The Study Of Metabolism Of Carbon And Sulfur In Triterpenoids Accumulation Induced By CO And H₂S

In order to explore the mechanism of triterpenoids accumulation induced by CO and H2S signaling molecules. The mycelium of Phellinus igniarius and suspension cells of birch were used as experimental materials to detect the content of triterpenoids treated by different concentration of CO and H2S donor, hemin and sodium hydrosulfide. On the base of it, we analyzed the cross-talk of endogenous CO and H2S and the effects of sulfur and carbon metabolism by pharmacological experiments, namely adding CO scavenger hemoglobin (Hb) and H2S scavengers Asia taurine (HT). The main findings are as follows:1. The effect of CO treatment on the accumulation of triterpenoidsDifferent concentrations of CO donor hemin were added to the 8-day-old Phellinus mycelium and birch suspension cells for 6h-8d. The results showed that CO treatment increased the content of triterpenoids. Among of them,30μmol/L CO treatment for Id had the best inducing effects in Phellinus mycelium, the triterpenoid content were enhanced by 31.1% comparing with the control. Its synthetic key genes of triterpenoid also increased after CO treatment; 20 mmol/L and 10mmol/L CO had the best inducing effects in birch suspension cells, the triterpenoid content were enhanced by 31%and 29.6%comparing with the control, respectively.2. The effect of H2S treatment on the accumulation of triterpenoidsDifferent concentrations of H2S donor odium hydrosulfide were added to the 8-day-old Phellinus mycelium for 6h-4d. The results found that H2S improved the accumulation of triterpenoids and betulin. Among of them,2mmol/L H2S treatment for 1d had the best inducing effects, the content of triterpenoid and betulin were enhanced by 17.1%and 40%comparing with the control, respectively.3. The cross-talk of CO and H2S in the accumulation of triterpenoidsTo clear the relationship between triterpenoids accumulation and the cross-talk of CO and H2S. CO and H2S scavengers HT, H2S and CO scavenger Hb were added to 8-day-old Phellinus mycelium for 12h. The results showed that CO treatment increased H2S content, the activity of key enzyme in the synthesis of H2S, and triterpenoids and betulin content. The above increased value induced by CO treatment was suppressed under the CO and H2S scavengers HT treatment, but the above value was still higher than the control. H2S treatment increased CO content, the activity of key enzyme in the synthesis of H2S, and triterpenoids and betulin content. The above increased value induced by H2S treatment was suppressed under the H2S and CO scavengers Hb treatment, but the above value was still higher than the control. From these results, it can be inferred that the cross-talk of endogenous CO and H2S involved in the triterpenoids accumulation.4. Carbon metabolism in triterpenoids accumulation under CO and H2S treatmentCO or H2S treatment increased the content of sucrose and fructose, the activity of sucrose phosphate synthase, had no effect on the levels of glucose in Phellinus mycelium. The treatment of CO with sucrose, fructose and glucose, respectively, enhanced triterpenoids accumulation than that of CO or sucrose, fructose and glucose. However, the treatment of H2S with sucrose, fructose and glucose, respectively, had the opposite results. The above results could be inferred that carbon metabolism played important role in triterpenoids accumulation under CO treatment.5. Sulfur metabolism in triterpenoids accumulation under CO and H2S treatmentCO or H2S treatment increased the content of soluble thiol, total sulfur, glutathione content and gene expression of glutathione reductase. The above increased value induced by CO or H2S treatment was suppressed under the CO and H2S scavengers HT treatment or H2S and CO scavenger, but the above value was still higher than the control. CO treatment increased the content of pyruvic acid, while H2S treatment decreased it. From the above results, we known CO and H2S treatment changed the metabolism of sulfur in Phellinus mycelium, but the relationship between sulfur metabolism and triterpenoids accumulation will be carried out in our next plan.