摘要
为获得无任何抗生素抗性基因的抗病转基因水稻新品系,将克隆自甜椒的两亲性蛋白AP1基因与潮霉素抗性选择标记基因(HPT)分别构建位于同一农杆菌双元载体上的两个独立的T-DNA区中,并经农杆菌介导,将其导入江苏省推广的两个粳稻品种广陵香粳和武香粳9号中,获得了一批转基因水稻植株。PCR和Southern杂交分析表明,AP1基因和HPT基因已同时导入受体基因组中,并从共转化植株的自交后代中筛选到了无HPT基因的转AP1基因水稻植株。结合田间农艺性状考察,选育了多个无抗性选择标记基因的纯合转AP1基因水稻新品系;抗病性鉴定结果表明,转AP1基因水稻对稻白叶枯病的抗性与未转化对照相比有显著提高,部分转基因水稻对纹枯病的抗性与未转化对照相比也有一定程度的提高。
In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene (HPT) were cloned into the two separate T-DNA regions of the binary vector pSB 130, respectively, and introduced into the calli derived from immature seeds of two elite Japonica rice varieties, Guanglingxiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many co-transgenic rice lines, containing both the AP1 gene and the marker gene, were regenerated, and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable-marker free transgenic rice plants were subsequently obtained from the progeny of the co-transformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.
出处
《中国农业科学》
CAS
CSCD
北大核心
2005年第12期2373-2379,共7页
Scientia Agricultura Sinica
基金
国家植物转基因专项(JY03-B-10)
国家自然科学基金项目(30170567)
江苏省高新技术项目(BG2002301)
江苏省教委基金项目(01KJB210003)资助
关键词
水稻
抗病
AP1基因
无抗性选择标记
白叶枯病
纹枯病
Rice (Oryza sativa L.)
Disease resistance
AP1 gene
Selectable marker-free
Bacterial leaf blight
Sheath blight