水飞蓟发状根培养体系的建立及其水飞蓟宾含量的测定(4)
[6] Huang Y， Su C Y， Kuo H J， et al. A comparison of strategies for multiple-gene co-transformation via hairy root induction[J]. Appl Microbiol Biot，2013，6(30)：643.
[7] 张海弢，包京姗，杨世海，等. 王不留行毛状根的诱导及培养[J].中国实验方剂学杂志， 2013，19(6)：141.
Establishment of culture system of Silybum marianum hairy roots and
determination of silybin
ZHANG Shu-li ZHANG Tian-zhu YANG Shi-hai
(1.College of Traditional Chinese Herbs， Jilin Agricultural University， Changchun 130118， China；
2.School of Pharmceutical Sciences， Changchun University of Chinese Medicine， Changchun 130000， China)
[Abstract] This research uses six Agrobacterium rhizogenes R1601， R15384， R1000， A4， R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A.rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time， pre-culturing， co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification， it can be seen that the DNA plasmids in the A.rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography， it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant. In this research， the silymarin hairy roots culturing system is established， which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
[Key words] Silybum marianum； Agrobacterium rhizogenes； hairy root； silybin
doi：10.4268/cjcmm20141111
[责任编辑 吕冬梅], 百拇医药(张淑丽 张天柱 杨世海)
[7] 张海弢，包京姗，杨世海，等. 王不留行毛状根的诱导及培养[J].中国实验方剂学杂志， 2013，19(6)：141.
Establishment of culture system of Silybum marianum hairy roots and
determination of silybin
ZHANG Shu-li ZHANG Tian-zhu YANG Shi-hai
(1.College of Traditional Chinese Herbs， Jilin Agricultural University， Changchun 130118， China；
2.School of Pharmceutical Sciences， Changchun University of Chinese Medicine， Changchun 130000， China)
[Abstract] This research uses six Agrobacterium rhizogenes R1601， R15384， R1000， A4， R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A.rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time， pre-culturing， co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification， it can be seen that the DNA plasmids in the A.rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography， it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant. In this research， the silymarin hairy roots culturing system is established， which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
[Key words] Silybum marianum； Agrobacterium rhizogenes； hairy root； silybin
doi：10.4268/cjcmm20141111
[责任编辑 吕冬梅], 百拇医药(张淑丽 张天柱 杨世海)