悬浮培养瑞香狼毒不定芽过程中黄酮和香豆素类物质的累积  

Accumulation of flavonoids and coumarins in the shoots of Stellera chamaejasme L. by suspension culture

作　　者：夏美玲 姜红 郭雯华 官旭杰 由香玲[1] XIA Mei-Ling;JIANG Hong;GUO Wen-Hua;GUAN Xu-Jie;YOU Xiang-Ling(College of Life Sciences,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《植物生理学报》2019年第4期457-465,共9页Plant Physiology Journal

基　　金：国家自然科学基金(30972390)~~

摘　　要：本文以瑞香狼毒不定芽为材料,研究了在250 mL培养瓶悬浮培养过程中,影响其生长及活性物质(伞形花内酯、东莨菪内酯、总黄酮)积累的四类因子(蔗糖浓度、培养基强度、接种量、接种长度),初步建立了不定芽的悬浮培养体系。在此基础上,研究了3 L生物反应器内,影响不定芽生长及活性物质积累的因素(蔗糖浓度、6-BA浓度、初始接种量)。结果显示,不定芽在250 mL培养瓶中悬浮培养周期是15 d,最佳蔗糖浓度、培养基强度、接种量、接种长度分别是30 g·L^(-1)、MS培养基、1.0 g、3 cm。在此培养条件下,不定芽中伞形花内酯、东莨菪内酯和黄酮累积量都在15 d达到最大值,分别为0.351、0.035和2.459 mg。3 L生物反应器内不定芽悬浮培养周期为24 d,最佳培养条件为蔗糖30 g·L^(-1)、6-BA 0.5 mg·L^(-1)和初始接种量15 g。此条件下,伞形花内酯、东莨菪内酯和黄酮累积量分别为6.041、0.763和51.334 mg。本研究建立了瑞香狼毒不定芽的小型生物反应器培养体系,为进一步大规模培养,高效生产上述三种活性物质奠定了基础。In this study, during the suspension culture of the shoots of Stellera chamaejasme in 250 mL culture flask, four kinds of factors(sucrose concentration, medium strength, quantity and length of inoculum) affecting the growth of shoots and the active substances(umbelliferone, scopoletin, total flavonoids) in them were studied, and the culture-flask suspension culture system of adventitious shoots was established. On this basis, the 3 L bioreactor culture system was established, by analyzing the effects of sucrose concentration, 6-BA concentration, initial inoculum quantity on shoots growth and accumulation of active substances. The results showed that the suspension culture period of adventitious shoots in a 250 mL culture flask was 15 d. The optimal sucrose concentration, medium strength, inoculum quantity and length were 30 g·L^-1, MS medium, 1.0 g and 3 cm, respectively. Under the optimal culture conditions, the accumulation of coumarins and flavonoids in adventitious shoots reached the maximum on the 15 th day, i.e., umbelliferone 0.351 mg, scopoletin 0.035 mg and flavonoid 2.459 mg, respectively. The suspension culture period of adventitious shoots in the 3 L bioreactor was 24 d. The optimal culture conditions were sucrose 30 g·L^-1, 6-BA 0.5 mg·L^-1 and initial quantity of inoculum 15 g. Under these conditions, the cumulative amounts of umbelliferone, scopoletin and flavonoids were 6.041, 0.763 and 51.334 mg, respectively. In this study, a small bioreactor culture system for adventitious shoots of Stellera chamaejasme was established, which laid a foundation for further large-scale cultivation and production of the above active substances.

关 键 词：瑞香狼毒 悬浮培养 黄酮 香豆素 

分 类 号：S567.239[农业科学—中草药栽培]