【作者】 冯圆圆；

【导师】 季静；

【作者基本信息】 天津大学 ， 遗传学， 2009， 硕士

【摘要】 在花卉的基因工程育种中,花期调控是重要的一个研究方向。本研究的主要目标是利用农杆菌介导的基因转化方法,采用热激诱导的组成型表达载体,将大豆热激蛋白启动子连接拟南芥或杨树的FT基因对安祖花进行转化,获得转化植株,待转化植株生长到一定阶段后诱导FT基因的表达,从而对外源FT基因对安祖花开花的影响进行研究,并探索利用FT基因控制花卉开花时间的方法。主要研究结果:建立了根癌农杆菌介导的安祖花遗传转化体系。以安祖花叶柄为转化受体,在OD600=0.6的菌液中浸染15min,再在黑暗条件下于含100μmol/L乙酰丁香酮的共培养基中培养3d可获得最佳的T-DNA转化效率。利用此优化的转化系统进行农杆菌的浸染,并在含卡那霉素100mg/L和头孢霉素300mg/L的筛选培养基上进行筛选,得到抗性愈伤组织,待其增殖到一定大小,分化成苗,再进行生根壮苗培养,最后对这些抗性苗进行PCR及GFP荧光检测。本实验分别获得了pJCGLOX-AtFT转化植株8个细胞系,pJCGLOX-PttFT1转化植株1个细胞系,为进一步的研究奠定了基础。更多还原

【Abstract】 Genetic engineering in the breeding of flowers, the flowering control is an important research direction.The main objective of this study is the use of Agrobacterium-mediated genetic transformation methods, the use of heat-shock-induced constitutive expression vector, soybean heat shock protein promoter of Arabidopsis thaliana or poplar to connect the FT gene conversion andraeanum flowers, was transformed into plants, applied to convert to a certain stage of plant growth after the induction of the expression of FT, FT exogenous gene which andraeanum flowers bloom on the impact of research, and to explore the use of FT genes control flower flowering time.The main findings: The establishment of the Agrobacterium tumefaciens-mediated genetic transformation system andraeanum flowers. Petiole to spend for the conversion andraeanum receptors, OD600 = 0.6 in the bacteria in the dip 15min, and then under the conditions in the dark with 100μmol/L AS were cultured in 3d the best available T-DNA transformation efficiency. Using this optimized transformation system of Agrobacterium tumefaciens disseminated and containing Km 100 mg/L and Cef 300 mg/L of the filter medium should be screened for resistant calli have been proliferating to a certain size, differentiate into seedlings and then cultured seedling root, the last of these resistant seedlings for PCR and GFP fluorescence detection. Respectively in this experiment pJCGLOX-AtFT plants into eight lines, pJCGLOX-PttFT1 strain into a plant for further research laid the foundation.更多还原