白花丹参毛状根诱导体系的建立与其内生真菌提高毛状根中丹酚酸含量的研究  

Hairy root induction of Salvia miltiorrhiza Bge. f. alba and fungal endophytes enhanced salvianolic acid contents in hairy root cultures

作　　者：陶如[1] 冯蕾[1] 赵法兴[2] 李艳玲[1] 冀海伟[1] 郝岗平[1] 

机构地区：[1]泰山医学院,山东泰安271000 [2]江西中医药高等专科学校,江西抚州344000

出　　处：《时珍国医国药》2015年第6期1469-1473,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81173489);山东省自然科学基金(No.ZR2010CM063);中国博士后科学基金项目(No.20110490051)

摘　　要：目的建立白花丹参毛状根诱导体系及检测其丹酚酸的合成,研究内生真菌对毛状根中丹酚酸生物合成的影响。方法通过发根农杆菌ACCC10060浸染白花丹参叶片,成功诱导出毛状根,毛状根在6,7-V液体培养基中扩大培养60d,并测定毛状根中丹酚酸的含量。白花丹参毛状根在6,7-V培养基中培养12 d后,添加从白花丹参分离的不同浓度的内生真菌诱导子,测定毛状根丹酚酸含量。结果发根农杆菌ACCC10060能够成功从白花丹参叶片诱导出毛状根,毛状根培养40 d和50 d时丹酚酸B和丹参素的最高产量可达9.38mg·g-1DW和1.281 mg·g-1DW。60(μg glc equ).ml-1内生真菌诱导子是促进丹酚酸合成的最佳浓度,内生真菌处理后30 d后,丹酚酸B和丹参素的产量分别增加1.55和2.68倍。结论白花丹参毛状根诱导培养可以做为一种生产丹酚酸的有效途径,60(μg glc equ)·ml-1内生真菌诱导子可以显著提高毛状根中丹酚酸含量。Objective To establish hairy root cultures of Salvia miltiorrhiza Bge. f. alba and test the salvianolic acid production of these cultures,and investigate the influence of fungal endophytes on the synthesis of salvianolic acid in hairy root culture of Salvia miltiorrhiza Bge. f. alba. Methods To induce hairy root of Salvia miltiorrhiza Bge. f. alba,leaf explants were infected with Agrobacterium rhizogenes ACCC10060. Hairy roots were cultured in 6,7- V liquid medium for 60 d and tested the salvianolic acid production of these cultures. To investigate the influence of fungal endophytes on the synthesis of salvianolic acid in hairy root culture of Salvia miltiorrhiza Bge. f. alba,various concentrations of fungal endophytes isolated from Salvia miltiorrhiza Bge were added to the hairy root cultures grown for 12 d. After cultured for 30 d,salvianolic acids content were tested. Results Hairy root cultures of Salvia miltiorrhiza Bge. f. alba were successfully established by infecting leaf explants with Agrobacterium rhizogenes ACCC10060. Hairy roots were cultured in 6,7- V liquid medium and Salvianolic acid B and Salvianic acid A were detected in hairy root cultures. After culture for 40 d and 50 d,maximum production of Salvianolic acid B and Salvianic acid A（ 9. 38 mg. g^- 1DW and 1. 281 mg·g^- 1DW respectively） were respectively attained. Salvianolic acid B and Salvianic acid A contents increased 1. 55 and 2. 68 times respectively after 60（ μg glc equ）. m L^- 1fungal endophytes elicitor treated for 30 d. Conclusion Hairy root culture of Salvia miltiorrhiza Bge. f. alba is a valuable alternative approach for the production of salvianolic acid,60（ μg glc equ）.m L^- 1fungal endophytes elicitor is the optimal concentration for salvianolic acid production.

关 键 词：白花丹参 毛状根诱导 丹酚酸B 丹参素 内生真菌 

分 类 号：R284.2[医药卫生—中药学] S567[医药卫生—中医学]