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目的建立发根农杆菌介导的垂序商陆遗传转化体系及毛状根株系生物量分析,为利用基因工程技术开发商陆奠定研究基础。方法活化发根农杆菌R1601,利用其不同浓度的菌液对垂序商陆的叶片和茎段进行转化,利用PCR技术鉴定毛状根;随机选取毛状根培养,鉴别不同株系间的生物量差异。结果垂序商陆对发根农杆菌R1601侵染敏感,4个浓度都能高效诱导垂序商陆形成毛状根,其中A600为1.0时,茎段和叶片的遗传转化效率都能达到100%;PCR鉴定表明毛状根中已经整合进了T-DNA序列;不同毛状根株系生物量分析表明毛状根株系生物量呈极显著差异。结论建立了发根农杆菌R1601介导的垂序商陆的高效遗传转化体系,不同毛状根株系间生物量存在极显著差异。
OBJECTIVE: To establish Agrobacterium -mediated Agrobacterium -mediated genetic transformation system of Pseudostellaria fortunei and biomass analysis of hairy-rooted strains, and to lay the foundation for the development of genetic engineering technology. Methods Agrobacterium rhizogenes R1601 was activated, and the leaves and stem segments of Pseudostellaria tabaci were transformed with different concentrations of bacterial broth. Hairy roots were identified by PCR. Hairy roots were randomly selected to identify the different organisms Differences in quantity. Results Agonis nigra was susceptible to Agrobacterium rhizogenes R1601 infection. All the four concentrations were highly effective in inducing hairy roots with the order Phytolacca aculeatus. When the A600 was 1.0, the genetic transformation efficiency of the stem segments and leaves could reach 100% ; PCR identification showed that the hairy roots had been integrated into the T-DNA sequence; the biomass analysis of different hairy rooted lines showed that the hairy rooted lines showed extremely significant differences in biomass. Conclusion A highly efficient genetic transformation system was established in Agrobacterium rhizogenes R1601. The biomass of different hairy root strains was significantly different.