hpt与bar基因作为水稻转基因筛选标记的比较研究  

A comparison study of hpt and bar as selection marker gene of transgenic rice

作　　者：张春雨[1] 李宏宇[2] 刘斌[2] 

机构地区：[1]沈阳农业大学农学院,沈阳110161 [2]中国农业科学院作物科学研究所,北京100081

出　　处：《遗传》2012年第12期1599-1606,共8页Hereditas（Beijing)

基　　金：国家转基因生物新品种培育重大专项(编号:2010ZX08010-002)资助

摘　　要：标记基因的选择是影响植物遗传转化和转基因后代筛选成败的关键因素。hpf与bar作为两种常用的水稻转化筛选标记被广泛应用于水稻的转化。为比较两者在实际应用中的效果,文章首先对比了在潮霉素和除草剂(Bialaphos)两种筛选剂下水稻遗传转化的情况。研究表明,hpf基因的转化筛选体系相对于bar基因在转化效率上提高近两倍,转化周期提前至少10d,且插入基因拷贝数更低。随后,文章分析了利用潮霉素浸种法在田间筛选转基因水稻的可行性,研究显示当潮霉素浓度大于167mg/L时,可以对以水稻品种kitaake为亲本的转基因材料进行有效筛选,达到常规除草剂的筛选效果。但与除草剂相比较,潮霉素的田间筛选成本却处于劣势。文章研究和讨论了hpt和bar基因在遗传转化和后代田间筛选中的优缺点,并提供了一种利用潮霉素浸种法筛选转基因后代阳性植株的手段,为将来在水稻转基因研究工作中根据实际需求选择合适的遗传转化、筛选体系提供参考。The decision of using selection marker is one of the key factors for success of plant genetic transformation and offspring screening.As two commonly used selection markers,hpt and bar genes are widely used in tissue culture-based rice transformation.To experimentally compare their performance,we investigated the efficiency of two transformation systems using Hygromycin and Bialaphos as the selection agents,respectively.The result indicated that the system using hpt gene as the selection marker saved 10 days and had double transformation efficiency and lower transgene copy number in comparison to the system using bar gene.Then,we assessed the feasibility of screening transgenic rice in the field by soaking the wild-type and transgenic seeds in a series of solutions containing step diluted hygromycin for two days.We targeted the suitable concentration for distinguishing the transgenic seeds from WT Kitaake seeds was 167 mg L-1.However,the cost of screening by hygromycin is still much higher than that of Basta in field test.Therefore,this study experimentally demonstrated the advantages and disadvantages of the hpt and bar gene as the selection markers and thus provided a reference for choose of an appropriate selection marker according to the practical applications.

关 键 词：hpt基因 BAR基因 转基因筛选 潮霉素浸种 水稻 

分 类 号：S511[农业科学—作物学]