亚磷酸脱氢酶作为筛选标记基因在水稻遗传转化中的应用  

Using phosphite dehydrogenase as a screening marker gene in rice genetic transformation

作　　者：刘同同 刘诗琦 袁丽丽 王创 LIU Tongtong;LIU Siqi;YUAN Lili;WANG Chuang(Microelement Research Center/Ministry of Agriculture and Rural Affairs Key Laboratory of Arable Land Conservation(Middle and Lower Reaches of Yangtse River)/College of Resources and Environment,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学资源与环境学院/微量元素研究中心/华中农业大学农业农村部长江中下游耕地保育重点实验室,武汉430070

出　　处：《华中农业大学学报》2022年第2期98-104,共7页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金面上项目(31772378)。

摘　　要：为筛选标记基因用于植物遗传转化,采用组织培养、营养液筛选和叶面喷施等手段,综合评价了具有高催化活性的亚磷酸脱氢酶基因ptxDQ作为筛选标记基因用于水稻遗传转化的潜力。结果显示,ptxDQ基因作为筛选标记基因时,抗性愈伤的筛选效率为44.37%~47.28%,显著高于潮霉素和草胺磷等抗性基因的筛选效率。获得的转基因阳性幼苗,可利用以亚磷酸盐为唯一磷源的培养液,通过简单培养即可筛选出阳性转基因幼苗。此外,还建立了通过叶面喷施亚磷酸盐筛选转基因幼苗的方法。相比目前的抗生素和除草剂类筛选系统,亚磷酸盐更廉价且不会造成环境污染。同时,作为一种正向筛选系统具有较高的筛选效率,在农作物转基因研究和农业可持续发展中具有潜在价值。Screening marker genes play a crucial role in plant genetic transformation,and the development of safe and efficient screening marker genes is of great significance for transgenic crops.The potential of the phosphite dehydrogenase gene ptxDQ with high catalytic activity as a screening marker gene for rice genetic transformation was comprehensively evaluated by means of tissue culture,nutrient solution screening and foliar spraying.The results showed that when ptxDQ gene was used as a screening marker,the screening efficiency of resistant callus was about 44.37%-47.28%,significantly higher than that of the resistance genes including hygromycin and glufosinate.The positive transgenic seedlings can be screened by simply culturing transformed seedlings in the culture medium with phosphite as the only phosphorus source.In addition,a method for screening transgenic seedlings by foliar spraying of phosphite was established.Phosphite is less expensive and less polluting than the current antibiotic and herbicide screening systems.As a forward screening system,it has high screening efficiency and has potential value in studying transgenic crops and developing sustainable agriculture.

关 键 词：遗传转化 筛选标记 亚磷酸脱氢酶 亚磷酸 水稻 

分 类 号：Q785[生物学—分子生物学]