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林业科学 ›› 2010, Vol. 46 ›› Issue (5): 64-69.doi: 10.11707/j.1001-7488.20100511

• 论文 • 上一篇    下一篇

培养基及培养条件对雷公藤愈伤组织生长和次生代谢产物含量的影响

李琰1,2,冯俊涛1,王永宏2,李玉平2,张兴1   

  1. 1.西北农林科技大学无公害农药研究服务中心陕西省生物农药工程技术研究中心杨凌712100;2.西北农林科技大学生命科学学院 杨凌712100
  • 收稿日期:2009-04-17 修回日期:1900-01-01 出版日期:2010-05-25 发布日期:2010-05-25
  • 通讯作者: 张兴

Effects of Basic Media and Culture Conditions on Callus Growth and Secondary Metabolite Content of Tripterygium wilfordii

Li Yan1,2;Feng Juntao1;Wang Yonghong2;Li Yuping2;Zhang Xing1     

  1. 1.Research and Development Center of Biorational Pesticide,Northwest A&F University Shaanxi Province Technology and Engineering Center of BiopesticideYangling 712100;2.College of Life Science,Northwest A&F UniversityYangling 712100
  • Received:2009-04-17 Revised:1900-01-01 Online:2010-05-25 Published:2010-05-25

摘要:

以MS,B5,NT,H,1/2MS,6,7-V及White为基本培养基,分别添加1.0 mg·L-12,4-D和0.5 mg·L-1KT,分析不同类型培养基对雷公藤愈伤组织生长及次生代谢产物含量的影响,并以NT为基本培养基进行光照条件、pH值、接种量、不同外植体以及培养时间试验,探讨雷公藤愈伤组织生长及次生代谢产物含量的最佳培养条件。结果表明:6,7-V培养基有利于愈伤组织生长和继代保存,White培养基上雷公藤内酯醇及雷公藤总生物碱含量最高,NT培养基适合进一步继代培养。pH5.8时,最有利于愈伤组织生长,pH6.7时雷公藤内酯醇的含量最高,而雷公藤总生物碱含量在pH5.2时最高;光照处理中,暗光不仅有利于愈伤组织生长,也有利于雷公藤内酯醇及雷公藤总生物碱的积累,且褐化程度较轻。根愈伤组织增长量、雷公藤内酯醇含量最高,而叶愈伤组织中雷公藤总生物碱的含量最高。愈伤组织最适继代时间为40~45天,雷公藤内酯醇及雷公藤总生物碱的产量在第50天时最高。

关键词: 雷公藤, 愈伤组织, 组织培养, 次生代谢产物

Abstract:

MS,B5,NT,H,1/2MS,6,7-V and White were used as basic media,supplemented with 1.0 mg·L-12,4-D and 0.5 mg·L-1 KT, to study their effects on callus growth and secondary metabolite content of Tripterygium wilfordii.Light,pH,inoculating quantity,explants and culture time were studied with NT as basic media to explore the best culture conditions of the callus growth and secondary metabolite content.The results showed that 6,7-V medium was best for callus growth and preservation.White was the best suitable for the accumulation of triptolide and total alkaloids.NT medium was suitable for subculture.The optimal pH for the callus growth was 5.8.The optimal pH for the accumulation of triptolide was 6.7 and the optimal pH for the total alkaloids accumulation was 5.2.Dark condition was best not only for callus growth,but also for the triptolide and total alkaloids accumulation,with lesser browning callus.The root callus growth and triptolide content were significantly higher than that of the stem and leaf callus.Alkaloid content of leaf callus was significantly higher than that of the root and stem callus.The optimal duration for callus subculture was around 40-45 days.The triptolide and total alkaloids yield was the highest in 50 days.

Key words: Tripterygium wilfordii, callus, tissue culture, secondary metabolites