水飞蓟发状根培养体系的建立及其水飞蓟宾含量的测定

实验采用6种发根农杆菌R1601，R15834，R1000，A4，R1025，R1感染水飞蓟植体，诱导水飞蓟发状根及其水飞蓟宾的产生。6种发根农杆菌均能诱导水飞蓟产生发状根，A4表现出较强的对水飞蓟的感染能力。实验通过侵染时间、预培养、共培养、pH等因素确定了诱导水飞蓟发状根产生的条件，确定MS液体培养基适合水飞蓟发状根的增殖。经过PCR分子鉴定，发根农杆菌中的DNA质粒成功的整合到转化根的基因组中。使用液相色谱测定了水飞蓟发状根中水飞蓟宾含量是水飞蓟植物中的2.5倍。实验所建立的水飞蓟发状根培养系统，为研究水飞蓟发状根大量培养生产水飞蓟宾奠定了基础。

Establishment of culture system of Silybum marianum hairy roots and determination of silybin

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.