A high taxol yield cell line of Taxus yunnanensis Cheng et L. K. Fu keeps a high taxol_producing level after successive subcultures for more than eight years. In this study, eight taxanes were isolated from the su...A high taxol yield cell line of Taxus yunnanensis Cheng et L. K. Fu keeps a high taxol_producing level after successive subcultures for more than eight years. In this study, eight taxanes were isolated from the suspension cell cultures of this cell line. Based on NMR and MS analyses, and comparison with literature data and standards, their structures were determined to be 2α,5α,10β_triacetoxy_14β_propionyloxy_4(20),11_taxadiene (1), 2α,5α,10β_triacetoxy_14β_(2′_methyl)_butyryloxy_4(20),11_taxadiene (2), 2α,5α,10β_14β_tetra_acetoxy_4 (20),11_taxadiene (3, taxuyunnanine C), 2α,5α,10β_triacetoxy_14β_(2′_methyl_3′_hydroxy)_butyryloxy_4(20),11_taxadiene (4, yunnanxane) and its 3′_epimer (5), baccatin Ⅳ (6), baccatin Ⅲ (7) and taxol (8), respectively. Among those compounds, 3, 5, 6 and 7 were reported to be isolated from the suspension cell cultures of T. yunnanensis for the first time. TLC and HPLC analyses indicated that the chemical constituents of the culture solution were similar to those of cultured cells. Moreover, the highest taxol content of this cell line reached 0.3% and the cell line could be applied for a large_scale culture.展开更多
Conditions have been established for the callus initiation and subculture of T chinensis. The calliwere induced by the explants cultured first on the medium MS supplemented with 1 .0 mg/L 2,4-D, 2g/L CH, and 25g/L suc...Conditions have been established for the callus initiation and subculture of T chinensis. The calliwere induced by the explants cultured first on the medium MS supplemented with 1 .0 mg/L 2,4-D, 2g/L CH, and 25g/L sucrose, then on The medium f MS+1 .0 mg/L NAA+0.5 mg/L BA+2 g/L CH+25 g/L sucrose. When the callus was Subcultured and tamed several times, it could grow fast and stable on the medium : MS+0.2mg/L 2,4-D+0.5mg/L NAA+0.5 mg/L BA+2 g/LCH+25 g/L sucrose. The contamination of explants was a result of endophytic microbes of T Chinensis. This could be avoided by adopting the tender shoots 3-5 cm long collected in early spring as the source of explants. The browning of the Cultures could be prevented and controlled by means of the selection of a suitable explants, hormonal regime in the medium, culture methods and the use of antioxidants.展开更多
The effects of the cultures of Penicilliun citrinum,Absidia glauca,Mucor rouxianus and Botrytis cinerea on the growth and taxol production in suspension cells of Taxus yunnanensis were investigated.The results showed ...The effects of the cultures of Penicilliun citrinum,Absidia glauca,Mucor rouxianus and Botrytis cinerea on the growth and taxol production in suspension cells of Taxus yunnanensis were investigated.The results showed that the addition of the cultures of Penicilliun citrinum or Botrytis cinerea into the suspension cell cultures of T.yunnanensis at the final stage of fast growing phase(the 12th day of culture) did not inhibited the cell growth,but dramatically increased taxol yield;The best result was obtained in the elicitation with Penicilliun citrinum (3.1 g/L dry mycelium) for 3 days,in which the taxol yield was increased by 1～2 times.展开更多
文摘A high taxol yield cell line of Taxus yunnanensis Cheng et L. K. Fu keeps a high taxol_producing level after successive subcultures for more than eight years. In this study, eight taxanes were isolated from the suspension cell cultures of this cell line. Based on NMR and MS analyses, and comparison with literature data and standards, their structures were determined to be 2α,5α,10β_triacetoxy_14β_propionyloxy_4(20),11_taxadiene (1), 2α,5α,10β_triacetoxy_14β_(2′_methyl)_butyryloxy_4(20),11_taxadiene (2), 2α,5α,10β_14β_tetra_acetoxy_4 (20),11_taxadiene (3, taxuyunnanine C), 2α,5α,10β_triacetoxy_14β_(2′_methyl_3′_hydroxy)_butyryloxy_4(20),11_taxadiene (4, yunnanxane) and its 3′_epimer (5), baccatin Ⅳ (6), baccatin Ⅲ (7) and taxol (8), respectively. Among those compounds, 3, 5, 6 and 7 were reported to be isolated from the suspension cell cultures of T. yunnanensis for the first time. TLC and HPLC analyses indicated that the chemical constituents of the culture solution were similar to those of cultured cells. Moreover, the highest taxol content of this cell line reached 0.3% and the cell line could be applied for a large_scale culture.
文摘Conditions have been established for the callus initiation and subculture of T chinensis. The calliwere induced by the explants cultured first on the medium MS supplemented with 1 .0 mg/L 2,4-D, 2g/L CH, and 25g/L sucrose, then on The medium f MS+1 .0 mg/L NAA+0.5 mg/L BA+2 g/L CH+25 g/L sucrose. When the callus was Subcultured and tamed several times, it could grow fast and stable on the medium : MS+0.2mg/L 2,4-D+0.5mg/L NAA+0.5 mg/L BA+2 g/LCH+25 g/L sucrose. The contamination of explants was a result of endophytic microbes of T Chinensis. This could be avoided by adopting the tender shoots 3-5 cm long collected in early spring as the source of explants. The browning of the Cultures could be prevented and controlled by means of the selection of a suitable explants, hormonal regime in the medium, culture methods and the use of antioxidants.
文摘The effects of the cultures of Penicilliun citrinum,Absidia glauca,Mucor rouxianus and Botrytis cinerea on the growth and taxol production in suspension cells of Taxus yunnanensis were investigated.The results showed that the addition of the cultures of Penicilliun citrinum or Botrytis cinerea into the suspension cell cultures of T.yunnanensis at the final stage of fast growing phase(the 12th day of culture) did not inhibited the cell growth,but dramatically increased taxol yield;The best result was obtained in the elicitation with Penicilliun citrinum (3.1 g/L dry mycelium) for 3 days,in which the taxol yield was increased by 1～2 times.
