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目的研究B5培养基主要成分对云南红豆杉Taxus yunnanensis细胞生长和产生10-去乙酰巴卡亭Ⅲ的影响。方法利用植物组织培养技术结合HPLC分析手段,通过改变B5培养基中CaCl2、NaH2PO4、(NH4)2SO4和KNO3的质量浓度,研究其对云南红豆杉细胞生长及10-去乙酰巴卡亭Ⅲ量的影响。结果试验组与对照品相比,B5培养基中高质量浓度的NaH2PO4能够显著促进云南红豆杉细胞生长,CaCl2、(NH4)2SO4和KNO3浓度对细胞生长无显著影响;B5培养基中低质量浓度的CaCl2、NaH2PO4、(NH4)2SO4和KNO3能够显著促进10-去乙酰巴卡亭Ⅲ合成与释放。结论B5培养基中NaH2PO4为300mg/L时,能够显著促进云南红豆杉细胞生长,细胞生长量是对照的1.2倍,CaCl2、(NH4)2SO4和KNO3分别在75~300、134~268和2500~5000mg/L对细胞生长无显著影响;B5培养基中CaCl2、NaH2PO4、(NH4)2SO4和KNO3的质量浓度分别为75、75、33.5、625mg/L时,能够显著促进细胞中10-去乙酰巴卡亭Ⅲ合成与释放,10-去乙酰巴卡亭Ⅲ的量分别是对照的1.5、1.4、2.4倍。
Objective To study the effects of main components of B5 medium on Taxus yunnanensis growth and 10-deacetylbaccatin III production in Taxus yunnanensis. Methods The plant tissue culture technique and HPLC analysis were used to study the effects of different concentrations of CaCl2, NaH2PO4, (NH4) 2SO4 and KNO3 on the growth of Taxus yunnanensis and the amount of 10-deacetyl-baccatin III influences. Results Compared with the control group, the high concentration of NaH2PO4 in B5 medium could significantly promote the growth of Taxus yunnanensis cells. The concentration of CaCl2, (NH4) 2SO4 and KNO3 had no significant effect on the cell growth. In B5 medium, CaCl2, NaH2PO4, (NH4) 2SO4 and KNO3 can significantly promote the synthesis and release of 10-deacetyl baccatin III. Conclusion When NaH2PO4 is 300mg / L in B5 medium, the growth of Taxus yunnanensis can be significantly promoted. The cell growth rate is 1.2 times of that of control. The contents of CaCl2, (NH4) 2SO4 and KNO3 are 75 ~ 300, 134 ~ 268 and 2500 ~ 5000mg / L had no significant effect on the cell growth. The concentration of CaCl2, NaH2PO4, (NH4) 2SO4 and KNO3 in B5 medium were 75, 75, 33.5 and 625mg / L respectively, Synthesis and release of kiosk Ⅲ, the amount of 10-deacetylbaccatin Ⅲ were 1.5, 1.4, 2.4 times of the control.