Adventitious Roots Culture In Bioreactor Of Medicinal Plants Of Panax Quinquefolium L. And Pseudostellaria Heterophylla

Panax quinquefolium L. and Pseudostellaria heterophylla are famous and used herbs, which have a wide range of applications in the medicine, food, cosmetics and other fields. So, they have a large market demand in domestic and international. In this study, adventitious roots of P. quinquefolium and P. hetropphylla provided by the laboratory, we conducted studies on bioreactor culture.In this study, P. quinquefolium adventitious roots were subjected to different concentration (0-20 mg/L) of methyl jasmonate (MJ) elicitation for 5 days before harvest. The level of total saponins in P. quinquefolium adventitious root increased much higher than that observed in the control, especially with 5 mg·L-1 MJ elicitation treatment, level of total saponins was 4.6-fold higher than the control group. It had been determined that the total RNA should be abstracted after 5 days when MJ added. The RNA sequencing technology was applied to analysis and screening the different fragment. Through this study, MJ could promote ginsenosides accumulation because MJ made ginsenoside pathway genes differentially expressed in P. quinquefolium adventitious roots. Gradually scale-up culture from 0.5 L shake flask to 3 Land 5 L balloon-type bubble bioreactor (BTBB) was observed. The biomass and the contents of active components of adventitious roots in bioreactor were significantly higher than that in shake flask. Furthermore, we optimized the bubble bioreactor with three different bioreactors (60°,90°,180°) and the culture conditions of P. quinquefolium adventious roots in bubble bioreactor culture. The results showed that, the most efficient bioreactor for cultivation of the P. quinquefolium adventitious roots was the cone-type bioreactors (with the 90°), and the optimal culture conditions we obtained in experiments were 0.5 vvm aeration rate,3% sucrose,1 MS salt strength and 1.5 g/L inoculum size. Using a fed-batch cultivation strategy, ginsenosides production and polysaccharide production were enhanced to 30.96 mg/L,673.43 mg/L, which were 5.12-fold,2.66-fold greater than with batch cultivation. Two-stage cultivation caused a significant increase in total saponins yield (102.37 mg/L) adventitious roots cultures. This value was higher than 3.31-fold and 16.95-fold compared to the batch cultivation and fed-batch cultivation. The antioxidant potential of methanolic extract [1, 1-diphenyl-2-picrylhydrazyl (DPPH)] of P. quinquefolium adventitious roots was also assessed. DPPH inhibition of ginsenoside and polysaccharide were higher than in adventitious root that in native root.Taking advantage of the established system for cultivating adventitious roots of P. heterophylla, we optimized the bubble bioreactor with three different angles (60°,90°, 180°) and the culture conditions of P. heterophylla adventious root in bubble bioreactor culture. The results showed that, the most efficient bioreactor for cultivation of the P. heterophylla adventitious roots was the cone-type biooreactors (with the 90°), and the optimal culture conditions we obtained in experiments were:0.3 vvm aeration rate,1.5 g/L inoculum size,3/4 MS salt strength.Our study provided theoretical reference and technical support for large-scale culture of P. quinquefolium and P. heterophylla adventitious roots.