CRISPR/Cas9介导靶向敲除水稻过氧化氢酶基因OsCAT  

The Targeted Knockout of Rice Catalase OsCAT3 Gene Mediated by CRISPR/Cas9

作　　者：曾嘉丽 欧阳林娟[1,2,3] 刘家林 贺浩华 朱昌兰[1,2,3] 彭小松 贺晓鹏[1,2,3] 傅军如 陈小荣[1,2,3] 边建民 徐杰[1,2,3] 孙晓棠 周大虎[1,2,3] 胡丽芳 Zeng Jiali;Ouyang Linjuan;Liu Jialin;He Haohua;Zhu Changlan;Peng Xiaosong;He Xiaopeng;Fu Junru;Chen Xiaorong;Bian Jianmin;Xu jie;Sun Xiaotang;Zhou Dahu;Hu Lifang(Key Laboratory of CropPhysiology, Ecology and Genetic Breeding, Ministry of Education, Jiangxi Agricultural University, Nanchang, 330045;Re- search Center of Super Rice Engineering and Techonology, Jiangxi Province, Nanchang, 330045;Collaboration Center for Double Cropping Rice Modermization Production, Nanchang, 330045)

机构地区：[1]江西农业大学作物生理生态与遗传育种教育部重点实验室,南昌330045 [2]江西省超级稻工程技术研究中心,南昌330045 [3]双季稻现代化生产协同中心,南昌330045

出　　处：《分子植物育种》2018年第12期3926-3930,共5页Molecular Plant Breeding

基　　金：江西农业大学研究生创新项目(NDYC2016-S002);国家自然科学基金项目(31160062;31660385);江西省教育厅科学基金项目(GJJ12242);江西省自然科学基金项目(20122BAB214017)共同资助

摘　　要：过氧化氢酶(catalase,CAT)作为重要的抗氧化酶,可催化细胞内过氧化氢的分解。为了研究水稻过氧化氢酶基因OsCAT3的功能,选择目的基因第二外显子上的"CTCGTTCAACGGCCCGCTGTGG"作为目标靶序列,构建了OsCAT3的CRISPR/CAS9载体,将目的载体转化水稻粳稻‘9522’后获得在PAM附近发生编辑的独立转基因株系,编辑方式主要表现为缺失和错配。与野生型相比,转基因植株呈现生长矮小、不分蘖、叶片呈明显的病叶状表型,这为深入研究水稻OsCAT3基因的作用机制提供了帮助。Catalase, as the important antioxidant enzyme, can catalyze the decomposition of hydrogen peroxide in cells. In order to study the function of rice catalase gene OsCAT3, the＂CTCGTTCAACGGCCCGCTGTGG＂sequ-ence on the second exon of target gene was selected as the target sequence and was applied to construct the CRISPR/CAS9 vector of OsCAT3. Then, the target vector was transformed into rice japonica 9522, and we obtained the independent transgenic plants editing near PAM. The way for target sequence editing was mainly manifested as missing and mismatched. Compared with the wild type, the transgenic plants displayed dwarf, no-tilling and obvious disease spot leaf phenotype, which might provide help for further studying the mechanism of OsCAT3 gene in rice.

关 键 词：水稻 过氧化氢酶 载体 转基因 

分 类 号：S511[农业科学—作物学]