发根农杆菌介导芸芥的遗传转化及其次生代谢物的研究

[目的]研究发根农杆菌介导芸芥的遗传转化条件，并测定次生代谢物的含量。[方法]通过发根农杆菌R1000介导的遗传转化，建立芸芥发状根的诱导和培养体系，采用氯化钯分光光度法和高效液相色谱法对该发根培养物合成硫苷的能力进行初步分析，并分析乙酰丁香酮（As）、浸染液pH值和浸染时间等因素对发状根转化频率的影响。[结果]子叶的转化率明显高于下胚轴；AS最佳浓度为100mg／L，浸染液最佳pH值为5．4，最佳浸染时间为20min。PCR的鉴定结果表明，发根农杆菌R1000的rolB基因已成功地转入并整合到该发根培养物的基因组中。[结论]该方法建立了芸芥发状根培养体系，并初步分析了该发状根中硫苷的含量，为以后的研究奠定了一定的基础。

Agrobacterium rhizogene Mediated Genetic Transformation of Eruca Sativa Mill and Analysis of Its Secondary Metabolites

[ Objective ] To study genetic transformation conditions of Agrobacterium rhizogene mediated Eruca Sativa Mill, and determine sec- ondary metabolite content. [ Method] Using hairy roots （HR） culture, an effective system for HR induction and culture of Eruca sativa was first established via Agrobacterium rhizogenes R1000 mediated genetic transformation, and both PdC12-spectrophotometer and high performance liquid chromatography （HPLC） analysis primarily revealed that the obtained HR culture had certain capacity of biosynthesizing Glucosinolate. Moreo- ver, the effect of explants types, 3.5-methoxy-4- ydroxyace- tophenone （AS） hconeentration, pH and infection times on transformation rate of Eruca sativa was analyzed. I Result] The result indicated that cotyledon is remarkably supper to hypocotyls, and optimal AS concentration, pH and infection times were 100 mg/L, 5.4 and 20 min, respectively, for increased transformation rate. Also polymerase chain reaction （PCR） a- nalysis demonstrated that rolB gene of T-DNA from Ri plasmid had been integrated into the genome of the transformed roots. [ Conclusion] The study established hairy roots cultivation system for Eruea sativa, the content of glucosinolate in hairy roots was determined, which will lay a foun- dation for future reserches.