Establisment Of Adventitious Roots Culture System Of Panax Ginseng And Analysis Of Panax Ginseng And Panax Notoginseng Soil Microorganism And Studies On Metabolic

Panax ginseng C.A.Mey.is a famous traditional medicinal herb all over the world and widely used..In order to improve the content of ginsenosides in the advancer roots of P.ginseng,the P.ginseng root lines were screened,the analysis of the dominant bacteria of soil microorganisms and endophytic bacteria of P.ginseng and P.notoginseng.The metabolic regulation of the adventitious root of P.ginseng was studied by adding the P.notoginseng dominant strain Chaetomium globosum.Firstly,according to the age of ginseng explants and the adventitious roots morphology(main roots and rootlet roots),six P.ginseng adventitious root culture systems were established.The content of ginsenoside and expression level of functional genes in adventitious roots of P.ginseng were compared with those of 5-year-old cultivated P.ginseng(5Y).The results showed that the content of ginsenoside in AR-5R(5-year-old Panax ginseng induced adventitious root rootlet root)was the closest to that in 5Y(31.88 mg/g),which was consistent with the level of gene expression.Thus AR-5R was selected as the optimal root system for adventitious roots of P.ginseng.In this study,the rhizosphere soil and root samples from different P.ginseng producing areas(Fusong,Jingyu,Tonghua)and P.notoginseng producing areas(Asalong,Huilong,demonstration garden)were used as research objects.High throughput sequencing was carried out in the bacterial 16S rDNA and fungal ITS1 regions respectively.Based on the highest active ingredient in Huilong P.notoginseng and Fusong 6-year-old ginseng,the dominant strains were predicted.The results showed that 16 strains of the dominant bacteria were found in the rhizosphere soil of P.ginseng,such as Pseudomonas citronellolis et al.15 strains of the dominant fungi were found in the rhizosphere soil of P.ginseng,such as Trichoderma spirale et al.30 strains of the dominant endophytic bacteria were found in ginseng root,such as Delftia tsuruhatensis et al,20 strains of the dominant endophytic fungi were found in ginseng root,such as Clonostachys divergens et al.18 strains of the dominant bacteria were found in the rhizosphere soil of P.notoginseng,such as bacterium Ellin5102 et al.32 strains of the dominant fungi were found in the rhizosphere soil of P.notoginseng,such as Chaetomium globosum et al.19 strains of the dominant endophyte bacteria were found in the P.notoginseng root,such as Sphingobium fuliginis et al.10 strains of the dominant endophyte fungi were found in the P.notoginseng root,such as Microidium phyllanthi et al.A total of 36 strains of bacteria,40 strains of fungi were isolated from the roots and soil of P.notoginseng by traditional microbial isolation method,and 15 strains were identified,such as Chaetomium globosum et al.C.globosum was selected as the following experimental strain because it was consistent with the predominant strain of P.notoginseng predicted by ITS1.In order to improve the content of ginsenosides,the effects of strain C.globosum on adventitious roots of P.ginseng under different treatments(sterilized C.globosum mycelium powders,suspension of spores,immobilized spores and immobilized mycelium)were investigated.It was found that the content of ginsenosides was significantly increased in the immobilized spore group.The optimum conditions were liquid-solid ratio of 10:1(mL/g),initial pH of 6.5,co-culture time of 24 h,and total saponins(56.29 mg/g)were 3.2 times of the blank group.By investigating the induction mechanism,it was found that elicitors could increase the accumulation of resistance-related signaling molecules such as Ca2+,NO and H2O2,and the expressions of defense genes(PAL and PR10)and functional genes(GPS,FPS,SS,SE,DS,CYP716A47,CYP716A53v2,UGT74AE2,UGT94Q2,UGTPgl and UGTPg100)were up-regulated significantly in the ginsenosides biosynthesis pathway.In order to determine the effective components in C.globosum,the co-culture fermentation broth was analyzed by GC-MS.6 specific metabolites were identified in C.globosum fermentation broth,of which L-aspartic acid may play a major role.The content of ginsenoside was increased obviously by co-culturing the optimal strains of C.globosum and the optimal P.ginseng root system,which provided theoretical basis and technical support for the industrialized production of ginseng active ingredients.