应用CRISPR/Cas9基因编辑技术获得高直链淀粉水稻种质  

CRISPR/Cas9 Technology-generated High-amylose Rice Varieties

作　　者：吴敏 黄娟 石桃雄 朱丽伟 邓娇 梁成刚 汪燕 刘飞 李荣 蔡芳 陈庆富[1] WU Min;HUANG Juan;SHI Taoxiong;ZHU Liwei;DENG Jiao;LIANG Chenggang;WANG Yan;LIU Fei;LI Rong;CAI Fang;CHEN Qingfu(School of Life Science,Guizhou Normal University/Research Center of Guizhou Buckwheat Engineering Industry Technology,Guizhou,Guiyang 550025,China)

机构地区：[1]贵州师范大学生命科学学院/贵州省荞麦工程技术研究中心,贵州贵阳550025

出　　处：《福建农业学报》2024年第1期17-24,共8页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(32060508);贵州省科技计划项目(黔科合基础-ZK[2021]一般109,黔科合基础-ZK[2023]一般278);贵州师范大学学术新苗基金项目(黔师新苗[2021]A16号);云南省重大科技专项与重点研发计划项目(202202AE090020)。

摘　　要：【目的】通过CRISPR/Cas9基因编辑技术遗传改良水稻种质,创制高直链淀粉新材料。【方法】以水稻品种中花11为试验材料,利用CRISPR/Cas9编辑系统对水稻淀粉分支酶(Starch branching enzymes,SBE)基因OsSBE3进行靶向编辑,利用PCR技术鉴定无标记纯合突变体,并测定其淀粉含量。【结果】T_(0)代获得10株突变体株系,T1代获得5个无标记纯合突变株系,其中4个株系(sbe3-22-6、sbe3-25-3、sbe3-25-4、sbe3-25-6)的直链淀粉含量和淀粉直支比显著高于野生型。【结论】本研究创制了高直链淀粉含量的水稻新种质,为水稻品质改良提供了参考。【Objective】A series of new rice germplasms with high content of amylose derived from Zhonghua 11 using the CRISPR/Cas9 technology was generated.【Method】The gene of starch branching enzyme in rice,OsSBE3,was targeted for the genetic editing by CRISPR/Cas9.Homozygous T-DNA-free mutants were verified by PCR with starch content measured.【Result】Ten mutant lines were obtained from the T_(0) generation.From the T1 generation,5 homozygous T-DNA-free lines were obtained that included 4 lines,i.e.,sbe3-22-6,sbe3-25-3,Sbe3-25-4,and sbe3-25-6,showing significantly increased amylose content and amylose/amylopectin ratio over the wild type.【Conclusion】A series of new rice germplasms with high amylose content was created.

关 键 词：水稻 直链淀粉 支链淀粉 淀粉直支比 基因编辑 CRISPR/Cas9 

分 类 号：S511[农业科学—作物学]