蒙古黄芪毛状根的高效诱导及毛状根生物量和总黄酮含量的比较分析  

The High Efficient Induction of Hairy Roots of Astragalus membranaceus var. mongholicus and Comparative Analysis the Biomass and Total Flavonoids Content of Hairy Roots

作　　者：姚庆收[1] 陈向明[1] 马玉娥 刘晓云 林婷婷 

机构地区：[1]滨州医学院药学院,山东烟台264003

出　　处：《中药材》2017年第8期1753-1757,共5页Journal of Chinese Medicinal Materials

基　　金：山东省高校科技计划资助项目(J15LE51)

摘　　要：目的:解决蒙古黄芪毛状根诱导率低的问题,并分析毛状根的生物量和总黄酮含量,以期为蒙古黄芪毛状根的开发利用奠定基础。方法:用不同浓度的发根农杆菌R1601侵染自然生长的蒙古黄芪幼嫩的叶片、叶柄和茎段,诱导毛状根;用PCR技术鉴定毛状根,统计毛状根的诱导率;用紫外分光光度法测定蒙古黄芪毛状根中总黄酮的含量。结果:在四个不同发根农杆菌R1601浓度侵染下,三种外植体的诱导率在79.3%~94.9%之间;当发根农杆菌R1601菌液A600的值为1.5时,叶片的毛状根诱导率最高,达到94.9%;PCR鉴定表明诱导得到的毛状根为发根农杆菌R1601侵染所得;不同毛状根株系间生物量有显著差异;不同毛状根株系间总黄酮含量差异不明显,但均极显著高于对照品中的总黄酮含量。结论:建立了高效的蒙古黄芪毛状根诱导体系,揭示在利用蒙古黄芪毛状根合成药用成分时要对毛状根株系进行筛选。Objective： To solve the low induction rate of Astragalus membranaceus var. mongholicus hairy roots problem,and to analyze the biomass and the total flavonoids content of hairy roots,in order to lay the foundation for the utilization of Astragalus membranaceus var. mongholicus hairy roots. Methods： Hairy roots induction from leaves,petiole and stem of the natural growth of the young Astragalus membranaceus var. mongholicus were implemented through using Agrobacterium rhizogenes R1601 with different concentrations. Hairy roots were identified by PCR analysis,the induction rate was counted. The content of total flavonoids of hairy roots was determined by using ultraviolet spectrophotometric method. Results： The induction rate of three kinds of explants reached 79. 3% ~ 94. 9%with infection of four different concentrations of Agrobacterium rhizogenes R1601. The most suitable A600 value of Agrobacterium rhizogenes R1601 for the infection was 1. 5,which could reach a maximum induction rate of hairy roots from leaves as 94. 9%. PCR identification showed that the hairy roots were induced by Agrobacterium rhizogenes R1601. The biomass of different hairy root strains had significant difference. The difference of the total flavonoids content is not obvious in different strains of hairy root,but the total flavonoids of every strain was significantly higher than that of control. Conclusion： The efficient hairy roots induction system of Astragalus membranaceus var. mongholicus is established,and shows that the hairy root lines are screened by using the hairy roots of Astragalus membranaceus var. mongholicus for the synthesis of medicinal composition.

关 键 词：蒙古黄芪 毛状根 生物量 总黄酮 

分 类 号：R282.2[医药卫生—中药学]