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牛蒡毛状根培养体系的建立及次生代谢产物的测定

发表时间:2015-10-15  浏览量:1549  下载量:834
全部作者: 李国新,程义婷,张悦怡,杨世海
作者单位: 吉林农业大学中药材学院
摘 要: 目的:建立牛蒡(Arctium lappa L.)毛状根的培养体系,对诱导出毛状根的次生代谢产物进行含量测定。方法:选取3种发根农杆菌R1000,R1,R15834,分别对牛蒡叶片、茎段、根段进行侵染,诱导毛状根的产生,对诱导出的毛状根进行PCR分子鉴定,并进一步扩大培养。通过高效液相色谱(high performance liquid chro-matography,HPLC)测定牛蒡毛状根、原植物牛蒡子、原植物牛蒡根的牛蒡苷含量并进行对比。结果:R1对牛蒡根段的感染能力最强;经过PCR分子鉴定,发根农杆菌中的DNA质粒成功地整合到转化根的基因组中;进一步液体培养28 d,干重、鲜重及生物量积累最高;测得牛蒡毛状根中次生代谢产物牛蒡苷含量最高,是原植物牛蒡子的1.16倍。结论:初步建立了牛蒡毛状根的培养体系,为进一步研究牛蒡毛状根的大量培养,以及生产更高含量的牛蒡苷奠定了基础。
关 键 词: 植物学;牛蒡;发根农杆菌;毛状根;牛蒡苷
Title: Establishment of culture system of Arctium lappa L. hairy roots and determination of the secondary metabolites
Author: LI Guoxin, CHENG Yiting, ZHANG Yueyi, YANG Shihai
Organization: School of Chinese Medicine Material, Jilin Agricultural University
Abstract: Objective: To establish the hairy roots culture system of Arctium lappa L. and study the hair roots growth and analyze secondary metabolites, which is polysaccharide content in hair roots culturing system. Methods: Leaf blades, stems and roots of A. lappa L. were infected by Agrobacterium rhizogenes R1000,R1 and R15834, respectively. Hairy root was optimized to expand the training system and detected on PCR. The arctinin of different parts of A. lappa L. were determined by high performance liquid chromatography (HPLC) and the results were compared with each other. Results: The strain R1 could make root inductivity, the PCR examination result showed that genes was inserted into the hairy roots of root by A. lappa L.. Hairy roots were further cultured 28 days, dry weight, fresh weight and biomass accumulation all went to the highest. Secondary metabolite, arctinin, in hairy root of A. lappa L. was 1.16 times higher than in seeds. Conclusion: Through these experiments, hairy root of A. lappa L. was established preliminarily, which laid a foundation for the massive cultivation and producing higher content of arctinin.
Key words: botany; Arctium lappa L.; Agrobacterium rhizogenes; hairy roots; arctinin
发表期数: 2015年10月第19期
引用格式: 李国新,程义婷,张悦怡,等. 牛蒡毛状根培养体系的建立及次生代谢产物的测定[J]. 中国科技论文在线精品论文,2015,8(19):2059-2066.
 
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