. PLS-DA was used to search the critical components contributing the differences of Methyl Jasmonate influenced adventitious roots and adventitious roots of P. notoginseng. The results showed that most of 20(s)-protopanaxadiol moieties and some 20(s)-protopanaxatriol moieties were biosynthesized more in the Methyl Jasmonate influenced adventitious roots. However, some ginsenosides (notoginsenoside R1, ginsenoside Re) were less biosynthesized at the same time. The ratio of main ginsenosides in the Methyl Jasmonate influenced adventitious roots was close to that of the native plant root of P. notoginseng. It suggested that the Methyl Jasmonate influenced adventitious roots is a potential valuable elicitor to be further studied for the production of ginsenosides.