天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (7): 1182-1189.doi: 10.16333/j.1001-6880.2020.7.013

• 研究简报 • 上一篇    下一篇

凤丹愈伤组织培养体系建立及丹皮酚含量测定

王若馨1,2，纪楠楠1,2，王华芳1,2*   

  1. 1北京林业大学林木育种国家工程实验室；2北京林业大学生物科学与技术学院，北京 100083
  • 出版日期:2020-07-28 发布日期:2020-08-03
  • 基金资助:
    国家自然科学基金重点项目（U1804233）；北京林业大学重大科研成果培育项目（2017CGP008）

Callus culture system and paeonol determination of Paeonia ostii T.Hong et J.X.Zhang var. lishizhenii B.A.Shen

WANG Ruo-xin1,2,JI Nan-nan1,2,WANG Hua-fang1,2 *   

  1. 1National Engineering Laboratory for Tree Breeding,Beijing Forestry University; 2College of Biological Sciences and Technology,Beijing Forestry University,Beijing 100083,China

  • Online:2020-07-28 Published:2020-08-03

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摘要： 本文旨在建立凤丹（Paeonia ostii var. lishizhenii）愈伤组织培养及制取丹皮酚的技术体系，为细胞工程制药提供技术基础。凤丹去皮种子以70%乙醇（V/V）浸泡2.0 min和有效氯1.0%的二氯异氰尿酸钠溶液浸泡20 min，去污染率高达97.78%；剥取种胚接种在含6-苄氨基嘌呤1.5 mg/L、2，4-二氯苯氧乙酸2.0 mg/L的MS培养基上，愈伤组织诱导率达90.00%；愈伤组织在含6-苄氨基嘌呤 1.8 mg/L、萘乙酸0.6 mg/L、蔗糖30 g/L、水解酪蛋白0.4 g/L、琼脂5 g/L、pH 7.3的改良WPM培养基上暗培养25天，增殖倍数达3.53，褐化率为11.78%；基于天然产物丹皮酚含量和愈伤组织生长曲线，培养25天时丹皮酚含量达到最高，为最佳收获期。


关键词: 凤丹, 愈伤组织, 增殖, 次生代谢物, 丹皮酚

Abstract:

The paper aims to establish a technical system for the cultivation of paeonol in P. ostii var. lishizhenii callus,and to provide a technical basis for cell engineering pharmacy.The mature and plump seeds of P. ostii var. lishizhenii were peeled and soaked in 70% ethanol(V/V) for 2.0 min and then soaked in 1.0% sodium dichloroisocyanurate solution for 20 minutes.The seed decontamination rate was 97.78%.Place sterilized on MS medium and then add 6-BA 1.5 mg/L and 2,4-D 2.0 mg/L.The callus induced was 90.00%.Optimized medium for cell culture was the modified WPM with 6-BA 1.8 mg/L,NAA 0.6 mg/L,sucrose 30 g/L,hydrolyzed casein 0.4 g/L,agar 5 g/L and pH 7.3.In the conditions,cell proliferation ratio was 3.53 and the browning rate was 11.78% on the 25th day.Based on the natural product paeonol content and callus growth curve,paeonol content reached the highest when cultured for 25 days,which was the best harvest time.

Key words: Paeonia ostii , var.lishizhenii, callus, proliferation, secondary metabolites, paeonol

中图分类号:  S685.11

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