Study on content of terpenoid indole alkaloids and transcriptional differences for covalent orca3/g10h in double-gene transgenic hairy roots of Catharanthus roseus / 中草药

Objective: In order to improve the content of terpenoid indole alkaloids (TIAs), orca3/g10h genes were introduced to the hairy roots in Catharanthus roseus. Methods: Bivalent expression vector CAMBIA1304+ +orca3 + g10h was constructed and introduced into Agrobacterium rhizogenes strain and transformed into C. roseus to obtain transgenic hairy roots. RT-qPCR was used to study the transcriptional differences of relative genes referred to the biosynthesis pathway of TIAs. Then HPLC was used to study TIAs content in the transgenic hairy roots of C. roseus, including vinblastine, vincristine, and ajmalicine. Results: The transcriptional level of genes that linked to biosynthesis of TIAs in the transgenic hairy roots of C. roseus, asα, ggpps, g10h, str, tdc, cpr, sgd, and dat, were all expressed higher than those of the nontransgenic roots. HPLC results showed that modified hairy root of C. roseus owned more total TIAs production, 58.23 mg/g, the number was larger than that of common roots in C. roseus as many as 27.5 times. On the other hand, the average content of vinblastine and vincristine was also more than the common roots in C. roseus. Among them, vinblastine content was the most. The number of production got 51.30 mg/g, which was as many as 197.3 times of the common root of C. roseus. Conclusion: Orca3/g10h double-gene transgenic hairy root of C. roseus can increase TIAs content efficiently.

Physiological characters of Catharanthus roseus mutant cells / 中草药

Objective To investigate the characteristics of growth, accumulation of indole alkaloid, and the optical concentration of the nutrition components in the Catharanthus roseus mutant cells induced by the colchicine, looking forward to getting the ideal material producing alkaloids which was suited for industrialized cell cultivation. Methods Mutant cells cultivated on the MS medium were harvested according to experiment designs,and the fresh weight was got.The indole alkaloid as extracted by organic solvent. The ajmalicine and catharanthine extracted from the samples were analyzed by RP-HPLC. Results The growth rate was faster and the indole alkaloid accumulation was more when the mutant cells were cultivated generations 30 and the both were sharply decreased at generations 45, while the contents of medical compounds amounted to the peak at the generations 20. The content of medical components and alkaloid accumulation were evidently prompted by the tryptophan added in the media, especially at the concentration of Ca2+ 1 760 and Zn2+ 12.6 mg/L, respectively. Conclusion C.roseus mutant cells probably is an ideal material for industrialized cell cultivation.

Enhancing accumulation of indole alkaloids in Catharanthus roseus leaves / 中草药

Objective To study the effect of three kinds of reagent: ethrel, Aspirin, tryptophan on the accumulation of medicinal component in Catharanthus roseus leaves. Methods To treat these potplants of C. roseus in green house with the three reagents in different concentrations, respectively. Then, two days later, the contents of catharanthine and vinblastine in leaves were determined by RP-HPLC. Results The three reagents can obviously improve the accumulation of indole alkaloids, including catharanthine and vinblastine in C. roseus leaves. Their optimum treatment concentration was finally comfirmed. Conclusion These three kinds of reagent apparently promote the accumulation of total indole alkoloids in the leaves of C. roseus.

Preliminary study of a vincristine-producing endophytic fungus isolated from leaves of Catharanthus roseus / 中草药

Object To select the endophytic fungi which produce vincristine by isolating fungi from the leaves of Catharanthus roseus (L.) G. Don. Methods The endophytic fungi were isolated from the leaves of C. roseus and the zymotic extracts were analyzed by TLC and HPLC. Results An endophytic fungus which is Mycelia sterilia 97CY_3 can produce vincristine. The content of vincristine in the fungus was determined as 0.205 ?g/L by HPLC. Conclusion Some endophytic fungi isolated from C. roseus can produce the anticancer substance vincristine which is the same as that of host plant producing.

Effects of fungal elicitors on accumulation of indole alkaloids in Catharanthus roseus calli / 中草药

Object To investigate the effects of fungal elicitors derived from the fungi Fusarium solani and Aspergillum niger on the accumulation of indole alkaloids in Catharanthus roseus calli. Methods The total indole alkaloid was extracted after the calli were treated with fungal elicitors. Then, the determination of ajmalicine and catharanthine was carried out by RP HPLC. Results The two fungal elicitors stimulate the accumulation not only the total indole alkaloid but the ajmalicine and catharanthine. The optimal exposure time of the two fungal elicitors for different kinds of indole alkaloid was investigated. Conclusion The two fungal elicitors have obvious effect on the accumulation of indole alkaloid in C. roseus calli.