Effect of carbon monoxide on growth and triterpenoid production in Betula platyphylla suspension cells / 中草药

ABSTRACT

Objective: To analyze the effect of carbon monoxide (CO) on the growth of Betula platyphylla suspension cells and triterpenoid production. Methods: CO donor hematin of 5, 10, 20, and 30 μmol/L were added to eight-day-old B. platyphylla suspension cells, and the change of triterpenoid content and gene expression of FPPS, LUS, CAS1, CAS2, and β-AS related to the triterpenoid synthesis were analyzed by chemical colorimetry, high performance liquid chromatography and real-time PCR, respectively. Results: CO treatment did not have a significant effect on dry weight of B. platyphylla suspension cells except for 2-8 d at 20 μmol/L and 30 μmol/L CO treatment. CO treatment also did not have a significant effect on pH value and malondialdehyde content except for 30 μmol/L CO treatment. CO treatment significantly increased the content of triterpenoid, betulin and oleanolic acid. the content of triterpenoid, betulin and oleanolic acid were the highest after 1 d at 30 μmol/L CO treatment, 4 d at 10 μmol/L CO treatment and 2 d at 10 μmol/L CO treatment, which was 1.3, 1.5, and 4.2 times of the control, respectively. Triterpenoid production in B. platyphylla suspension cells induced by CO was further confirmed by key enzyme genes of FPPS, LUS, CAS1, CAS2, and β-AS related to the triterpenoid synthesis by RT-PCR. Conclusion: CO treatment could effectively enhance the triterpenoid production in B. platyphylla suspension cell culture.