利用CRISPR/Cas9技术编辑OsRR22基因创制耐盐水稻种质资源  

Creation of Salt-Tolerant Rice Germplasm by Editing OsRR22 Gene via CRISPR/Cas9 Technique

作　　者：牛淑琳 鞠培娜 周冠华 戴南平 周晋军[4] 谢先芝[1] 郑崇珂 Niu Shulin;Ju Peina;Zhou Guanhua;Dai Nanping;Zhou Jinjun;Xie Xianzhi;Zheng Chongke(Institute of Wetland Agriculture and Ecology,Shandong Academy of Agricultural Sciences,Jinan 250100,China;College of Life Sciences,Shandong Normal University,Jinan 250014,China;College of Life Sciences,Shandong Agricultural University,Taian 271018,China;Institute of Crop Germplasm Resources,Shandong Academy of Agricultural Sciences,Jinan 250100,China)

机构地区：[1]山东省农业科学院湿地农业与生态研究所,山东济南250100 [2]山东师范大学生命科学学院,山东济南250014 [3]山东农业大学生命科学学院,山东泰安271018 [4]山东省农业科学院农作物种质资源研究所,山东济南250100

出　　处：《山东农业科学》2023年第2期30-35,共6页Shandong Agricultural Sciences

基　　金：山东省农业科学院农业科技创新工程项目(CXGC2022A11);山东省重点研发计划(重大科技创新工程)项目(2021TZXD005)。

摘　　要：水稻是重要的粮食作物之一,在盐碱地开发和改良中发挥着重要作用。为了创制高耐盐水稻种质资源,利用CRISPR/Cas9基因编辑技术,在水稻品种盐丰47中对OsRR22设计3个靶位点进行基因编辑。通过PCR和测序鉴定,在T_(0)代获得16个突变单株,其中靶位点1和靶位点2均有纯合突变,靶位点3没有检测到突变。在T_(1)代转基因株系中,获得3株纯合突变且无T-DNA插入的纯合突变体。对T_(2)代进行农艺性状分析发现,与野生型相比,突变体的株高显著降低,产量相关农艺性状均无显著变化。苗期耐盐性鉴定结果表明,在200 mmol/L NaCl处理7天后,突变株的存活率比野生型提高30%以上。综上,利用CRISPR/Cas9技术对水稻品种盐丰47进行了耐盐性改良,为耐盐水稻新品种的培育提供了理论和材料基础。Rice is one of the most important food crops and plays an important role in the development and improvement of saline-alkali lands.In order to create high salt-tolerant rice germplasms,three target sites were designed for OsRR22 gene editing in rice variety Yanfeng 47 by using CRISPR/Cas9 technology.Through PCR and sequencing methods,16 mutant individual plants were obtained in T_(0)generation,in which,target site 1 and target site 2 had homozygous mutations,and no mutation was detected at target site 3.Three homozygous mutants without T-DNA insertion were obtained from T_(1)transgenic lines.Agronomic character analysis of T_(2)generation showed that compared with the wild type,plant height of the mutant was significantly reduced,and there was no significant change in yield-related agronomic characters.The results of salt tolerance identification at seedling stage showed that the survival rate of mutants was over 30%higher than that of the wild type after 7 days of 200 mmol/L NaCl treatment.In conclusion,the salt tolerance of Yanfeng 47 mutants was improved by CRISPR/Cas9 technology,which could provide theoretical and material bases for the cultivation of new salt-tolerant rice varieties.

关 键 词：水稻 耐盐性 OsRR22 基因编辑 

分 类 号：S511[农业科学—作物学] Q78[生物学—分子生物学]