【摘要】 花青素是一种水溶性的黄酮类物质,可使植物呈现出红、蓝、紫和红紫等颜色.外源花青素代谢调控基因的表达可使花青素在植物细胞内积累,使植物体外观上表现出色彩的变化,易于观察,因此可作为报告基因用于植物转基因研究,快速报告细胞、组织、器官或植株是否被转化.本研究用花青素代谢调控基因Bi和C1作为报告基因,以epsp作为筛选标记基因,构建了植物表达载体pBAC9009.基因枪转化玉米自交系501的幼胚,经过除草剂草甘膦抗性筛选和分化再生,获得了转化再生植株75株,其中43株获得结实种子.T0代植株有18株在苗期或生长阶段表现局部或全紫色,8个结实果穗有零星的紫色种子,从外观上可直接确认为转化植株.结合PCR和RT-PCR的分子检测及花青素含量分析,表明叶片和籽粒为紫色的玉米植株中外源目的基因已整合并且高效表达,即紫色表型与分子检测的结果高度一致.该结果表明我们成功建立了以花青素基因作为报告基因的植物转基因可视化跟踪表达系统.该系统的应用可以大大提高工作效率,节约检测成本,对于植物转基因研究具有重要意义.更多还原

【Abstract】 Anthocyanin is a kind of water-soluble flavonoids,which make the plants showing red,blue,purple,red purple or other colors.Expression of the metabolic regulation gene causes the accumulation of anthocyanins in plant cells,and makes the plant organs change color,easy to observe.In this paper,using Bi and Cl gene which involved in anthocyanin synthesis pathway as a visual reporter,and glyphosate resistant gene epsp,encoding 5-enolpyruvylshikimate-3 phosphate synthase,as a selection marker,plant expression vector pBAC9009 was constructed.Immature embryos and embryogenic callus of several excellent maize inbred lines were transformed with biolistic bombardment.Undertaking in vitro culture steps of callus induction,glyphosate resistance selection,differentiation and plant regeneration,75 transgenic plants were regenerated,and 43 ears of corn were harvested.There are 18 T0 lines expressed parts or whole purple on the seedling or the growth stage,8 ears of these lines with scattered purple seeds.Analysis of PCR,RT-PCR and anthocyanin content showed that exogenous genes were well integrated in the genome of maize and efficiently expressed in the purple leaves and seeds.In conclusion,we successfully constructed a visual tracking system for transgene which make the transformed plants can be easily distinguished from their appearance,only by its color appearance at the early stage.Application of the system can greatly improve efficiency of transgene,and save costs.It is very important for plant transgenic research.更多还原