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| Preliminary Study on Triptolide and Total Alkaloid Production of Tripterygium wilfordii Hook. f. Embryoid Culture |
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Abstract Tripterygium wilfordii Hook. f. is a kind of perennial woody vine plant, and it's period of growth is very long and the content of secondary metabolites is low. The natural resources can’t supply enough needs in medicine and agriculture. The way that modern cell engineering technology which produces the secondary metabolites can solve the natural resource shortage and protect the ecological environment for wild T. wilfordii. The leaves of one-year-old T. wilfordii were used to induce calli and suspension embryoid was established using the loose calli. The results showed that green embryoid could be easily induced from calli in 1/2MS+60 mL/L coconut juice liquid medium. The separate embryoid would grow and propagate in 1/2MS+0.5 mg/L NAA. The suitable condition for embryoid culture was 1/2MS basal medium with 1.0 mg/L NAA and 30 g/L sucrose, the initial inoculum was 1.5 g/100mL, pH 5.8, and the medium volume per flask was 120 mL /250 mL after screening inoculum size, pH, the medium volume per flask. The two culture systems were obtained according to the biomass accumulation after adding endophyte fungal elicitors. The triptolide was 1.07 and 1.44 times in embryoid than that in the leaves and in the natural root bark in 1/2MS+0.5 mg/ L NAA+30 g/L sucrose+Y1(leaf endophytes)，pH 5.8; the total alkaloids were 17.02 and 1.46 times in embryoid than that in the leaves and in the natural root bark in 1/2MS+0.5 mg/L NAA+30 g/L sucrose+G4(root endophytes)，pH 5.8. The death rate of embryoid extracts was over 90% after 72 h on Plutella xylostella L. 3rd, the biological activity of embryoid extracts were the same as root bark. The content of triptolide and total alkaloid in embryoid which was induced from leaf calli matched the wild leaf and root bark, there was no significant difference of insecticidal activities between the embryoid extract and root bark. These results show that the culture of embryoid may be one of the pathways to solve the shortage of natural resource in T. wilfordii.
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Received: 17 September 2012
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