水稻新型核转化筛选标记基因的鉴定及应用  

Identification and application of new selectable marker genes for rice nuclear transformation

作　　者：张娟娟[1,2] 林拥军[1,2] 周菲[1,2] 

机构地区：[1]华中农业大学作物遗传改良国家重点实验室 [2]华中农业大学国家植物基因研究中心(武汉),武汉430070

出　　处：《华中农业大学学报》2015年第1期1-8,共8页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金青年项目(31100945);教育部博士点新教师项目(20110146120016)

摘　　要：将人工合成的来自Escherichia coli的dsdA基因与来自Schizosaccharomyces pombe的dao1基因,通过农杆菌侵染方式转化到水稻中花11中。对转化植株进行分子生物学检测表明,目的基因整合到了水稻核基因组中,其中获得转化dsdA的单拷贝植株7个,转化dao1的单拷贝植株6个。通过对单拷贝转基因家系表达量检测及T0代种子萌发测验发现,dao1基因在2个(编号6,7)转基因家系中表达量显著,T0代种子在含D-Ala的培养基上,具有明显抗性。dsdA基因在所有的转基因家系中表达量相对偏低,在含D-Ser的培养基上,T0代种子有一定抗性。Two new selectable marker gene dsdA and daol from Escherichia coli and Schizosac- charomyces were synthesized by codon-optimization, and then transformed into japonica rice cultivar zhonghua 11 through Agrobacterium mediated gene transformation. Southern hybridization of the DNA transformants showed that 7 single-copy plants ofdsdA gene and 6 single-copy plants of daol gene were obtained. Results of detecting expression levels of the single-copy transformants and the germination texts of To generation seeds showed that there were two single-copy gene transformants （numbered as 6 and 7） of daol gene,indicating high expression levels and consistent with seed germination testing on D-Ala. In all single-copy transformants of dsdA gene,the expression levels of dsdA were relatively low. Test of seed germination showed that there was the resistance to D-Ser. This study initially explored the application of dsdA,daol as new selectable marker genes for rice nuclear transformation, and will lay the foundation for its extensive applications.

关 键 词：水稻 转基因 筛选标记基因 D-氨基酸 生物安全 

分 类 号：S511[农业科学—作物学]