| The culture system of Arctium lappa L. hairy roots was established and efficient secondary metabolites were extracted. In this paper, Agrobacterium rhizogenes R1000, R1, R15834 were selected to induce the hairy roots, the selection of the implant, the infection time, culture and co-culture time, different bacteria liquid concentration, medium pH value detection were inspected, optimizing the best induction system of Arctium lappa L. hairy roots. Then the Arctium lappa L. hairy roots were tested by PCR, T-DNA from Ri plasmid was confirmed to integrate into the genome. The best hairy roots were chosen to further expand by liquid cultivation and the condition was optimized. By hairy root growth curve, screening of liquid medium, and researching on the effects of different carbon source, the optimal liquid medium of Arctium lappa L. hairy roots growth was selected. The content of arctiin of Arctium lappa L. hairy roots was detected by high performance liquid chromatography(HPLC), the conclusion was showed that the higher content of hairy roots than Arctium lappa L.seeds and roots. The best extraction conditions were optimized through orthogonal design.Test results show that the hairy roots were induced by choosing Agrobacterium rhizogenes R1. The roots were infected of 12 min, cultured of 48 h and co-culture of 36 h, the concentration of the cultivation was 0.8 and the pH value of 6.5. The PCR conclusion was integrated into the genome DNA. Further expand liquid cultivation, White medium was chosen, carbon source was added, and was trained by 28 days. The HPLC detection comparison results show that the highest arcttin of Arctium lappa L. hairy roots, was higher 1.16 times than the Arctium lappa L.seeds, the arcttin content was optimized by the orthogonal experiment, the best condition: ethanol volume fraction is 80%, and solid-liquid ratio of 1:20, ultrasonic extraction time for 30 min, under this condition, the extraction yield of arcttin was 7.345%. |
PDF Full Text Request