浙江农林大学学报2017，Vol.34Issue(1)：129-136,8.DOI:10.11833/j.issn.2095-0756.2017.01.018
农杆菌介导bar基因转化水稻胚性愈伤组织的研究
Transformation of the bar gene into embryogenic calli of rice via Agrobacterium tumefaciens
摘要
Abstract
To establish the embryogenic calli (EC) transformation system of rice with Basta as a selectable a-gent, the glufosinate-resistant gene (bar gene) was transformed into EC of rice cultivars, ‘Nipponbare’ ,‘Xiushui 134’ and ‘Zhonghua 11’ by Agrobacterium-mediated transformation. After 3 cycles of glufosi-nate-resistant screening with 15, 10 and 10 mg·L-1 glufosinate, transgenic EC was detected by PCR. The trans-formation efficiency of ‘Nipponbare’ , ‘Xiushui 134’ and ‘Zhonghua 11’ was 77.9%, 67.5% and 92.2% respectively. The results showed that the selectable concentrations were effective for resistant EC screening and the resistant EC subsequently differentiated into seedlings. The frequency of EC differentiation of‘Zhonghua 11’ was 86.7% and the seedling rate was 30.7%; the frequency of EC differentiation of ‘Xiushui 134’ was 44.4%, and the seedling rate was 66.7%;the frequency of EC differentiation of ‘Nipponbare’ was 76.1%; and the seedling rate was 65.6%. Therefore, of the 3 cultivars, the EC of ‘Nipponbare’ for genetic transformation with glufosinate as a selectable agent was most efficient. The glufosinate selection transformation system not only provides a technical basis for rice genetic transformation and functional validation, but also provides transgenic materials with herbicide resistance for rice breeding.关键词
分子生物学/草丁膦/bar基因/胚性愈伤组织/粳稻/农杆菌介导法转化Key words
molecular biology/glufosinate/bar gene/embryogenic calli(EC)/Oryza sativa/Agrobacterium-me-diated transformatiom分类
农业科技引用本文复制引用
何少海,姚盛存,丰宇凯,汪轲,李飞飞..农杆菌介导bar基因转化水稻胚性愈伤组织的研究[J].浙江农林大学学报,2017,34(1):129-136,8.基金项目
国家自然科学基金青年基金资助项目（31401461）；浙江省大学生科技创新活动计划（新苗人才计划）项目（2015R412002）；2015年浙江农林大学农学院大学生创新训练计划项目 （）
