利用CRISPR∕Cas9技术编辑水稻丙酮酸磷酸双激酶基因PPDK  

Editing rice pyruvate phosphodouble kinase gene(PPDK)by CRISPR∕Cas9 technology

作　　者：孙佳 吴清清 周文昊 林冬枝 宋忠明 董彦君 SUN Jia;WU Qingqing;ZHOU Wenhao;LIN Dongzhi;SONG Zhongming;DONG Yanjun(College of Life Science,Shanghai Normal University,Shanghai Key Laboratory of Plant Molecular Sciences,Shanghai 200234,China;Shanghai Seed Management Station,Shanghai 201103,China)

机构地区：[1]上海师范大学生命科学学院,上海市植物分子科学重点实验室,上海200234 [2]上海市种子管理总站,上海201103

出　　处：《上海农业学报》2021年第3期1-6,共6页Acta Agriculturae Shanghai

基　　金：国家自然科学基金项目(30971552);国家重点研发项目(2016YFD0100902);上海市科委项目(19391900200);上海市教委项目(2017-01-07-00-02-E00039)。

摘　　要：采用CRISPR∕Cas9技术,以水稻丙酮酸磷酸双激酶基因PPDK为靶基因,构建载体,用农杆菌介导的方法转化粳稻品种‘嘉花1号’,获得1个碱基A缺失的编辑突变植株ppdk3。该突变体米粒的胚乳为白色粉质,其淀粉颗粒变小,结构松散,I-KI染色溶液颜色明显比野生型浅,直链淀粉含量下降约5%,抗性淀粉含量略有增加。与野生型相比,该突变体的千粒重和单株穗数下降明显,水稻淀粉合成相关基因的转录表达水平也有大幅下降。试验表明:利用CRISPR∕Cas9技术可以有效地对水稻淀粉合成相关基因进行编辑,为稻米品质改良提供材料。The rice pyruvate phosphodouble kinase gene(PPDK)was used as the target gene to construct vector by CRISPR∕Cas9 technology.The vector was transformed into Japonica rice variety‘Jiahua 1’by Agrobacterium-mediated method,and an editing mutant plant ppdk3 with A-base deletion was obtained.The endosperm of ppdk3 mutant rice was white and powdery,its starch granule became smaller and the structure was loose,the color of I-KI staining solution was lighter than that of the wild type,the amylose content decreased by about 5%,and the resistant starch content increased slightly.Compared with the wild type,the 1000-grain weight and panicle number per plant of the mutant decreased significantly,and the transcription levels of starch synthesis related genes decreased significantly also.The results showed that CRISPR∕Cas9 technology could effectively edit rice starch synthesis related genes and provide materials for rice quality improvement.

关 键 词：水稻 CRISPR∕Cas9技术 PPDK基因 淀粉合成相关基因 

分 类 号：S511.03[农业科学—作物学]