| 摘要: |
| 【目的】建立曼地亚红豆杉的愈伤组织细胞悬浮培养体系，为紫杉醇生产提供参考。【方法】以曼地亚红豆杉幼茎诱导的愈伤组织为材料,以B5 培养基为基本培养基,利用单因素试验或正交试验，研究愈伤组织接种量、蔗糖质量浓度、激素类型及配比和防褐化试剂对曼地亚红豆杉细胞生长的影响。【结果】曼地亚红豆杉细胞悬浮培养的最佳培养基和培养条件为：B5+2，4-D 0.6 mg/L+NAA 0.8 mg/L+KT 1.0 mg/L+柠檬酸220 mg/L+PVP 350 mg/L+水解乳蛋白 450 mg/L+蔗糖30 g/L(pH=5.8)，接种量 0.1 g/mL，在培养箱中110 r/mn、25 ℃暗培养。【结论】曼地亚红豆杉愈伤组织细胞在所建立的培养体系中经过多次继代后，生长状况良好，表明该细胞悬浮体系比较稳定，可以用于大规模培养红豆杉细胞。 |
| 关键词: 曼地亚红豆杉 愈伤组织 细胞悬浮培养 植物生长激素 防褐化试剂 |
| DOI： |
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| 基金项目:陕西省教育厅资助项目（017K107）；陕西省科学技术研究发展计划项目(2009K01-11) |
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| Establishment of cell suspension culture system for Taxus media |
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ZHAO Ji-peng,YANG Shu-shen
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| Abstract: |
| 【Objective】This study aimed to establish a suspension culture system for Taxus media cell so as to improve the production of taxol.【Method】The callus was extracted from stems of young Taxus media.Effects of sugar,callus inoculum size,phytohormones and anti-browning reagents on the growth of cells were studied by orthogonal test design with basic medium B5.【Result】The best culturing medium for Taxus media cells was B5+2,4-D 0.6 mg/L+NAA 0.8 mg/L+KT 1.0 mg/L+citric acid 220 mg/L+PVP 350 mg/L+LH 450 mg/L+sucrose 30 g/L (pH=5.8).The best culturing conditions were:Inoculum size 0.1 g/mL,incubator rotation speed 110 r/min,temperature 25 ℃,and in dark environment.【Conclusion】 Cells grew well in the proposed culturing system,indicating that the stable cell suspension system was suitable for large-scale cultivation of Taxus cells. |
| Key words: Taxus media callus cell suspension cultures plant growth hormone anti-browning reagent |
