紫花苜蓿UFO基因克隆及转化

紫花苜蓿(Medicago sativa L.)是多年生豆科牧草。为研究UFO(Unusual Floral Organs)基因相关功能,以紫花苜蓿中苜1号为试验材料,利用PCR技术在紫花苜蓿中克隆出UFO基因。通过DNA重组技术成功构建3302-3flag-UFO植物表达载体,利用农杆菌介导法对紫花苜蓿进行转基因操作,最终获得转基因紫花苜蓿。试验中激素浓度配比与植物表达载体的改良,提高了紫花苜蓿转基因过程中的转化率,而激素种类与配比则影响抗性愈伤的形成与愈伤组织的玻璃化程度。本试验共获得67个抗性愈伤,最终有56个分化成幼嫩紫花苜蓿转基因植株,炼苗入土获得17株转基因植株。遗传转化过程中获得的转基因植株为进一步研究UFO基因功能提供材料。

Cloning and Transformation of UFO Gene in Medicago sativa L.

Medicago sativa L.is a perennial legume forage.In order to analyze the function of UFO gene from Medicago sativa L.,we isolated the full-length coding sequence by PCR amplification in alfalfa variety‘Zhongmu No.1’.The plant expression vector 3302-3flag-UFO was constructed by DNA recombination technology,and transgenic alfalfa was obtained with deployment of Agrobacterium tumefaciens(Smith&Towns.)Conn mediated transformation method.The transgenic efficiency of alfalfa was improved by changing the ratio of hormone concentrations and improving plant expression vector.The type and ratio of hormones have resulted in a difference on the formation of resistant calli and the degree of vitrification of calli.By taking use of optimal transformation strategy,a total of 67 resistant calli were generated,while 56 young alfalfa regenerated plants were obtained.Finally,17 mature regenerated plants were obtained for seed propagation.Taking together,this study generated the transgenic plants with UFO over-expression vector,which might provide materials for further functional study of UFO gene.