稻香相关基因OsBADH2在“吉粳88”中的基因编辑研究  

Editing of Fragrant Rice Related Gene OsBADH2 in ‘Jijing 88’

作　　者：周岩 郭嘉[2,3] 胡玉锋 魏健 李毅丹[2,3] ZHOU Yan;GUO Jia;HU Yu-feng;WEI Jian;LI Yi-dan(Changchun Normal University,Changchun 130032;Agro-Biotechnology Research Institute,Jilin Academy of Agricultural Sciences,Changchun 130033;Jilin Provincial Key Laboratory of Agricultural Biotechnology,Changchun 130033;School of Life Science,Jilin Normal University,Siping 136000)

机构地区：[1]长春师范大学生命科学学院,长春130032 [2]吉林省农业科学院农业生物技术研究所,长春130033 [3]吉林省农业生物技术重点实验室吉林省农业科学院,长春130033 [4]吉林师范大学生命科学学院,四平136000

出　　处：《生物技术通报》2020年第3期88-94,共7页Biotechnology Bulletin

基　　金：吉林省科技发展计划项目(20170101014);吉林省农业创新工程项目(CXGC-2017TD004,C92070502)

摘　　要：近年来稻米的香味品质受到消费市场的格外重视。水稻OsBADH2基因是影响稻米香味的重要基因,选择优质粳稻品种,针对该基因进行基因编辑,有望创制出香味品质优异的粳稻材料。本研究通过分析OsBADH2基因外显子序列的保守性,选择了3个特异性靶点,分别构建了U6启动子驱动的包含不同靶点的gRNA表达盒,并将其连入植物基因编辑表达载体。利用基因枪介导的瞬时转化体系,对非香型粳稻品种"吉粳88"进行遗传转化,T_0代共获得847株再生材料。经高分辨率溶解曲线(High Resolution Melting,HRM)分析和测序验证,24株T_0代材料的OsBADH2基因编辑靶点发生了不同类型的编辑。在T_1代材料中获得了7类不含转基因成分,且OsBADH2基因靶点纯合的基因编辑后代材料。本研究结果表明基于基因枪介导的瞬时转化体系,转化再生过程中省略了抗性筛选过程,并辅以高效的HRM检测,能够在T_1代即获得位点发生纯合编辑且无转基因成分的基因编辑后代材料。本研究获得的OsBADH2基因编辑的"吉粳88"后代材料也为培育香味品质改良的粳稻品种提供了新材料。The fragrant quality of rice is an important agronomic trait concerned by consumers.Gene OsBADH2 in rice is the key one that affects the fragrant of rice,selecting high-quality elite rice varieties and conducting gene editing based on the gene is expected to create japonica rice materials with excellent fragrance.In this study,we analyzed the conservation of the exon sequence of OsBADH2 gene,selected three specific targets,and constructed the gRNA expression cassette driven by U6 promoter containing different targets and ligated into plant gene editing expression vector.Then,we used gene gun-mediated transient expression system to have genetic transformation of non-fragrant elite rice variety“Jijing 88”and obtained 847 regenerated materials of T0.Finally we used high-resolution melting(HRM)to analyze and verify by sequencing,and there were different types of editing of OsBADH2 gene editing targets in 24 strains of T0 generation materials.Seven T1 lines were confirmed to be non-transgenic and homozygous in target sites.Our results showed that the resistance screening process was omitted in the process of transformation and regeneration based on the transient transformation system mediated by gene gun,and the gene-edited progenies with homozygous editing and non-transgenic components could be obtained in T1 generation while supplemented by efficient HRM detection.The“Jijing 88”progenies via editing OsBADH2 gene also provides new materials for the cultivation of elite rice varieties with improved fragrance quality.

关 键 词：基因组编辑 高分辨率溶解曲线 OsBADH2 水稻 

分 类 号：S511.22[农业科学—作物学]